< References on T.cruzi in 1994

References on T.cruzi in 1994

Record 1 of 131 - MEDLINE (R) 1994 TI: Encephalomyelitis due to a Sarcocystis neurona-like protozoan in a rhesus monkey (Macaca mulatta) infected with simian immunodeficiency virus. AU: Klumpp-SA; Anderson-DC; McClure-HM; Dubey-JP AD: Yerkes Regional Primate Research Center, Emory University, Atlanta, Georgia. SO: Am-J-Trop-Med-Hyg. 1994 Sep; 51(3): 332-8 ISSN: 0002-9637 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: A captive-born rhesus monkey (Macaca mulatta) experimentally infected with simian immunodeficiency virus developed neurologic abnormalities approximately seven months postinoculation. A chronic necrotizing encephalomyelitis with intralesional protozoal schizonts was diagnosed histologically. The protozoa was identified as Sarcocystis neurona based on its morphologic characteristics by light and electron microscopic examination, the developmental stages of the schizonts, and positive staining with antisera against Sarcocystis cruzi by immunocytochemical techniques. Although S. neurona may be confused with Toxoplasma gondii by light microscopy, the former lacks rhoptries, is in direct contact with the host cell cytoplasm, and divides by endopolygeny. Sarcocystis neurona has recently been identified as an etiologic agent of encephalomyelitis in horses, raccoons, and mink. MIME: Brain-pathology; Brain-parasitology; Encephalomyelitis-complications; Encephalomyelitis-pathology; Microscopy,-Electron; Sarcocystis-isolation-and-purification; Sarcocystis-ultrastructure; Sarcocystosis-complications; Sarcocystosis-pathology; Simian-Acquired-Immunodeficiency-Syndrome-pathology; Spinal-Cord-pathology; Spinal-Cord-parasitology MJME: *Encephalomyelitis-veterinary; *Macaca-mulatta; *Sarcocystosis-veterinary; *Simian-Acquired-Immunodeficiency-Syndrome-complications; *SIV- TG: Animal; Case-Report; Female; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: RR00165RRNCRR AN: 95030140 UD: 9501 SB: AIM Record 2 of 131 - MEDLINE (R) 1994 TI: Production and characterization of monoclonal antibodies against the major cysteine proteinase of Trypanosoma cruzi. AU: Gonzalez-G; Orn-A; Cazzulo-JJ; Gronvik-KO AD: Catedra de Inmunologia, Facultad de Quimica, Universidad de la Republica, Montevideo, Uruguay. SO: Scand-J-Immunol. 1994 Oct; 40(4): 389-94 ISSN: 0300-9475 PY: 1994 LA: ENGLISH CP: ENGLAND AB: In the present study we describe the production and characterization of a panel of monoclonal antibodies (MoAbs) directed against cruzipain (Crz), the major cysteine proteinase from Trypanosoma cruzi. The five MoAbs, BD6, BF2, CG2, CH8, and DC10 were analysed with respect to affinity and specificity. None of the MoAbs cross-reacted with papain, which has regions of high homology with Crz. Treatment of the antigen with periodate did not affect the binding of the MoAbs, suggesting that they bind to the polypeptide moiety of Crz. CH8 recognized a continuous epitope located at the C-terminal extension of the proteinase that appeared to be highly immunogenic. Although the rest of the MoAbs recognized epitopes located in the catalytic domain, the enzymatic activity of Crz was not impaired by the binding of the MoAbs. Characterization of the antibody-binding sites revealed the presence of at least four separate epitopes. MIME: Antibodies,-Monoclonal-immunology; Antibodies,-Protozoan-immunology; Antibody-Affinity; Antibody-Specificity; Enzyme-Linked-Immunosorbent-Assay; Epitope-Mapping; Mice-; Mice,-Inbred-BALB-C; Precipitin-Tests; Trypanosoma-cruzi-enzymology MJME: *Antibodies,-Monoclonal-biosynthesis; *Antibodies,-Protozoan-biosynthesis; *Cysteine-Proteinases-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 3.4.22; EC 3.4.22.-; 0; 0 NM: Cysteine-Proteinases; cruzipain; Antibodies,-Monoclonal; Antibodies,-Protozoan AN: 95025638 UD: 9501 Record 3 of 131 - MEDLINE (R) 1994 TI: [Experimental Trypanosoma cruzi infection via contaminated water and food] TO: Infeccion experimental con Trypanosoma cruzi a traves de agua y alimentos contaminados. AU: Calvo-Mendez-ML; Nogueda-Torres-B; Alejandre-Aguilar-R; Cortes-Jimenez-M AD: Departamento de Parasitologia, Escuela Nacional de Ciencias Biologicas del I.P.N., Mexico, D.F., Mexico. SO: Rev-Latinoam-Microbiol. 1994 Jan-Mar; 36(1): 67-9 ISSN: 0187-4640 PY: 1994 LA: SPANISH; NON-ENGLISH CP: MEXICO AB: In order to determine the efficiency of different foods and water to maintain the infectivity of T. cruzi, the percentage of animals that resulted infected when they were ingested was registered. The materials were contaminated with metacyclic trypomastigotes from triatomine bugs feces, the infection in the mice were registered by directed observation of the parasite in the blood and corroborate by xenodiagnosis. Pasteurized milk infected the highest number of mice and the infectivity lasted longer than any other item tested. The efficacy of infectivity of fresh cheese and rice lasted after three hours and the percentage of infected mice was lower than with milk. Cooked and raw beefmeal and water resulted in the lowest, although similar number of infected mice. The infective capacity lasted only for a short time. It appears that the main differences obtained in infectivity depended on the different contents of moisture and nutrients in the solution. MIME: Animal-Feed; Cattle-; Dairy-Products-parasitology; English-Abstract; Food-Handling; Humidity-; Meat-parasitology; Mice-; Rice-parasitology; Time-Factors; Triatoma-parasitology; Trypanosoma-cruzi-growth-and-development; Trypanosoma-cruzi-pathogenicity MJME: *Chagas-Disease-transmission; *Food-Contamination; *Food-Parasitology; *Trypanosoma-cruzi; *Water-Pollution TG: Animal; Male PT: JOURNAL-ARTICLE AN: 95025155 UD: 9501 Record 4 of 131 - MEDLINE (R) 1994 TI: [Inhibition of oxidative phosphorylation in Crithidia fasciculata and Trypanosoma cruzi by lipophilic o-quinones and nifurtimox] TO: Inhibicion de la oxidacion fosforilante en Crithidia fasciculata y Trypanosoma cruzi por o-quinonas lipofilicas y nifurtimox. AU: Biscardi-AM; Fernandez-Villamil-SH; Stoppani-AO AD: Centro de Investigaciones Bioenergeticas, Facultad de Medicina, Universidad de Buenos Aires, Argentina. SO: Rev-Argent-Microbiol. 1994 Apr-Jun; 26(2): 72-86 ISSN: 0325-7541 PY: 1994 LA: SPANISH; NON-ENGLISH CP: ARGENTINA AB: ATP and ADP levels were determined in Crithidia fasciculata and Trypanosoma cruzi. The nucleotide levels in crithidia or epimastigotes at the stationary phase of growth were, in nmol/10(8) cells, 15-40, and 3-7, for ATP and ADP, respectively. Incubation with the lipophilic o-naphthoquinones CG 8-935, CG 9-442 and CG 10-248 or the anti-chagasic nitrofuran nifurtimox, significantly decreased ATP level, with non-significant variations of the ADP level. The kinetics of ATP variation showed an initial 1-2 h lag and the diminution of the ATP level reached maximum value after 4-6 h incubation. Addition of L-glutamate or D-glucose as energy sources produced 2- or 3-fold increase of ATP level, after incubation the protozoa for 4-6 h with the corresponding substrates. Quinones and nifurtimox strongly depressed D-glucose or L-glutamate effects. Buthionine sulfoximine an inhibitor of glutathione biosynthesis, enhanced the effect of nifurtimox on ATP level in Crithidia fasciculata. It is concluded that by inhibiting ATP synthesis, either directly or-through oxygen radicals, the assayed drugs produced their cytotoxic action. MIME: Adenosine-Diphosphate-metabolism; Adenosine-Triphosphate-metabolism; Crithidia-fasciculata-metabolism; English-Abstract; Naphthoquinones-; Trypanosoma-cruzi-metabolism MJME: *Crithidia-fasciculata-drug-effects; *Nifurtimox-pharmacology; *Oxidative-Phosphorylation-drug-effects; *Quinones-pharmacology; *Trypanosoma-cruzi-drug-effects TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 23256-30-6; 56-65-5; 58-64-0 NM: Naphthoquinones; Quinones; Nifurtimox; Adenosine-Triphosphate; Adenosine-Diphosphate AN: 95024689 UD: 9501 Record 5 of 131 - MEDLINE (R) 1994 TI: An acidic component of the heterogeneous Tc-85 protein family from the surface of Trypanosoma cruzi is a laminin binding glycoprotein. AU: Giordano-R; Chammas-R; Veiga-SS; Colli-W; Alves-MJ AD: Departamento de Bioquimica, Universidade de Sao Paulo, Brazil. SO: Mol-Biochem-Parasitol. 1994 May; 65(1): 85-94 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Successful infection of mammalian host by trypomastigotes of Trypanosoma cruzi is a complex event, involving host receptors and parasite ligands. Interaction of the trypomastigote stage with laminin, a component of specialized extracellular matrices, as basement membranes, is studied in this report. Binding of 125I-laminin to trypomastigotes is specific and 2-5 x 10(3) laminin binding sites were calculated to be present on the surface of live trypomastigotes. Anti-laminin antibodies were able to inhibit the invasion of cultured cells by trypomastigotes (75-62%), suggesting that laminin may be involved in the adhesion of the parasite to host cells. By affinity chromatography, an 85-kDa glycoprotein was isolated (laminin binding glycoprotein, LBG) from trypomastigote lysates, but not from epimastigote lysates. It is suggested that at least fragment E8 (but not E1') from laminin could be involved in the reaction which is independent of the carbohydrate moieties from both ligand and receptor, as suggested by glycosidase or tunicamycin treatments. It is also shown that LBG is an acidic component of the polymorphic Tc-85 protein family, a trypomastigote-specific surface membrane glycoprotein which contains several polypeptides recognized by the monoclonal antibody H1A10, and previously related with the invasion process of the parasite. MIME: Antibodies,-Monoclonal; Binding-Sites; Carrier-Proteins-isolation-and-purification; Carrier-Proteins-metabolism; Glycoproteins-isolation-and-purification; Glycoproteins-metabolism; Laminin-antagonists-and-inhibitors; Laminin-immunology; Molecular-Weight; Peptide-Fragments-isolation-and-purification; Peptide-Fragments-metabolism; Protozoan-Proteins-isolation-and-purification; Trypanosoma-cruzi-pathogenicity MJME: *Laminin-metabolism; *Protozoan-Proteins-metabolism; *Trypanosoma-cruzi-metabolism TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0; 0 NM: Antibodies,-Monoclonal; Carrier-Proteins; Glycoproteins; Laminin; Peptide-Fragments; Protozoan-Proteins AN: 95021530 UD: 9501 Record 6 of 131 - MEDLINE (R) 1994 TI: Mediation of Trypanosoma cruzi invasion by heparan sulfate receptors on host cells and penetrin counter-receptors on the trypanosomes. AU: Herrera-EM; Ming-M; Ortega-Barria-E; Pereira-ME AD: Department of Medicine, Tufts-New England Medical Center Hospitals, Boston, MA 02111. SO: Mol-Biochem-Parasitol. 1994 May; 65(1): 73-83 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Trypanosoma cruzi attaches and invades a large variety of mammalian cells by receptor-mediated interactions, one of them involving the binding of parasite trans-sialidase to host sialyl receptors. Three proteoglycan-deficient mutants of Chinese hamster ovary (CHO) cells were used to probe the role of host heparin and heparan sulfate glycosaminoglycans (GAG) in T. cruzi invasion. All three mutants supported adhesion and infection to a much lower extent than the parental CHO cells. One of the mutants, pgsD-677, did not express heparan sulfate while containing three- to four-fold excess chondroitin sulfate, yet the cell line was a poor substrate for T. cruzi adhesion. Proteoglycan-deficient cells obtained by inhibiting GAG synthesis in parental cells with p-nitrophenyl-beta-D-xyloside, were also poor hosts for T. cruzi invasion. Furthermore, digestion of parental cells with heparinase and heparitinase, two lyases that specifically depolymerize heparin and heparan sulfate, reduced the potential of the cells to support T. cruzi adhesion and growth. Lyases that digested chondroitin sulfate and other GAGs did not affect T. cruzi invasion. These results suggest that heparin/heparan sulfate epitopes are receptors for T. cruzi invasion. The corresponding counter-receptor on T. cruzi appears to be penetrin, a heparin-binding protein that promotes trypanosome penetration into cells. Purified penetrin caused agglutination of red blood cells, and the hemagglutination was exquisitely sensitive to heparin and heparan sulfate. However, sialic acid and sialyl compounds did not inhibit penetrin-induced hemagglutination. Recombinant penetrin competitively inhibited T. cruzi invasion of proteoglycan-containing parental cells, but not of proteoglycan-deficient mutants nor of heparitinase-treated cells. Furthermore, consistent with the sugar specificity of penetrin as a hemagglutinin, recombinant penetrin competed for trypanosome invasion of a CHO cell mutant (Lec2) that expresses heparan sulfate but not sialyl residues. Given that the release of sialic acid from the proteoglycan-deficient mutants further reduced T. cruzi invasion, as did the removal of heparan sulfate from the Lec2 mutant, and given that penetrin does not bind to sialic acid with high affinity, the results indicate that the penetrin-heparan sulfate pathway for T. cruzi invasion is distinct from the trans-sialidase-sialic acid route. MIME: Cell-Adhesion-physiology; CHO-Cells; Glycoproteins-pharmacology; Glycoproteins-physiology; Glycosaminoglycans-genetics; Glycosaminoglycans-physiology; Hamsters-; Hemagglutination-physiology; Heparin-genetics; Heparin-physiology; Heparitin-Sulfate-genetics; Mutation-; Neuraminidase-pharmacology; Neuraminidase-physiology; Receptors,-Cell-Surface-genetics; Sialic-Acids-physiology; Trypanosoma-cruzi-pathogenicity MJME: *Heparitin-Sulfate-physiology; *Protozoan-Proteins-physiology; *Receptors,-Cell-Surface-physiology; *Trypanosoma-cruzi-physiology TG: Animal; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI18102AINIAID RN: EC 3.2.1.-; EC; 0; 0; 0; 0; 0; 9005-49-6; 9050-30-0 NM: trans-sialidase,-Trypanosoma-cruzi; Neuraminidase; Glycoproteins; Glycosaminoglycans; Protozoan-Proteins; Receptors,-Cell-Surface; Sialic-Acids; Heparin; Heparitin-Sulfate AN: 95021529 UD: 9501 Record 7 of 131 - MEDLINE (R) 1994 TI: An approach to functional complementation by introduction of large DNA fragments into Trypanosoma cruzi and Leishmania donovani using a cosmid shuttle vector. AU: Kelly-JM; Das-P; Tomas-AM AD: Department of Medical Parasitology, London School of Hygiene and Tropical Medicine, UK. SO: Mol-Biochem-Parasitol. 1994 May; 65(1): 51-62 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: To extend the range of genetic tools available for the functional analysis of trypanosomatid genes we have constructed a cosmid shuttle vector (pcosTL) which facilitates the introduction of large DNA fragments into Trypanosoma cruzi and Leishmania donovani. The vector contains several features to simplify library construction and insert mapping and transformed cells can be selected on the basis of G418 resistance. To evaluate the vector and to determine the fidelity of replication we first constructed cosmid libraries and isolated clones containing the T. cruzi major cysteine protease genes (a tandemly repeated array) and the L. donovani trypanothione reductase gene (a single copy gene). T. cruzi and L. donovani cells transfected with their respective cosmids were characterised by the presence of multiple copies of cosmid DNA and by a considerable over-expression of the corresponding enzyme activity. Rearrangements or deletions of insert sequences were not detected. These findings and the observation that cosmid DNA can be rescued unaltered from transformed parasites suggest that the pcosTL vector will be ideally suited for studies involving functional complementation. MIME: Base-Sequence; Chromosome-Mapping; Cosmids-; Cysteine-Proteinases-genetics; DNA-Primers-genetics; DNA,-Protozoan-genetics; Gene-Library; Genes,-Protozoan; Genetic-Complementation-Test; Genetic-Vectors; Leishmania-donovani-enzymology; Molecular-Sequence-Data; NADH,-NADPH-Oxidoreductases-genetics; Transfection-; Trypanosoma-cruzi-enzymology MJME: *DNA,-Recombinant-genetics; *Leishmania-donovani-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 1.6.; EC 1.6.4.-; EC 3.4.22; 0; 0; 0; 0; 0 NM: NADH,-NADPH-Oxidoreductases; trypanothione-reductase; Cysteine-Proteinases; Cosmids; DNA-Primers; DNA,-Protozoan; DNA,-Recombinant; Genetic-Vectors AN: 95021527 UD: 9501 Record 8 of 131 - MEDLINE (R) 1994 TI: Cloning and characterization of a gene for the stage-specific 82-kDa surface antigen of metacyclic trypomastigotes of Trypanosoma cruzi. AU: Araya-JE; Cano-MI; Yoshida-N; da-Silveira-JF AD: Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina, Sao Paulo, Brazil. SO: Mol-Biochem-Parasitol. 1994 May; 65(1): 161-9 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: We have cloned and sequenced a cDNA clone coding for a metacyclic trypomastigote-specific surface glycoprotein with a molecular mass of 82 kDa (MTS-gp82). By immunoblotting and immunoprecipitation, antibodies against the recombinant protein recognized an 82-kDa protein of metacyclic trypomastigotes, without any detectable reaction towards amastigotes, epimastigotes or tissue culture-derived trypomastigotes. The insert of the MTS-gp82 cDNA clone strongly hybridized with a single 2.2-kb metacyclic trypomastigote mRNA, suggesting that the steady-state levels of mRNAs for MTS-gp82 are developmentally regulated. MTS-gp82 is encoded by a multigene family whose members are distributed in several chromosomes. Sequence analysis revealed 40-56% identity at amino acid level between MTS-gp82 and members of Trypanosoma cruzi gp85/sialidase family (TSA-1, Tt34c1, SA85-1.1). MTS-gp82 showed several amino acid motifs that are characteristic of gp85/sialidase family, such as the Asp box (SxDxGxTW), the subterminal (VTVxNVFLYNR) motif and the putative GPI-anchor sequence. On the basis of its structural features, the MTS-gp82 gene could be included in the T. cruzi gp85/sialidase family, but constituting a distinct group which is preferentially expressed in metacyclic trypomastigotes. MIME: Amino-Acid-Sequence; Antigens,-Protozoan-chemistry; Antigens,-Surface-chemistry; Cloning,-Molecular; DNA,-Complementary-genetics; DNA,-Protozoan-genetics; Genes,-Reiterated; Glycoproteins-genetics; Molecular-Sequence-Data; Molecular-Weight; Neuraminidase-genetics; RNA,-Messenger-genetics; RNA,-Messenger-metabolism; RNA,-Protozoan-genetics; RNA,-Protozoan-metabolism; Sequence-Homology,-Amino-Acid; Trypanosoma-cruzi-growth-and-development MJME: *Antigens,-Protozoan-genetics; *Antigens,-Surface-genetics; *Genes,-Protozoan; *Trypanosoma-cruzi-genetics; *Trypanosoma-cruzi-immunology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 3.2.1.-; EC; 0; 0; 0; 0; 0; 0; 0; 138820-03-8 NM: trans-sialidase,-Trypanosoma-cruzi; Neuraminidase; Antigens,-Protozoan; Antigens,-Surface; DNA,-Complementary; DNA,-Protozoan; Glycoproteins; RNA,-Messenger; RNA,-Protozoan; trypomastigote-specific-surface-antigen AN: 95021521 UD: 9501 SI: GENBANK/L14824 Record 9 of 131 - MEDLINE (R) 1994 TI: A recombinant Trypanosoma cruzi trans-sialidase lacking the amino acid repeats retains the enzymatic activity. AU: Campetella-OE; Uttaro-AD; Parodi-AJ; Frasch-AC AD: Instituto de Investigaciones Bioquimicas, Fundacion Campomar, Buenos Aires, Argentina. SO: Mol-Biochem-Parasitol. 1994 Apr; 64(2): 337-40 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS MIME: Amino-Acid-Sequence; Antigens,-Protozoan-genetics; Base-Sequence; DNA-Primers-genetics; DNA,-Protozoan-genetics; Genes,-Protozoan; Glycoproteins-immunology; Glycoproteins-metabolism; Molecular-Sequence-Data; Neuraminidase-immunology; Neuraminidase-metabolism; Protozoan-Proteins-genetics; Protozoan-Proteins-immunology; Protozoan-Proteins-metabolism; Recombinant-Proteins-genetics; Recombinant-Proteins-immunology; Recombinant-Proteins-metabolism; Repetitive-Sequences,-Nucleic-Acid; Trypanosoma-cruzi-immunology MJME: *Glycoproteins-genetics; *Neuraminidase-genetics; *Trypanosoma-cruzi-enzymology; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't GS: TS PT: JOURNAL-ARTICLE RN: EC 3.2.1.-; EC; 0; 0; 0; 0; 0; 0 NM: trans-sialidase,-Trypanosoma-cruzi; Neuraminidase; Antigens,-Protozoan; DNA-Primers; DNA,-Protozoan; Glycoproteins; Protozoan-Proteins; Recombinant-Proteins AN: 95021510 UD: 9501 SI: GENBANK/L26499 Record 10 of 131 - MEDLINE (R) 1994 TI: A short interspersed repetitive element provides a new 3' acceptor site for trans-splicing in certain ribosomal P2 beta protein genes of Trypanosoma cruzi. AU: Vazquez-MP; Schijman-AG; Levin-MJ AD: Instituto de Investigaciones en Ingenieria Genetica y Biologia Molecular (INGEBI), Buenos Aires, Argentina. SO: Mol-Biochem-Parasitol. 1994 Apr; 64(2): 327-36 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Four Trypanosoma cruzi genomic DNA fragments carrying different TcP2 beta genes have been isolated and sequenced. Three of them had a single TcP2 beta gene, while the 3.8-kb-long DNA segment encoding the TcP2 beta-H1.8 locus showed two TcP2 beta genes arranged in tandem. These genes were physically connected by a 428-bp-long DNA sequence that was also located immediately 5' to the first gene and immediately 3' to the second. Comparison of the 4 TcP2 beta gene loci, suggested that the insertion of this repeated element originated the duplication of its target sequence, a poly(dT) stretch. Approximately 1200 copies of this short sequence, named short interspersed repetitive element (SIRE), were found scattered in the genome. Analysis of the 5' non-coding regions of different TcP2 beta mRNAs, and RNA-PCR experiments suggested that the insertion of a SIRE upstream of a TcP2 beta-H1.8 gene introduced a new 3' spliced leader (SL) acceptor site in the TcP2 beta-H1.8 pre-mRNAs, encoded within the SIRE. Consequently, in the mature H1.8 mRNA the SL sequence is followed by 38 bases directly transcribed from the SIRE. Structural and functional features of this repeated element reveal similarity to the short interspersed repetitive DNA sequences detected in the genomes of several microorganisms. MIME: Base-Sequence; Binding-Sites; DNA,-Protozoan-genetics; Genes,-Protozoan; Genes,-Reiterated; Molecular-Sequence-Data; RNA-Splicing; RNA,-Messenger-genetics; RNA,-Messenger-metabolism; RNA,-Protozoan-genetics; RNA,-Protozoan-metabolism; Trypanosoma-cruzi-metabolism MJME: *Phosphoproteins-genetics; *Repetitive-Sequences,-Nucleic-Acid; *Ribosomal-Proteins-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't GS: TcP2beta PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0; 0 NM: phosphoprotein-P2; DNA,-Protozoan; Phosphoproteins; Ribosomal-Proteins; RNA,-Messenger; RNA,-Protozoan AN: 95021509 UD: 9501 SI: GENBANK/X75031; GENBANK/X75033; GENBANK/X75030; GENBANK/X75032 Record 11 of 131 - MEDLINE (R) 1994 TI: The structure, organization, and expression of the Leishmania donovani gene encoding trypanothione reductase. AU: Taylor-MC; Kelly-JM; Chapman-CJ; Fairlamb-AH; Miles-MA AD: Department of Medical Parasitology, London School of Hygiene and Tropical Medicine, UK. SO: Mol-Biochem-Parasitol. 1994 Apr; 64(2): 293-301 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Trypanothione reductase (TR) is an NADPH-dependent flavoprotein oxidoreductase central to thiol metabolism in the trypanosomatids. We report here the cloning by expression of the Leishmania donovani gene. It is single copy, expresses a 2.6-kb transcript and a 52-kDa protein and is located on a 1.1-Mbp chromosome. The 491 amino acid sequence has 76% similarity to Crithidia fasciculata and 67% similarity to Trypanosoma cruzi, Trypanosoma congolense and Trypanosoma brucei TR. Residues recognising the adenosine pyrophosphate moiety of NADPH and FAD, and residues in the catalytic site segment (A47-A67) involving electron transfer from TR to trypanothione disulphide (T(S)2) were completely conserved. Thus inhibitors of TR are likely to be active against the enzyme from all the parasitic trypanosomatids. Two peptide inserts (39-47, 131-140) seen in other TR genes and a C-terminal extension of 19 residues were also present. When the gene was introduced back into L. donovani at high copy number using the pTEX expression vector, we detected elevated expression of TR RNA and a 14-fold increase in TR activity. Transfection and overexpression of the TR gene will facilitate studies of gene function and of the dependence of trypanosomatids on TR for protection against oxidative stress. MIME: Amino-Acid-Sequence; Base-Sequence; Chromosome-Mapping; Cloning,-Molecular; DNA,-Complementary-genetics; DNA,-Protozoan-genetics; Gene-Expression; Molecular-Sequence-Data; Sequence-Homology,-Amino-Acid; Transfection- MJME: *Genes,-Protozoan; *Leishmania-donovani-enzymology; *Leishmania-donovani-genetics; *NADH,-NADPH-Oxidoreductases-genetics TG: Animal; Support,-Non-U.S.-Gov't GS: TR PT: JOURNAL-ARTICLE RN: EC 1.6.; EC 1.6.4.-; 0; 0 NM: NADH,-NADPH-Oxidoreductases; trypanothione-reductase; DNA,-Complementary; DNA,-Protozoan AN: 95021506 UD: 9501 SI: GENBANK/Z23135 Record 12 of 131 - MEDLINE (R) 1994 TI: Combined occurrence of trypanosomal sialidase/trans-sialidase activities and leishmanial metalloproteinase gene homologues in Endotrypanum sp. AU: Medina-Acosta-E; Paul-S; Tomlinson-S; Pontes-de-Carvalho-LC AD: Rockefeller University, New York, NY. SO: Mol-Biochem-Parasitol. 1994 Apr; 64(2): 273-82 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Endotrypanum (order Kinetoplastida: family Trypanosomatidae) is a parasite of forest dwelling tree sloths (Edentata: genera Choleopus and Bradypus). Unique among the haemoflagellates, this protozoan has an intraerythrocytic phase in the mammalian host. Nevertheless, many striking similarities exist between Endotrypanum and the human pathogen Leishmania that make it a useful model for epidemiological and evolutionary aspects of the biology of trypanosomatids. Importantly, Endotrypanum species share both the insect vector and host reservoir with certain species of Leishmania (subgenus Viannia). Because mixed infections with Endotrypanum and Leishmania are common in sloths and, therefore, likely to occur in the sandfly vector, there is a need for adequate biochemical markers to distinguish Endotrypanum from Leishmania infections. In this paper we show that Endotrypanum promastigotes possess sialidase and trans-sialidase activities, which are absent from Leishmania, and which are not closely related to the previously described trypanosomal sialidase/trans-sialidase enzyme. We also document the occurrence in Endotrypanum of homologues of the leishmanial surface metalloproteinase gp63 genes. The combined occurrence of sialidase/trans-sialidase activities and gp63 gene homologues in a unique organism has important ramifications for both field and laboratory studies on the biology of trypanosomatids, especially those related to host infection and evolution. MIME: DNA,-Protozoan-genetics; Evolution-; Genes,-Protozoan; Glycoproteins-genetics; Leishmania-enzymology; Leishmania-genetics; Nucleic-Acid-Hybridization; Species-Specificity; Trypanosoma-enzymology; Trypanosoma-genetics; Trypanosomatina-classification MJME: *Metalloproteinases-genetics; *Neuraminidase-genetics; *Trypanosomatina-enzymology; *Trypanosomatina-genetics TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI22229AINIAID; AI21539AINIAID; K11AI0107102AINIAID RN: EC 3.2.1.-; EC; EC 3.4.24; EC 3.4.24.-; 0; 0 NM: trans-sialidase,-Trypanosoma-cruzi; Neuraminidase; Metalloproteinases; glycoprotein-gp63,-Leishmania; DNA,-Protozoan; Glycoproteins AN: 95021504 UD: 9501 Record 13 of 131 - MEDLINE (R) 1994 TI: Dihydrolipoamide dehydrogenase in the trypanosoma subgenus, trypanozoon. AU: Else-AJ; Clarke-JF; Willis-A; Jackman-SA; Hough-DW; Danson-MJ AD: Department of Biochemistry, University of Bath, England. SO: Mol-Biochem-Parasitol. 1994 Apr; 64(2): 233-9 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: The enzyme dihydrolipoamide dehydrogenase has been discovered and characterised in four salivarian trypanosomes of the subgenus trypanozoon: Trypanosoma brucei brucei, T. b. gambiense, T. b. rhodesiense, and Trypanosoma evansi. The three T. brucei species, which have insect procyclic forms biochemically distinct from their mammalian bloodstream forms, express dihydrolipoamide dehydrogenase in both cell types, but have higher levels in the procyclic forms. Determination of Michaelis constants for the enzyme from each of the three T. brucei species did not reveal any significant kinetic differences between the bloodstream and procyclic enzymes. On Western blots, antibodies raised against dihydrolipoamide dehydrogenase from the stereorarian trypanosome, Trypanosoma cruzi, cross-react strongly with the dihydrolipoamide dehydrogenase from all three T. brucei species; by this method, the relative molecular masses of their dihydrolipoamide dehydrogenases are indistinguishable. Dihydrolipoamide dehydrogenase was purified from both the bloodstream and the procyclic forms of T. b. brucei, and the N-terminal have been sequenced. These sequences are identical to the derived protein sequence of the cloned gene (Else et al., Eur. J. Biochem. 212 (1993) 423-429), but have a nine amino acid N-terminal truncation, giving an N-terminus equivalent to that of T. cruzi dihydrolipoamide dehydrogenase. The T. b. brucei dihydrolipoamide dehydrogenase gene has been expressed in Escherichia coli and the resultant protein purified; its N-terminus is processed in a similar fashion to that in the trypanosome, but with reduced specificity. MIME: Amino-Acid-Sequence; Antibodies,-Protozoan; Antigens,-Protozoan-genetics; Cloning,-Molecular; Cross-Reactions; Escherichia-coli-genetics; Lipoamide-Dehydrogenase-genetics; Lipoamide-Dehydrogenase-immunology; Molecular-Sequence-Data; Rats-; Rats,-Wistar; Species-Specificity; Trypanosoma-genetics; Trypanosoma-immunology; Trypanosomiasis-parasitology MJME: *Lipoamide-Dehydrogenase-metabolism; *Trypanosoma-enzymology TG: Animal; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; 0; 0 NM: Lipoamide-Dehydrogenase; Antibodies,-Protozoan; Antigens,-Protozoan AN: 95021500 UD: 9501 Record 14 of 131 - MEDLINE (R) 1994 TI: Cytosolic-free calcium elevation in Trypanosoma cruzi is required for cell invasion. AU: Moreno-SN; Silva-J; Vercesi-AE; Docampo-R AD: Department of Veterinary Pathobiology, University of Illinois at Urbana-Champaign, Illinois 61801. SO: J-Exp-Med. 1994 Oct 1; 180(4): 1535-40 ISSN: 0022-1007 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: To replicate, the trypomastigote form of Trypanosoma cruzi must invade host cells. Since a role for Ca2+ in the process of cell invasion by several intracellular parasites has been postulated, changes in the intracellular Ca2+ concentration in T. cruzi trypomastigotes and in tissue culture L6E9 myoblasts during their interaction were studied at the single cell level using digital imaging fluorescence microscopy or in cell suspensions by fluorescence spectrophotometry. An increase in cytosolic Ca2+ in T. cruzi trypomastigotes was detected at the single cell level after association of the parasites with the myoblasts. Ca2+ mobilization in the host cells was also detected upon contact with trypomastigotes either at the single cell level or in cells grown in coverslips and exposed to suspensions of trypomastigotes. Pretreatment of the parasites with the Ca2+ chelators quin 2 (50 microM) or bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA, 50 microM) decreased the trypomastigotes' association to myoblasts by approximately 40 and 63%, respectively, thus indicating that an increase in intracellular Ca2+ concentration in the parasites is required for cell invasion in addition to Ca2+ mobilization in the host cells. MIME: Cell-Line; Cytosol-metabolism; Egtazic-Acid-analogs-and-derivatives; Egtazic-Acid-pharmacology; Muscles-parasitology; Trypanosoma-cruzi-pathogenicity MJME: *Calcium-metabolism; *Trypanosoma-cruzi-metabolism TG: Animal; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI23259AINIAID; GM11223GMNIGMS RN: 67-42-5; 73630-08-7; 7440-70-2 NM: Egtazic-Acid; 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic-acid; Calcium AN: 95016445 UD: 9501 Record 15 of 131 - MEDLINE (R) 1994 TI: Nitric oxide is involved in control of Trypanosoma cruzi-induced parasitemia and directly kills the parasite in vitro. AU: Vespa-GN; Cunha-FQ; Silva-JS AD: Department of Immunology, Faculty of Medicine of Ribeirao Preto, Sao Paulo, Brazil. SO: Infect-Immun. 1994 Nov; 62(11): 5177-82 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: This study was carried out to determine the role of reactive nitrogen intermediates in Trypanosoma cruzi infection. In vitro, splenocytes obtained during the acute phase of infection produced elevated amounts of nitric oxide (NO) that were correlated with the resistance or susceptibility of the animals. In vivo, the levels of NO2- plus NO3- in plasma during the later phase of infection were higher in C57BL/6 mice than in BALBL/c mice. The treatment of infected C57BL/6 mice with inhibitors of NO synthase increased parasitemia and mortality. Finally, we found that the NO donor drug S-nitroso-acetyl-penicillamine is able to kill trypomastigotes in vitro in the absence of any other cells, suggesting a direct NO-mediated killing of T. cruzi. MIME: Amino-Acid-Oxidoreductases-antagonists-and-inhibitors; Arginine-analogs-and-derivatives; Arginine-pharmacology; Chagas-Disease-blood; Mice-; Mice,-Inbred-BALB-C; Mice,-Inbred-C57BL; Nitrates-blood; Nitric-Oxide-pharmacology; Nitrites-blood; Spleen-parasitology; Trypanosoma-cruzi-drug-effects MJME: *Chagas-Disease-immunology; *Nitric-Oxide-metabolism; *Trypanosoma-cruzi-growth-and-development TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; EC 1.4.; 0; 0; 10102-43-9; 17035-90-4; 2149-70-4; 7004-12-8 NM: nitric-oxide-synthase; Amino-Acid-Oxidoreductases; Nitrates; Nitrites; Nitric-Oxide; omega-N-methylarginine; nitroarginine; Arginine AN: 95012744 UD: 9501 Record 16 of 131 - MEDLINE (R) 1994 TI: The wisent (Bison bonasus, Bovidae) as an intermediate host of three Sarcocystis species (Apicomplexa: Sarcocystidae) of cattle. AU: Odening-K; Wesemeier-HH; Walter-G; Bockhardt-I AD: Institute for Zoo Biology and Wildlife Research, Berlin, Germany. SO: Folia-Parasitol-Praha. 1994; 41(2): 115-21 ISSN: 0015-5683 PY: 1994 LA: ENGLISH CP: CZECH-REPUBLIC AB: Sarocysts were found in muscle tissue of a wisent (Bison bonasus) which was born and kept in Germany. Light microscopic and TEM examination revealed all the three named species known from cattle: Sarcocystis cruzi ("thin-walled", with longer hair-like villar protrusions of the primary cyst wall); S. hirsuta ("thick-walled", with tongue-like protrusions of the cyst wall arising with very short and narrow stalklets from the surface of the cyst and containing rows of electron-dense granules in the core); and S. hominis ("thick-walled", with finger-like protrusions of the cyst wall not constricted at their base and containing few or no electron-dense granules). So far, only S. cruzi was known to occur in Bison bison in North America. The findings in the wisent strikingly support a modified conception of the intermediate host specificity in Bovinae. In this connection the identity of S. cruzi and S. poephagicanis is suggested as well as that of S. hirsuta and S. poephagi. MIME: Animals,-Zoo-parasitology; Cattle-Diseases-transmission; Germany-; Microscopy,-Electron; Muscle,-Skeletal-pathology; Muscle,-Smooth-parasitology; Organ-Specificity; Sarcocystis-ultrastructure; Sarcocystosis-transmission; Sarcocystosis-veterinary; Species-Specificity MJME: *Bison-parasitology; *Cattle-parasitology; *Disease-Vectors; *Sarcocystis-isolation-and-purification TG: Animal; Female PT: JOURNAL-ARTICLE AN: 95011891 UD: 9501 Record 17 of 131 - MEDLINE (R) 1994 TI: Trans-sialidase and sialidase activities discriminate between morphologically indistinguishable trypanosomatids. AU: Medina-Acosta-E; Franco-AM; Jansen-AM; Sampol-M; Neves-N; Pontes-de-Carvalho-L; Grimaldi-Junior-G; Nussenzweig-V AD: New York University Medical Center, Michael Heidelberger Division of Immunology, New York 10016. SO: Eur-J-Biochem. 1994 Oct 1; 225(1): 333-9 ISSN: 0014-2956 PY: 1994 LA: ENGLISH CP: GERMANY AB: The expression of trans-sialidase and sialidase activities in the kinetoplastid protozoa was explored as a potential marker to discriminate between the morphologically indistinguishable flagellates isolated from human, insects and vertebrate reservoir hosts. By virtue of the differences observed in the ratios of these enzyme activities, a collection of 52 species and strains comprising the major taxa of these parasites could be separated into four expression types. Type-I parasites express comparable levels of both trans-sialidase and sialidase activities (Endotrypanum species and Trypanosoma lewisi). Type-II parasites express predominantly trans-sialidase activity (Trypanosoma cruzi and Trypanosoma conorhini). Type-III parasites express sialidase activity exclusively (Trypanosoma rangeli and Trypanosoma leeuwenhoeki). Type-IV parasites do not express either activity (Leishmania species and Trypanoplasma borreli). The measurement of trans-sialidase and sialidase activities thus permits the differentiation of parasites frequently found in the same insect vectors that are difficult to distinguish, such as T. cruzi and T. rangeli, or in the same sylvatic vertebrate and invertebrate hosts, such as Leishmania and Endotrypanum. MIME: Disease-Reservoirs; DNA,-Protozoan-isolation-and-purification; Insects-; Neuraminidase-biosynthesis; Polymerase-Chain-Reaction; Sialyltransferases-biosynthesis; Trypanosoma-isolation-and-purification; Trypanosomatina-enzymology; Trypanosomatina-isolation-and-purification; Vertebrates- MJME: *Neuraminidase-analysis; *Sialyltransferases-analysis; *Trypanosoma-classification; *Trypanosoma-enzymology; *Trypanosomatina-classification TG: Animal; Comparative-Study; Human; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI32966AINIAID RN: EC; 0; 0 NM: Neuraminidase; DNA,-Protozoan; Sialyltransferases AN: 95010122 UD: 9501 Record 18 of 131 - MEDLINE (R) 1994 TI: Experimental Chagas' disease: electrophysiology and cell composition of the neuromyopathic inflammatory lesions in mice infected with a myotropic and a pantropic strain of Trypanosoma cruzi. AU: Mirkin-GA; Jones-M; Sanz-OP; Rey-R; Sica-RE; Gonzalez-Cappa-SM AD: Departamento de Microbiologia, Facultad de Medicina, Universidad de Buenos Aires, Argentina. SO: Clin-Immunol-Immunopathol. 1994 Oct; 73(1): 69-79 ISSN: 0090-1229 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: C3H/HeN mice infected with the pantropic/reticulotropic Trypanosoma cruzi RA strain disclosed electromyographic signs (EMG) of neuropathic damage, while those infected with the myotropic CA-I strain showed EMG suggestive of primary muscle involvement. Although both strains induced inflammatory infiltrates in hamstring muscles (HM), damage was more severe in mice infected with CA-I. In sciatic nerves (SN) of mice infected with the RA strain, increased inflammatory changes, amastigote nests, and myelin digestion chambers were consistently found during the course of infection. On the other hand, the CA-I strain produced minor inflammatory changes without detectable amastigotes in such tissue. The RA strain induced chronic leptomeningitis in spinal cord (SC), while infiltrates were limited to spinal roots and dorsal ganglia in animals infected with CA-I. In mice infected with RA, phenotypic analysis of inflammatory lesions showed a consistent predominance of CD8+ T cells in nervous tissue throughout the course of infection and in HM during the chronic phase whereas natural killer cells were detected at 120 and 270 days pi. In mice infected with CA-I, a predominance of CD8+ cells in SN was only detected during the acute phase and in HM during the late chronic phase; B lymphocytes bearing surface IgM were present in all studied tissues at 270 days pi. In addition, positive fluorescence for mouse IgG was observed at 120 days pi in muscle interstitium. These results strongly suggest that T. cruzi strain-dependent mechanisms are involved in the development of neuromyopathic damage. MIME: Antigens,-Protozoan-analysis; CD8-Positive-T-Lymphocytes; Electromyography-; Mice-; Mice,-Inbred-C3H; Muscle,-Skeletal-parasitology; Neuromuscular-Junction-pathology; Neuromuscular-Junction-parasitology; Sciatic-Nerve-parasitology; Spinal-Cord-parasitology; Trypanosoma-cruzi-immunology MJME: *Chagas-Disease-pathology; *Chagas-Disease-physiopathology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0 NM: Antigens,-Protozoan AN: 95008384 UD: 9501 Record 19 of 131 - MEDLINE (R) 1994 TI: Immunoassay with recombinant Trypanosoma cruzi antigens potentially useful for screening donated blood and diagnosing Chagas disease. AU: Pastini-AC; Iglesias-SR; Carricarte-VC; Guerin-ME; Sanchez-DO; Frasch-AC AD: Instituto de Investigaciones Bioquimicas Fundacion Campomar, Buenos Aires, Argentina. SO: Clin-Chem. 1994 Oct; 40(10): 1893-7 ISSN: 0009-9147 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: A new enzyme immunoassay (EIA), the Dia Kit Bio-Chagas assay (Gador S.A.), is potentially useful for detecting antibodies to Trypanosoma cruzi in the diagnosis of infected individuals and the screening of blood in blood banks. The EIA is carried out on test strips of plastic backing covered with a nitrocellulose membrane to which a mixture of recombinant T. cruzi antigens 1, 2, shed acute-phase antigen, 13, and 30 has been applied as a horizontal line. A horizontal line of human IgG is included to monitor the test procedure. The test strips exhibited homogeneity in the adsorption of the mixture of recombinant antigens (CV = 6.0%) and the human IgG (CV = 8.6%). The EIA results obtained with sera positive by xenodiagnosis showed 100% agreement between both types of tests; tested against sera with positive and negative matched results of indirect hemagglutination, immunofluorescence, and ELISA, the EIA results agreed for 99.1% (347 of 350) and 99.6% (299 of 300) of the samples, respectively. MIME: beta-Galactosidase-genetics; Adsorption-; Antigens,-Protozoan-genetics; Cloning,-Molecular; Immunoenzyme-Techniques-statistics-and-numerical-data; Reagent-Kits,-Diagnostic-statistics-and-numerical-data; Recombinant-Fusion-Proteins; Recombinant-Proteins-immunology; Sensitivity-and-Specificity MJME: *Antibodies,-Protozoan-blood; *Antigens,-Protozoan-immunology; *Chagas-Disease-diagnosis; *Immunoenzyme-Techniques; *Trypanosoma-cruzi-immunology TG: Animal; Comparative-Study; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; 0; 0; 0; 0; 0 NM: beta-Galactosidase; Antibodies,-Protozoan; Antigens,-Protozoan; Reagent-Kits,-Diagnostic; Recombinant-Fusion-Proteins; Recombinant-Proteins AN: 95008216 UD: 9501 Record 20 of 131 - MEDLINE (R) 1994 TI: Presence of antiheart and antiskeletal muscle glycolipid autoantibodies in the sera of patients with chagasic cardiopathy. AU: Laguens-RP; Argel-MI; Chambo-J; Storino-R; Cabeza-Meckert-PM AD: Catedra de Patologia B, Facultad de Ciencias Medicas de la Universidad Nacional de La Plata, Argentina. SO: Can-J-Cardiol. 1994 Sep; 10(7): 769-76 ISSN: 0828-282X PY: 1994 LA: ENGLISH CP: CANADA AB: OBJECTIVE: To characterize biochemically and isolate the skeletal and heart muscle cell epitope recognized by the autoantibodies present in the serum of chronically infected Trypanosoma cruzi patients. Secondly, to use that epitope in an immunoenzymatic assay for determining differences in antibody titre among Chagas' and other protozoan and heart diseases and between asymptomatic and cardiopathic chagasic patients. DESIGN: Isolated human skeletal and heart muscle cells were treated with organic solvents, pronase, neuraminidase and sodium metaperiodate before immunofluorescence assay. Glycolipids were extracted from human skeletal muscle for ELISA. PATIENTS: Sera were collected from 155 patients with positive serology for T cruzi infection; 44 healthy blood bank donors; and from patients after heart transplantation (16 patients), during the first month after cardiac infarction (eight) or cardiotomy (10), dilated myocardiopathy (21), leishmaniasis (12), acute toxoplasmosis (four) and hyperthyroid ophthalmopathy (five). MAIN RESULTS: Immunofluorescence assay revealed that the chagasic sera recognized epitopes that appeared to be glycolipid in nature. ELISA showed that the chagasic sera contained a higher titre of antiskeletal muscle glycolipid antibodies than the control sera and that, in the chagasic population, antibody titre was significantly higher in patients with heart failure than in asymptomatic subjects or in those presenting only electrocardiographic abnormalities. CONCLUSIONS: The skeletal and heart muscle epitope recognized by antibodies present in the sera of chagasic patients has the characteristics of a glycolipid. ELISA with glycolipids extracted from human skeletal muscle indicated that chagasic patients presented a higher antibody titre and that patients with heart failure showed a titre significantly higher than those who were asymptomatic or with electrocardiographic abnormalities, suggesting that those antibodies could be immunological markers and even predictors of heart failure in Chagas' disease. MIME: Cardiomyopathy,-Congestive-immunology; Enzyme-Linked-Immunosorbent-Assay; Fluorescent-Antibody-Technique; Leishmaniasis-immunology; Myocardial-Infarction-immunology; Toxoplasmosis-immunology MJME: *Autoantibodies-blood; *Chagas-Cardiomyopathy-immunology; *Glycolipids-immunology; *Muscle,-Skeletal-immunology; *Myocardium-immunology TG: Human; In-Vitro PT: JOURNAL-ARTICLE RN: 0; 0 NM: Autoantibodies; Glycolipids AN: 95007192 UD: 9501 Record 21 of 131 - MEDLINE (R) 1994 TI: In vitro culture of cardiac mast cells from mice experimentally infected with Trypanosoma cruzi. AU: Postan-M; Correa-R; Ferrans-VJ; Tarleton-RL AD: Department of Zoology, University of Georgia, Athens 30602. SO: Int-Arch-Allergy-Immunol. 1994 Nov; 105(3): 251-7 ISSN: 1018-2438 PY: 1994 LA: ENGLISH CP: SWITZERLAND AB: In this study we describe a mast cell population obtained by in vitro culture of hearts of Trypanosoma-cruzi-infected mice. Heart tissue was placed in culture and observed daily for the efflux of cells. Mast cells appeared in the medium during the 1st week of culture in the area immediately surrounding the tissues and increased in numbers over time. The cells were identified as mast cells by electron microscopy and by positive staining of granules with Giemsa, toluidine blue, and berberine sulfate techniques. Histopathological analysis of the cultured heart fragments from infected mice showed numerous mast cells, located mostly in areas where the histologic structure was abnormal and surrounded by fibrous connective tissue. This is the first report on in situ proliferation and migration of mast cells form inflamed heart tissues. In situ grown mast cells might be useful in developing in vitro models to study the role of these cells in T. cruzi-induced and other myocardiopathies. MIME: Cells,-Cultured; Mast-Cells-ultrastructure; Mice-; Mice,-Inbred-C3H; Microscopy-; Microscopy,-Electron; Microscopy,-Phase-Contrast; Myocardium-ultrastructure MJME: *Chagas-Disease-pathology; *Mast-Cells-pathology; *Myocardium-pathology TG: Animal; Male; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI22070AINIAID AN: 95003755 UD: 9501 Record 22 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi: sequence of the ATP-binding site of a P-glycoprotein gene. AU: Dallagiovanna-B; Castanys-S; Gamarro-F AD: Instituto de Parasitologia y Biomedicina Lopez-Neyra, Consejo Superior de Investigaciones Cientificas, Granada, Spain. SO: Exp-Parasitol. 1994 Aug; 79(1): 63-7 ISSN: 0014-4894 PY: 1994 LA: ENGLISH CP: UNITED-STATES MIME: Amino-Acid-Sequence; Base-Sequence; Binding-Sites; Blotting,-Northern; Blotting,-Southern; Carrier-Proteins-chemistry; Carrier-Proteins-metabolism; Conserved-Sequence; Drug-Resistance-genetics; DNA-Primers-chemistry; DNA,-Protozoan-analysis; Genes,-Protozoan; Membrane-Glycoproteins-chemistry; Membrane-Glycoproteins-metabolism; Molecular-Sequence-Data; Polymerase-Chain-Reaction; RNA,-Protozoan-analysis; Sequence-Alignment; Trypanosoma-cruzi-chemistry MJME: *Adenosine-Triphosphate-metabolism; *Carrier-Proteins-genetics; *Membrane-Glycoproteins-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't GS: tcpgp1; ltpgpA; HuMRP; HuCFTR; Ehpgp1; Humdr1; Pfmdr1; ldmdr1 PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0; 0; 56-65-5 NM: Carrier-Proteins; DNA-Primers; DNA,-Protozoan; Membrane-Glycoproteins; P-Glycoprotein; RNA,-Protozoan; Adenosine-Triphosphate AN: 94326875 UD: 9411 SI: GENBANK/L20818 Record 23 of 131 - MEDLINE (R) 1994 TI: Effect of protein kinase inhibitors on the invasion process of macrophages by Trypanosoma cruzi. AU: Vieira-MC; de-Carvalho-TU; de-Souza-W AD: Laboratorio de Ultraestrutura Celular Hertha Meyer, Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Brasil. SO: Biochem-Biophys-Res-Commun. 1994 Sep 15; 203(2): 967-71 ISSN: 0006-291X PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The interaction of Trypanosoma cruzi with different vertebrate cells involves two distinct steps, attachment and internalization. Genistein and staurosporine, drugs which inhibit protein kinases, specially tyrosine kinase, are able to block the infection of macrophages by T. cruzi, suggesting that protein phosphorylation is a key event on this process. MIME: Alkaloids-pharmacology; Isoflavones-pharmacology; Mice- MJME: *Macrophages,-Peritoneal-parasitology; *Protein-Kinases-antagonists-and-inhibitors; *Trypanosoma-cruzi-drug-effects; *Trypanosoma-cruzi-physiology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; 0; 0; 446-72-0; 62996-74-1 NM: Protein-Kinases; Alkaloids; Isoflavones; genistein; staurosporine AN: 94380083 UD: 9412 Record 24 of 131 - MEDLINE (R) 1994 TI: Identification of immunodominant epitopes in Trypanosoma cruzi trypomastigote surface antigen-1 protein that mask protective epitopes. AU: Wrightsman-RA; Dawson-BD; Fouts-DL; Manning-JE AD: Department of Molecular Biology and Biochemistry, University of California, Irvine 92717. SO: J-Immunol. 1994 Oct 1; 153(7): 3148-54 ISSN: 0022-1767 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The gene that encodes trypomastigote surface Ag-1 (TSA-1), a major surface Ag of the bloodstream trypomastigote stage of Trypanosoma cruzi, was expressed in a baculovirus expression system. To determine the epitope(s) in TSA-1 that was recognized during T. cruzi infection and after immunization with TSA-1, subregions of the TSA-1 gene were expressed in a bacterial expression system. As seen by Western blotting, both mice and rabbits immunized with recombinant TSA-1 protein, as well as T. cruzi-infected mice, developed strong immune responses to the carboxyl-proximal region of TSA-1, but show no reaction to the amino-proximal portion of TSA-1. When mice were immunized with either recombinant TSA-1 protein or the carboxyl-proximal region of TSA-1, they did not survive challenge with 10(3) bloodstream trypomastigotes. However, 70% of the mice immunized with the amino-proximal portion of TSA-1 survived challenge with 10(3) bloodstream trypomastigotes. Thus, the immune responses elicited by recombinant TSA-1 or the carboxyl-proximal portion of TSA-1 are nonprotective during T. cruzi infection. In contrast, vaccination with the amino proximal region of TSA-1 elicits a protective immune response. These results suggest that responses to immunodominant epitope(s) within the carboxyl-proximal portion of TSA-1 mask epitopes within the amino-proximal portion that are capable of stimulating host-protective immune responses. It is suggested that immunodominant regions in surface molecules such as TSA-1 may provide a mechanism for the parasite to evade the host immune response by directing the response away from epitopes that have the potential to elicit a reaction that is damaging to the parasite. MIME: Antigenic-Determinants; Immunization-; Lymphocyte-Transformation; Mice-; Recombinant-Proteins-immunology; T-Lymphocytes-immunology MJME: *Antigens,-Protozoan-immunology; *Antigens,-Surface-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI18873AINIAID RN: 0; 0; 0; 0; 138820-03-8 NM: Antigenic-Determinants; Antigens,-Protozoan; Antigens,-Surface; Recombinant-Proteins; trypomastigote-specific-surface-antigen AN: 94375878 UD: 9412 SB: AIM Record 25 of 131 - MEDLINE (R) 1994 TI: Role of sialic acid in the resistance of Trypanosoma cruzi trypomastigotes to complement. AU: Tomlinson-S; Pontes-de-Carvalho-LC; Vandekerckhove-F; Nussenzweig-V AD: Michael Heidelberger Division of Immunology, Kaplan Cancer Center, Department of Pathology, New York University Medical Center, NY 10016. SO: J-Immunol. 1994 Oct 1; 153(7): 3141-7 ISSN: 0022-1767 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Trypomastigotes of Trypanosoma cruzi, mammalian infective forms of the parasite, express an unusual cell surface trans-sialidase. This enzyme enables the parasite to rapidly sialylate its surface when supplied with alpha(2,3)-linked sialic acid from glycoconjugates in serum or on cell surfaces. Here we used a novel fluorescence-based, trypomastigote lysis assay to evaluate the role of sialic acid on the parasite's plasma membrane in providing protection against the complement cascade. Trypomastigotes were desialylated, and sialic acid removal was confirmed by a chemical assay and also by flow cytometry with the use of a mAb that recognizes a T. cruzi-sialylated epitope. Compared with sialylated trypomastigotes, which were completely refractory to lysis by human serum, only about 5% of the desialylated trypomastigotes were lysed by complement. However, further analysis revealed that the desialylated parasites had been resialylated during exposure to serum complement. Next we incubated desialylated trypomastigotes with samples of desialylated human serum. Although the sialidase-treated serum retained its full hemolytic activity, lysis of trypomastigotes increased only from 5 to 24%. This increase correlated with an enhanced deposition of complement protein C3 on the parasite surface. The ratio of C3b to lytically inactive iC3b was increased for desialylated, compared with sialylated, parasites. We conclude that although parasite sialic acid promotes C3b cleavage into iC3b, this mechanism alone does not account for the robust resistance of these parasites to complement lysis. MIME: Complement-3-metabolism; Neuraminidase-metabolism; Trypsin-pharmacology MJME: *Complement-3-immunology; *Sialic-Acids-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Human; In-Vitro; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI32966AINIAID; AI08499AINIAID RN: EC; EC; 0; 0 NM: Neuraminidase; Trypsin; Complement-3; Sialic-Acids AN: 94375877 UD: 9412 SB: AIM Record 26 of 131 - MEDLINE (R) 1994 TI: IL-10 mediates susceptibility to Trypanosoma cruzi infection. AU: Reed-SG; Brownell-CE; Russo-DM; Silva-JS; Grabstein-KH; Morrissey-PJ AD: Infectious Disease Research Institute, Seattle, WA 98109. SO: J-Immunol. 1994 Oct 1; 153(7): 3135-40 ISSN: 0022-1767 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: IL-10 has been shown to inhibit some aspects of macrophage activation, including the in vitro IFN-gamma-mediated intracellular killing of the protozoan parasite Trypanosoma cruzi. We have previously shown that genetically susceptible mice produced more IL-10 during T. cruzi infection than resistant mice, suggesting an association between IL-10 production and disease susceptibility. In the present study, such an association was documented. IL-10 mRNA was present in the spleens of susceptible C57BL/6 mice, but not in resistant (C57BL/6 x DBA/2) F1 mice, as early as 2 days after infection with T. cruzi. In susceptible mice, IL-10 mRNA was found in enriched populations of splenic T cells and peritoneal macrophages by 4 days after infection. By 14 days after infection, IL-10 mRNA was detected in enriched populations of splenic T cells and peritoneal macrophages, as well as by splenic B cells and macrophages. In SCID mice infected with T. cruzi, IL-10 mRNA was detected in peritoneal cells 2 days after infection. The IL-10 mRNA production was not abolished by treatment with anti-asialo GM-1 Ab before infection, which is consistent with its production by macrophages. Finally, the role of endogenous IL-10 production in the susceptibility to T. cruzi infection was demonstrated by the protection of highly susceptible C57BL/6 mice against acute disease and death from T. cruzi by the administration of neutralizing anti-IL-10 mAb. This study demonstrated an important and perhaps essential role of IL-10 in mediating in vivo susceptibility to T. cruzi infection. MIME: Base-Sequence; DNA-Primers-chemistry; Gene-Expression; Interferon-Type-II-genetics; Killer-Cells,-Natural-immunology; Mice-; Mice,-Inbred-BALB-C; Mice,-Inbred-C57BL; Mice,-Inbred-DBA; Mice,-SCID; Molecular-Sequence-Data; RNA,-Messenger-genetics; Spleen-cytology MJME: *Chagas-Disease-immunology; *Interleukin-10-physiology; *Trypanosoma-cruzi-immunology TG: Animal; Female; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI22726AINIAID; AI16282AINIAID; TW04346TWFIC RN: 0; 0; 130068-27-8; 82115-62-6 NM: DNA-Primers; RNA,-Messenger; Interleukin-10; Interferon-Type-II AN: 94375876 UD: 9412 SB: AIM Record 27 of 131 - MEDLINE (R) 1994 TI: Changes in Trypanosoma cruzi kinetoplast DNA minicircles induced by environmental conditions and subcloning. AU: Alves-AM; De-Almeida-DF; von-Kruger-WM AD: Laboratorio de Fisiologia Celular, Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Brazil. SO: J-Eukaryot-Microbiol. 1994 Jul-Aug; 41(4): 415-9 ISSN: 1066-5234 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Reversible changes in kinetoplast DNA (kDNA) minicircles sequences were observed in clones of Trypanosoma cruzi strain Y, following a number of passages during exponential growth phase or after subcloning in blood-free medium. kDNA restriction patterns of clones were similar to those of the original uncloned strain, while subclones presented distinct kDNA restriction patterns. Homology experiments demonstrated strong hybridization between kDNA with the same electrophoretic mobility patterns while only weak signals were observed with kDNA of different patterns. The changes observed, which are unprecedented in T. cruzi clones, characterize transkinetoplastidy, and seem to be associated with similarly reversible changes both in zymodeme and in infectivity. MIME: Blotting,-Southern; Clone-Cells; DNA-Restriction-Enzymes; DNA,-Kinetoplast-genetics; Electrophoresis,-Polyacrylamide-Gel; Mice-; Nucleic-Acid-Hybridization; Trypanosoma-cruzi-growth-and-development MJME: *DNA,-Kinetoplast-analysis; *Trypanosoma-cruzi-genetics TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 3.1.21; 0 NM: DNA-Restriction-Enzymes; DNA,-Kinetoplast AN: 94372954 UD: 9412 Record 28 of 131 - MEDLINE (R) 1994 TI: Intracellular localization of a Trypanosoma cruzi kDNA minicircle transcript using RNA: RNA in situ hybridization. AU: Thertulien-R; Simpson-Haidaris-PJ; Haidaris-CG AD: Department of Microbiology & Immunology, University of Rochester School of Medicine and Dentistry, New York 14642. SO: J-Eukaryot-Microbiol. 1994 Jul-Aug; 41(4): 402-7 ISSN: 1066-5234 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Using RNA: RNA in situ hybridization, the intracellular location of a transcript encoded by and spanning the entire length of a Trypanosoma cruzi kinetoplast DNA minicircle was determined. In axenically cultured T. cruzi epimastigotes, the hybridization signal was restricted to the kinetoplast, which was situated in the perinuclear region of the cell. Following conversion of epimastigotes to culture-derived metacyclic trypomastigotes, the kinetoplast moved to an acentric position in the metacyclic trypomastigote. Again, the hybridization signal co-localized with the position of the kinetoplast. These results suggested that the transcript remained closely associated with the T. cruzi kinetoplast within the mitochondrion in each of the morphological forms. Using specific oligonucleotide probes derived from a cDNA encoding the transcript, the entire native kDNA minicircle encoding the transcript was cloned and its nucleotide sequence was determined. The nucleotide sequence of the intact native minicircle was identical to that of the full-length cDNA corresponding to the minicircle transcript, indicating that the transcript was not modified prior to the time of cDNA synthesis and cloning. MIME: Base-Sequence; Cloning,-Molecular; DNA,-Complementary-genetics; In-Situ-Hybridization; Molecular-Sequence-Data; RNA,-Messenger-genetics; RNA,-Protozoan-genetics; Trypanosoma-cruzi-growth-and-development MJME: *DNA,-Kinetoplast-genetics; *RNA,-Messenger-analysis; *RNA,-Protozoan-analysis; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: GM14943GMNIGMS; HL30616HLNHLBI RN: 0; 0; 0; 0 NM: DNA,-Complementary; DNA,-Kinetoplast; RNA,-Messenger; RNA,-Protozoan AN: 94372952 UD: 9412 SI: GENBANK/X56188 Record 29 of 131 - MEDLINE (R) 1994 TI: Trans-sialidase, SAPA amino acid repeats and the relationship between Trypanosoma cruzi and the mammalian host. AU: Frasch-AC AD: Instituto de investigaciones Bioquimicas Fundacion Campomar, Buenos Aires, Argentina. SO: Parasitology. 1994; 108 Suppl: S37-44 ISSN: 0031-1820 PY: 1994 LA: ENGLISH CP: ENGLAND AB: During invasion of multicellular organisms, protozoan parasites expose functional molecules that become targets for the host immune response. Recent research on Trypanosoma cruzi, the agent of Chagas' disease, suggests a new model of how the parasite might deal with this problem. Several antigens of T. cruzi have tandemly repeated amino acid motifs in molecules with as yet unknown functions. In two cases, these repeats are in molecules with a defined structure or function. Both proteins are implicated in the invasion of host-cells by the parasite. One of these is the core protein of a putative mucin-like glycoprotein that has Thr/Pro-rich repeats which, by themselves, might define the structure of a highly O-glycosylated molecule. The other protein is SAPA/trans-sialidase/neuraminidase, a molecule able to transfer sialic acid, that has so far only been described in trypanosomes. The amino acid repeats present in SAPA/transsialidase/neuraminidase are unrelated to the enzymic activity and constitute an immunodominant C-terminal domain. The N-terminal domain of SAPA/trans-sialidase/neuraminidase controls the enzymic activity since a recombinant molecule lacking the repeats conserves trans-sialidase activity. That both domains are functionally independent is also indicated by experiments that show that antibodies directed against the amino acid repeats are unable to inhibit trans-sialidase activity. A large number of proteins having trans-sialidase related sequences but lacking enzymic activity are also present in the surface membrane of the parasite. The immunodominant SAPA/trans-sialidase/neuraminidase repeats, together with the complex network of cross-reacting epitopes present in related but enzymatically inactive proteins might contribute to the delay in mounting an effective antibody response.(ABSTRACT TRUNCATED AT 250 WORDS) MIME: Amino-Acid-Sequence; Antigens,-Protozoan-chemistry; Glycoproteins-chemistry; Host-Parasite-Relations; Molecular-Sequence-Data; Neuraminidase-chemistry; Protozoan-Proteins-chemistry; Sequence-Homology,-Amino-Acid; Trypanosoma-cruzi-enzymology; Trypanosoma-cruzi-immunology MJME: *Antigens,-Protozoan-genetics; *Glycoproteins-genetics; *Neuraminidase-genetics; *Protozoan-Proteins-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE; REVIEW; REVIEW,-TUTORIAL RN: EC 3.2.1.-; EC; 0; 0; 0; 141176-81-0 NM: trans-sialidase,-Trypanosoma-cruzi; Neuraminidase; Antigens,-Protozoan; Glycoproteins; Protozoan-Proteins; shed-acute-phase-antigen,-Trypanosoma AN: 94366802 UD: 9412 Record 30 of 131 - MEDLINE (R) 1994 TI: Disposition of nifurtimox and metabolite activity against Trypanosoma cruzi using rat isolated perfused liver. AU: Gonzalez-Martin-G; Paulos-C; Guevara-A; Ponce-G AD: Department of Pharmacy, Faculty of Chemistry, Pontifical Catholic University of Chile, Santiago. SO: J-Pharm-Pharmacol. 1994 May; 46(5): 356-9 ISSN: 0022-3573 PY: 1994 LA: ENGLISH CP: ENGLAND AB: Nifurtimox disposition was investigated using the rat isolated perfused-liver method after administration of 25 micrograms mL-1 nifurtimox, and its disappearance was monitored by analysing the perfusate sample at various times. Biliary excretion was also measured. The drug concentration profile underwent a biexponential decline over the 2-h study period, with a terminal half-life of 62.76 +/- 17.56 min. Nifurtimox is a high clearance compound (15.23 +/- 5.53 mL min-1). The extraction ratio was 0.621 +/- 0.159. Biliary excretion accounted for 0.05% of the dose, the remainder consisting of highly polar metabolites. By 2 h, a minimal fraction of unchanged nifurtimox was recovered from the perfusate. Nifurtimox activity against Trypanosoma cruzi (clone CA-1) during the perfusion was also determined. Epimastigotes isolated from continuous culture were exposed to the samples of perfusate at different perfusion times in a microtitre plate. After an incubation time of 72 h at 27 degrees C, the parasite number in each well was counted under a microscope. From 0 to 75 min after the perfusion, the anti-trypanosomal activity decreased, but an increase in activity was observed at the later times. These findings show that active metabolites are formed during the perfusion. MIME: Bile-metabolism; Half-Life; Liver-drug-effects; Nifurtimox-pharmacology; Perfusion-; Rats-; Rats,-Sprague-Dawley MJME: *Liver-metabolism; *Nifurtimox-pharmacokinetics; *Trypanosoma-cruzi-drug-effects TG: Animal; Human; In-Vitro; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 23256-30-6 NM: Nifurtimox AN: 94365754 UD: 9412 Record 31 of 131 - MEDLINE (R) 1994 TI: Hydroxyurea-induced synchrony of DNA replication in the Kinetoplastida. AU: Galanti-N; Dvorak-JA; Grenet-J; McDaniel-JP AD: Department of Cell Biology and Genetics, School of Medicine, University of Chile, Santiago. SO: Exp-Cell-Res. 1994 Sep; 214(1): 225-30 ISSN: 0014-4827 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: We have developed a reliable and reproducible method to induce synchrony of the DNA synthetic cycle in the Kinetoplastida. The method involves treatment of cultures with 20 mM hydroxyurea (HU) and fetal bovine serum. Both stationary-phase and exponential-phase cultures can be synchronized. However, in the case of exponential-phase cultures the population doubling time and rate of DNA synthesis of the population influenced the time of exposure to HU. The treatment of kinetoplastids with 20 mM HU did not adversely affect the cells as judged by oxygen consumption, RNA, and protein content. We postulate that the requirement for high HU levels, which would be toxic to vertebrate cells, may be due to a lower affinity of kinetoplastid ribonucleotide reductase, the target enzyme for HU. Some of the kinetoplastids are pathogens of man and his food chain. Consequently, the development of a reliable technique for synchronization of the kinetoplastids should not only permit a detailed analysis of their cellular and molecular biology but provide a means to collect and characterize biochemical and immunochemical substances relevant to the infectious process. MIME: Cell-Division-drug-effects; Crithidia-drug-effects; Flow-Cytometry; Oxygen-Consumption; Protozoan-Proteins-analysis; RNA,-Protozoan-analysis; Trypanosoma-cruzi-drug-effects MJME: *DNA-Replication-drug-effects; *DNA,-Protozoan-biosynthesis; *Hydroxyurea-pharmacology; *Kinetoplastida-drug-effects TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-Non-P.H.S. PT: JOURNAL-ARTICLE RN: 0; 0; 0; 127-07-1 NM: DNA,-Protozoan; Protozoan-Proteins; RNA,-Protozoan; Hydroxyurea AN: 94364422 UD: 9412 Record 32 of 131 - MEDLINE (R) 1994 TI: Cross-reactivity studies and differential serodiagnosis of human infections caused by Trypanosoma cruzi and Leishmania spp; use of immunoblotting and ELISA with a purified antigen (Ag163B6). AU: Malchiodi-EL; Chiaramonte-MG; Taranto-NJ; Zwirner-NW; Margni-RA AD: Instituto de Estudios de la Inmunidad Humoral (IDEHU-CONICET), Catedra de Inmunologia, FFyB-UBA, Buenos Aires. SO: Clin-Exp-Immunol. 1994 Sep; 97(3): 417-23 ISSN: 0009-9104 PY: 1994 LA: ENGLISH CP: ENGLAND AB: Results of our studies on the reactivity of chagasic and leishmaniasic sera with the purified T. cruzi-specific antigen 163B6, as assessed by ELISA, and with complex antigenic mixtures from T. cruzi and Leishmania mexicana, by immunoblotting, are presented here. Our objective was to identify the antigens responsible for the exhibited cross-reactivity between trypanosomiasis and leishmaniasis, and to find a specific reactivity pattern corresponding to each parasitosis. In spite of the high cross-reactivity observed with the immunoblotting, the use of 7.5% A-B gels made it possible to identify a characteristic pattern for each parasitosis, that could be distinguished by the naked eye. The characteristic pattern corresponding to chagasic patients was ascribed to reactivity with T. cruzi bands of mol. wts 131, 125, 116, 111, 51-45 and 43 kD, that were not recognized by leishmaniasic sera. Trypanosoma cruzi antigens of mol. wts 85, 81, 70, 65-60, 37 and 32 kD were considered as crossing antigens, since they were recognized by leishmaniasis sera. With L. mexicana, most of the chagasic patients presented reaction with antigen of mol. wts 124, 107, 92, 59 and 32 kD, while bands of mol. wts 155, 140, 73, 56 and 48 kD were recognized only by leishmaniasic sera. In this study we found 12 out of 45 sera of patients with leishmaniasis, from a region endemic for both parasitoses, which exhibited a pattern of bands very similar to those corresponding to chagasic individuals, strongly suggesting a mixed infection. This hypothesis was verified by using a purified specific antigen of T. cruzi, Ag163B6, which would be the major cysteine proteinase of this specie (cruzipain). By ELISA, these 12 sera showed a positive reaction with this purified antigen, as those of chagasic patients, thus leading to the confirmation of the presence of a mixed infection. MIME: Antibodies,-Protozoan-analysis; Antigens,-Protozoan-isolation-and-purification; Chagas-Disease-immunology; Electrophoresis,-Polyacrylamide-Gel; Enzyme-Linked-Immunosorbent-Assay; Immunoblotting-; Leishmaniasis,-Cutaneous-immunology; Molecular-Weight; Serodiagnosis- MJME: *Antigens,-Protozoan-immunology; *Chagas-Disease-diagnosis; *Cross-Reactions-immunology; *Leishmania-mexicana-immunology; *Leishmaniasis,-Cutaneous-diagnosis; *Trypanosoma-cruzi-immunology TG: Animal; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0 NM: Antibodies,-Protozoan; Antigens,-Protozoan AN: 94363905 UD: 9412 Record 33 of 131 - MEDLINE (R) 1994 TI: Ca2+ homeostasis and mitochondrial bioenergetics in Trypanosoma cruzi. AU: Vercesi-AE AD: Departamento de Bioquimica, Universidade Estadual de Campinas, Brasil. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 501-3 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: Digitonin can be used to selectively permeabilize the plasma membrane of Trypanosoma cruzi without significantly affecting the functional integrity of mitochondria or the endoplasmic reticulum. This permits the study of functional properties of these subcellular organelles in situ, such as respiration, oxidative phosphorylation and Ca2+ transport. The ability of these cells to take up and hydrolyze Fura-2/AM, followed by the retention of the fluorescent Ca2+ indicator Fura-2, permits the determination of cytosolyc free Ca2+ concentrations in different T. cruzi stages. MIME: Digitonin-metabolism; Fura-2-metabolism; Homeostasis-; Membrane-Potentials MJME: *Calcium-metabolism; *Mitochondria-metabolism; *Trypanosoma-cruzi-metabolism TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 11024-24-1; 7440-70-2; 96314-98-6 NM: Digitonin; Calcium; Fura-2 AN: 94362579 UD: 9412 Record 34 of 131 - MEDLINE (R) 1994 TI: The alpha-mannosidase of Trypanosoma cruzi: structure and function. AU: Oeltmann-T; Carter-C; Merkle-R; Moreman-K AD: Vanderbilt University, Nashville, TN 37232. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 483-8 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: Trypanosoma cruzi alpha-mannosidase has been purified to apparent homogeneity. It is a 240,000-Da tetramer composed of four identical subunits (58,000 Da). Each subunit contains one N-linked high-mannose oligosaccharide. Based on pH optimum and sensitivity to inhibition by swainsonine, we suggest it to be lysosomal, but this has yet to be demonstrated directly. The enzyme appears to be developmentally regulated and may be a key enzyme in the degradation of the lipopeptidophosphoglycan (LPPG) during transformation from epimastigote to trypomastigote. Preliminary experiments suggest T. cruzi does not utilize the mannose 6-phosphate recognition system for sorting alpha-mannosidase (or other acid hydrolases) to the lysosome. To clone the alpha-mannosidase from T. cruzi we have used the same approach that has been used for other alpha-mannosidases. The cDNA amplification product was subcloned and sequenced. Comparison of the T. cruzi alpha-mannosidase sequence with the alpha-mannosidases that were used in the original primer design demonstrated a greater similarity to murine lysosomal and Dictyostelium alpha-mannosidases than to Golgi alpha-mannosidases. MIME: Base-Sequence; Carbohydrate-Sequence; Mannosidases-isolation-and-purification; Mannosidases-metabolism; Molecular-Sequence-Data; Polymerase-Chain-Reaction; Structure-Activity-Relationship MJME: *Mannosidases-chemistry; *Trypanosoma-cruzi-enzymology TG: Animal PT: JOURNAL-ARTICLE RN: EC 3.2.1. NM: Mannosidases AN: 94362576 UD: 9412 Record 35 of 131 - MEDLINE (R) 1994 TI: Identification of a putative pore-forming hemolysin active at acid pH in Leishmania amazonensis. AU: Noronha-FS; Ramalho-Pinto-FJ; Horta-MF AD: Departamento de Bioquimica-Imunologia, Universidade Federal de Minas Gerais, Belo Horizonte, Brasil. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 477-82 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: Several organisms, including the protozoa Entamoeba histolytica and Trypanosoma cruzi, have been shown to contain pore-forming proteins (PFP) thought to play a role in the pathogenesis of the diseases they generate. In the present report, we show that promastigotes of Leishmania amazonensis express a hemolysin that appears to cause colloid-osmotic lysis, typical of pore formation. This hemolysin affects red blood cells of different species at 37 degrees C, but not at 0 degrees C, with maximum activity at pH 5.0. The hemolytic activity is heat-labile, but lysis is not affected by protease inhibitors. These results suggest the involvement of a protein with no proteolytic or detergent activity. Hemolysis is inhibited by polyethyleneglycol, suggesting its colloid-osmotic nature. Hemolytic extracts of the parasite contain a polypeptide that reacts with antibodies to perforin from mouse cytotoxic T lymphocytes or to C9 from human complement. In addition, genomic DNA of L. amazonensis contains a fragment that hybridizes to a perforin cDNA probe. The circumstantial evidence suggests that the L. amazonensis hemolytic activity may be mediated by a PFP homologous to perforin and C9. MIME: Hydrogen-Ion-Concentration; Leishmania-physiology; Peptides-physiology MJME: *Hemolysins-metabolism; *Hemolysis-physiology; *Leishmania-chemistry; *Peptides-metabolism TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0 NM: Hemolysins; Peptides AN: 94362575 UD: 9412 Record 36 of 131 - MEDLINE (R) 1994 TI: From lysosomes into the cytosol: the intracellular pathway of Trypanosoma cruzi. AU: Andrews-NW AD: Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 471-5 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: The protozoan parasite Trypanosoma cruzi invades a wide variety of vertebrate cells, by a mechanism distinct from phagocytosis. No pseudopods or other host cell surface alterations are observed during trypanosome entry, and invasion is enhanced after actin filaments are disrupted with cytochalasin D. These observations created a puzzle; what is the origin of the membrane required to form the intracellular vacuoles, if it is not originated from the host cell plasma membrane? Recent observations provided the answer: during cell invasion T. cruzi recruits host lysosomes, which gradually fuse with the plasma membrane at the site of parasite entry. The membrane of the parasitophorous vacuole is, therefore, very similar if not identical to the membrane of lysosomes. Lgps, major glycoproteins from mammalian lysosomes, are desialylated by a glycosylphosphatidylinositol (GPI)-anchored trans-sialidase present on the surface of trypomastigote forms. Lack of sialic acid on lgps facilitates membrane lysis by a parasite-secreted molecule, Tc-TOX, that has membrane pore-forming activity at acidic pH. We propose a sequential model in which these trypanosome products would promote parasite entry into host cells and their subsequent liberation into the cytosol. MIME: Cell-Membrane-parasitology; Cell-Membrane-ultrastructure; Trypanosoma-cruzi-chemistry; Vacuoles-parasitology MJME: *Lysosomes-parasitology; *Sialic-Acids-physiology; *Trypanosoma-cruzi-physiology TG: Animal; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: R29AI27260AINIAID; RO1AI32056AINIAID RN: 0 NM: Sialic-Acids AN: 94362574 UD: 9412 Record 37 of 131 - MEDLINE (R) 1994 TI: GPI-anchored glycoconjugates from Trypanosoma cruzi trypomastigotes are recognized by lytic anti-alpha-galactosyl antibodies isolated from patients with chronic Chagas' disease. AU: Almeida-IC; Ferguson-MA; Schenkman-S; Travassos-LR AD: Disciplina de Biologia Celular, Escola Paulista de Medicina, Sao Paulo, SP, Brasil. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 443-7 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: The target molecules on the cell surface of Trypanosoma cruzi trypomastigotes reacting with lytic anti-alpha-galactosyl antibodies from chronic patients with Chagas' disease (Ch anti-Gal) have been purified by solvent extraction and identified as glycoconjugates migrating in the 74-96-kDa range (F2 antigen) and in the 120-200-kDa range (F3 antigen) on SDS-PAGE. The F3 antigen was tested for binding to Ch and normal human serum (NHS) anti-Gal and to MoAb 3C9. We observed that Ch anti-Gal and MoAb 3C9, but not NHS anti-Gal, bind strongly to the trypomastigote glycoconjugates. These antibodies, however, did not compete with each other for binding to F3 molecules, indicating that they are recognizing different epitopes. Binding of Ch anti-Gal to F3 antigen is abolished by treatment of these molecules with alpha- but not beta-galactosidase. Binding of 3C9 MoAb is abolished by treatment of F3 with sialidase. F2/F3 antigens absorbed Ch anti-Gal as well as lytic antibodies from total chagasic sera. These antigens also specifically discriminate between the serum reactivity of patients with active Chagas' disease and those of sera from cured patients, drug-treated patients with dissociated serology (positive conventional serology, negative trypanolytic activity), healthy individuals, and patients with several other infectious diseases. We also observed that F2/F3 antigens are anchored to the parasite membrane via glycosylphosphatidylinositol (GPI). The alpha-galactosyl epitopes recognized by Ch anti-Gal are present in a series of O-linked oligosaccharide chains in the mucin-like glycoprotein component of the complex. MIME: Blotting,-Western; Chronic-Disease; Enzyme-Linked-Immunosorbent-Assay; Glycoconjugates-chemistry; Glycoconjugates-immunology; Oligosaccharides-chemistry; Oligosaccharides-immunology MJME: *Antibodies,-Protozoan-immunology; *Antigens,-Protozoan-analysis; *Chagas-Disease-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0 NM: Antibodies,-Protozoan; Antigens,-Protozoan; Glycoconjugates; Oligosaccharides AN: 94362569 UD: 9412 Record 38 of 131 - MEDLINE (R) 1994 TI: Sialic acid acceptors of different stages of Trypanosoma cruzi are mucin-like glycoproteins linked to the parasite membrane by GPI anchors. AU: Acosta-A; Schenkman-RP; Schenkman-S AD: Disciplina de Biologia Celular, Escola Paulista de Medicina, Sao Paulo, Brasil. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 439-42 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: Trans-sialidase catalyzes the transference of sialic acid from host to the Trypanosoma cruzi surface. Here, we characterize the sialic acid acceptors of this protozoan parasite as mucin-like molecules, which are anchored to the membrane by glycosylphosphatidylinositol. The mucins isolated from the insect stages differ from the mucins isolated from the mammalian stages in size and reactivity to monoclonal antibodies, suggesting that they are formed by variable polypeptide chains and/or O-linked carbohydrate structures. MIME: Chromatography,-Agarose; Glycoproteins-isolation-and-purification; Mucins-metabolism; Trypanosoma-cruzi-physiology MJME: *Glycoproteins-metabolism; *Neuraminidase-metabolism; *Protozoan-Proteins-metabolism; *Sialic-Acids-metabolism; *Trypanosoma-cruzi-enzymology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; 0; 0; 0; 0 NM: Neuraminidase; Glycoproteins; Mucins; Protozoan-Proteins; Sialic-Acids AN: 94362568 UD: 9412 Record 39 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi: identification of a membrane cysteine proteinase linked through a GPI anchor. AU: Fresno-M; Hernandez-Munain-C; de-Diego-J; Rivas-L; Scharfstein-J; Bonay-P AD: Centro de Biologia Molecular, Universidad Autonoma de Madrid, Spain. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 431-7 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: The biochemical and functional properties of T. cruzi GP50/55, a novel glycosylphosphatidylinositol (GPI)-anchored membrane antigen have been investigated. A 50-52-kDa thiol proteinase activity could be immunoprecipitated with monoclonal antibodies (mAb) directed against GP50/55 (mAb C10), different from the one reactive with mAbs against lysosomal cysteine proteinase GP57/51. Furthermore, the mAb C10-reactive proteinase corresponded to the GPI-anchored surface antigen since the proteolytic and antigenic activity partitioned to the aqueous phase after Triton X114 phase separation of phosphatidylinositol specific phospholipase C (PI-PLC)-treated parasites. Of several proteins immunoprecipitated by a polyclonal anti-lysosomal cysteine proteinase, an mAb to GP57/51 recognized a 60-kDa protein, whereas mAb C10 recognized antigens ranging between 52 and 50 kDa. The GP50/55 antigen detected by mAb C10 is expressed on the parasite surface whereas the GP57/51 antigen is mainly intracellular. The internal peptide sequence obtained from purified GP50/55 showed that it is more homologous to the prototype of the cysteine proteinases superfamily, papain, than to the two T. cruzi lysosomal cysteine proteinases so far described. Our data indicate that the T. cruzi GP50/55 is a novel GPI-anchored cysteine proteinase and may represent another isoform of this heterogeneous group of proteinases. MIME: Antigens,-Protozoan-metabolism; Cysteine-Proteinases-metabolism; Flow-Cytometry; Glycoproteins-metabolism; Glycosylphosphatidylinositols-metabolism; Precipitin-Tests; Trypanosoma-cruzi-metabolism MJME: *Antigens,-Protozoan-chemistry; *Cysteine-Proteinases-chemistry; *Glycoproteins-chemistry; *Glycosylphosphatidylinositols-chemistry; *Trypanosoma-cruzi-chemistry TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 3.4.22; 0; 0; 0 NM: Cysteine-Proteinases; Antigens,-Protozoan; Glycoproteins; Glycosylphosphatidylinositols AN: 94362567 UD: 9412 Record 40 of 131 - MEDLINE (R) 1994 TI: Free and protein-linked glycoinositolphospholipids in Trypanosoma cruzi. AU: de-Lederkremer-RM AD: Departamento de Quimica Organica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 239-42 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: Two glycoinositol phospholipids (GIPL A and GIPL B) have been purified from epimastigotes of Trypanosoma cruzi at the logarithmic phase of growth (2 days). The GIPLs differ mainly in the lipid moiety and are similar to the lipopeptidophosphoglycan (LPPG) previously isolated from epimastigotes at the stationary phase (4-5 days). [3H]-palmitic acid was incorporated into 1-O-hexadecyl-2-O-palmitoylglycerol in GIPL A and into a sphinganine ceramide with palmitic acid and lignoceric acid as the fatty acids in GIPL B. The lipids could be released by incubation with phosphatidylinositol-specific phospholipase C (PI-PLC) or glycosylphosphatidylinositol phospholipase D (GPI-PLD) from rat serum. The oligosaccharides share the common core structure of the glycosylphosphatidylinositol (GPI) membrane anchors. Microheterogeneity was demonstrated, as well as substitution by galactose, which is mainly in the furanose configuration as was previously described for the LPPG. However, methylation analysis indicated that 20% of the galactose is present as terminal pyranose units. In infective trypomastigotes, [3H]-palmitic acid was incorporated into the anchor of the Tc-85 glycoprotein. The lipid cleaved by phospholipase C digestion was identified as 1-O-hexadecylglycerol and the main oligosaccharide has the structure of the conserved core of all GPI anchors. [3H]-palmitic acid-labelled Tc-85 released into the culture medium as membrane vesicles showed 80% resistance to the action of PI-PLC. However, after mild alkaline hydrolysis, part of the radioactivity was released by the enzyme. MIME: Carbohydrate-Sequence; Fatty-Acids-metabolism; Glycolipids-metabolism; Glycosylphosphatidylinositols-metabolism; Molecular-Sequence-Data; Phosphatidylinositols-metabolism; Protozoan-Proteins-metabolism; Trypanosoma-cruzi-metabolism MJME: *Glycolipids-chemistry; *Glycosylphosphatidylinositols-chemistry; *Phosphatidylinositols-chemistry; *Protozoan-Proteins-chemistry; *Trypanosoma-cruzi-chemistry TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0 NM: Fatty-Acids; Glycolipids; Glycosylphosphatidylinositols; Phosphatidylinositols; Protozoan-Proteins AN: 94362538 UD: 9412 Record 41 of 131 - MEDLINE (R) 1994 TI: Characterization of a phospholipase C-resistant inositol containing glycolipid from Trypanosoma cruzi. AU: Heise-N; Raper-J; Cardoso-de-Almeida-ML AD: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Sao Paulo, Brasil. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 233-8 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: Since glycosylphosphatidylinositol is the most common form of attachment of proteins to membranes in T. cruzi, and that this parasite depends on surface-mediated interactions for survival within the vector and mammalian host, it is probable that a drug which interfers with the metabolism of glycosylphosphatidylinositol (GPI) could be successfully employed in chemotherapy. Over the last few years several groups have been characterizing this mode of attachment in T. cruzi and more recently we have been concentrating our efforts on the identification of candidate precursors for protein anchors in metacyclic trypomastigotes. Previously detected GPI heterogeneity regarding solubilization of a major stage-specific antigen (1G7-Ag) by phospholipase C led us to investigate whether biosynthetic precursors with similar properties could also be identified. Two glycolipid species whose migration properties resemble glycolipids A and C of T. brucei were amenable to biosynthetic radiolabelling with palmitic acid, inositol, ethanolamine, glucosamine and mannose. Following purification, these species were submitted to classical GPI diagnostic treatments. In both cases digestion with GPI-specific phospholipase D (GPIPLD) produced phospatidic acid and treatment with either mild base or phospholipase A2 (PLA2) produced free fatty acid, indicating an acylation at least at position 2 of the glycerol. The glycolipid A-like species proved to be susceptible to solubilization by PIPLC of B. thuringiensis and by GPIPLC of T. brucei and the glycolipid C-like material proved to be fully resistant to both lipases.(ABSTRACT TRUNCATED AT 250 WORDS) MIME: Chromatography,-Thin-Layer; Fatty-Acids-metabolism; Glycolipids-metabolism; Glycosylphosphatidylinositols-metabolism; Phospholipase-C-metabolism; Protein-Precursors-metabolism; Protozoan-Proteins-metabolism; Trypanosoma-cruzi-metabolism MJME: *Fatty-Acids-chemistry; *Glycolipids-chemistry; *Glycosylphosphatidylinositols-chemistry; *Phospholipase-C-chemistry; *Protein-Precursors-chemistry; *Protozoan-Proteins-chemistry; *Trypanosoma-cruzi-chemistry TG: Animal PT: JOURNAL-ARTICLE RN: EC; 0; 0; 0; 0; 0 NM: Phospholipase-C; Fatty-Acids; Glycolipids; Glycosylphosphatidylinositols; Protein-Precursors; Protozoan-Proteins AN: 94362537 UD: 9412 Record 42 of 131 - MEDLINE (R) 1994 TI: Structural features of LPPG and related compounds isolated from Trypanosoma cruzi trypomastigotes. AU: Casal-OL; Zingales-B; Colli-W AD: Departamento de Quimica Organica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina. SO: Braz-J-Med-Biol-Res. 1994 Feb; 27(2): 227-31 ISSN: 0100-879X PY: 1994 LA: ENGLISH CP: BRAZIL AB: A hydrophobic fraction isolated from trypomastigotes of Trypanosoma cruzi is being characterized using immunological and chemical techniques. The lipopeptidophosphoglycan (LPPG) was identified in this fraction since it gave a positive reaction with anti-LPPG rabbit serum and had similar structural features such as the presence of ceramide as the lipid moiety, furanoic galactose, and a glycan moiety consistent with that obtained from an authentic sample of epimastigote LPPG, as judged by thin-layer chromatography. Furthermore, the hydrophobic fraction contained other glycolipids with different structural features. The lipid moiety of these compounds is alkylglycerol rather than a ceramide, the carbohydrate chain appears to be less complex than that in LPPG and no reactivity was observed towards an anti-LPPG serum. MIME: Carbohydrate-Sequence; Chromatography,-Thin-Layer; Glycoconjugates-chemistry; Molecular-Sequence-Data MJME: *Peptidoglycan-chemistry; *Phospholipids-chemistry; *Protozoan-Proteins-chemistry; *Trypanosoma-cruzi-chemistry TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0 NM: Glycoconjugates; Peptidoglycan; Phospholipids; Protozoan-Proteins AN: 94362536 UD: 9412 Record 43 of 131 - MEDLINE (R) 1994 TI: Isolation and characterization of the gene encoding histone H2A from Trypanosoma cruzi. AU: Puerta-C; Martin-J; Alonso-C; Lopez-MC AD: Instituto de Parasitologia y Biomedicina, Consejo Superior de Investigaciones Cientificas, Granada, Spain. SO: Mol-Biochem-Parasitol. 1994 Mar; 64(1): 1-10 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: In the present paper we report the isolation and characterization of the sequence of two genomic DNA fragments coding for the histone H2A of Trypanosoma cruzi. An analysis of the predicted amino acid sequence shows the presence of the amino-terminal motif characteristic of the H2A histones proteins and the Lys-Lys motif reported to be the site for the ubiquitin attachment. Southern blots of total parasite DNA probed with the H2A sequence suggested that the T. cruzi histone H2A gene is encoded in two independent gene clusters. The molecular karyotyping of the parasite indicated that these two clusters locate in a single chromosome of about 700 kb in length. The T. cruzi H2A mRNA is polyadenylated as are the basal histone mRNAs of higher eukaryotes and the histone mRNAs of yeast. By polymerase chain reaction amplification and sequencing and by S1 mapping we determined respectively the 5' and 3' end of the gene showing that the miniexon is added to the mRNA 71 nucleotides upstream of the ATG initiation codon and that the polyadenylation site locates in nucleotide position 773-775 close to invert repeats. MIME: Amino-Acid-Sequence; Base-Sequence; DNA,-Protozoan-genetics; Molecular-Sequence-Data; Nucleic-Acid-Conformation; Protozoa-genetics; Saccharomyces-cerevisiae-genetics; Sequence-Alignment; Sequence-Homology,-Amino-Acid; Species-Specificity MJME: *Genes,-Structural,-Protozoan; *Histones-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0 NM: DNA,-Protozoan; Histones AN: 94359525 UD: 9412 SI: GENBANK/X67287 Record 44 of 131 - MEDLINE (R) 1994 TI: Characterization of a short interspersed reiterated DNA sequence of Trypanosoma cruzi located at the 3'-end of a poly(A)+ transcript. AU: Requena-JM; Martin-F; Soto-M; Lopez-MC; Alonso-C AD: Centro de Biologia Molecular Severo Ochoa, CSIC-UAM, Madrid, Spain. SO: Gene. 1994 Sep 2; 146(2): 245-50 ISSN: 0378-1119 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: We have carried out the molecular characterization of a highly repeated DNA element, called E12, from Trypanosoma cruzi, which has been found to be interspersed along its genome. The E12 element, repeated about 5.6 x 10(3) times, is found in most of the chromosomal bands of the parasite. Three subregions may be defined within the element on the basis of sequence similarities with other trypanosome genomic sequences. Northern blot analysis demonstrated that sequences of the E12 element are present in several polyadenylated RNA species of T. cruzi. The isolation and characterization of a cDNA clone, pSPFM55, which showed hybridization with the E12 probe, indicated that only one of the E12 subregions, E12A, is found in the cDNA and that it is located at the 3'-end providing the site of polyadenylation addition. The location and high degree of nucleotide conservation of E12A suggest a possible functional role of this sequence in gene expression. MIME: Amino-Acid-Sequence; Base-Sequence; Blotting,-Southern; DNA,-Protozoan-isolation-and-purification; Molecular-Sequence-Data; Sequence-Homology,-Nucleic-Acid; Transcription,-Genetic MJME: *DNA,-Protozoan-genetics; *Repetitive-Sequences,-Nucleic-Acid-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0 NM: DNA,-Protozoan AN: 94357444 UD: 9412 SI: GENBANK/L22304 Record 45 of 131 - MEDLINE (R) 1994 TI: Animal reservoirs for Trypanosoma cruzi infection in an endemic area in Paraguay. AU: Fujita-O; Sanabria-L; Inchaustti-A; De-Arias-AR; Tomizawa-Y; Oku-Y AD: Department of Parasitology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan. SO: J-Vet-Med-Sci. 1994 Apr; 56(2): 305-8 ISSN: 0916-7250 PY: 1994 LA: ENGLISH CP: JAPAN AB: Animal reservoirs for Trypanosoma cruzi infection were investigated in 5 communities in the Department of San Pedro, currently one of Paraguay's most highly endemic areas. A total of 112 domestic animals (37 cattle, 2 horses, 1 ass, 20 pigs, 44 dogs and 8 cats) and 4 wild animals (1 white-eared opossum, 2 yellow armadillos and 1 common long-nosed armadillo) were examined for blood. Although no trypomastigotes were found by 2 direct observation methods, the microhaematocrit and Giemsa stained thick and thin smears methods, several forms of trypanosoma flagellates morphologically identical to T. cruzi were detected in the liver infusion tryptose (LIT) medium from a single sample taken from a yellow armadillo, Euphractus sexicintus. When serum samples of all the animals were examined for antibody to T. cruzi by direct agglutination (DA) test, 3 cattle, 2 pigs, 16 dogs and 3 cats had positive titers (1:32 or greater), but no wild animals showed positive reactions. T. cruzi was not found by culture nor microscopic examination of samples from any of the seropositive animals. However, domestic animals such as cattle, pigs, dogs and cats which were found to be seropositive in this study, possibly act as an animal reservoir in this endemic area as well as armadillos in which T. cruzi was observed. MIME: Animals,-Domestic; Animals,-Wild; Armadillos-; Cats-; Cattle-; Chagas-Disease-epidemiology; Demography-; Dogs-; Geography-; Horses-; Opossums-; Paraguay-; Swine- MJME: *Cat-Diseases; *Cattle-Diseases; *Chagas-Disease-veterinary; *Disease-Reservoirs; *Dog-Diseases; *Horse-Diseases; *Swine-Diseases TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE AN: 94355455 UD: 9412 Record 46 of 131 - MEDLINE (R) 1994 TI: [Detection of Trypanosoma cruzi in experimental samples by the DNA polymerase chain reaction method] TO: Deteccion de Trypanosoma cruzi en muestras experimentales por el metodo de reaccion en cadena de la ADN polimerasa. AU: Monteon-Padilla-VM; Reyes-Lopez-PA; Rosales-Encina-JL AD: Departamento de Inmunologia, Instituto Nacional de Cardiologia Ignacio Chavez, Mexico, D.F. SO: Arch-Inst-Cardiol-Mex. 1994 Mar-Apr; 64(2): 135-43 ISSN: 0020-3785 PY: 1994 LA: SPANISH; NON-ENGLISH CP: MEXICO AB: Chagas' disease (American Trypanosomiasis) affects more than 20 million people in Latin America. Almost 30% of those people may develop a chronic disease, which is expressed mainly as a chronic chagasic cardiopathy (CCC). Recent studies in Mexico have shown that 40% of patients suffering dilated cardiomyopathy do have serum antibodies against Trypanosoma cruzi. It is well known the difficulties of parasitologic diagnosis of CCC, which in less extent does exist for serologic diagnosis. Here we report a diagnostic method based on a molecular approach. It is able to recognize parasite DNA, and may have a clinical application. Two oligonucleotides (KNS1 and KNS2) designed from kinetoplast minicircle DNA, were used to amplify the hypervariable region by PCR technology. The method allowed an amplification of 0.8 to 1.5 minicircle DNA molecules, which equals 1/12,000 of parasite. When tissue DNA samples from mice infected with T. cruzi were subjected to amplification, a product was obtained that was recognized by a DNA probe specific for minicircle. These results correlate with immunohistochemical studies showing tissular parasites. Molecular diagnosis of American Trypanosomiasis, could be applied in human studies. MIME: Base-Sequence; Chagas-Cardiomyopathy-diagnosis; Chronic-Disease; DNA-Probes; DNA,-Kinetoplast-genetics; English-Abstract; Immunohistochemistry-; Mice-; Molecular-Sequence-Data; Trypanosoma-cruzi-genetics MJME: *Chagas-Cardiomyopathy-parasitology; *Polymerase-Chain-Reaction; *Trypanosoma-cruzi-isolation-and-purification TG: Animal; Comparative-Study; Human PT: JOURNAL-ARTICLE RN: 0; 0 NM: DNA-Probes; DNA,-Kinetoplast AN: 94354724 UD: 9412 Record 47 of 131 - MEDLINE (R) 1994 TI: [Acute-phase reaction and parasitism in the central adrenal vein in Chagas' disease patients] TO: Reacao de fase aguda e parasitismo na veia central da supra-renal de chagasicos cronicos. AU: da-Cunha-DF; Vieira-C-de-O; de-Paula-e-Silva-G; Eredia-GR; Teixeira-V-de-P AD: Disciplina de Nutrologia, Faculdade de Medicina do Triangulo Mineiro (FMTM), Uberaba, MG. SO: Rev-Soc-Bras-Med-Trop. 1994 Apr-Jun; 27(2): 83-6 ISSN: 0037-8682 PY: 1994 LA: PORTUGUESE; NON-ENGLISH CP: BRAZIL AB: The systemic reaction to severe trauma and/or infection, acute phase response (APR), are often associated with immunosuppression and reactivation of chronic latent infection. Our main purpose was to verify, in a group of 71 autopsied chronic chagasic with or without APR, the frequency of T. cruzi nests in the central vein of adrenal gland (CVAG). APR, defined by: 1) death secondary to sepsis and/or trauma plus, 2) bleeding stress gastric ulcerations or 3) spleen reactional state or 4) liver steatosis, was observed in 30 chronic chagasic (APR+). Weight, height and body mass index (BMI) were obtained. APR(+) chronic chagasic had worse nutritional status than APR(-) ones: weight = 49.0 vs 54.5 kg; BMI = 17.5 vs 20.6 kg/m2 (median p < 0.05). CVAG T. cruzi nests frequency were similar (43.3% and 43.9%, respectively) between both Groups. We conclude that APR(+) chronic chagasic had worse nutritional status than APR(-) ones, and that APR development did not change the CVAG T. cruzi nests frequency. MIME: Adult-; Aged-; Aged,-80-and-over; Chronic-Disease; English-Abstract; Immune-Tolerance; Middle-Age; Nutritional-Status; Trypanosoma-cruzi-isolation-and-purification; Veins-parasitology MJME: *Acute-Phase-Reaction-immunology; *Acute-Phase-Reaction-parasitology; *Adrenal-Glands-blood-supply; *Adrenal-Glands-parasitology; *Chagas-Disease-immunology; *Chagas-Disease-parasitology TG: Animal; Comparative-Study; Female; Human; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE AN: 94353095 UD: 9412 Record 48 of 131 - MEDLINE (R) 1994 TI: [The neurons of the cardiac plexus in acute Trypanosoma cruzi infection in white rats] TO: Estudo dos neuronios do plexo cardiaco na infeccao aguda pelo Trypanosoma cruzi em ratos albinos. AU: Chapadeiro-E; Beraldo-PS; Jesus-PC; Fernandes-PD; Junqueira-Junior-LF AD: Laboratorio Cardiovascular, Faculdade de Ciencias da Saude, Universidade de Brasilia, Brasil. SO: Rev-Soc-Bras-Med-Trop. 1994 Apr-Jun; 27(2): 79-81 ISSN: 0037-8682 PY: 1994 LA: PORTUGUESE; NON-ENGLISH CP: BRAZIL MIME: Acute-Disease; English-Abstract; Ganglia,-Parasympathetic-pathology; Myocardium-pathology; Rats- MJME: *Chagas-Cardiomyopathy-pathology; *Heart-innervation; *Neurons-pathology TG: Animal; Comparative-Study PT: JOURNAL-ARTICLE AN: 94353094 UD: 9412 Record 49 of 131 - MEDLINE (R) 1994 TI: Glycolipid and protein profiles in trypanosomatids. AU: Branquinha-MH; Bergter-EB; de-Meirelles-MN; Vermelho-AB AD: Departamento de Microbiologia Geral, Universidade Federal do Rio de Janeiro, Brasil. SO: Parasitol-Res. 1994; 80(4): 336-41 ISSN: 0932-0113 PY: 1994 LA: ENGLISH CP: GERMANY AB: A comparative study of glycolipid and protein composition in Trypanosoma cruzi and non-pathogenic trypanosomatids was carried out using Triton X-114 extraction. Protein profiles were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulphate (SDS-PAGE), and glycolipids were detected using high-performance thin-layer chromatography (HPTLC). Hydrophilic protein profiles were similar in non-pathogenic protozoa. Endotrypanum schaudinni, Crithidia luciliae and T. mega showed five characteristic protein bands ranging between 30 and 66 kDa. In the hydrophobic phase, a band of 50 kDa was present only in T. mega. Strain-specific protein distribution was detected in T. cruzi clone Dm28c and T. cruzi G and Y strains; clone Dm28c had five typical hydrophilic proteins at between 24 and 45 kDa, the G strain had two bands at 45 kDa in the hydrophilic phase and the Y strain had a major protein band at 24 kDa in both phases. T. dionisii and T. cruzi clone Dm28c showed a characteristic distribution of three hydrophilic proteins of approx. 45 kDa. Qualitative analysis of glycolipid composition showed that the T. cruzi strains and Dm28c clone and T. dionisii had four orcinol-positive spots, whereas in the other non-pathogenic trypanosomatids only three glycolipids were detected. MIME: Crithidia-chemistry; Electrophoresis,-Polyacrylamide-Gel; Glycolipids-isolation-and-purification; Molecular-Weight; Polyethylene-Glycols-chemistry; Protozoan-Proteins-isolation-and-purification; Trypanosoma-cruzi-chemistry MJME: *Glycolipids-analysis; *Protozoan-Proteins-analysis; *Trypanosomatina-chemistry TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 9036-19-5 NM: Glycolipids; Polyethylene-Glycols; Protozoan-Proteins; Nonidet-P-40 AN: 94352951 UD: 9412 Record 50 of 131 - MEDLINE (R) 1994 TI: Expression of a novel cell surface lipophosphoglycan-like glycoconjugate in Trypanosoma cruzi epimastigotes. AU: Singh-BN; Lucas-JJ; Beach-DH; Costello-CE AD: Department of Microbiology and Immunology, State University of New York Health Science Center, Syracuse 13210. SO: J-Biol-Chem. 1994 Sep 2; 269(35): 21972-82 ISSN: 0021-9258 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The lipophosphoglycan (LPG)-like glycoconjugate expressed on the cell surface of Trypanosoma cruzi epimastigotes was isolated, purified, and partially characterized. The glycoconjugate migrated as a homogeneous band (42 kDa) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Matrix-assisted laser desorption ionization mass spectral analysis of the native molecule indicated the presence of two major components whose molecular masses were about 18.4 and 22.5 kDa. The LPG could be metabolically labeled with [3H]galactose, [3H]mannose, [14C]glucose, or [3H]palmitic acid. Monosaccharide compositional analysis of the LPG indicated that galactose, glucosamine, and sialic acid predominate over mannose, galactosamine, and inositol. A peptide associated with the LPG molecule contained about 40 amino acid residues per inositol and had threonine as the predominant amino acid. The LPG showed strong binding to Ricinus communis agglutinin-1 and Tritium vulgare wheat germ agglutinin, indicating the presence of terminal beta 1,4-linked galactosyl residue(s) and N-acetylglucosamine, respectively. Lectin binding studies also suggested the presence of a terminal beta-galactose and GlcNAc in the glycan-inositol lipid core of LPG. Virtually all of the sialic acids appeared to be located in the saccharide portion of the molecule. Treatment of the LPG with phosphatidylinositol-specific phospholipase C liberated an alkylacylglycerol. Structural analysis of the alkylacylglycerol and its acidic methanolysis products by gas-liquid chromatography/mass spectrometry indicated that the glycerol substituents were primarily the C16 1-alkyl group and C16 2-acyl group. The ratio of inositol to 1-O-alkyl-2-O-acylglycerol was 1:1. Treatment of the glycoconjugate with nitrous acid released a major phospholipid product that migrated close to the phosphatidylinositol standard on thin layer chromatography. This result implied that phosphatidylinositol was glycosidically linked to the nonacetylated amino sugar. Furthermore, the LPG was found to contain phosphate and was labile to mild acid hydrolysis, strongly suggesting that the intact molecule is related to Leishmania LPG. The most striking and unique feature of T. cruzi LPG is the presence of large amounts of glucosamine and sialic acid as well as galactosamine. These results indicate that the glycoconjugate expressed on the T. cruzi cell surface is a new type of LPG-like molecule anchored on the cell surface via an alkylacylphosphatidylinositol. MIME: Carbohydrate-Sequence; Carbohydrates-metabolism; Cell-Membrane-metabolism; Chromatography,-Affinity; Chromatography,-Gas; Chromatography,-Ion-Exchange; Chromatography,-Thin-Layer; Deamination-; Electrophoresis,-Polyacrylamide-Gel; Hydrogen-Ion-Concentration; Hydrolysis-; Mass-Fragmentography-methods; Methane-metabolism; Molecular-Sequence-Data; Phosphoric-Diester-Hydrolases-metabolism MJME: *Glycoconjugates-biosynthesis; *Glycosphingolipids-biosynthesis; *Trypanosoma-cruzi-metabolism TG: Animal; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: NIAIDAI05802AINIAID; RR00317RRNCRR RN: EC 3.1.4; EC; 0; 0; 0; 0; 74-82-8 NM: Phosphoric-Diester-Hydrolases; 1-phosphatidylinositol-phosphodiesterase; lipophosphonoglycan; Carbohydrates; Glycoconjugates; Glycosphingolipids; Methane AN: 94350939 UD: 9412 Record 51 of 131 - MEDLINE (R) 1994 TI: Humoral immune response to the Trypanosoma cruzi complement regulatory protein as an indicator of parasitologic clearance in human Chagas' disease. AU: Norris-KA; Galvao-LM; Schrimpf-JE; Cancado-JR; Krettli-AU AD: Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pennsylvania 15261. SO: Infect-Immun. 1994 Sep; 62(9): 4072-4 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Immunoprecipitation of the purified 160-kDa complement regulatory protein of Trypanosoma cruzi by Chagas' disease patient sera was examined as a possible correlate of the complement-mediated lysis test and as an indicator of parasite clearance. The results presented demonstrate that assessment of the humoral response to this antigen is a useful indicator of parasite clearance and may be particularly helpful in the assessment of some patients for whom other serological tests produce ambiguous results. MIME: Chagas-Disease-parasitology MJME: *Antibodies,-Protozoan-blood; *Chagas-Disease-immunology; *Complement-immunology; *Protozoan-Proteins-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Human; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI32719AINIAID RN: 0; 0; 9007-36-7 NM: Antibodies,-Protozoan; Protozoan-Proteins; Complement AN: 94341922 UD: 9411 Record 52 of 131 - MEDLINE (R) 1994 TI: Trypanocidal effect of boldine and related alkaloids upon several strains of Trypanosoma cruzi. AU: Morello-A; Lipchenca-I; Cassels-BK; Speisky-H; Aldunate-J; Repetto-Y AD: Department of Biochemistry, Faculty of Medicine, University of Chile, Santiago. SO: Comp-Biochem-Physiol-Pharmacol-Toxicol-Endocrinol. 1994 Mar; 107(3): 367-71 PY: 1994 LA: ENGLISH CP: ENGLAND AB: The alkaloids boldine, glaucine, predicentrine, apomorphine, coclaurine, norarmepavine and codeine were tested against the epimastigotes of the Tulahuen and LQ strains and the DM 28c clone of Trypanosoma cruzi. The micromolar concentration to inhibit 50% of the culture growth (Tulahuen strain) for apomorphine, glaucine, predicentrine, boldine, norarmepavine, coclaurine and codeine were 29, 90, 85, 110, 310, 580 and > 1000 respectively. Similar values were obtained with the LQ strain and the DM 28c clone. The most active compounds in inhibiting culture growth also inhibited cell respiration, suggesting that these drugs may act by blocking mitochondrial electron transport. The trypanocidal effects of these alkaloids appear to be correlated with their antioxidative activities. MIME: Antioxidants-pharmacology; Electron-Transport-drug-effects; Mitochondria-drug-effects; Trypanosoma-cruzi-growth-and-development MJME: *Alkaloids-pharmacology; *Aporphines-pharmacology; *Trypanocidal-Agents-pharmacology; *Trypanosoma-cruzi-drug-effects TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 476-70-0 NM: Alkaloids; Antioxidants; Aporphines; Trypanocidal-Agents; boldine AN: 94340231 UD: 9411 Record 53 of 131 - MEDLINE (R) 1994 TI: Characterization of glucose transport and cloning of a hexose transporter gene in Trypanosoma cruzi. AU: Tetaud-E; Bringaud-F; Chabas-S; Barrett-MP; Baltz-T AD: Laboratoire Biologie Moleculaire et Immunologie de Protozoaires Parasites, Universite Bordeaux II, France. SO: Proc-Natl-Acad-Sci-U-S-A. 1994 Aug 16; 91(17): 8278-82 ISSN: 0027-8424 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: A gene from Trypanosoma cruzi, TcrHT1, which encodes a member of the glucose transporter superfamily has been cloned. The gene is similar in sequence to the T. brucei hexose transporter THT1 and the Leishmania transporter Pro-1 and is present in the T. cruzi genome as a cluster of at least eight tandemly reiterated copies. Northern blot analysis revealed two mRNA transcripts which differ in size with respect to their 3' untranslated regions. When injected with in vitro transcribed TcrHT1 mRNA, Xenopus oocytes express a hexose transporter with properties similar to those of T. cruzi. Glucose transport in T. cruzi is mediated via a carrier with unique properties when compared with the other glucose transporters already characterized among the Kinetoplastida. It is a facilitated transporter with a high affinity for D-glucose (Km = 84.1 +/- 7.9 microM and Vmax = 46 +/- 9.4 nmol/min per mg of protein) that shares with other kinetoplastid hexose transporters the ability to recognize D-fructose, which distinguishes these carriers from the human erythrocyte glucose transporter GLUT1. MIME: Amino-Acid-Sequence; Base-Sequence; Biological-Transport,-Active-drug-effects; Blotting,-Northern; Cloning,-Molecular; DNA-Primers; DNA,-Complementary-metabolism; Genes,-Protozoan; Kinetics-; Leishmania-metabolism; Molecular-Sequence-Data; Monosaccharide-Transport-Proteins-genetics; Monosaccharide-Transport-Proteins-isolation-and-purification; Monosaccharides-pharmacology; Polymerase-Chain-Reaction; Protozoan-Proteins-genetics; Protozoan-Proteins-isolation-and-purification; Recombinant-Proteins-biosynthesis; Recombinant-Proteins-isolation-and-purification; Recombinant-Proteins-metabolism; RNA,-Protozoan-biosynthesis; Sequence-Homology,-Amino-Acid; Sequence-Homology,-Nucleic-Acid; Transcription,-Genetic; Trypanosoma-cruzi-genetics MJME: *Genes,-Reiterated; *Glucose-metabolism; *Monosaccharide-Transport-Proteins-metabolism; *Protozoan-Proteins-metabolism; *Trypanosoma-cruzi-metabolism TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't GS: TcrHT1 PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0; 0; 0; 0; 50-99-7 NM: trypanosome-hexose-transporter-1; DNA-Primers; DNA,-Complementary; Monosaccharide-Transport-Proteins; Monosaccharides; Protozoan-Proteins; Recombinant-Proteins; RNA,-Protozoan; Glucose AN: 94336729 UD: 9411 SI: GENBANK/U05588 Record 54 of 131 - MEDLINE (R) 1994 TI: Release of nitric oxide during the experimental infection with Trypanosoma cruzi. AU: Petray-P; Rottenberg-ME; Grinstein-S; Orn-A AD: Laboratorio de Virologia, Hospital de Ninos Dr. Ricardo Gutierrez, Buenos Aires, Argentina. SO: Parasite-Immunol. 1994 Apr; 16(4): 193-9 ISSN: 0141-9838 PY: 1994 LA: ENGLISH CP: ENGLAND AB: We analysed the production of nitric oxide (NO) intermediates by cells from BALB/c mice infected with either virulent (Tulahuen or RA) or avirulent (CA-1) strains of Trypanosoma cruzi. Peritoneal or spleen cells from mice infected with T. cruzi released NO when incubated without further stimuli. Cells from mice during the acute stage of infection accumulated higher levels of inducible NO synthase mRNA and produced both, before and after lypopolysaccharide stimulation, higher amounts of NO than cells from mice chronically infected with T. cruzi. NO synthesis showed similar kinetics in connection with all three strains of T. cruzi, but cells from mice inbred with the Tulahuen or RA strains released higher levels of IFN-gamma, an activator of the NO pathways, than cells from mice infected with the CA-1 strain. In vivo administration of L-Ng-monomethyl-L-arginine (L-NMMA), a competitive inhibitor of NO synthase, increased the susceptibility of mice to T. cruzi. We conclude that infection with T. cruzi induces NO production, and suggest that NO plays a role in the resistance against the parasite. MIME: Amino-Acid-Oxidoreductases-metabolism; Arginine-analogs-and-derivatives; Arginine-pharmacology; Base-Sequence; Disease-Models,-Animal; DNA-Primers; Interferon-Type-II-metabolism; Mice-; Mice,-Inbred-BALB-C; Molecular-Sequence-Data; Nitric-Oxide-antagonists-and-inhibitors; Peritoneal-Cavity-cytology; RNA,-Messenger-metabolism; Spleen-cytology; Trypanosoma-cruzi-pathogenicity; Virulence-drug-effects MJME: *Chagas-Disease-metabolism; *Nitric-Oxide-biosynthesis TG: Animal; Female; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; EC 1.4.; 0; 0; 10102-43-9; 17035-90-4; 7004-12-8; 82115-62-6 NM: nitric-oxide-synthase; Amino-Acid-Oxidoreductases; DNA-Primers; RNA,-Messenger; Nitric-Oxide; omega-N-methylarginine; Arginine; Interferon-Type-II AN: 94336251 UD: 9411 Record 55 of 131 - MEDLINE (R) 1994 TI: Assays of hemolytic toxins. AU: Rowe-GE; Welch-RA AD: Department of Medical Microbiology and Immunology, University of Wisconsin-Madison 53706. SO: Methods-Enzymol. 1994; 235: 657-67 ISSN: 0076-6879 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The ability to produce a cytolytic toxin contributes to the success of many organisms in a particular niche by such diverse means as lysis of a phagolysosomal membrane of the macrophage by hemolysin from the intracellular parasite Trypanosoma cruzi, disruption of leukocyte activity by the Escherichia coli hemolysin, and destruction of invading bacteria by hemolysin from the annelid Glycera dibranchiata. The relative contribution of erythrocyte lysis to survival of the cytolysin producer is still under investigation. Nevertheless, the hemolytic phenotype is both a powerful tool for identifying novel cytolysins and a convenient marker for studying cytolytic activity in established toxins. MIME: Bacterial-Toxins-analysis; Bacterial-Toxins-pharmacology; Cell-Membrane-Permeability-drug-effects; Chromates-metabolism; Culture-Media; Erythrocyte-Membrane-drug-effects; Hemolysins-classification; Hemolysins-pharmacology; Hemolysis-; Invertebrates-metabolism; Microscopy,-Phase-Contrast; Neutral-Red; Phospholipases-analysis; Phospholipases-pharmacology; Sodium-Compounds-metabolism; Sphingomyelin-Phosphodiesterase-analysis; Sphingomyelin-Phosphodiesterase-pharmacology; Stains-and-Staining; Substrate-Specificity; Surface-Active-Agents-analysis; Surface-Active-Agents-pharmacology MJME: *Hemolysins-analysis TG: Animal PT: JOURNAL-ARTICLE; REVIEW; REVIEW,-TUTORIAL RN: EC 3.1.-; EC; 0; 0; 0; 0; 0; 0; 553-24-2; 7775-11-3 NM: Phospholipases; Sphingomyelin-Phosphodiesterase; Bacterial-Toxins; Chromates; Culture-Media; Hemolysins; Sodium-Compounds; Surface-Active-Agents; Neutral-Red; sodium-chromate(VI) AN: 94335733 UD: 9411 Record 56 of 131 - MEDLINE (R) 1994 TI: Intermediate metabolism in Trypanosoma cruzi. AU: Cazzulo-JJ AD: Instituto de Investigaciones Bioquimicas Lus F. Leloir. Fundacion Campomar, CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina. SO: J-Bioenerg-Biomembr. 1994 Apr; 26(2): 157-65 ISSN: 0145-479X PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Epimastigotes of Trypanosoma cruzi, the causative agent of Chagas disease, catabolize proteins and amino acids with production of MH3, and glucose with production of reduced catabolites, chiefly succinate and L-alanine, even under aerobic conditions. This "aerobic fermentation of glucose" is probably due to both the presence of low levels of some cytochromes, causing a relative inefficiency of the respiratory chain for NADH, reoxidation during active glucose catabolism, and the lack of NADH dehydrogenase and phosphorylation site I, resulting in the entry of reduction equivalents into the chain mostly as succinate. Phosphoenol pyruvate carboxykinase and pyruvate kinase may play an essential role in diverting glucose carbon to succinate or L-alanine, and L-malate seems to be the major metabolite for the transport of glucose carbon and reduction equivalents between glycosome and mitochondrion. The parasite contains proteinase and peptidase activities. The major lysosomal cysteine proteinase, cruzipain, has been characterized in considerable detail, and might be involved in the host/parasite relationship, in addition to its obvious role in parasite nutrition. Among the enzymes of amino acid catabolism, two glutamate dehydrogenases (one NADP- and the other NAD-linked), alanine aminotransferase, and the major enzymes of aromatic amino acid catabolism (tyrosine aminotransferase and aromatic alpha-hydroxy acid dehydrogenase), have been characterized and proposed to be involved in the reoxidation of glycolytic NADH. MIME: Aerobiosis-; Amino-Acids-metabolism; Ammonia-metabolism; Biological-Transport,-Active; Carbohydrates-metabolism; Fermentation-; Glucose-metabolism; Oxidation-Reduction; Protozoan-Proteins-metabolism MJME: *Trypanosoma-cruzi-metabolism TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE; REVIEW; REVIEW,-TUTORIAL RN: 0; 0; 0; 50-99-7; 7664-41-7 NM: Amino-Acids; Carbohydrates; Protozoan-Proteins; Glucose; Ammonia AN: 94334314 UD: 9411 Record 57 of 131 - MEDLINE (R) 1994 TI: trans-sialidase and sialic acid acceptors from insect to mammalian stages of Trypanosoma cruzi. AU: Briones-MR; Egima-CM; Acosta-A; Schenkman-S AD: Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Sao Paulo, Brazil. SO: Exp-Parasitol. 1994 Sep; 79(2): 211-4 ISSN: 0014-4894 PY: 1994 LA: ENGLISH CP: UNITED-STATES MIME: Chagas-Disease-enzymology; Chagas-Disease-metabolism; Gene-Expression-Regulation,-Enzymologic; Insect-Vectors-enzymology; Insect-Vectors-metabolism; Mammals-; Neuraminidase-biosynthesis; Neuraminidase-genetics; Trypanosoma-cruzi-genetics; Trypanosoma-cruzi-metabolism MJME: *Chagas-Disease-parasitology; *Insect-Vectors-parasitology; *Neuraminidase-metabolism; *Sialic-Acids-metabolism; *Trypanosoma-cruzi-enzymology TG: Animal PT: JOURNAL-ARTICLE; REVIEW; REVIEW,-TUTORIAL RN: EC 3.2.1.-; EC; 0; 131-48-6 NM: trans-sialidase,-Trypanosoma-cruzi; Neuraminidase; Sialic-Acids; N-acetylneuraminic-acid AN: 94333538 UD: 9411 Record 58 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi cDNA encodes a tandemly repeated domain structure characteristic of small stress proteins and glutathione S-transferases. AU: Schoneck-R; Plumas-Marty-B; Taibi-A; Billaut-Mulot-O; Loyens-M; Gras-Masse-H; Capron-A; Ouaissi-A AD: Research Laboratory on Trypanosomatids, INSERM U415, Lille, France. SO: Biol-Cell. 1994; 80(1): 1-10 ISSN: 0248-4900 PY: 1994 LA: ENGLISH CP: FRANCE AB: The isolation, characterization, and expression of a novel cDNA encoding a Trypanosoma cruzi polypeptide (TcAc2), homologous to various small stress proteins and glutathione S-transferases, are described. The deduced amino-acid sequence revealed two domains sharing 27% identity and an additional 27% similarity to each other suggesting that the molecule may have evolved from a single domain by a process of gene duplication and fusion. The TcAc2 cDNA was subcloned into the pGEX-2T vector for expression in E coli. In vitro translation products of epimastigote mRNA, immunoprecipitated with anti-TXepi serum, showed a major radioactive band of 52 kDa. Immunoprecipitation of [35S]methionine labelled epimastigote and trypomastigote antigens after pulse chase experiments, using anti-TcAc2 fusion protein antibodies, showed that the protein is released into the culture medium. Moreover, Western blot analysis revealed a single band of 52 kDa with epimastigote, trypomastigote and amastigote antigens. Primary structure homology searches revealed that each TcAc2 domain contained within its N-terminus significant homology to Solanum tuberosum pathogenesis-related protein PR1, soybean heat shock protein 26-A, auxin regulated clone pCNT103 from Nicotiana tabacum and Drosophila melanogaster glutathione S-transferase 27 (GST27). This finding was supported by a comparison of hydrophobicity profiles of TcAc2 and these proteins. Most of them play a central role in protection mechanisms against stress. Based on the homology between TcAc2, glutathione S-transferases (GST) and small stress proteins, it is likely that the TcAc2 gene product may play a crucial role in parasite's adaptation to its microenvironment. These molecules could be considered as members of the GST superfamily, where the T cruzi protein may take a particular place because of its internal gene duplication. MIME: Amino-Acid-Sequence; Base-Sequence; Blotting,-Western; Cloning,-Molecular; Drosophila-melanogaster-genetics; DNA-Primers; DNA-Probes; DNA,-Complementary; Escherichia-coli; Gene-Library; Glutathione-Transferases-genetics; Glutathione-Transferases-isolation-and-purification; Heat-Shock-Proteins-genetics; Heat-Shock-Proteins-isolation-and-purification; Methionine-metabolism; Molecular-Sequence-Data; Plants-genetics; Polymerase-Chain-Reaction; Restriction-Mapping; RNA,-Messenger-biosynthesis; Sequence-Homology,-Amino-Acid; Sulfur-Radioisotopes MJME: *DNA,-Protozoan-genetics; *Glutathione-Transferases-biosynthesis; *Heat-Shock-Proteins-biosynthesis; *Repetitive-Sequences,-Nucleic-Acid; *Trypanosoma-cruzi-genetics TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; 0; 0; 0; 0; 0; 0; 0; 7005-18-7 NM: Glutathione-Transferases; DNA-Primers; DNA-Probes; DNA,-Complementary; DNA,-Protozoan; Heat-Shock-Proteins; RNA,-Messenger; Sulfur-Radioisotopes; Methionine AN: 94332034 UD: 9411 SI: GENBANK/L07519 Record 59 of 131 - MEDLINE (R) 1994 TI: A differentially expressed gene family encoding "amastin," a surface protein of Trypanosoma cruzi amastigotes. AU: Teixeira-SM; Russell-DG; Kirchhoff-LV; Donelson-JE AD: Howard Hughes Medical Institute, Iowa City, Iowa 52242. SO: J-Biol-Chem. 1994 Aug 12; 269(32): 20509-16 ISSN: 0021-9258 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: A new family of closely related glycoproteins, collectively called amastins, has been found on the surface of the amastigote form of Trypanosoma cruzi. The gene family encoding these amastigote-specific proteins was identified by differentially screening an amastigote cDNA library with reverse transcribed poly(A)+ RNA from amastigote and epimastigote stages of the parasite. Amastins are encoded by eight or more tandem genes, at least five of which are distinguished by nucleotide point changes. The 1.4-kilobase amastin mRNAs are 50 times more abundant in amastigotes than in epimastigotes or trypomastigotes. The amastin genes are transcribed to an equal extent in both amastigotes and epimastigotes, indicating that the stage-specific amastin mRNA levels are determined by a post-transcriptional mechanism. Sequence determination of full-length cDNAs reveals an open reading frame encoding 174 amino acids and a 700-base pair 3'-untranslated region. Nascent amastins contain four distinct hydrophobic regions of 20-30 amino acids each, 2 at internal locations and 1 each at the N and C termini. MIME: Amino-Acid-Sequence; Base-Sequence; DNA,-Complementary; Molecular-Sequence-Data; Sequence-Homology,-Amino-Acid; Sequence-Homology,-Nucleic-Acid; Transcription,-Genetic MJME: *Genes,-Reiterated; *Membrane-Glycoproteins-genetics; *Protozoan-Proteins-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-Non-P.H.S.; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI24711AINIAID; AI26889AINIAID; AI34207AINIAID RN: 0; 0; 0; 0 NM: amastin; DNA,-Complementary; Membrane-Glycoproteins; Protozoan-Proteins AN: 94327626 UD: 9411 SI: GENBANK/U04337; GENBANK/U04338; GENBANK/U04339; GENBANK/U04340; GENBANK/U04341 Record 60 of 131 - MEDLINE (R) 1994 TI: Randomly amplified polymorphic DNA (RAPD) and isoenzyme analysis of Trypanosoma rangeli strains. AU: Steindel-M; Dias-Neto-E; Pinto-CJ; Grisard-EC; Menezes-CL; Murta-SM; Simpson-AJ; Romanha-AJ AD: Departmento de Microbiologia e Parasitologia Universidade Federal de Santa Catarina C.P. Brazil. SO: J-Eukaryot-Microbiol. 1994 May-Jun; 41(3): 261-7 ISSN: 1066-5234 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Sixteen Trypanosoma rangeli strains were compared by isoenzyme and randomly amplified polymorphic DNA (RAPD) analysis. Eight strains were isolated from either Rhodnius prolixus or Homo sapiens from Honduras, Colombia and Venezuela. Another eight strains were isolated from either Panstrongylus megistus or the rodent Echimys dasythrix from the State of Santa Catarina, southern Brazil. All six T. rangeli strains isolated from P. megistus were co-infections with Trypanosoma cruzi, demonstrating an overlap of the sylvatic cycles of these parasites and that the accurate identification of species is of utmost importance. Both isoenzyme and RAPD analysis revealed two distinct groups of T. rangeli strains, one formed by the strains from Santa Catarina and the other, by the strains from Honduras, Colombia and Venezuela. With the five enzymes used, all the strains from Santa Catarina had identical profiles which overlapped with those of the other regions only in the pattern obtained with malic enzyme. Analysis of 138 RAPD bands by means of an unweighted pair group method analysis (UPGMA) phenogram using the Dice similarity coefficient allowed the separation of the two groups based on their divergence at a lower level of similarity than the phenon line. We show that the identification of T. cruzi and T. rangeli in naturally mixed infections is readily achieved by either RAPD or isoenzyme analysis. MIME: Alanine-Aminotransferase-analysis; Aspartate-Aminotransferase-analysis; Base-Sequence; Glucosephosphate-Isomerase-analysis; Honduras-; Malate-Dehydrogenase-analysis; Molecular-Sequence-Data; Panstrongylus-; Phosphoglucomutase-analysis; Phylogeny-; Rhodnius-; Rodentia-; South-America; Trypanosoma-enzymology; Trypanosoma-genetics; Trypanosomiasis-parasitology MJME: *DNA,-Protozoan-analysis; *Isoenzymes-analysis; *Polymorphism-Genetics; *Trypanosoma-classification TG: Animal; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; EC; EC; EC; EC; EC; 0; 0 NM: Malate-Dehydrogenase; malate-dehydrogenase-(decarboxylating); Aspartate-Aminotransferase; Alanine-Aminotransferase; Glucosephosphate-Isomerase; Phosphoglucomutase; DNA,-Protozoan; Isoenzymes AN: 94325882 UD: 9411 Record 61 of 131 - MEDLINE (R) 1994 TI: The different behavior of diphtheria toxin, modeccin and ricin in HeLa cells infected with Trypanosoma cruzi. AU: Osuna-A; Rodriguez-Cabezas-N; Gamarro-F; Mascaro-C AD: Departamento de Parasitologia, Facultad de Ciencias, Universidad de Granada, Spain. SO: J-Eukaryot-Microbiol. 1994 May-Jun; 41(3): 231-6 ISSN: 1066-5234 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: We have studied the action of diphtheria toxin, modeccin and ricin on HeLa cells infected by Trypanosoma cruzi. Parasitized HeLa cells were resistant to diphtheria toxin and modeccin, whereas non-parasitized cells from the same cultures and control cultures showed cytopathological alterations. Protein synthesis, assayed by the incorporation of labelled methionine, diminished in toxin-treated control cultures but remained unaltered in the infected ones, compared to synthesis by untreated infected cells. Ricin, on the other hand, is a toxin that enters the cytoplasm by endocytosis. It has greater cytopathological effects in parasitized cells than in non-parasitized ones from the same cultures or uninfected control cells. Protein synthesis was inhibited in infected cultures treated with ricin. MIME: Cell-Survival-drug-effects; Drug-Resistance; Hela-Cells-drug-effects; Hela-Cells-parasitology MJME: *Diphtheria-Toxin-toxicity; *Lectins-toxicity; *Proteins-biosynthesis; *Ricin-toxicity; *Toxins-pharmacology; *Trypanosoma-cruzi-physiology TG: Animal; Comparative-Study; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 65988-88-7; 9009-86-3 NM: Diphtheria-Toxin; Lectins; Proteins; Toxins; modeccin; Ricin AN: 94325881 UD: 9411 Record 62 of 131 - MEDLINE (R) 1994 TI: Screening of natural and synthetic drugs against Trypanosoma cruzi. 1. Establishing a structure/activity relationship. AU: de-Castro-SL; Pinto-MC; Pinto-AV AD: Departamento de Ultra-estrutura e Biologia Celular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil. SO: Microbios. 1994; 78(315): 83-90 ISSN: 0026-2633 PY: 1994 LA: ENGLISH CP: ENGLAND AB: The activity of 45 compounds against bloodstream forms of Trypanosoma cruzi was investigated. The aim was to consider new agents which might subsequently be assayed for chemoprophylaxis in donated blood. In a preliminary screening the drugs were assayed (50 to 1,000 microM at 29 degrees C) and those active against bloodstream forms at concentrations below 600 microM were selected for further assays under blood-bank conditions (4 degrees C/24 h). Three compounds isolated from natural sources and six synthetic agents were selected. The active compounds of plant origin included purpurin, a member of the trihydroxylated anthraquinone group, which is known to exhibit trypanocidal activity. Among the active synthetic compounds, five displayed a common structural feature in that they were potentially one-electron acceptors, via reductive functional groups. All five compounds form tricentered C or N intermediates, joined in a hypothetical 'Y' radical pattern. It is possible that the trypanocidal mechanisms initiated by these compounds are similar to those found with crystal violet, since this dye, which is already used in endemic areas for the treatment of banked blood, also conforms to this general Y structural pattern. MIME: Benzhydryl-Compounds-chemical-synthesis; Benzhydryl-Compounds-pharmacology; Drug-Screening; Dyes-chemical-synthesis; Dyes-pharmacology; Gentian-Violet-pharmacology; Hematoxylin-pharmacology; Lectins-pharmacology; Naphthols-chemical-synthesis; Naphthols-pharmacology; Phenolphthaleins-chemical-synthesis; Phenolphthaleins-pharmacology; Picrates-chemical-synthesis; Picrates-pharmacology; Structure-Activity-Relationship; Strychnine-pharmacology; Trypanocidal-Agents-chemistry MJME: *Trypanocidal-Agents-pharmacology; *Trypanosoma-cruzi-drug-effects TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0; 0; 0; 0; 131-73-7; 517-28-2; 548-62-9; 57-24-9; 603-45-2 NM: purpurin; Benzhydryl-Compounds; Dyes; Lectins; Naphthols; Phenolphthaleins; Picrates; Trypanocidal-Agents; dipicrylamine; Hematoxylin; Gentian-Violet; Strychnine; aurin AN: 94322722 UD: 9411 Record 63 of 131 - MEDLINE (R) 1994 TI: [Hemotherapy and transfusional Chagas' disease in Brazil] TO: Hemoterapia e doenca de Chagas transfusional no Brasil. AU: Moraes-Souza-H; Wanderley-DM; Brener-S; Nascimento-RD; Antunes-CM; Dias-JC AD: Faculdade de Medicina do Triangulo Mineiro, Brasil. SO: Bol-Oficina-Sanit-Panam. 1994 May; 116(5): 406-18 ISSN: 0030-0632 PY: 1994 LA: PORTUGUESE; NON-ENGLISH CP: UNITED-STATES AB: With the increased presence of Chagas' disease in urban areas and the rising importance of transfusional transmission of Trypanosoma cruzi, a proper and realistic approach to hemotherapeutic treatment has become crucial in Brazil. Bringing together data from various institutions, this study analyzed hemotherapy and the problem of transfusional Chagas' disease in 850 Brazilian municipalities from 1988 to 1989. It was found that some type of hemotherapy was practiced in 68.8% of these municipalities at the time, this practice being qualitatively and quantitatively proportional to the population size of the municipality. The official blood bank system supplied the blood used in 13% of these services. In relation to prevention of the main diseases transmissible by transfusion, prior screening of donors was carried out by 75.2% of the services for syphilis, 65.4% for hepatitis, 53.8% for AIDS, and 66.9% for Chagas' disease. These percentages vary by region and by size of the municipality. The majority of donors are classified as voluntary, with only 2% categorized as paid donors. In the case of Chagas' disease, most services used only one serologic technique to screen donors, most commonly hemagglutination or immunofluorescence, while only 10.3% of services had previous experience with chemoprophylaxis using gentian violet. The proportion of potential donors with positive serology for anti-Trypanosoma cruzi antibodies was around 1%. These data were confirmed by information from blood banks and Brazilian hemotherapy professionals. MIME: Blood-Banks-statistics-and-numerical-data; Blood-Donors-statistics-and-numerical-data; Blood-Transfusion-statistics-and-numerical-data; Brazil-epidemiology; Chagas-Disease-epidemiology; English-Abstract; Prevalence-; Seroepidemiologic-Methods; Urban-Population-statistics-and-numerical-data MJME: *Blood-Transfusion-adverse-effects; *Chagas-Disease-transmission TG: Human PT: JOURNAL-ARTICLE AN: 94318164 UD: 9411 Record 64 of 131 - MEDLINE (R) 1994 TI: Mice infected with Trypanosoma cruzi produce antibodies against the enzymatic domain of trans-sialidase that inhibit its activity. AU: Leguizamon-MS; Campetella-OE; Gonzalez-Cappa-SM; Frasch-AC AD: Departamento de Microbiologia, Facultad de Medicina, Universidad de Buenos Aires, Argentina. SO: Infect-Immun. 1994 Aug; 62(8): 3441-6 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: trans-Sialidase (TS) is an enzymatic activity described only for trypanosomes that is involved in the invasion of host cells by Trypanosoma cruzi. The enzyme that is shed by the parasite is made of two domains, the C-terminal region containing immunodominant amino acid repeats that define the SAPA antigen and the N-terminal domain that contains the putative region for enzymatic activity. The SAPA antigen induces a strong humoral response detected shortly after infection, both in humans and in mice. This response is directed to the immunodominant domain but is irrelevant in terms of neutralization of TS activity. We now show that TS activity can be detected in sera from acutely infected mice. However, mice infected with a T. cruzi strain whose growth can be controlled by the host did not have detectable levels of TS activity in sera. In fact, sera from these mice were able to abolish TS activity. This inhibition was due to the presence of specific antibodies directed against the enzymatic domain of the protein since they also abolish the activity of a recombinant molecule lacking the immunodominant amino acid repeats. The neutralizing antibodies were present from day 30 after the infection, while antibodies to the immunodominant repeats were detected by day 8 postinoculation, suggesting that the in vivo role of these repeats is to defect the humoral response to the repeat domain until the infection is established. MIME: Mice-; Mice,-Inbred-C3H; Neuraminidase-antagonists-and-inhibitors; Neuraminidase-blood MJME: *Antibodies,-Protozoan-blood; *Chagas-Disease-immunology; *Neuraminidase-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; 0 NM: Neuraminidase; Antibodies,-Protozoan AN: 94314465 UD: 9410 Record 65 of 131 - MEDLINE (R) 1994 TI: O-glycosidically linked N-acetylglucosamine-bound oligosaccharides from glycoproteins of Trypanosoma cruzi. AU: Previato-JO; Jones-C; Goncalves-LP; Wait-R; Travassos-LR; Mendonca-Previato-L AD: Instituto de Microbiolgia CCS-bloco I, Universidade Federal do Rio de Janeiro, Brasil. SO: Biochem-J. 1994 Jul 1; 301 ( Pt 1): 151-9 ISSN: 0264-6021 PY: 1994 LA: ENGLISH CP: ENGLAND AB: In this report we describe studies on the structures of the O-linked carbohydrate units in cell-surface glycoproteins of epimastigote forms of the G-strain of Trypanosoma cruzi. Mild alkaline reductive degradation of the 38/43 kDa glycoproteins resulted in beta-elimination of glycosylated threonine and/or serine residues, and the liberation of N-acetylglucosaminitol, galactobiosyl-, galactotriosyl-, galactotetraosyl- and galactopentaosyl-N-acetylglucosaminitol. The structures of these oligosaccharide alditols were established by n.m.r. spectroscopy and methylation analysis as: Galf beta 1-4(Galp beta 1-6)GlcNAc-ol; Galp beta 1-3Galp beta 1-6(Galf beta 1-4)GlcNAc-ol; [(Galp beta 1-3)(Galp beta 1-2)Galp beta 1-6](Galf beta 1-4)GlcNAc-ol; [(Galp beta 1-3)(Galp beta 1-2)Galp beta 1-6](Galp beta 1-2Galf beta 1-4)GlcNAc-ol. MIME: Carbohydrate-Conformation; Carbohydrate-Sequence; Glycoproteins-isolation-and-purification; Methylation-; Molecular-Sequence-Data; Molecular-Weight; Nuclear-Magnetic-Resonance; Oligosaccharides-isolation-and-purification MJME: *Acetylglucosamine-chemistry; *Glycoproteins-chemistry; *Oligosaccharides-chemistry; *Trypanosoma-cruzi-chemistry TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 7512-17-6 NM: Glycoproteins; Oligosaccharides; Acetylglucosamine AN: 94311825 UD: 9410 Record 66 of 131 - MEDLINE (R) 1994 TI: Serological evidence implicating Neospora species as a cause of abortion in British cattle. AU: Trees-AJ; Guy-F; Low-JC; Roberts-L; Buxton-D; Dubey-JP AD: Liverpool School of Tropical Medicine, University of Liverpool. SO: Vet-Rec. 1994 Apr 16; 134(16): 405-7 ISSN: 0042-4900 PY: 1994 LA: ENGLISH CP: ENGLAND AB: By means of an immunofluorescence antibody test (IFAT), using in vitro cultured parasites as antigen, antibodies to Neospora species at titres > or = 1/1280 were found in 11 of 120 Scottish cattle that had recently aborted but in only one of 97 cattle from herds in which there had been no recent abortions (P < 0.01). The specificity of the antibodies was confirmed by the lack of cross reactivity between samples with high titres to Neospora and toxoplasma antigen in a direct agglutination test, and by the absence of reactivity at > or = 1/640 in the IFAT of convalescent sera from cattle infected experimentally with Toxoplasma gondii, Sarcocystis cruzi, Eimeria bovis, E alabamensis, Cryptosporidium parvum and Babesia divergens. These results demonstrate that Neospora species infection occurs commonly in aborting cattle in Britain, and that the IFAT may be a useful tool for investigating the infection. MIME: Antibodies,-Protozoan-analysis; Apicomplexa-immunology; Cattle-; Cattle-Diseases-diagnosis; Coccidiosis-diagnosis; Fetal-Death-parasitology; Fetal-Death-veterinary; Fluorescent-Antibody-Technique-veterinary; Great-Britain; Pregnancy-; Pregnancy-Complications,-Parasitic-diagnosis MJME: *Abortion,-Veterinary-parasitology; *Apicomplexa-; *Cattle-Diseases-parasitology; *Coccidiosis-veterinary; *Pregnancy-Complications,-Parasitic-veterinary TG: Animal; Female; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0 NM: Antibodies,-Protozoan AN: 94310755 UD: 9410 Record 67 of 131 - MEDLINE (R) 1994 TI: Use of proteinase K in the excystation of Sarcocystis cruzi sporocysts for in vitro culture and DNA extraction. AU: Ndiritu-W; Cawthorn-RJ; Kibenge-FS AD: Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, Canada. SO: Vet-Parasitol. 1994 Mar; 52(1-2): 57-60 ISSN: 0304-4017 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Proteinase K was used for the cleaning of Sarcocystis cruzi (Apicomplexa) sporocysts prior to excystation. Bovine pulmonary endothelial cell cultures inoculated with the excysted sporozoites remained free of bacterial contamination for the duration of the experiment and had high yields of merozoites. The excysted sporozoites also yielded genomic DNA that could be labelled efficiently with 32P dATP by the random priming method. MIME: Cattle-; Cell-Line; Endothelium-cytology; Endothelium-parasitology; Lung-cytology; Lung-parasitology; Sarcocystis-growth-and-development; Sarcocystis-genetics MJME: *DNA,-Protozoan-isolation-and-purification; *Sarcocystis-isolation-and-purification; *Serine-Proteinases-metabolism TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 3.4.21; EC 3.4.21.-; 0 NM: Serine-Proteinases; Tritirachium-alkaline-proteinase; DNA,-Protozoan AN: 94303135 UD: 9410 Record 68 of 131 - MEDLINE (R) 1994 TI: Effects of immunosuppression and benzonidazole on Trypanosoma cruzi parasitism during experimental acute Chagas' disease. AU: Okumura-M; Mester-M; Iriya-K; Amato-Neto-V; Gama-Rodrigues-J AD: Department of Gastrointestinal Surgery, University of Sao Paulo, Brazil. SO: Transplant-Proc. 1994 Jun; 26(3): 1587-9 ISSN: 0041-1345 PY: 1994 LA: ENGLISH CP: UNITED-STATES MIME: Chagas-Disease-pathology; Heart-parasitology; Immunosuppression-methods; Mice-; Mice,-Inbred-BALB-C; Trypanosoma-cruzi-isolation-and-purification MJME: *Chagas-Disease-drug-therapy; *Chagas-Disease-immunology; *Cyclosporine-pharmacology; *Immunosuppression-; *Myocardium-pathology; *Nitroimidazoles-therapeutic-use; *Prednisone-pharmacology; *Trypanocidal-Agents-therapeutic-use; *Trypanosoma-cruzi-growth-and-development TG: Animal; Comparative-Study; Male PT: JOURNAL-ARTICLE RN: 0; 0; 22994-85-0; 53-03-2; 59865-13-3 NM: Nitroimidazoles; Trypanocidal-Agents; benzonidazole; Prednisone; Cyclosporine AN: 94302959 UD: 9410 Record 69 of 131 - MEDLINE (R) 1994 TI: Serodiagnosis of Chagas' disease by enzyme-linked immunosorbent assay using two synthetic peptides as antigens. AU: Peralta-JM; Teixeira-MG; Shreffler-WG; Pereira-JB; Burns-JM Jr; Sleath-PR; Reed-SG AD: Instituto de Microbiologia, UFRJ, Rio de Janerio, Brazil. SO: J-Clin-Microbiol. 1994 Apr; 32(4): 971-4 ISSN: 0095-1137 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: An enzyme-linked immunosorbent assay (ELISA) was developed for detecting antibodies against Trypanosoma cruzi. Two synthetic T. cruzi peptides, TcD and PEP2, were used. The specificity and sensitivity of the peptide ELISA were determined with 260 serum samples from individuals living in an area in which Chagas' disease is endemic. ELISAs were performed with the peptides singly or in combination. The evaluation of these tests showed that 168 (93.8%) of 179 serum samples from T. cruzi-infected patients were positive when TcD peptide was used as antigen; 164 (91.6%) samples were positive with PEP2, and 178 (99.4%) samples were positive when the two peptides were combined. Thus, the sensitivity of the ELISA using the two peptides exceeded 99%. The specificity was evaluated by using a panel of 118 serum samples that included samples from 81 individuals living in an area of endemicity with negative serology for Chagas' disease and from 37 patients from areas in which T. cruzi was not endemic but with other pathologies, such as leishmaniasis, tuberculosis, and leprosy. Only two false-positive serum samples were found in this group of individuals, giving a test specificity of more than 98%. Because these peptides can be synthesized and are very stable at room temperature, the use of such reagents can improve the standardization and reproducibility of ELISAs for the serodiagnosis of T. cruzi infection. MIME: Amino-Acid-Sequence; Antibodies,-Protozoan-blood; Antigens,-Protozoan-genetics; Chagas-Disease-immunology; Chagas-Disease-parasitology; Enzyme-Linked-Immunosorbent-Assay-statistics-and-numerical-data; Evaluation-Studies; False-Positive-Reactions; Molecular-Sequence-Data; Peptides-chemical-synthesis; Peptides-genetics; Peptides-immunology; Sensitivity-and-Specificity; Serodiagnosis-statistics-and-numerical-data; Trypanosoma-cruzi-genetics; Trypanosoma-cruzi-immunology MJME: *Antigens,-Protozoan; *Chagas-Disease-diagnosis; *Enzyme-Linked-Immunosorbent-Assay-methods; *Serodiagnosis-methods TG: Animal; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0 NM: Antibodies,-Protozoan; Antigens,-Protozoan; Peptides AN: 94299833 UD: 9410 Record 70 of 131 - MEDLINE (R) 1994 TI: Chagas' disease screening in blood bank employing enzyme immunoassay. AU: Knecher-LM; Rojkin-LF; Capriotti-GA; Lorenzo-LE AD: Wiener Laboratory, S.A.I.C., Rosario, Argentina. SO: Int-J-Parasitol. 1994 Apr; 24(2): 207-11 ISSN: 0020-7519 PY: 1994 LA: ENGLISH CP: ENGLAND AB: Because of the high prevalence of Trypanosoma cruzi infection in Latin America, antibody screening in blood banks is mandatory in this area. This screening may also become a concern in the U.S.A. considering the high frequency of Latin American donors. The tests usually employed (indirect hemagglutination, direct agglutination, immunofluorescence and latex agglutination assays) involve subjective interpretation of results and do not fit the automated procedures requirements of large laboratories. Thus, an enzyme immunoassay was developed using a mixture of antigens purified from the membrane and the cytoplasm of the parasite. Serum of plasma could be used as sample, in a procedure involving 90 min total incubation time and 2 washing steps. Results could be interpreted either spectrophotometrically or by the naked eye. The method was used to test 661 samples from patients undergoing different stages of Chagas' disease, 120 patients suffering other parasitosis and 880 normal subjects. Results were compared with those obtained with the methods mentioned above. The proposed test showed better reproducibility, specificity and sensitivity than those of reference methods, plus an objective interpretation of results and suitability to automation. MIME: Reproducibility-of-Results; Sensitivity-and-Specificity MJME: *Antibodies,-Protozoan-blood; *Blood-Banks; *Chagas-Disease-diagnosis; *Enzyme-Linked-Immunosorbent-Assay; *Trypanosoma-cruzi-immunology TG: Animal; Human PT: JOURNAL-ARTICLE RN: 0 NM: Antibodies,-Protozoan AN: 94299333 UD: 9410 Record 71 of 131 - MEDLINE (R) 1994 TI: Homologous cysteine proteinase genes located on two different chromosomes from Trypanosoma rangeli. AU: Tanaka-T; Kaneda-Y; Iida-A; Tanaka-M AD: Department of Infectious Diseases, Tokai University School of Medicine, Kanagawa, Japan. SO: Int-J-Parasitol. 1994 Apr; 24(2): 179-88 ISSN: 0020-7519 PY: 1994 LA: ENGLISH CP: ENGLAND AB: DNA fragments were obtained by the polymerase chain reaction (PCR) using genomic DNA from T. rangeli as template and oligonucleotide primers encoding the active site amino acids of cysteine proteinase. After amplification by PCR, several DNA products were observed. These were purified and used as templates for a second round of PCR. This resulted in two DNA products of 475 and 498 bp. The 498 bp DNA (Tr-CP) contained both the sense and antisense primer sequences, and encoded a polypeptide having substantial homology with eukaryotic cysteine proteinases. The other product (Tr-DMR), which lacked the antisense primer sequence, encoded a polypeptide having homology with the DNA mismatch repair gene from Saccharomyces cerevisiae. The overall homology of the Tr-CP-encoded polypeptide to the cysteine proteinases from other trypanosome species was 69% identity (T. cruzi) and 73% identity (T. brucei), respectively. Northern blot analysis revealed that the Tr-CP gene was specifically expressed in T. rangeli as a 1.7 kb mRNA. A karyotype map of the chromosomes was also performed using pulsed-field gradient gel electrophoresis and Southern blot hybridization with these genes. T. rangeli has 14 chromosome bands ranging from 350 kb to 1.6 Mb, which were fewer in number and smaller in size compared with those from T. cruzi. The Tr-CP fragment and the Tr-DMR fragment hybridized with equal intensity on chromosome 1 (350 kb) and chromosome 2 (470 kb), respectively. These results suggest that non-pathogenic T. rangeli contains a conserved gene corresponding to the cysteine proteinase or a closely related enzyme, and that there is more than one copy of this gene, each found on different chromosomes. MIME: Amino-Acid-Sequence; Base-Sequence; Consensus-Sequence; Cysteine-Proteinases-chemistry; DNA-Primers-chemistry; Electrophoresis,-Gel,-Pulsed-Field; Karyotyping-; Molecular-Sequence-Data; Polymerase-Chain-Reaction; Sequence-Homology,-Amino-Acid; Trypanosoma-enzymology MJME: *Cysteine-Proteinases-genetics; *DNA,-Protozoan-chemistry; *Genes,-Protozoan; *Trypanosoma-genetics TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 3.4.22; 0; 0 NM: Cysteine-Proteinases; DNA-Primers; DNA,-Protozoan AN: 94299330 UD: 9410 SI: GENBANK/M99495; GENBANK/M99496 Record 72 of 131 - MEDLINE (R) 1994 TI: Expression of the Leishmania tarentolae ubiquitin-encoding and mini-exon genes. AU: Fleischmann-J; Campbell-DA AD: Department of Microbiology and Immunology, University of California, Los Angeles 90024. SO: Gene. 1994 Jun 24; 144(1): 45-51 ISSN: 0378-1119 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: To develop models for transcription and trans-splicing in kinetoplastid protozoa, we have characterized ubiquitin (Ubi) gene organization and mRNA processing in Leishmania tarentolae (Lt). Three ubi loci were characterized: two discrete Ubi-extension protein 52 (EP52)-encoding genes (ubiA and ubiB) and a polymorphic polyubiquitin-encoding gene (ubiC). The three loci resided on chromosomes of 2.05 Mb, 630 kb and 2.9 Mb, respectively. On the basis of upstream flanking gene identity, ubiB appears to be the homologue of the tandemly repeated ubi-EP52/1 and 2 in Trypanosoma brucei (Tb). Similar to Trypanosoma cruzi, Lt did not contain a homologue of the ubi-EP76 that has been found in Saccharomyces cerevisiae and multicellular organisms. All three Lt ubi loci were transcribed. The primary transcripts from the ubi loci were processed at the 5'-end by trans-splicing with the mini-exon. A Lt mini-exon gene (min) that gave rise to a 95-nt primary transcript, which is the second template in the trans-splicing reaction, was also characterized. MIME: Amino-Acid-Sequence; Base-Sequence; Chromosome-Mapping; DNA,-Protozoan; Molecular-Sequence-Data; RNA-Splicing; RNA,-Messenger-genetics; Transcription,-Genetic MJME: *Exons-; *Gene-Expression; *Genes,-Protozoan; *Leishmania-genetics; *Ubiquitin-genetics TG: Animal; Support,-Non-U.S.-Gov't GS: ubiA; min; ubiB; ubiC PT: JOURNAL-ARTICLE RN: 0; 0; 0 NM: DNA,-Protozoan; RNA,-Messenger; Ubiquitin AN: 94299167 UD: 9410 SI: GENBANK/X73119; GENBANK/X73118; GENBANK/X73120; GENBANK/X73121 Record 73 of 131 - MEDLINE (R) 1994 TI: Comparison of chagasic and non-chagasic myocardiopathies by ELISA and immunoblotting with antigens of Trypanosoma cruzi and Trypanosoma rangeli. AU: O'Daly-JA; Carrasco-H; Fernandez-V; Rodriguez-MB AD: Instituto Venezolano de Investigaciones Cientificas (IVIC), Center of Microbiology and Cell Biology, Caracas. SO: Acta-Trop. 1994 Apr; 56(4): 265-87 ISSN: 0001-706X PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Trypanosoma cruzi associated myocardiopathy, or Chagas disease, continues to be a serious problem in Venezuela, for which there is neither a vaccine nor a cure. In order to learn more about the humoral immune response to trypanosomal antigens, and to try to identify dominant antigens, we used ELISA and immunoblotting to study the reactivity of sera from patients with chagasic and non-chagasic myocardiopathies, against surface and secreted proteins from T. cruzi and T. rangeli. Both species are found in the same insect vector, but only T. cruzi is thought to be pathogenic in vertebrates. The ELISA results fell into three patterns: (1) high reactivity values with both T. cruzi and T. rangeli surface and secreted proteins; (2) high values to T. cruzi but low values with T. rangeli; and (3) high values to T. rangeli and low values with T. cruzi. This finding that some chagasic sera react more strongly against T. rangeli than against T. cruzi is intriguing, and warrants further investigation. When chagasic sera were tested on Western blots of total extracts of T. cruzi and T. rangeli, the pattern of reactive bands was similar against both parasites, but no two sera showed an identical pattern. Furthermore, there was no correlation between a particular immunoblotting pattern and either the antibody titer, or the severity of the disease. Several T. cruzi and T. rangeli antigens were recognized by sera from healthy controls as well as from patients with other tropical diseases endemic in Venezuela. Overall, our results suggest that the humoral immune response to trypanosomal antigens is complex, and no single antigen may be the determining factor in the pathogenesis of chagasic myocardiopathy. MIME: Adult-; Antigens,-Protozoan-isolation-and-purification; Antigens,-Surface-immunology; Antigens,-Surface-isolation-and-purification; Blotting,-Western; Chagas-Cardiomyopathy-diagnosis; Chagas-Cardiomyopathy-physiopathology; Electrophoresis,-Polyacrylamide-Gel; Enzyme-Linked-Immunosorbent-Assay; Immune-Sera-immunology; Mice-; Mice,-Inbred-C3H; Mice,-Inbred-C57BL; Middle-Age; Myocardial-Diseases-diagnosis; Myocardial-Diseases-physiopathology; Protozoan-Proteins-immunology; Protozoan-Proteins-isolation-and-purification; Trypanosoma-immunology MJME: *Antibodies,-Protozoan-blood; *Antigens,-Protozoan-immunology; *Chagas-Cardiomyopathy-immunology; *Myocardial-Diseases-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Comparative-Study; Female; Human; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0 NM: Antibodies,-Protozoan; Antigens,-Protozoan; Antigens,-Surface; Immune-Sera; Protozoan-Proteins AN: 94295476 UD: 9410 Record 74 of 131 - MEDLINE (R) 1994 TI: Intermembrane lipid transfer during Trypanosoma cruzi-induced erythrocyte membrane destabilization. AU: Lujan-HD; Bronia-DH AD: Catedra de Quimica Biologica, Facultad de Ciencias Medicas, Universidad Nacional de Cordoba, Argentina. SO: Parasitology. 1994 Apr; 108 ( Pt 3): 323-34 ISSN: 0031-1820 PY: 1994 LA: ENGLISH CP: ENGLAND AB: The ability of Trypanosoma cruzi to induce erythrocyte membrane destabilization in vitro was studied. Epimastigote forms adhered to human erythrocytes and caused fusion or lysis of the red cells, depending on the conditions of the interaction. Red cells were fused in the presence of calcium, while haemolysis was induced in the absence of the cation. Dextran 60 C facilitated fusion but delayed lysis. Optimum pH and temperature for fusion were 7.4 and 37 degrees C, respectively. Lipid alterations were produced in the plasma membrane of the red cell during the interaction with the parasite. A Ca(2+)-independent increase of lysophospholipids and free fatty acids was common to both the lysis and fusion processes. A relative increase of 1,2-diacylglycerides was unique to the fusion process and these changes were dependent on Ca2+. The transfer of free fatty acids and lysophospholipids from T. cruzi to erythrocyte membranes was demonstrated using parasites pre-labelled with radioactive phospholipids. Pre-treatment of parasites with exogenous phospholipase A2 abolished the fusogenicity, while lysis was increased. Neither fusion nor haemolysis occurred when the parasites were pre-treated with fatty acid free albumin, phospholipase A2 inhibitors or when these compounds were present in the medium during the parasite-erythrocyte interaction. Our results suggest that T. cruzi induces erythrocyte membrane destabilization in vitro by transfer of lipid material in a calcium independent manner and that this ion is necessary for other membrane alterations that lead to erythrocyte fusion. MIME: Calcium-pharmacology; Cell-Fusion-drug-effects; Crithidia-fasciculata-physiology; Edetic-Acid-pharmacology; Erythrocyte-Membrane-drug-effects; Erythrocyte-Membrane-ultrastructure; Erythrocytes-cytology; Erythrocytes-ultrastructure; Fatty-Acids,-Nonesterified-metabolism; Hydrogen-Ion-Concentration; Leishmania-physiology; Lysophospholipids-metabolism; Magnesium-pharmacology; Membrane-Fusion-drug-effects; Temperature-; Time-Factors MJME: *Erythrocyte-Membrane-metabolism; *Erythrocytes-parasitology; *Membrane-Fusion; *Membrane-Lipids-metabolism; *Trypanosoma-cruzi-physiology TG: Animal; Comparative-Study; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 60-00-4; 7439-95-4; 7440-70-2 NM: Fatty-Acids,-Nonesterified; Lysophospholipids; Membrane-Lipids; Edetic-Acid; Magnesium; Calcium AN: 94294207 UD: 9410 Record 75 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi infection suppresses nuclear factors that bind to specific sites on the interleukin-2 enhancer. AU: Soong-L; Tarleton-RL AD: Department of Zoology, University of Georgia, Athens 30602. SO: Eur-J-Immunol. 1994 Jan; 24(1): 16-23 ISSN: 0014-2980 PY: 1994 LA: ENGLISH CP: GERMANY AB: Interleukin-2 (IL-2) gene expression, a critical early event during T lymphocyte activation, is severely suppressed in mice infected with the protozoan parasite Trypanosoma cruzi, the causative agent of human Chagas' disease. Our previous observation that reduction of IL-2 mRNA in T cells from T. cruzi-infected mice is not due to an increased degradation of the mRNA suggests a repression of the IL-2 gene at the transcriptional level. In this study, we have measured the level of nuclear factors that bind to specific sites on the IL-2 enhancer. Splenocytes and splenic T cells from acutely infected mice show a marked decrease in the level of AP-1, and a modest decrease in the level of NF-kappa B and nuclear factor of activated T cells (NF-AT). DNA-binding activity of Oct-1 was least affected in T cells from infected mice. Although the basal level of AP-1 activity is comparable in cells from uninfected and infected mice, mitogen-induced AP-1 activation is absent in the cells from T. cruzi-infected mice. Sodium deoxycholate treatment slightly enhances NF-kappa B-binding activity in splenocyte nuclear and whole-cell extracts from infected mice, suggesting that a blockage of the activation of NF-kappa B is only partially responsible for the decrease in the level of NF-kappa B in T cells from T. cruzi-infected mice. These data identify the molecular basis of IL-2 gene regulation in T. cruzi infection and suggest that T cells are anergized as a result of the infection. MIME: Base-Sequence; Blotting,-Northern; Cells,-Cultured; Mice-; Mice,-Inbred-C57BL; Molecular-Sequence-Data; NF-kappa-B-metabolism; RNA,-Messenger-metabolism; Spleen-cytology; T-Lymphocytes-metabolism MJME: *Chagas-Disease-genetics; *Enhancer-Elements-Genetics-genetics; *Interleukin-2-genetics; *Transcription-Factors-metabolism TG: Animal; Female; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: R01AI22070AINIAID RN: 0; 0; 0; 0 NM: Interleukin-2; NF-kappa-B; RNA,-Messenger; Transcription-Factors AN: 94291716 UD: 9410 Record 76 of 131 - MEDLINE (R) 1994 TI: Activated T and B lymphocytes in peripheral blood of patients with Chagas' disease. AU: Dutra-WO; Martins-Filho-OA; Cancado-JR; Pinto-Dias-JC; Brener-Z; Freeman-Junior-GL; Colley-DG; Gazzinelli-G; Parra-JC AD: Centro de Pesquisas Rene Rachou, FIOCRUZ, Belo Horizonte, Brazil. SO: Int-Immunol. 1994 Apr; 6(4): 499-506 ISSN: 0953-8178 PY: 1994 LA: ENGLISH CP: ENGLAND AB: Whole blood preparations from patients with either the indeterminate (asymptomatic) or cardiac clinical forms of chronic Trypanosoma cruzi infection were analyzed by flow cytometry using double-labeling to identify subsets of circulating lymphocytes. Several significant differences were demonstrated between the blood lymphocyte profiles of chagasic patients and non-chagasic controls. Clear increase in the percentages and actual numbers of double-positive cells of the phenotype CD3+/HLA-DR+, as well as decrease in the percentage of CD45RA+/CD4+ and CD45RA+/CD8+ T cells, indicate greater numbers of activated T cells circulating in the blood of infected patients. Consistent parallel increases were seen also in the B lymphocyte subset which stained double-positive for CD19/CD5. There were no significant differences in the circulation of these chronic chagasic patients in the CD4:CD8 ratios. Also, no substantive phenotypic differences were observed in the lymphocyte populations between the two ends of the clinical spectrum (indeterminate versus cardiac) in chronic human Chagas' disease. These observations demonstrate that increased levels of activated T cells and CD5+ B cells are present in the circulation of people with chronic Chagas' disease. These are cell phenotypes that have been associated in other conditions with autoimmune, polyclonal, and hyperimmune responses. The specificities of these activated cells and the roles they may play in resistance or pathogenesis during chronic Chagas' disease need now to be determined. MIME: Adult-; Aged-; Aged,-80-and-over; Antigens,-CD-blood; Chronic-Disease; CD4-CD8-Ratio; Flow-Cytometry; Fluorescent-Antibody-Technique; HLA-DR-Antigens-immunology; Middle-Age MJME: *B-Lymphocyte-Subsets-immunology; *Chagas-Disease-immunology; *Lymphocyte-Transformation-physiology; *T-Lymphocyte-Subsets-immunology TG: Human; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-Non-P.H.S.; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI26505AINIAID RN: 0; 0 NM: Antigens,-CD; HLA-DR-Antigens AN: 94289330 UD: 9410 Record 77 of 131 - MEDLINE (R) 1994 TI: Modulation of T-cell responsiveness during Trypanosoma cruzi infection: analysis in different lymphoid compartments. AU: Vandekerckhove-F; Darji-A; Rivera-MT; Carlier-Y; Vray-B; Billiau-A; De-Baetselier-P AD: University of Leuven, Rega Institute, Department of Immunobiology, Belgium. SO: Parasite-Immunol. 1994 Feb; 16(2): 77-85 ISSN: 0141-9838 PY: 1994 LA: ENGLISH CP: ENGLAND AB: Spleen and lymph node cells of Trypanosoma cruzi-infected mice were studied for mitogen-induced responsiveness in terms of proliferation and lymphokine production (IL-2, IFN-gamma). Splenocyte (SP) as well as lymph node cell (LN) proliferation and IL-2 production were depressed during the acute phase of the infection. Proliferative capacity of LN cells recovered completely and that of SP partially during the chronic phase. In contrast to these suppressive effects, the mitogen-induced IFN-gamma response was enhanced. In vitro co-incubation of normal SP or LN cells with trypomastigotes resulted in a reduced mitogen-induced cell proliferation and IL-2 secretion, similar to those seen with cells taken from infected mice. In contrast, trypomastigotes exerted a stimulatory activity on the mitogen-induced IFN-gamma response of both SP and LN cells. Addition of lymph node cells from T. cruzi-infected mice (LN-I) to lymph node cells of control mice (LN-C) suppressed strongly the mitogen-induced responsiveness of such cocultures. A marginal level of suppression was recorded in cocultures of spleen cells from infected mice (SP-I) and control spleen cells (SP-C). The potent suppressive cells within LN-I populations were identified as macrophage-like and such cells were absent in SP-C and peritoneal exudate cells from T. cruzi infected animals. MIME: Acute-Disease; Cells,-Cultured; Concanavalin-A-pharmacology; Lymphocyte-Transformation-immunology; Lymphokines-biosynthesis; Mice-; Mice,-Inbred-BALB-C MJME: *Chagas-Disease-immunology; *Immune-Tolerance; *Lymph-Nodes-immunology; *Spleen-immunology; *T-Lymphocytes-immunology TG: Animal; Female; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 11028-71-0 NM: Lymphokines; Concanavalin-A AN: 94286286 UD: 9409 Record 78 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi infection in mice induces a polyisotypic hypergammaglobulinaemia and parasite-specific response involving high IgG2a concentrations and highly avid IgG1 antibodies. AU: el-Bouhdidi-A; Truyens-C; Rivera-MT; Bazin-H; Carlier-Y AD: Laboratory of Parasitology, Faculty of Medicine, University of Brussels (ULB), Belgium. SO: Parasite-Immunol. 1994 Feb; 16(2): 69-76 ISSN: 0141-9838 PY: 1994 LA: ENGLISH CP: ENGLAND AB: Trypanosoma cruzi infection in BALB/c mice induced a reversible polyisotypic hypergammaglobulinaemia, with particularly high levels of IgG2a, IgM and IgE. Hypergammaglobulinaemia started during the acute phase of infection and persisted during chronic disease until 11-13 weeks post-infection (w.p.i.), when immunoglobulin levels, with the exception of IgE, returned near normal values. Parasite-specific antibodies counted for 14 to 23% of gammaglobulinaemia, in acute and chronic infection respectively. The titres of IgM antibodies rose from two w.p.i. IgA, IgE and IgG subclass antibodies built up gradually over the time of parasite clearance (i.e., between three and six w.p.i.). All antibody isotypes, including IgM reached significant and stable titres throughout chronic infection. IgG2a, IgG1 and IgM antibodies had constantly higher titres than the other antibody isotypes. The dominance of IgG2a antibodies was due to their high plasma concentrations, around 70% of all antibodies available in the chronic infection. IgG1 had the highest functional avidity, whereas its concentration corresponded to only 10% of the whole antibody fraction. These results indicate that T. cruzi infection in mice induces a polyisotypic humoral immune response, dominated by some antibody isotypes, with major differences in concentrations and functional avidities. This could be of crucial importance in determining the outcome of infection. MIME: Antibodies,-Protozoan-blood; Antibody-Affinity-immunology; Chagas-Disease-complications; Enzyme-Linked-Immunosorbent-Assay; Hypergammaglobulinemia-immunology; IgG-blood; Immunoglobulin-Isotypes-blood; Mice-; Mice,-Inbred-BALB-C MJME: *Antibodies,-Protozoan-biosynthesis; *Chagas-Disease-immunology; *Hypergammaglobulinemia-etiology; *IgG-biosynthesis; *Trypanosoma-cruzi-immunology TG: Animal; Female; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0 NM: Antibodies,-Protozoan; IgG; Immunoglobulin-Isotypes AN: 94286285 UD: 9409 Record 79 of 131 - MEDLINE (R) 1994 TI: Production of aromatic alpha-hydroxyacids by epimastigotes of Trypanosoma cruzi, and its possible role in NADH reoxidation. AU: Montemartini-M; Santome-JA; Cazzulo-JJ; Nowicki-C AD: IQUIFIB (UBA-CONICET), Facultad de Farmacia y Bioquimica, Universidad de Buenos Aires, Argentina. SO: FEMS-Microbiol-Lett. 1994 May 1; 118(1-2): 89-92 ISSN: 0378-1097 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Epimastigotes of Trypanosoma cruzi in culture produce and excrete into the medium small amounts of phenyllactic acid and p-hydroxyphenyllactic acids, presumably arising from the catabolism of the aromatic amino acids phenylalanine and tyrosine, respectively. This production might constitute a minor pathway for the reoxidation of cytosolic NADH, through the concerted action of tyrosine aminotransferase and aromatic alpha-hydroxyacid dehydrogenase. MIME: Indoles-metabolism; Models,-Biological; Oxidation-Reduction; Phenylpropionates-metabolism; Trypanosoma-cruzi-growth-and-development MJME: *Hydroxy-Acids-metabolism; *Lactates-metabolism; *NAD-metabolism; *Oxidoreductases-metabolism; *Trypanosoma-cruzi-metabolism TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 1.; 0; 0; 0; 0; 1821-52-9; 306-23-0; 53-84-9 NM: Oxidoreductases; Hydroxy-Acids; Indoles; Lactates; Phenylpropionates; indole-3-lactic-acid; 4-hydroxyphenyllactic-acid; NAD AN: 94283871 UD: 9409 Record 80 of 131 - MEDLINE (R) 1994 TI: Effects of severe protein restriction in levels of parasitemia and in mortality of mice acutely infected with Trypanosoma cruzi. AU: Gomes-NG; Pereira-FE; Domingues-GC; Alves-JR AD: Departamento de Patologia, Universidade Federal do Espirito Santo, Vitoria. SO: Rev-Soc-Bras-Med-Trop. 1994 Jan-Mar; 27(1): 19-24 ISSN: 0037-8682 PY: 1994 LA: ENGLISH CP: BRAZIL AB: Adult mice were submitted to different degrees of protein restriction for five weeks (4.75, 9.5, 14.25 and 19% of protein in isocaloric diets with normal content of mineral and vitamins), being subsequently infected with two strains of Trypanosoma cruzi: 10(5) trypomastigotes of Y strain or 10(4) trypomastigotes of CL strain. The same diet was maintained for all animals and the infection was followed up by evaluation of blood parasites, mortality and intensity of lesions in the heart and skeleton muscle. Only severe protein restriction (4.75%) induced decrease in resistance to the infection with both the Y and CL strains of T. cruzi, which resulted in higher parasitemia and mortality. The inflammatory lesions in heart and skeleton muscle were less extensive in groups with severe protein restriction despite the increased number of parasite in muscle cells. Depression of immune mechanisms could be responsible for the reduced resistance and reduced inflammatory reaction after T. cruzi infection in severely protein restricted animals. MIME: Acute-Disease; Chagas-Disease-blood; Chagas-Disease-mortality; Dietary-Proteins-administration-and-dosage; Mice-; Protein-Energy-Malnutrition-blood; Protein-Energy-Malnutrition-mortality; Time-Factors MJME: *Chagas-Disease-parasitology; *Protein-Energy-Malnutrition-parasitology TG: Animal; Comparative-Study; Male PT: JOURNAL-ARTICLE RN: 0 NM: Dietary-Proteins AN: 94278165 UD: 9409 Record 81 of 131 - MEDLINE (R) 1994 TI: Sera from chronic Chagasic patients and rodents infected with Trypanosoma cruzi inhibit trans-sialidase by recognizing its amino-terminal and catalytic domain. AU: Pereira-Chioccola-VL; Schenkman-S; Kloetzel-JK AD: Laboratorio de Xenodiagnostico, Instituto Dante Pazzanese de Cardiologia, Sao Paulo, Brazil. SO: Infect-Immun. 1994 Jul; 62(7): 2973-8 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: We investigated whether sera from chronic Chagasic patients and animals infected with Trypanosoma cruzi inhibit the removal of sialic acid from human erythrocytes and the transfer of sialic acid from sialyllactose to [14C]lactose in the reactions catalyzed by the parasite trans-sialidase. Sera from Swiss mice and Calomys callosus animals infected with three different T. cruzi strains inhibit both reactions. Inhibition increases during the infection, reaching maximal levels when the parasitemia decreases. Among 44 sera of untreated chronic Chagasic patients, 40 inhibit both reactions. Inhibition is observed with total, defatted sera or with purified immunoglobulins. Whereas most of the inhibitory antibodies from Chagasic patients react with the papain fragment of trans-sialidase in immunoblots, a few patients have noninhibitory antibodies that react only with the entire trans-sialidase. These findings may be relevant for the pathology of Chagas' disease. MIME: Amino-Acid-Sequence; Erythrocytes-metabolism; IgG-immunology; Lactose-analogs-and-derivatives; Lactose-metabolism; Mice-; Molecular-Sequence-Data; Papain-metabolism; Peptide-Fragments-immunology; Rodentia-; Sialic-Acids-metabolism; Trypanosoma-cruzi-enzymology MJME: *Antibodies,-Protozoan-blood; *Chagas-Disease-immunology; *Neuraminidase-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Comparative-Study; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 3.2.1.-; EC; EC; 0; 0; 0; 0; 35890-38-1; 63-42-3 NM: trans-sialidase,-Trypanosoma-cruzi; Neuraminidase; Papain; Antibodies,-Protozoan; IgG; Peptide-Fragments; Sialic-Acids; N-acetylneuraminoyllactose; Lactose AN: 94274316 UD: 9409 Record 82 of 131 - MEDLINE (R) 1994 TI: Detection of antibodies in sera from Chagas' disease patients using a Trypanosoma cruzi immunodominant recombinant antigen. AU: Paranhos-Bacalla-GS; Santos-MR; Cotrim-PC; Rassi-A; Jolivet-M; Camargo-ME; Da-Silveira-JF AD: Bio-Merieux, Lyon, France; Biolab Diagnostica, Sao Paulo, Brasil. SO: Parasite-Immunol. 1994 Mar; 16(3): 165-9 ISSN: 0141-9838 PY: 1994 LA: ENGLISH CP: ENGLAND AB: A Trypanosoma cruzi DNA fragment encoding an immunodominant repetitive antigen (H49) was subcloned into a protein purification and expressed system. Purified H49 peptide reacted specifically in an enzyme-linked immunosorbent assay (ELISA) with sera from T. cruzi-infected patients, but not with sera from patients with other parasitic diseases such as leishmaniasis and T. rangeli-infection. The H49 recombinant ELISA was able to detect specific antibodies in 84% of chronic chagasic serum samples tested. One of the major advantage of the recombinant ELISA for serodiagnosis of chronic Chagas' disease resides in its high specificity (100%). Our data suggest that recombinant peptides could provide a practical basis for specific diagnosis tests for Chagas' disease. MIME: Antigens,-Protozoan-immunology; Electrophoresis,-Polyacrylamide-Gel; Enzyme-Linked-Immunosorbent-Assay; Immunodominant-Epitopes-genetics; Protozoan-Infections-immunology; Serodiagnosis-; Trypanosoma-cruzi-genetics MJME: *Antibodies,-Protozoan-blood; *Chagas-Disease-immunology; *Immunodominant-Epitopes-immunology; *Recombinant-Proteins-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0 NM: Antibodies,-Protozoan; Antigens,-Protozoan; Immunodominant-Epitopes; Recombinant-Proteins AN: 94268886 UD: 9409 Record 83 of 131 - MEDLINE (R) 1994 TI: Inhibition of Trypanosoma cruzi-specific immune responses by a protein produced by T. cruzi in the course of Chagas' disease. AU: Kierszenbaum-F; Lopez-HM; Sztein-MB AD: Department of Microbiology, Michigan State University, East Lansing 48824-1101. SO: Immunology. 1994 Mar; 81(3): 462-7 ISSN: 0019-2805 PY: 1994 LA: ENGLISH CP: ENGLAND AB: Immunosuppression is readily demonstrable in the acute phase of Trypanosoma cruzi infection but subsides during the chronic phase. In vitro, living T. cruzi induces important alterations in mitogen-activated human T and B lymphocytes and inhibits their capacity to proliferate. These effects are reproduced by a protein spontaneously released by this parasite, termed trypanosomal immunosuppressive factor (TIF). In this study we asked whether TIF would also inhibit a T. cruzi-specific immune response and if it is produced in a mammalian host during infection. A significant reduction in the level of [3H]thymidine incorporation by spleen cells from chronically infected mice stimulated with a T. cruzi antigen preparation ensued when TIF was added to the cultures. Production of TIF in T. cruzi-infected individuals was denoted by the ability of serum IgG from either chronically infected patients or mice to abolish, in a concentration-dependent manner, the capacity of TIF to suppress interleukin-2 receptor expression by phytohaemagglutinin-stimulated human lymphocytes. This neutralizing activity was absent in the IgG fractions prepared from sera of healthy volunteers, noninfected mice or mice killed at different times during acute T. cruzi infection. Circulating anti-TIF antibodies represent indirect evidence of TIF production in vivo which, together with TIF-mediated inhibition of T. cruzi-specific lymphoproliferation, raise the possibility that TIF controls anti-parasite immune responses in vivo. The presence of TIF-neutralizing antibodies during chronic but not acute T. cruzi infection may be one of the reasons why immunosuppression is confined to the acute stage. MIME: Acute-Disease; Cell-Division-immunology; Chronic-Disease; IgG-immunology; Lymphocytes-immunology; Mice-; Mice,-Inbred-CBA; Mice,-Inbred-Strains; Receptors,-Interleukin-2-analysis; Spleen-immunology; Suppressor-Factors,-Immunologic-biosynthesis MJME: *Antigens,-Protozoan-immunology; *Chagas-Disease-immunology; *Immune-Tolerance-immunology; *Suppressor-Factors,-Immunologic-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI26542AINIAID RN: 0; 0; 0; 0 NM: Antigens,-Protozoan; IgG; Receptors,-Interleukin-2; Suppressor-Factors,-Immunologic AN: 94266333 UD: 9409 Record 84 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi: murine virus contaminant of the experimental infection. AU: Rangel-H-de-A; Verinaud-L; Camargo-IJ; Gilioli-R; Sakurada-JK AD: Department of Immunology, Institute of Biology, Sao Paulo, Brazil. SO: Exp-Parasitol. 1994 Jun; 78(4): 429-31 ISSN: 0014-4894 PY: 1994 LA: ENGLISH CP: UNITED-STATES MIME: Coronavirus-immunology; Coronavirus-Infections-immunology; Mice-; Mice,-Inbred-BALB-C; Mice,-Inbred-CBA; Serial-Passage MJME: *Chagas-Disease-complications; *Coronavirus-isolation-and-purification; *Coronavirus-Infections-complications TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE AN: 94265901 UD: 9409 Record 85 of 131 - MEDLINE (R) 1994 TI: Evidence for guanosine triphosphate--binding proteins in Trypanosoma cruzi. AU: Oz-HS; Huang-H; Wittner-M; Tanowitz-HB; Bilezikian-JP; Morris-SA AD: Department of Pathology, Albert Einstein College of Medicine, Bronx, New York. SO: Am-J-Trop-Med-Hyg. 1994 May; 50(5): 620-31 ISSN: 0002-9637 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The transformation of the parasite Trypanosoma cruzi from the blood-borne trypomastigote to the intracellular amastigote constitutes a key clinical feature in the pathophysiology of Chagas' disease. That this transition occurs without change in the integrity of the plasma membrane of the parasite suggests the presence of biochemical structures, i.e., signal transduction systems, that convey information regarding the external milieu of the host so as to facilitate this transformation. In higher eukaryotes, it has been found that a heterotrimeric GTP-binding protein (G-protein), composed of alpha beta gamma subunits, constitutes a critical component of this complex. Two closely related groups of G-proteins are substrates for cholera toxin (CT)- (Gs) and pertussis toxin (PT)- (Gi1-3 and Go) dependent ADP ribosylation. In concert, they link plasma membrane receptors to adenylate cyclase, resulting in the stimulation or inhibition, respectively, of cAMP generation. In this report, we demonstrate the presence of both groups of G-proteins. Cholera toxin-dependent ADP ribosylation of 42- and 45-kD proteins was demonstrable in amastigotes (AMAST), in the cytosol of epimastigotes (EPI), and weakly in trypomastigotes (TRYP), suggesting the presence of the stimulatory GTP-binding protein, Gs, in T. cruzi. Antisera generated against the alpha s subunit of the Gs heterotrimeric protein (anti-alpha s) bound to a 45-kD protein CT substrate in the rank order TRYP >> AMAST approximately EPI cytosol. Immunoprecipitation of CT-32P-ADP-ribosylated membranes with anti-alpha s resulted in 42- and 45-kD proteins. However, no Gs-mediated activation of adenylate cyclase was demonstrable in reconstitution studies using cyc- lymphoma cells, which lack a functional Gs but possess a beta-adrenergic receptor and adenylyl cyclase enzyme. Pertussis toxin-catalyzed ADP ribosylation was demonstrable in 39-40-kD particulate proteins of EPI, less strongly in AMAST, and least in TRYP, consistent with the presence of inhibitory (Gi) and Go GTP-binding proteins. In support of this observation, immunochemical analysis of the PT substrates identified the presence of alpha o and alpha i1-2-3 in EPI, AMAST and TRYP, although, with the exception of alpha i3, both toxin and associated immunochemical PT substrates are decreased in AMAST and TRYP relative to EPI. Although the functions of these putative G-proteins in T. cruzi are still unclear, their expression may be regulated by the state of parasite differentiation.(ABSTRACT TRUNCATED AT 400 WORDS) MIME: Adenosine-Diphosphate-metabolism; Amino-Acid-Sequence; Blotting,-Western; Cholera-Toxin-metabolism; Immune-Sera-chemistry; Molecular-Sequence-Data; Pertussis-Toxins-metabolism; Precipitin-Tests; Signal-Transduction; Trypanosoma-cruzi-metabolism MJME: *G-Proteins-analysis; *Protozoan-Proteins-analysis; *Trypanosoma-cruzi-chemistry TG: Animal; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI12770AINIAID; AI26368AINIAID; AI29749AINIAID RN: 0; 0; 0; 0; 58-64-0; 70323-44-3; 9012-63-9 NM: G-Proteins; Helminth-Proteins; Immune-Sera; Protozoan-Proteins; Adenosine-Diphosphate; Pertussis-Toxins; Cholera-Toxin AN: 94262998 UD: 9409 SB: AIM Record 86 of 131 - MEDLINE (R) 1994 TI: Didelphis marsupialis, an important reservoir of Trypanosoma (Schizotrypanum) cruzi and Leishmania (Leishmania) chagasi in Colombia. AU: Travi-BL; Jaramillo-C; Montoya-J; Segura-I; Zea-A; Goncalves-A; Velez-ID AD: Fundacion Centro Internacional de Entrenamiento e Investigaciones Medicas, Cali, Colombia. SO: Am-J-Trop-Med-Hyg. 1994 May; 50(5): 557-65 ISSN: 0002-9637 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The role of Didelphis marsupialis as a reservoir of zoonotic hemoflagellates was examined in two ecologically distinct settings in Colombia. While 72% (12 of 18) of the opossums collected in the tropical rain forest harbored Trypanosoma cruzi, other mammals in the area had lower infection rates: 1.3% (Proechymis semispinosus [spiny rat]; 13% Tylomys mirae [climbing rat]; and 6% Rattus rattus). Trypanosoma cruzi isolates from D. marsupialis were similar to zymodeme 1 (Z1), and two of four phenotypes were shared with Tylomys mirae, which is also predominantly arboreal. Terrestrial (P. semispinosus) and peridomestic (R. rattus) animals were infected with Z3 or other Z1 phenotypes, respectively. Schizodeme analysis showed polymorphisms among isolates from mammals, reflecting diverse modes of transmission, and a complex epidemiologic situation. Despite the lower infection rate of the opossum (14%) found in our study in the tropical dry forest as compared with the tropical wet forest, Chagas' disease has been reported only in the former area. This suggests that the lack of alternative blood sources for triatomines of the tropical dry forest, where mammals are less abundant than in the wet forest, may increase the risk of human infection. Among several species of mammals captured in the tropical dry forest, Leishmania chagasi was isolated from 22.7% (5 of 22) D. marsupialis. This finding confirms the important role of opossums in Colombian foci of visceral leishmaniasis, including those where the phlebotomine species involved in transmission is Lutzomyia evansi, an alternative vector to the more common Lutzomyia longipalpis. MIME: Colombia-; Isoenzymes-analysis; Polymorphism-Genetics; Rain-; Tropical-Climate; Trypanosoma-cruzi-classification; Trypanosoma-cruzi-enzymology; Trypanosoma-cruzi-isolation-and-purification MJME: *Chagas-Disease-transmission; *Disease-Reservoirs; *Leishmania-infantum-isolation-and-purification; *Leishmaniasis,-Visceral-transmission; *Opossums-parasitology; *Zoonoses- TG: Animal; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI16315AINIAID RN: 0 NM: Isoenzymes AN: 94262987 UD: 9409 SB: AIM Record 87 of 131 - MEDLINE (R) 1994 TI: Myocardial changes in acute Trypanosoma cruzi infection. Ultrastructural evidence of immune damage and the role of microangiopathy. AU: Andrade-ZA; Andrade-SG; Correa-R; Sadigursky-M; Ferrans-VJ AD: Goncalo Moniz Research Center (FIOCRUZ), Bahia, Brazil. SO: Am-J-Pathol. 1994 Jun; 144(6): 1403-11 ISSN: 0002-9440 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Histological and ultrastructural studies of the hearts of dogs sacrificed 18 to 26 days after intraperitoneal inoculation with 4 x 10(5) blood forms of the 12 SF strain of Trypanosoma cruzi/kg of body weight disclosed myocarditis characterized by parasitic invasion of some myocytes, damage and necrosis of nonparasitized myocytes, and interstitial infiltration by mononuclear cells. Nonparasitized myocytes showed alterations ranging from mild edema to severe myocytolysis. These changes often were accompanied by contacts of myocytes with lymphocytes (both granular and agranular) and macrophages. These contacts were characterized by focal loss of the myocyte basement membrane and close approximation of the plasma membranes of the two cells. Contacts between lymphocytes and capillary endothelial cells were also frequent. Platelet aggregates and fibrin microthrombi were observed in some capillaries. Our findings suggest that immune effector cells play a major role in the pathogenesis of the myocyte damage and the microangiopathy in acute Chagas' disease. MIME: Cell-Communication; Chagas-Cardiomyopathy-immunology; Dogs-; Endothelium,-Vascular-pathology; Endothelium,-Vascular-ultrastructure; Lymphocytes-pathology; Lymphocytes-ultrastructure; Macrophages-pathology; Macrophages-ultrastructure; Microcirculation-; Microscopy,-Electron; Necrosis- MJME: *Chagas-Cardiomyopathy-pathology; *Coronary-Vessels-pathology; *Heart-parasitology; *Myocardium-pathology; *Myocardium-ultrastructure; *Trypanosoma-cruzi-ultrastructure TG: Animal PT: JOURNAL-ARTICLE AN: 94262719 UD: 9409 SB: AIM Record 88 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi: studies on the reactivity of antibodies bound to the surface of blood forms at the early phase of infection. AU: De-Gaspari-EN; Stolf-AM; Umezawa-ES; Zingales-B; Abrahamsohn-IA AD: Secao de Imunologia, Instituto Adolfo Lutz, Sao Paulo, Brazil. SO: Acta-Trop. 1994 Feb; 56(1): 79-87 ISSN: 0001-706X PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: The specificity and reactivity of antibodies bound to the surface of Trypanosoma cruzi blood forms at the very early acute phase of murine infection was investigated. Surface-bound antibodies of the IgG and IgM isotypes were recovered from blood forms upon incubation at 37 degrees C. The eluted antibodies immunoprecipitated several trypomastigote surface polypeptides from 80 to 100 kDa. In contrast, for epimastigotes a very faint reactivity was detected only for antigens of 50 and 95 kDa. The shed antibodies promoted in vitro complement-mediated lysis of live blood forms and reacted with fixed trypomastigotes by immunofluorescence. Thus, blood forms are already coated with active trypomastigote-specific antibodies with a potential role in the host defense, although the low levels of serum antibodies have prevented the demonstration of humoral protection at the early stages of infection. MIME: Antigenic-Determinants; IgG-immunology; IgM-immunology; Mice-; Mice,-Inbred-BALB-C MJME: *Antibodies,-Protozoan-immunology; *Antigens,-Surface-immunology; *Chagas-Disease-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0 NM: Antibodies,-Protozoan; Antigenic-Determinants; Antigens,-Surface; IgG; IgM AN: 94262527 UD: 9409 Record 89 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi affects nitric oxide production by murine peritoneal macrophages. AU: Pakianathan-DR; Kuhn-RE AD: Department of Biology, Wake Forest University, Winston-Salem, North Carolina 27109. SO: J-Parasitol. 1994 Jun; 80(3): 432-7 ISSN: 0022-3395 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Macrophages from mice that are infected with various intracellular pathogens including Leishmania major, Trypanosoma cruzi, and Salmonella typhimurium are stimulated to produce large quantities of nitric oxide (NO). Both viable and heat-treated L. major amastigotes have been shown to be effective co-signals for NO production in vitro. NO produced by macrophages has anti-microbial and immunosuppressive functions in an immune response. We have shown previously that NO plays a complicated role in T. cruzi infections since macrophages are important both in mediating an immune response against the parasite as well as in mediating immunosuppression. In this study we examined how T. cruzi affects NO production by macrophages from C3HeB/FeJ and C57BL/6 mice in vitro. We found that live trypomastigotes neither stimulate nor decrease NO production by interferon (IFN)-gamma-activated macrophages. However, heat-treated or glutaraldehyde-fixed trypomastigotes of T. cruzi significantly decrease NO production by IFN-gamma-activated macrophages and as a result decrease macrophage-mediated trypanocidal and immunosuppressive activity. We have determined that this decrease in NO production by T. cruzi is not due to stimulation of transforming growth factor-beta production and involves tumor necrosis factor-alpha only in C3HeB/FeJ macrophages. This study demonstrates the complexity of the T. cruzi-macrophage interaction as well as confirms previously demonstrated differences between macrophages from 2 strains of mice. MIME: Interferon-gamma,-Recombinant-pharmacology; Lymphocyte-Transformation; Macrophages,-Peritoneal-immunology; Mice-; Mice,-Inbred-C3H; Mice,-Inbred-C57BL; Tumor-Necrosis-Factor-pharmacology MJME: *Chagas-Disease-immunology; *Macrophages,-Peritoneal-metabolism; *Nitric-Oxide-biosynthesis; *Trypanosoma-cruzi-immunology TG: Animal; Female PT: JOURNAL-ARTICLE RN: 0; 0; 10102-43-9 NM: Interferon-gamma,-Recombinant; Tumor-Necrosis-Factor; Nitric-Oxide AN: 94253984 UD: 9409 Record 90 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi ribosomal DNA: mapping of a putative distal promoter. AU: Martinez-Calvillo-S; Hernandez-R AD: Instituto de Investigaciones Biomedicas, Universidad Nacional Autonoma de Mexico, Mexico D.F. SO: Gene. 1994 May 16; 142(2): 243-7 ISSN: 0378-1119 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: The nucleotide (nt) sequence (2106 bp) of a cloned rDNA (encoding ribosomal RNA) spacer region from Trypanosoma cruzi was determined and a putative transcription start point (tsp) was mapped. The assigned length for the transcribed spacer is 1768 bp and its tsp is present 270-bp upstream from an alternative tsp published for the equivalent gene from another T. cruzi strain [Dietrich et al., Gene 125 (1993) 103-107]. Sequence comparisons of the nt flanking both T. cruzi tsp with the homologous regions from both other trypanosomatids, and other eukaryotes, indicate that these sequences are poorly conserved within the family Trypanosomatidae. This finding reinforces the proposal that the speciation of trypanosomatids may have occurred early in evolution. MIME: Base-Sequence; Conserved-Sequence; DNA,-Protozoan-chemistry; DNA,-Protozoan-genetics; DNA,-Ribosomal-chemistry; Genes,-Protozoan-genetics; Molecular-Sequence-Data; RNA-Processing,-Post-Transcriptional; RNA,-Protozoan-analysis; RNA,-Ribosomal-analysis; Sequence-Alignment; Sequence-Analysis,-DNA; Transcription,-Genetic MJME: *DNA,-Ribosomal-genetics; *Promoter-Regions-Genetics-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0 NM: DNA,-Protozoan; DNA,-Ribosomal; RNA,-Protozoan; RNA,-Ribosomal AN: 94252574 UD: 9409 SI: GENBANK/L13926 Record 91 of 131 - MEDLINE (R) 1994 TI: Identification of antibodies with muscarinic cholinergic activity in human Chagas' disease: pathological implications. AU: Goin-JC; Borda-E; Leiros-CP; Storino-R; Sterin-Borda-L AD: CEFYBO-CONICET, Buenos Aires, Argentina. SO: J-Auton-Nerv-Syst. 1994 Apr; 47(1-2): 45-52 ISSN: 0165-1838 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: We examined the possible role of altered humoral immunity in dysautonomic syndrome in Chagas' disease by analyzing the effect of sera and IgG on the binding of radioligand to heart muscarinic cholinergic receptors and on the contractility of myocardium. Human Chagasic IgG inhibited in a non-competitive manner the binding of [3H]quinuclidinyl benzilate to the cardiac cell membrane. Moreover, human Chagasic IgG behaved as a partial muscarinic cholinergic agonist, reducing heartcontractility and inhibiting the action of pilocarpine. The prevalence of the cholinergic antibody activity was higher in sera from T. cruzi-infected asymptomatic individuals with dysautonomic syndrome than in those without autonomic nervous system alterations. The presence of these antibodies could explain the progressive receptor blockade in the parasympathetic branch of the autonomic nervous system, leading to dysautonomia. MIME: Antibody-Formation-physiology; Autonomic-Nervous-System-Diseases-etiology; Autonomic-Nervous-System-Diseases-immunology; Autonomic-Nervous-System-Diseases-pathology; Cell-Membrane-metabolism; Chagas-Disease-complications; Chagas-Disease-pathology; Erythrocytes-immunology; Erythrocytes-metabolism; IgG-immunology; Myocardial-Contraction-drug-effects; Pilocarpine-pharmacology; Quinuclidinyl-Benzilate-diagnostic-use; Rats-; Turkeys- MJME: *Antibodies-physiology; *Chagas-Disease-immunology; *Receptors,-Muscarinic-immunology TG: Animal; Human; In-Vitro; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 6581-06-2; 92-13-7 NM: Antibodies; IgG; Receptors,-Muscarinic; Quinuclidinyl-Benzilate; Pilocarpine AN: 94246086 UD: 9408 Record 92 of 131 - MEDLINE (R) 1994 TI: Molecular cloning and characterization of the 78-kilodalton glucose-regulated protein of Trypanosoma cruzi. AU: Tibbetts-RS; Kim-IY; Olson-CL; Barthel-LM; Sullivan-MA; Winquist-AG; Miller-SD; Engman-DM AD: Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, Illinois. SO: Infect-Immun. 1994 Jun; 62(6): 2499-507 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The protozoan Trypanosoma cruzi is the etiologic agent of Chagas' disease, an illness responsible for morbidity and death among millions of Latin Americans. Mice also develop this disease when infected with T. cruzi and are a useful model organism for the study of parasite-specific immune responses. To identify immunogenic T. cruzi antigens, serum from an infected mouse was used to isolate clones from a T. cruzi epimastigote cDNA expression library. One of these clones was found to encode the 78-kDa glucose-regulated protein (grp78), the endoplasmic reticular member of the 70-kDa heat shock protein (hsp70) family. Like the mammalian and yeast grp78s, the T. cruzi protein contains an endoplasmic reticular leader peptide and a carboxyl-terminal endoplasmic reticular retention sequence. T. cruzi grp78 is encoded by a tandemly arranged family of three genes located on a chromosome of 1.6 Mb. The effects on grp78 expression of heat shock and tunicamycin treatment, the latter of which specifically stimulates mammalian grp78, were investigated. While the level of the grp78 protein remained constant under all circumstances, grp78 mRNA was unaffected by heat shock but induced fivefold by tunicamycin. Finally, we found that grp78 is the most immunogenic of the T. cruzi heat shock proteins we have characterized, reacting strongly in immunoblots with sera from infected mice. MIME: Amino-Acid-Sequence; Antibodies,-Protozoan-analysis; Base-Sequence; Carrier-Proteins-analysis; Carrier-Proteins-immunology; Chagas-Disease-immunology; Chromosome-Mapping; Cloning,-Molecular; Heat-; Mice-; Molecular-Sequence-Data; RNA,-Messenger-analysis MJME: *Carrier-Proteins-genetics; *Heat-Shock-Proteins-genetics; *Trypanosoma-cruzi-immunology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0 NM: immunoglobulin-heavy-chain-binding-protein; Antibodies,-Protozoan; Carrier-Proteins; Heat-Shock-Proteins; RNA,-Messenger AN: 94245361 UD: 9408 SI: GENBANK/L23420 Record 93 of 131 - MEDLINE (R) 1994 TI: A lytic monoclonal antibody to Trypanosoma cruzi bloodstream trypomastigotes which recognizes an epitope expressed in tissues affected in Chagas' disease. AU: Zwirner-NW; Malchiodi-EL; Chiaramonte-MG; Fossati-CA AD: Instituto de Estudios de la Inmunidad Humoral, Catedra de Inmunologia, Facultad de Farmacia y Bioquimica, Universidad de Buenos Aires, Argentina. SO: Infect-Immun. 1994 Jun; 62(6): 2483-9 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: It has been suggested that molecular mimicry between the antigens of Trypanosoma cruzi and the host could have a role in the onset of the chronic stage of Chagas' disease. In this article, we report on a monoclonal antibody (MAb), CAK20.12 (immunoglobulin G2b), which reacts with a polypeptidic epitope of a 150-kDa antigen expressed on the surface of several strains of T. cruzi. This MAb also causes lysis of bloodstream trypomastigotes. Serum samples from 30 of 30 patients with chronic and 11 of 13 patients with acute Chagas' disease present specific antibodies to this antigen. MAb CAK20.12 reacts, by indirect immunofluorescence, with human and syngeneic murine striated muscle tissue, with the smooth muscle layer of cardiac arteries, with the lamina muscularis mucosae and the external striated muscle layer of the esophagus, and with the smooth muscle cells of the colon from normal syngeneic mice. Reactivity with the small intestine was very weak, and no reactivity with ventricle or atrium tissue was detected. Adsorption with an antigenic fraction from normal murine striated muscle or from T. cruzi epimastigotes confirmed that MAb CAK20.12 recognizes a common epitope present in parasites and host tissues. MAb CAK20.12, lytic for the infective form of T. cruzi, recognizes an epitope expressed in striated and smooth muscle cells of the host tissues affected in the chronic stage of Chagas' disease. MIME: Blotting,-Western; Fluorescent-Antibody-Technique; Mice-; Mice,-Inbred-BALB-C MJME: *Antibodies,-Monoclonal-immunology; *Antigenic-Determinants-analysis; *Antigens,-Protozoan-immunology; *Chagas-Disease-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0 NM: Antibodies,-Monoclonal; Antigenic-Determinants; Antigens,-Protozoan AN: 94245359 UD: 9408 Record 94 of 131 - MEDLINE (R) 1994 TI: Skewed V beta TCR repertoire of CD8+ T cells in murine Trypanosoma cruzi infection. AU: Leite-de-Moraes-MC; Coutinho-A; Hontebeyrie-Joskowicz-M; Minoprio-P; Eisen-H; Bandeira-A AD: Unite d'Immunoparasitologie, Institut Pasteur, Paris, France. SO: Int-Immunol. 1994 Mar; 6(3): 387-92 ISSN: 0953-8178 PY: 1994 LA: ENGLISH CP: ENGLAND AB: We have followed CD4 and CD8 TCR V beta repertoires during the acute phase of Trypanosoma cruzi infection in a resistant mouse strain (C57BL/6). No major changes were found in the V beta TCR distributions analyzed (covering roughly 40% of the TCR repertoire) in peripheral CD4 T lymphocytes, confirming the polyclonal nature of CD4 T cell responses. In contrast, in most animals, an over-representation of V beta 5 and V beta 14 TCR families was disclosed in the CD8 T cell compartment, superimposed on a predominantly polyclonal response. The preferential expansion of V beta 5+CD8+ T cells was also observed after infection of sensitive (C3H/HeJ, BALB/c) mouse strains. These observations suggest the existence of CD8 T cell-directed superantigenic activities associated with parasites. MIME: Cells,-Cultured; Flow-Cytometry; Mice-; Mice,-Inbred-BALB-C; Mice,-Inbred-C3H; Mice,-Inbred-C57BL; Spleen-cytology MJME: *Antigens,-CD8-analysis; *Chagas-Disease-immunology; *Receptors,-Antigen,-T-Cell,-alpha-beta-analysis; *Receptors,-Antigen,-T-Cell,-alpha-beta-genetics; *T-Lymphocytes-immunology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0 NM: Antigens,-CD8; Receptors,-Antigen,-T-Cell,-alpha-beta AN: 94242716 UD: 9408 Record 95 of 131 - MEDLINE (R) 1994 TI: Analyzing expression of the calmodulin and ubiquitin-fusion genes of Trypanosoma cruzi using simultaneous, independent dual gene replacements. AU: Chung-SH; Gillespie-RD; Swindle-J AD: Department of Microbiology and Immunology, University of Tennessee, Memphis 38163. SO: Mol-Biochem-Parasitol. 1994 Jan; 63(1): 95-107 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: We describe here a strategy for introducing simultaneous, independent gene replacements into the Trypanosoma cruzi chromosome. The goal of this study was to use two linear DNA fragments to simultaneously replace the CalA2 calmodulin and FUS1 ubiquitin-fusion genes with the neomycin resistance (neo(r)) and chloramphenicol acetyltransferase (CAT) genes, respectively. One clone (D6), of thirty G418-resistant clones analyzed, carried the desired dual gene replacement. CDNA sequence analysis indicated that the CAT mRNA was accurately trans-spliced using the previously identified FUS1 mini-exon addition site. However, DNA sequence analysis of the intergenic sequence immediately upstream of the neo(r) gene in clone D6 identified a mutation which altered the pattern of trans-splicing of the neo(r) mRNA. Possible effects of this mutation on 3' splice acceptor site selection are discussed. MIME: Base-Sequence; Calmodulin-genetics; Chloramphenicol-Acetyltransferase-genetics; Cloning,-Molecular; Drug-Resistance-genetics; DNA,-Protozoan-genetics; DNA,-Recombinant-genetics; Gene-Expression-Regulation; Genetic-Vectors; Molecular-Sequence-Data; Neomycin-pharmacology; RNA-Splicing-genetics; RNA,-Messenger-genetics; RNA,-Protozoan-genetics; Transformation,-Genetic; Trypanosoma-cruzi-drug-effects; Ubiquitin-genetics MJME: *Genes,-Protozoan; *Trypanosoma-cruzi-genetics TG: Animal; Support,-U.S.-Gov't,-P.H.S. GS: CalA2; FUS1; neor; CAT PT: JOURNAL-ARTICLE CN: AI26578AINIAID; RR05423RRNCRR RN: EC; 0; 0; 0; 0; 0; 0; 0; 1404-04-2 NM: Chloramphenicol-Acetyltransferase; Calmodulin; DNA,-Protozoan; DNA,-Recombinant; Genetic-Vectors; RNA,-Messenger; RNA,-Protozoan; Ubiquitin; Neomycin AN: 94239476 UD: 9408 Record 96 of 131 - MEDLINE (R) 1994 TI: Characterization of kinetoplast DNA minicircles in Trypanosoma rangeli. AU: Recinos-RF; Kirchhoff-LV; Donelson-JE AD: Department of Biochemistry, University of Iowa, Iowa City. SO: Mol-Biochem-Parasitol. 1994 Jan; 63(1): 59-67 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Kinetoplast DNA, the mitochondrial DNA of kinetoplastid protozoans, is a network of interlocked minicircles and maxicircles. We analyzed the sequence organization of minicircle DNAs in the El Tocuyo strain and the San Augustin clone B6 of Trypanosoma rangeli. The frequencies of different minicircle types, as defined by the number of 136-bp conserved regions (CRs), are different in the two strains. About half of the 1.7-kb T. rangeli El Tocuyo minicircles have 1 CR and most of the others have 2. In contrast, most of the 1.6-kb T. rangeli San Augustin minicircles have 2 CRs, while some have four. The CR contains a replication origin at one end and is conserved both within and between the two strains. Comparisons of the T. rangeli El Tocuyo and T. rangeli San Augustin minicircle CRs with minicircle CRs of other kinetoplastid species reveal that they are most similar to those of Trypanosoma cruzi. MIME: Base-Sequence; Cloning,-Molecular; Conserved-Sequence; DNA,-Kinetoplast-ultrastructure; Microscopy,-Electron; Models,-Genetic; Molecular-Sequence-Data; Sequence-Homology,-Nucleic-Acid; Species-Specificity; Trypanosoma-cruzi-genetics MJME: *DNA,-Kinetoplast-genetics; *Trypanosoma-genetics TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI2471AINIAID; DK25295DKNIDDK; GM07337GMNIGMS RN: 0 NM: DNA,-Kinetoplast AN: 94239472 UD: 9408 SI: GENBANK/L19388; GENBANK/L19389; GENBANK/L19390; GENBANK/L19391; GENBANK/L19392; GENBANK/L19393; GENBANK/L19394; GENBANK/L19395; GENBANK/M18814; GENBANK/M18815; GENBANK/M18816; GENBANK/M19174; GENBANK/M19175; GENBANK/M19176; GENBANK/M19177; GENBANK/M19178; GENBANK/M19179; GENBANK/M19180; GENBANK/M19181; GENBANK/M19185; GENBANK/M19186; GENBANK/M19187; GENBANK/M19188; GENBANK/M19189; GENBANK/M19190; GENBANK/M19191; GENBANK/M19192; GENBANK/J01454; GENBANK/J01455; GENBANK/M15322; + Record 97 of 131 - MEDLINE (R) 1994 TI: Expression and evolution of members of the Trypanosoma cruzi trypomastigote surface antigen multigene family. AU: Ruef-BJ; Dawson-BD; Tewari-D; Fouts-DL; Manning-JE AD: Department of Molecular Biology and Biochemistry, University of California, Irvine 92717. SO: Mol-Biochem-Parasitol. 1994 Jan; 63(1): 109-20 ISSN: 0166-6851 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: The trypomastigote specific surface antigens of Trypanosoma cruzi are encoded by a supergene family which includes the TSA family. The TSA family is characterized by the presence of a 27-bp tandem repeat array in the coding region. Here, we report the characterization and analysis of the three TSA family members in the Esmeraldo strain of the parasite. In this strain 2 distinct telomeric members are expressed abundantly as 3.7-kb mRNAs, while the remaining member is located at an internal chromosomal site and is expressed at less than 2% of the level seen for the telomeric members. Based on hybridization to DNA separated by PFGE, 3 chromosomes of sizes 1.8 Mb, 0.98 Mb, and 0.90 Mb each contain one of the telomeric members. In addition, the two smaller chromosomes also contain the single internal member. Since both chromosomes contain similar TSA family members, and vary only slightly in size, we suggest that they are homologues. Comparisons of the nucleotide sequences of the different members of the family show that the internal gene differs from the telomeric genes primarily in sequences found 3' of the repeat array. These comparisons also reveal that the three genes are analogous, supporting the hypothesis that short segments between the family members are exchanged by gene conversion events. We propose that similar conversion events between members of different gene families may generate some of the diversity found within the supergene family. MIME: Amino-Acid-Sequence; Antigens,-Surface-genetics; Base-Sequence; DNA,-Protozoan-genetics; Evolution-; Gene-Conversion; Gene-Expression; Molecular-Sequence-Data; Repetitive-Sequences,-Nucleic-Acid; Restriction-Mapping; Sequence-Homology,-Amino-Acid; Sequence-Homology,-Nucleic-Acid; Trypanosoma-cruzi-growth-and-development; Variant-Surface-Glycoproteins,-Trypanosoma-genetics; Variant-Surface-Glycoproteins,-Trypanosoma-immunology; Variation-Genetics MJME: *Antigens,-Protozoan-genetics; *Genes,-Protozoan; *Genes,-Reiterated; *Trypanosoma-cruzi-genetics; *Trypanosoma-cruzi-immunology TG: Animal; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI18873AINIAID RN: 0; 0; 0; 0; 138820-03-8 NM: Antigens,-Protozoan; Antigens,-Surface; DNA,-Protozoan; Variant-Surface-Glycoproteins,-Trypanosoma; trypomastigote-specific-surface-antigen AN: 94239457 UD: 9408 SI: GENBANK/U02613; GENBANK/U02614; GENBANK/U02615 Record 98 of 131 - MEDLINE (R) 1994 TI: Transfer of modified sialic acids by Trypanosoma cruzi trans-sialidase for attachment of functional groups to oligosaccharide [published erratum appears in Anal Biochem 1994 May 1;218(2):484] AU: Lee-KB; Lee-YC AD: Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218. SO: Anal-Biochem. 1994 Feb 1; 216(2): 358-64 ISSN: 0003-2697 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Donor specificity of Trypanosoma cruzi trans-sialidase (TcTs) has been investigated with modified 2-[4-methylumbelliferone]-alpha- ketoside of N-acetyl-D-neuraminic acid (4MU-NANA) as donor and lactose as acceptor. 4MU-NANA was treated with periodate under mild conditions to generate an aldehyde on the exocyclic side chain. The oxidized 4MU-NANA was derivatized with various primary amines by reductive amination to yield potential donors. High-performance anion-exchange chromatography equipped with pulsed amperometric detector was used to assay the transglycosylation activity of TcTs. Several modified 4MU-NANA derivatives served as substrates by TcTs and they may be utilized to make valuable intermediates, including those for fluorescence energy transfer measurement or photoaffinity labeling experiment. MIME: Amination-; Carbohydrate-Sequence; Chromatography,-High-Pressure-Liquid; Chromatography,-Ion-Exchange-methods; Fibrinogen-metabolism; Fluorescent-Dyes-metabolism; Glycopeptides-metabolism; Hymecromone-analogs-and-derivatives; Hymecromone-metabolism; Molecular-Sequence-Data; Oxidation-Reduction; Periodic-Acid; Substrate-Specificity MJME: *Neuraminidase-metabolism; *Oligosaccharides-metabolism; *Sialic-Acids-metabolism; *Trypanosoma-cruzi-enzymology TG: Animal PT: JOURNAL-ARTICLE RN: EC; 0; 0; 0; 0; 0; 10450-60-9; 59322-44-0; 90-33-5; 9001-32-5 NM: Neuraminidase; Bacterial-Proteins; Fluorescent-Dyes; Glycopeptides; Oligosaccharides; Sialic-Acids; Periodic-Acid; 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic-acid; Hymecromone; Fibrinogen AN: 94234563 UD: 9408 Record 99 of 131 - MEDLINE (R) 1994 TI: Evaluation of a serological test system for the diagnosis of Sarcocystis cruzi infection in cattle using S. cruzi merozoite antigen. AU: Savini-G; Dunsmore-JD; Robertson-ID AD: School of Veterinary Studies, Murdoch University, Australia. SO: Vet-Parasitol. 1994 Feb; 51(3-4): 181-9 ISSN: 0304-4017 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Fifty serum samples were examined by enzyme-linked immunosorbent assay (ELISA) for antibodies directed against crude antigens isolated from cystozoites and merozoites of Sarcocystis cruzi and Sarcocystis tenella/Sarcocystis arieticanis and merozoites of Toxoplasma gondii. Of these 50 samples, 25 were from cattle originating from an area where the prevalence of S. cruzi was found to be very low (9%) and in which no cystozoites were detected, and 25 were from cattle which were found by a digestion method to be heavily infected with S. cruzi. A very high correlation was observed between the parasitological data and the results obtained from the serological assays which used antigens from either cystozoites or merozoites of S. cruzi. The assay using the antigen derived from merozoites provided the best result for discriminating infected and non-infected animals. There was some cross-reactivity between the antigen derived from cystozoites of heterologous species of Sarcocystis and S. cruzi antibodies, and some cross-reactivity between antigen of T. gondii and antibodies to S. cruzi. The reproducibility of the assays was found to be high and similar results were observed when the sera were tested on two separate occasions. The unpurified S. cruzi merozoite antigen produced in vitro is relatively accurate in discriminating positive and negative animals and may be used for diagnosis in economically important hosts such as cattle and sheep. MIME: Cattle-; Cross-Reactions; Enzyme-Linked-Immunosorbent-Assay; False-Negative-Reactions; Reproducibility-of-Results; Sarcocystosis-diagnosis; Toxoplasma-immunology MJME: *Antibodies,-Protozoan-blood; *Antigens,-Protozoan-immunology; *Cattle-Diseases-diagnosis; *Sarcocystis-immunology; *Sarcocystosis-veterinary TG: Animal; Comparative-Study PT: JOURNAL-ARTICLE RN: 0; 0 NM: Antibodies,-Protozoan; Antigens,-Protozoan AN: 94225769 UD: 9408 Record 100 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi glycoprotein of M(r) 56,000 characterization and assessment of its potential to protect against fatal parasite infections. AU: Harth-G; Mills-AA; Lin-T; Araujo-FG AD: Department of Medicine, University of California Los Angeles 90024-1680. SO: Mol-Microbiol. 1994 Jan; 11(2): 261-71 ISSN: 0950-382X PY: 1994 LA: ENGLISH CP: ENGLAND AB: A approximately 56,000 Da membrane glycoprotein purified from epimastigotes of Trypanosoma cruzi was characterized biochemically and tested for its efficacy to induce protection in mice from a lethal challenge with this protozoan parasite. Immunofluorescence assays with live and formalin-fixed epimastigotes and trypomastigotes localized the glycoprotein to the flagellum, the body of the parasite, and the cell membrane. Immunoblotting demonstrated the glycoprotein's presence in nearly equal amounts in all developmental stages of several T. cruzi isolates. Mice immunized with the purified glycoprotein and challenged with 10,000 infectious trypomastigote forms of isolate Y survived the controls by up to four days. This significant protection makes this antigen a potential candidate for a multi-subunit vaccine against T. cruzi. MIME: Antigens,-Protozoan-administration-and-dosage; Antigens,-Protozoan-isolation-and-purification; Chagas-Disease-prevention-and-control; Electrophoresis,-Polyacrylamide-Gel; Flagella-ultrastructure; Fluorescent-Antibody-Technique; Membrane-Glycoproteins-administration-and-dosage; Membrane-Glycoproteins-isolation-and-purification; Mice-; Molecular-Weight; Trypanosoma-cruzi-pathogenicity; Trypanosoma-cruzi-ultrastructure; Vero-Cells MJME: *Antigens,-Protozoan-immunology; *Chagas-Disease-immunology; *Membrane-Glycoproteins-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Female; Male; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI04717AINIAID RN: 0; 0 NM: Antigens,-Protozoan; Membrane-Glycoproteins AN: 94224144 UD: 9408 Record 101 of 131 - MEDLINE (R) 1994 TI: Leishmanicidal and trypanocidal activities of Bolivian medicinal plants. AU: Fournet-A; Barrios-AA; Munoz-V AD: Institut Francais de Recherche pour le Developpement en Cooperation (ORSTOM), Departement Sante, Paris, France. SO: J-Ethnopharmacol. 1994 Jan; 41(1-2): 19-37 ISSN: 0378-8741 PY: 1994 LA: ENGLISH CP: IRELAND AB: Cutaneous and mucocutaneous leishmaniasis are endemic diseases in South America, especially in the subandean areas of the humid lowlands of Bolivia. Fourteen plants used topically in folk medicine to treat cutaneous leishmaniasis were collected in the tropical regions of colonization and in the rain forest occupied by Chimane Indians. Three of four plants used by the Chimane Indians exhibited an in vitro activity against three species of Leishmania. Two of ten plants used by the colonists showed an in vitro activity. We have also included results obtained with extracts from 53 Bolivian medicinal plants used for other diseases and from 43 plants collected with basis of chemotaxonomic criteria from all parts of Bolivia. All extracts were also screened in vitro against three strains of Trypanosoma cruzi (Trypanosomatidae), the causative agent of Chagas' disease. MIME: Antiprotozoal-Agents-isolation-and-purification; Antiprotozoal-Agents-therapeutic-use; Bolivia-; Chagas-Disease-drug-therapy; Indians,-South-American; Leishmaniasis,-Cutaneous-drug-therapy; Medicine,-Traditional; Plant-Extracts-isolation-and-purification; Plant-Extracts-therapeutic-use MJME: *Antiprotozoal-Agents-pharmacology; *Leishmania-drug-effects; *Plant-Extracts-pharmacology; *Plants,-Medicinal; *Trypanosoma-cruzi-drug-effects TG: Animal; Human PT: JOURNAL-ARTICLE RN: 0; 0 NM: Antiprotozoal-Agents; Plant-Extracts AN: 94223897 UD: 9408 Record 102 of 131 - MEDLINE (R) 1994 TI: Depletion of T-cell subpopulations results in exacerbation of myocarditis and parasitism in experimental Chagas' disease. AU: Tarleton-RL; Sun-J; Zhang-L; Postan-M AD: Department of Zoology, University of Georgia, Athens 30602. SO: Infect-Immun. 1994 May; 62(5): 1820-9 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The contribution of T-cell subpopulations to immunopathology in murine Trypanosoma cruzi infection was studied by using in situ localization of lymphocytes and in vivo depletion of T-lymphocyte populations. CD8+ T cells were the major lymphocyte population in the inflamed hearts of C3H/HeSnJ mice infected with the Sylvio X10/4 clone of T. cruzi at all time points of the acute and chronic phases of the infection examined. Depletion of CD8+ and/or CD4+ T cells beginning on day 20 of the infection resulted in a moderate decrease in the inflammation and an increase in parasite burden in the hearts of mice at day 30 of infection. Longer-term depletion, beginning at day 20 and extending as long as 200 days of infection, resulted in an increased inflammatory response in the heart. A large proportion of the inflammatory cells in the hearts of anti-CD8- or anti-CD4- and anti-CD8-treated mice were Thy1+ and CD4- CD8-. At 200 days of infection, the increased inflammation was accompanied by an increase in the parasite load in the heart. These results show that T-cell subset depletion does not prevent the inflammatory response associated with acute and chronic T. cruzi infection. The increased parasite load in T-cell-depleted mice also demonstrates the participation of these T-cell subsets in regulation of parasite load throughout the course of the infection. The increased inflammatory response despite T-cell depletion and in association with increased numbers of tissue parasites suggests that intracellular parasites are a driving force behind the inflammatory response in chronic murine T. cruzi infection. MIME: Chagas-Cardiomyopathy-immunology; Chagas-Cardiomyopathy-parasitology; Mice-; Mice,-Inbred-C3H; Mice,-Inbred-C57BL MJME: *Chagas-Cardiomyopathy-etiology; *Heart-parasitology; *Lymphocyte-Depletion; *Myocarditis-etiology; *T-Lymphocyte-Subsets-physiology TG: Animal; Female; Male; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI22070AINIAID AN: 94222549 UD: 9408 Record 103 of 131 - MEDLINE (R) 1994 TI: Antigens shared by Leishmania species and Trypanosoma cruzi: immunological comparison of the acidic ribosomal P0 proteins. AU: Skeiky-YA; Benson-DR; Elwasila-M; Badaro-R; Burns-JM Jr; Reed-SG AD: Infectious Disease Research Institute, Seattle, Washington 98109. SO: Infect-Immun. 1994 May; 62(5): 1643-51 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Patients with visceral leishmaniasis produce high levels of immunoglobulin, but the specificities of antibodies produced are not well characterized. In an effort to identify leishmania antigens that are specific to Leishmania species or are cross-reactive with other parasitic protozoa, we have cloned and characterized full-length genomic and cDNA clones encoding a Leishmania chagasi acidic ribosomal antigen, LcP0, recognized during human infections. The protein is homologous to the Trypanosoma cruzi and human ribosomal proteins TcP0 and HuP0, respectively. Unlike most higher eukaryotes, but similar to TcP0, LcP0 has a C-terminal heptapeptide sequence resembling those of the archaebacterial acidic (P-like) proteins. The highly charged C-terminal acidic domain of LcP0 contains a serine residue typically found in most eukaryotes but lacking in all T. cruzi P proteins we have characterized thus far. L. chagasi-infected individuals as well as those with T. cruzi infections have antibodies cross-reactive with recombinant LcP0 and TcP0 as well as HuP0. However, the properties of anti-P0 antibodies in T. cruzi and L. chagasi infection sera are quite different. Through the use of synthetic peptides, we showed that while T. cruzi infection anti-TcP0 antibodies are exclusively directed against the C-terminal domain of TcP0, L. chagasi infection sera contain antibodies reactive with epitopes other than the C-terminal sequence of LcP0. Thus, anti-LcP0 antibodies in L. chagasi infection sera represent the first characterized deviation from the restricted immunodominant C-terminal epitope involved in the generation of anti-P0 antibodies following infection or autoimmune diseases. MIME: Amino-Acid-Sequence; Antigenic-Determinants; Base-Sequence; Blotting,-Southern; Cloning,-Molecular; Leishmaniasis-immunology; Mice-; Mice,-Inbred-BALB-C; Molecular-Sequence-Data; Phosphoproteins-genetics; Rabbits- MJME: *Antigens,-Protozoan-immunology; *Leishmania-immunology; *Phosphoproteins-immunology; *Ribosomal-Proteins-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Comparative-Study; Human; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI25038AINIAID; AI22726AINIAID; AI30639AINIAID RN: 0; 0; 0; 0; 0 NM: phosphoprotein-P0; Antigenic-Determinants; Antigens,-Protozoan; Phosphoproteins; Ribosomal-Proteins AN: 94222525 UD: 9408 SI: GENBANK/L29300 Record 104 of 131 - MEDLINE (R) 1994 TI: Infection with Trypanosoma cruzi during pregnancy in rats and a decrease in chronic myocardial lesions in their infected offspring. AU: Davila-HO; Revelli-SS; Moreno-HS; Valenti-JL; Musso-OC; Poli-HO; Morini-JC; Bottasso-OA AD: Division Inmunologia, Facultad de Ciencias Medicas, Universidad Nacional de Rosario, Argentina. SO: Am-J-Trop-Med-Hyg. 1994 Apr; 50(4): 506-11 ISSN: 0002-9637 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: To ascertain whether maternal infection with Trypanosoma cruzi may influence the course of the parasitic infection in offspring, two groups of female 1 rats were mated with syngeneic sires. One group of females was infected with 10(6) trypomastigotes of T. cruzi three times at weekly intervals. All offspring were nursed by their mothers until weaning and then separated into two groups of young, one to be infected with the same dose of T. cruzi, and the other to remain uninfected. Infection of pregnant rats caused no aggravated disease but resulted in a self-controlled infection that did not cause any deaths or affect their reproductive capacity. The number of young delivered, litter size, fertility coefficient, and offspring weights at weaning were also unaffected by maternal infection; however, the survival coefficient decreased in comparison with values recorded in the offspring of uninfected mothers. The latter finding is likely due to neonatal transmission, since bloodstream forms of T. cruzi were observed in a few offspring of infected mothers. While infected offspring whose mothers had been inoculated with T. cruzi during pregnancy were not protected from acute infection, the occurrence of chronic focal myocarditis was less prevalent when compared with that recorded in chronically infected offspring born to uninfected mothers. MIME: Acute-Disease; Antibodies,-Protozoan-blood; Chagas-Disease-blood; Chronic-Disease; Disease-Models,-Animal; Kinetics-; Pregnancy-; Pregnancy-Complications,-Parasitic-blood; Rats-; Rats,-Inbred-Strains; Trypanosoma-cruzi-immunology MJME: *Chagas-Cardiomyopathy-pathology; *Chagas-Disease-pathology; *Myocardium-pathology; *Pregnancy-Complications,-Parasitic-pathology TG: Animal; Female; Male PT: JOURNAL-ARTICLE RN: 0 NM: Antibodies,-Protozoan AN: 94219710 UD: 9407 SB: AIM Record 105 of 131 - MEDLINE (R) 1994 TI: The nuclear genomes of African and American trypanosomes are strikingly different. AU: Musto-H; Rodriguez-Maseda-H; Bernardi-G AD: Laboratoire de Genetique Moleculaire, Institut Jacques Monod, Paris, France. SO: Gene. 1994 Apr 8; 141(1): 63-9 ISSN: 0378-1119 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: We have investigated the compositional distributions of exons and their different codon positions, as well as the codon usage and amino-acid (aa) composition of the nuclear genomes of the African and American trypanosomes Trypanosoma brucei and T. cruzi. Very large differences between the two species were found in all the properties investigated. The most striking differences concern the compositional distributions of third codon positions and the extremely large nucleotide divergence of third codon position for homologous genes encoding proteins that are highly conserved in their aa sequences. Moreover, if coding sequences from each species are divided into two groups according to the GC levels in third codon positions, very different codon usages and aa compositions are found. This indicates a compositional compartmentalization in both genomes which had previously been detected in T. brucei (and T. equiperdum) by compositional fractionation. MIME: Base-Composition; Codon-genetics; Exons-genetics; Sequence-Analysis,-DNA; Sequence-Homology,-Nucleic-Acid MJME: *DNA,-Protozoan-chemistry; *Genes,-Protozoan; *Genome-; *Trypanosoma-brucei-brucei-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0 NM: Codon; DNA,-Protozoan AN: 94215889 UD: 9407 Record 106 of 131 - MEDLINE (R) 1994 TI: [The first 13 years of controlling Chagas' disease in Mambai, Goias, Brazil 1980-1992] TO: Los 13 primeros anos del control de la enfermedad de Chagas en Mambai, Goias, Brasil, 1980-1992. AU: Marsden-P; Garcia-Zapata-MT; Castillo-EA; Prata-AR; Macedo-VO AD: Universidade de Brasilia, Nucleo de Medicina Tropical e Nutricao, D.F., Brasil. SO: Bol-Oficina-Sanit-Panam. 1994 Feb; 116(2): 111-7 ISSN: 0030-0632 PY: 1994 LA: SPANISH; NON-ENGLISH CP: UNITED-STATES AB: The Mambai Project was launched in 1980 for the purpose of making a longitudinal clinical-epidemiologic study of Chagas' disease and of serving as a pilot program for the Ministry of Health of Brazil. At the beginning of the project, a census was carried out, the housing units were evaluated, and clinical examination and laboratory tests were performed on the population. After a phase of massive attack with insecticides, ongoing epidemiologic surveillance was instituted in order to detect residual foci of triatomine bugs in all the housing units in the municipality. The campaign included a program of health education combined with community participation. All infested housing units were selectively fumigated. In 1988 a new census was carried out, together with a serologic survey of children born after the control program was initiated. This article describes the results of epidemiologic surveillance during the first 13 years of the program. The prevalence of T. infestans in the housing units diminished to levels that suggest that vector transmission of the disease in Mambai has been stopped. However, the risk of reinfestation from neighboring areas without control programs and the risk of colonization by secondary T. sordida vectors are factors that should be carefully monitored. MIME: Brazil-epidemiology; Chagas-Disease-epidemiology; Chagas-Disease-parasitology; Chagas-Disease-transmission; Child-; English-Abstract; Housing-; Insect-Control-statistics-and-numerical-data; Insect-Vectors-parasitology; Longitudinal-Studies; Population-Surveillance; Prevalence-; Rural-Population-statistics-and-numerical-data; Triatoma-parasitology; Trypanosoma-cruzi-pathogenicity MJME: *Chagas-Disease-prevention-and-control TG: Animal; Comparative-Study; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE AN: 94213696 UD: 9407 Record 107 of 131 - MEDLINE (R) 1994 TI: The structure of Trypanosoma cruzi trypanothione reductase in the oxidized and NADPH reduced state. AU: Lantwin-CB; Schlichting-I; Kabsch-W; Pai-EF; Krauth-Siegel-RL AD: Abteilung Biophysik, Max-Planck-Institut fur Medizinische Forschung, Heidelberg, Germany. SO: Proteins. 1994 Feb; 18(2): 161-73 ISSN: 0887-3585 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The three-dimensional structure of trypanothione reductase (TR) (EC from Trypanosoma cruzi has been solved at 0.33 nm resolution by molecular replacement using the structure of C. fasciculata TR as a starting model. Elucidation of the T. cruzi TR structure represents the first step in the rational design of a drug against Chagas' disease. The structure of T. cruzi TR is compared with those of C. fasciculata TR as well as human and E. coli glutathione reductase (GR). In the FAD-binding domain, TR has two insertions, each about 10 residues long, which do not occur in GR. The first one is a rigid loop stabilizing the position of helix 91-117 which is responsible for the wider active site of TR as compared to GR. The second insertion does not occur where it is predicted by sequence alignment; rather the residues extend three strands of the 4-stranded beta-sheet by one or two residues each. This increases the number of hydrogen bonds within the sheet structure. The structure of the NADPH.TR complex has been solved at 0.33 nm resolution. The nicotinamide ring is sandwiched between the flavin ring and the side chain of Phe-198 which undergoes the same conformational change upon coenzyme binding as Tyr-197 in GR. In addition to Arg-222 and Arg-228, which are conserved in TR and GR, Tyr-221--the last residue of the second beta-sheet strand of the beta alpha beta dinucleotide binding fold--is in hydrogen bonding distance to the 2' phosphate group of NADPH. MIME: Amino-Acid-Sequence; Binding-Sites; Chagas-Disease-drug-therapy; Crithidia-fasciculata-enzymology; Crithidia-fasciculata-genetics; Crystallography,-X-Ray; Drug-Design; FAD-chemistry; Glutathione-Reductase-chemistry; Glutathione-Reductase-genetics; Molecular-Sequence-Data; Molecular-Structure; NADH,-NADPH-Oxidoreductases-antagonists-and-inhibitors; NADH,-NADPH-Oxidoreductases-genetics; NADP-chemistry; Oxidation-Reduction; Protein-Conformation; Protein-Structure,-Secondary; Trypanosoma-cruzi-genetics MJME: *NADH,-NADPH-Oxidoreductases-chemistry; *Trypanosoma-cruzi-enzymology TG: Animal; Comparative-Study; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 1.6.; EC 1.6.4.-; EC; 146-14-5; 53-59-8 NM: NADH,-NADPH-Oxidoreductases; trypanothione-reductase; Glutathione-Reductase; FAD; NADP AN: 94211757 UD: 9407 Record 108 of 131 - MEDLINE (R) 1994 TI: Pathology of patients with Chagas' disease and acquired immunodeficiency syndrome. AU: Rocha-A; de-Meneses-AC; da-Silva-AM; Ferreira-MS; Nishioka-SA; Burgarelli-MK; Almeida-E; Turcato-Junior-G; Metz-K; Lopes-ER AD: Centro de Ciencias Biomedicas, Universidade Federal de Uberlandia, Minas Gerais, Brazil. SO: Am-J-Trop-Med-Hyg. 1994 Mar; 50(3): 261-8 ISSN: 0002-9637 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The main pathologic findings in 23 patients with acquired immunodeficiency syndrome (AIDS) and Chagas' disease are reviewed; five are from our own experience and 18 from the literature. The presence of Trypanosoma cruzi parasites and/or T. cruzi antibodies in blood and cerebrospinal fluid was recorded and computerized tomograms of the brain were evaluated. Twenty (87%) of the 23 subjects developed severe, multifocal or diffuse meningoencephalitis with necrosis and hemorrhage associated with numerous tissue parasites. The second most severely affected site was the heart. Seven (30.4%) of the 23 cases had myocarditis on pathologic examination. It was acute in four patients, chronic in two, and simultaneously acute and chronic in one. Acute myocarditis and meningoencephalitis are interpreted as being caused by relapses of chronic T. cruzi infections. An AIDS permissive role is suggested for these conditions since immunologic defense against T. cruzi is mediated mainly by T lymphocytes, whose CD4 subpopulation is depleted in patients with this disease. Consequently, AIDS is a factor that may favor the reactivation of T. cruzi infections. The lesions reported in the association of Chagas' disease with AIDS were compared with those reported from patients without AIDS having fatal, acute, vector-transmitted infections, contaminated blood transfusions, or accidental exposures in the laboratory. For the latter three, meningoencephalitis is uncommon. Only immunosuppressed cases of Chagas' disease have been described as having a pseudotumoral presentation that shows expanding lesions with a mass effect in the cranial cavity that causes intracranial hypertension and simulates neoplasms (tumors such as gliomas, lymphomas, metastases, etc.). MIME: Acquired-Immunodeficiency-Syndrome-complications; Adult-; Brain-parasitology; Chagas-Cardiomyopathy-complications; Chagas-Cardiomyopathy-pathology; Chagas-Disease-complications; Heart-parasitology; Meningoencephalitis-complications; Meningoencephalitis-pathology; Middle-Age; Trypanosoma-cruzi-isolation-and-purification MJME: *Acquired-Immunodeficiency-Syndrome-pathology; *Brain-pathology; *Chagas-Disease-pathology; *Myocardium-pathology TG: Animal; Case-Report; Female; Human; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE; REVIEW; REVIEW-OF-REPORTED-CASES AN: 94197294 UD: 9407 SB: AIM Record 109 of 131 - MEDLINE (R) 1994 TI: Antileishmanial activity of a tetralone isolated from Ampelocera edentula, a Bolivian plant used as a treatment for cutaneous leishmaniasis. AU: Fournet-A; Barrios-AA; Munoz-V; Hocquemiller-R; Roblot-F; Cave-A AD: Institut Francais de Recherche Scientifique pour le Developpement en Cooperation (ORSTOM), Departement Sante, Paris, France. SO: Planta-Med. 1994 Feb; 60(1): 8-12 ISSN: 0032-0943 PY: 1994 LA: ENGLISH CP: GERMANY AB: The stem bark of Ampelocera edentula Kuhlm. (Ulmaceae) is used by the Chimanes Indians from Bolivia for the treatment of cutaneous leishmaniasis caused by the protozoan Leishmania braziliensis. A chloroform extract of the stem barks was found to be active against extracellular forms of Leishmania ssp. and Trypanosoma cruzi at 50 micrograms/ml. Bioassay-guided fractionation of this extract allowed us to isolate one active compound. Its structure was elucidated by spectral and chemical studies as 4-hydroxy-1-tetralone. BALB/c mice infected with L. amazonensis (PH8) or L. venezuelensis were treated one day after the parasitic infection with 4-hydroxy-1-tetralone (25 mg/kg/day) or with reference drug, Glucantime (56 mg Sbv/kg/day) for 14 days. Lesion development was the criteria used to evaluate the disease severity. 4-Hydroxy-1-tetralone was slightly less effective than the reference drug against L. amazonensis or L. venezuelensis. Single treatment near the site of infection, 14 days after infection with L. amazonensis, with 4-hydroxy-1-tetralone (50 mg/kg) was more effective than Glucantime (112 mg/kg). This study is, to our knowledge, the first to show the activity of a tetralone for the experimental treatment of New World cutaneous leishmaniasis. MIME: Antiprotozoal-Agents-isolation-and-purification; Leishmania-drug-effects; Mice-; Mice,-Inbred-BALB-C; Molecular-Structure; Trypanosoma-cruzi-drug-effects MJME: *Antiprotozoal-Agents-pharmacology; *Leishmania-braziliensis-drug-effects; *Leishmaniasis,-Cutaneous-drug-therapy; *Plants,-Medicinal-chemistry; *Tetrahydronaphthalenes-pharmacology TG: Animal; Female; Male PT: JOURNAL-ARTICLE RN: 0; 0; 21032-12-2 NM: Antiprotozoal-Agents; Tetrahydronaphthalenes; 4-hydroxy-1-tetralone AN: 94181617 UD: 9406 Record 110 of 131 - MEDLINE (R) 1994 TI: A proteolytic fragment of Trypanosoma cruzi trans-sialidase lacking the carboxyl-terminal domain is active, monomeric, and generates antibodies that inhibit enzymatic activity. AU: Schenkman-S; Chaves-LB; Pontes-de-Carvalho-LC; Eichinger-D AD: Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina, Sao Paulo, Brazil. SO: J-Biol-Chem. 1994 Mar 18; 269(11): 7970-5 ISSN: 0021-9258 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: trans-Sialidase isolated from trypomastigote forms of Trypanosoma cruzi, the protozoan parasite that causes Chagas' disease, is multimeric and heterogeneous in size. We show here that limited proteolysis of tans-sialidase with papain yields a single monomeric polypeptide chain of 70 kDa that conserves full enzymatic activity on soluble and membrane-bound substrates. The papain fragment lacks most of the 12-amino acid repeats of the carboxyl-terminal domain that comprises about 50% of the native trans-sialidase. When injected into rabbits, the papain-generated fragment induces antibodies that inhibit trans-sialidase activity and trypomastigote sialylation. The repeats are also not required for the stability of the enzyme or for the correct folding during the biosynthesis in Escherichia coli, but seem essential for trans-sialidase oligomerization. We conclude that trans-sialidase is composed of two structurally and functionally independent domains. MIME: Amino-Acid-Sequence; Base-Sequence; Chromatography,-Affinity; DNA-Primers; Immunoblotting-; Kinetics-; Molecular-Sequence-Data; Neuraminidase-isolation-and-purification; Peptides-chemical-synthesis; Peptides-immunology; Plasmids-; Rabbits-immunology; Recombinant-Proteins-antagonists-and-inhibitors; Recombinant-Proteins-isolation-and-purification; Recombinant-Proteins-metabolism; Restriction-Mapping; Sequence-Deletion MJME: *Antibodies-pharmacology; *Neuraminidase-antagonists-and-inhibitors; *Neuraminidase-metabolism; *Peptide-Fragments-immunology; *Peptide-Fragments-metabolism; *Trypanosoma-cruzi-enzymology TG: Animal; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: RO3TW0022701TWFIC RN: EC; 0; 0; 0; 0; 0; 0 NM: Neuraminidase; Antibodies; DNA-Primers; Peptide-Fragments; Peptides; Plasmids; Recombinant-Proteins AN: 94179162 UD: 9406 Record 111 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi-induced immunosuppression: blockade of costimulatory T-cell responses in infected hosts due to defective T-cell receptor-CD3 functioning. AU: Lopes-MF; dos-Reis-GA AD: Department of Immunology, Federal University of Rio de Janeiro, Brazil. SO: Infect-Immun. 1994 Apr; 62(4): 1484-8 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: A model of experimental Trypanosoma cruzi murine infection with chemically induced metacyclic forms (opossum clone Dm28c) showed a marked state of T-cell unresponsiveness during acute phase, but lacked evidence of suppressor cell activity. Spleen cells from infected mice were suppressed in vitro in responses to T-cell activators concanavalin A, anti-Thy1 monoclonal antibody (MAb), and anti-CD3 MAb compared with spleen cells from control littermates. Activation with accessory cell-independent stimulus provided by immobilized anti-CD3 was defective in splenic CD4-positive T cells from infected mice, but not in such cells from control mice. No evidence of splenic suppressor cell activity was found in cell-mixing experiments using nylon-passed T cells from control and infected donors. Kinetic experiments showed that there was a discrete stage in infection when T cells were already suppressed in response to anti-CD3 but still responded to anti-CD69 MAb. In these T cells, immobilized anti-CD3 failed to enhance simultaneous CD69 responses, although anti-CD3 enhanced CD69 responses in control T cells from uninfected donors. These results demonstrate an intrinsic defect in T-cell receptor-mediated T-cell activation, which could be a mechanism generating T-cell suppression during infection by T. cruzi. MIME: Antigens,-CD-physiology; Antigens,-Differentiation,-T-Lymphocyte-physiology; Mice-; Mice,-Inbred-BALB-C MJME: *Chagas-Disease-immunology; *Immune-Tolerance; *Lymphocyte-Transformation; *Receptor-CD3-Complex,-Antigen,-T-Cell-physiology; *T-Lymphocytes-immunology TG: Animal; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0 NM: Antigens,-CD; Antigens,-Differentiation,-T-Lymphocyte; Leu-23-antigen; Receptor-CD3-Complex,-Antigen,-T-Cell AN: 94178964 UD: 9406 Record 112 of 131 - MEDLINE (R) 1994 TI: Identification and detection of Trypanosoma cruzi by using a DNA amplification fingerprint obtained from the ribosomal intergenic spacer. AU: Gonzalez-N; Galindo-I; Guevara-P; Novak-E; Scorza-JV; Anez-N; Da-Silveira-JF; Ramirez-JL AD: Facultad de Ciencias, Universidad de Los Andes, Merida. SO: J-Clin-Microbiol. 1994 Jan; 32(1): 153-8 ISSN: 0095-1137 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: We designed a PCR assay targeted on repeated elements of the ribosomal intergenic spacer which produces highly polymorphic DNA band patterns for different strains of Trypanosoma cruzi. By labeling the PCR products with digoxigenin and by chemiluminescence detection, we improved the assay sensitivity by three orders of magnitude to get T. cruzi strain fingerprints in feces of the trypanosome-infected triatomine bug vector. We also developed a capture assay for the digoxigenin-labeled PCR products that allowed us to detect T. cruzi in triatomine bug vector feces and in human serum samples with a solid support. MIME: Base-Sequence; Chemiluminescence-; Cloning,-Molecular; Digoxigenin-; DNA-Fingerprinting; DNA-Probes; Feces-microbiology; Insect-Vectors-microbiology; Molecular-Sequence-Data; Repetitive-Sequences,-Nucleic-Acid; Rhodnius-parasitology; Sensitivity-and-Specificity; Species-Specificity; Trypanosoma-cruzi-genetics MJME: *DNA,-Protozoan-genetics; *DNA,-Ribosomal-genetics; *Polymerase-Chain-Reaction-methods; *Trypanosoma-cruzi-isolation-and-purification TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 1672-46-4 NM: DNA-Probes; DNA,-Protozoan; DNA,-Ribosomal; Digoxigenin AN: 94172012 UD: 9406 SI: GENBANK/M63895 Record 113 of 131 - MEDLINE (R) 1994 TI: An unusually small gene encoding a putative mucin-like glycoprotein in Trypanosoma cruzi. AU: Reyes-MB; Pollevick-GD; Frasch-AC AD: Instituto de Investigaciones Bioquimicas Fundacion Campomar, Buenos Aires, Argentina. SO: Gene. 1994 Mar 11; 140(1): 139-40 ISSN: 0378-1119 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: The gene encoding a putative core protein of a mucin-like glycoprotein was identified in Trypanosoma cruzi. It contains five repeats of eleven amino acids each, eight of which are Thr and two of which are Pro residues. These Thr-Pro-rich repeats resemble the ones in the human MUC2 gene encoding mucin. MIME: Amino-Acid-Sequence; Base-Sequence; Consensus-Sequence; DNA,-Protozoan; Genes,-Protozoan; Molecular-Sequence-Data; Trypanosoma-cruzi-metabolism MJME: *Mucins-genetics; *Trypanosoma-cruzi-genetics TG: Animal; Human; Support,-Non-U.S.-Gov't GS: MUC2 PT: JOURNAL-ARTICLE RN: 0; 0 NM: DNA,-Protozoan; Mucins AN: 94171072 UD: 9406 SI: GENBANK/L20809 Record 114 of 131 - MEDLINE (R) 1994 TI: Distribution of alpha-galactosyl-containing epitopes on Trypanosoma cruzi trypomastigote and amastigote forms from infected Vero cells detected by Chagasic antibodies. AU: Souto-Padron-T; Almeida-IC; de-Souza-W; Travassos-LR AD: Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Brazil. SO: J-Eukaryot-Microbiol. 1994 Jan-Feb; 41(1): 47-54 ISSN: 1066-5234 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Reactivity of different Trypanosoma cruzi developmental forms with purified Chagasic anti-alpha-galactosyl antibodies (anti-Gal) was studied using epimastigotes from axenic cultures, trypomastigotes and amastigotes from infected Vero cell cultures, and an immunogold labeling method as observed by electron microscopy. Epimastigotes were poorly labeled, whereas extracellular trypomastigotes and amastigotes bound heterogeneously to the antibody with many cells being intensely labeled at the cell surface, including the membrane lining the cell body, the flagellum and the flagellar pocket. Parasites with poor labeling at the cell surface generally had several gold particles within the cell, mostly in cytoplasmic vacuoles. The Golgi complex of trypomastigotes was strongly labeled. Intracellular parasites were labeled at the parasite cell surface or within vacuolar structures. The expression in T. cruzi-infected Vero cells of alpha-galactosyl antigenic structures acquired from the parasite was shown by moderate labeling with Chagasic anti-Gal of the membrane lining parasite-free outward cell projections. The reactivity with purified anti-Gal from healthy individuals at the same concentrations of Chagasic anti-Gal was poor, with gold particles appearing in the nucleus and cytoplasm but not at the cell surface. It paralleled the labeling with Bandeireae simplicifolia IB-4 lectin. The results provide a basis for autoimmune reactions involving anti-Gal from chronic Chagasic patients. MIME: Antigenic-Determinants-chemistry; Carbohydrate-Sequence; Cercopithecus-aethiops; Lectins-chemistry; Molecular-Sequence-Data; Trisaccharides-chemistry; Trypanosoma-cruzi-chemistry; Trypanosoma-cruzi-ultrastructure; Vero-Cells MJME: *Antigenic-Determinants-analysis; *Lectins-analysis; *Trisaccharides-analysis; *Trypanosoma-cruzi-immunology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0 NM: Antigenic-Determinants; Euonymus-europaeus-lectin; Lectins; Trisaccharides AN: 94169886 UD: 9406 Record 115 of 131 - MEDLINE (R) 1994 TI: Chemoenzymic preparation of a glycoconjugate polymer having a sialyloligosaccharide: Neu5Ac alpha (2-->3)Gal beta (1-->4)GlcNAc. AU: Nishimura-S; Lee-KB; Matsuoka-K; Lee-YC AD: Division of Biological Science, Graduate School of Science Hokkaido University, Sapporo, Japan. SO: Biochem-Biophys-Res-Commun. 1994 Feb 28; 199(1): 249-54 ISSN: 0006-291X PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Water-soluble polyacrylamide having 3'-sialyl N-acetyl-lactosamine [Neu5Ac alpha (2-->3)Gal beta (1-->4)GlcNAc] was enzymatically prepared by stepwise sugar-elongation on a water-soluble GlcNAc-bearing polyacrylamide. It was demonstrated that the flexible GlcNAc branches of the polymer chains allow quantitative galactosylation with bovine galactosyl transferase and partial sialylation by Trypanosoma cruzi trans-sialidase. Unsialylated N-acetyl-lactosamine side chains can be removed with beta-D-galactosidase and N-acetyl-beta-D-glucosaminidase to afford the targeted polymer containing 3'-sialyl N-acetyl-lactosamine. MIME: Acetylgalactosamine-chemistry; Carbohydrate-Sequence; Glycoconjugates-chemistry; Nuclear-Magnetic-Resonance; Oligosaccharides-chemistry; Sialic-Acids-chemistry MJME: *Glycoconjugates-chemical-synthesis; *Oligosaccharides-chemical-synthesis PT: JOURNAL-ARTICLE RN: 0; 0; 0; 31022-50-1 NM: Glycoconjugates; Oligosaccharides; Sialic-Acids; Acetylgalactosamine AN: 94168582 UD: 9406 Record 116 of 131 - MEDLINE (R) 1994 TI: Possible integration of Trypanosoma cruzi kDNA minicircles into the host cell genome by infection. AU: Teixeira-AR; Arganaraz-ER; Freitas-LH Jr; Lacava-ZG; Santana-JM; Luna-H AD: Department of Pathology, Faculty of Health Sciences, University of Brasilia, Brazil. SO: Mutat-Res. 1994 Mar 1; 305(2): 197-209 ISSN: 0165-1110 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Infection with Trypanosoma cruzi is known to induce the division of peritoneal macrophages in BALB/c mice. We have demonstrated, by cytogenetic analysis, that accessory DNA elements are associated with the metaphase macrophage chromosomes of such infected macrophages. The identification of these accessory DNA elements with T. cruzi DNA is strongly supported by the association of 3H-label with some chromatids in macrophages previously infected with T. cruzi which had been labelled with 3H-methyl-thymidine. The karyotyping consistently showed preferential associations of T. cruzi DNA with chromosomes 3, 6 and 11. A conclusive demonstration of the parasite origin of the integrated DNA came from fluorescein in situ hybridization studies using specific parasite DNAs as probes. In order to determine the identity of the inserted DNA and to investigate the nature of the integration mechanism, Southern blot analyses were performed on DNA extracted from both uninfected and infected (but parasite-free) macrophages. Hybridizations of BamHI, EcoRI and TaqI digests of DNA from T. cruzi-infected host cells all revealed the presence of a 1.7-kb DNA fragment when probed with kDNA. The covalent association of kDNA with that of the host was confirmed by the fact that AluI and Hinf-I digests of DNA from infected host cells produced a number of bands, in a size range of 0.8-3.6 kb, which hybridized with kDNA minicircles. None of these bands was found in DNA purified from cell-free preparations of the parasite and thus it must be concluded that they represent insertion fragments between parasite and host cell DNA. These results strongly suggest that kDNA minicircles from T. cruzi have been integrated into the genome of the host cell following infection. MIME: Cells,-Cultured; Chagas-Disease-parasitology; Chromatids-ultrastructure; Chromatin-ultrastructure; DNA-Probes; DNA,-Circular-metabolism; DNA,-Kinetoplast-metabolism; In-Situ-Hybridization; Macrophages,-Peritoneal-pathology; Macrophages,-Peritoneal-parasitology; Metaphase-; Mice-; Mice,-Inbred-BALB-C; Trypanosoma-cruzi-pathogenicity MJME: *DNA,-Circular-genetics; *DNA,-Kinetoplast-genetics; *Macrophages,-Peritoneal-physiology; *Trypanosoma-cruzi-genetics TG: Animal; Female; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0 NM: Chromatin; DNA-Probes; DNA,-Circular; DNA,-Kinetoplast AN: 94166808 UD: 9406 Record 117 of 131 - MEDLINE (R) 1994 TI: Chagas' disease: carcinogenic activity of the antitrypanosomal nitroarenes in mice. AU: Teixeira-AR; Calixto-MA; Teixeira-ML AD: Laboratory for Multidisciplinary Research in Chagas' Disease, Faculty of Health Sciences, University of Brasilia, Brazil. SO: Mutat-Res. 1994 Mar 1; 305(2): 189-96 ISSN: 0165-1110 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: The carcinogenic activity of antitrypanosomal 2-nitroimidazole, 5-nitroimidazole and 5-nitrofuran derivatives was assessed in female Swiss mice of the same age group. A statistically significantly higher incidence of growths was seen in mice into which 2-nitro had been injected than in mice receiving 5-nitro derivatives intraperitoneally. A histologic type of lymphoblastic lymphoma that invades lymph nodes, spleen, liver, lungs and lymphatic tissue elsewhere was frequently found in nitroarene-treated mice. Further, it is shown that the potency of the drug, rather than the duration of its administration, was usually associated with the growth of lymphomas. The 2-nitro derivative which induced the highest incidence of lymphomas significantly decreased the survival of treated mice; this probably occurred because it undergoes enzymatic reduction of the nitro group more efficiently than the 5-nitro compounds used. The differences of incidence of lymphomas in mice receiving any of these nitroarenes and in control mice that received daily injections of 0.15 M saline were statistically significant (alpha = 0.05). The indiscriminate use of these nitroarenes to treat Trypanosoma cruzi infections in man could therefore induce a significant number of lymphomas. MIME: Antiprotozoal-Agents-therapeutic-use; Cell-Division-drug-effects; Liver-drug-effects; Liver-pathology; Lymph-Nodes-drug-effects; Lymph-Nodes-pathology; Lymphoma-pathology; Mice-; Molecular-Structure; Nifurtimox-toxicity; Nifurtimox-therapeutic-use; Nitroimidazoles-toxicity; Nitroimidazoles-therapeutic-use MJME: *Antiprotozoal-Agents-toxicity; *Carcinogens-toxicity; *Chagas-Disease-drug-therapy; *Lymphoma-chemically-induced TG: Animal; Female; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 22994-85-0; 23256-30-6; 33450-08-7 NM: Antiprotozoal-Agents; Carcinogens; Nitroimidazoles; benzonidazole; Nifurtimox; MK-436 AN: 94166807 UD: 9406 Record 118 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi: effects of infection on receptor-mediated chronotropy and Ca2+ mobilization in rat cardiac myocytes. AU: Bergdolt-BA; Tanowitz-HB; Wittner-M; Morris-SA; Bilezikian-JP; Moreno-AP; Spray-DC AD: Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 00461-1602. SO: Exp-Parasitol. 1994 Mar; 78(2): 149-60 ISSN: 0014-4894 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Acute Trypanosoma cruzi infection is commonly associated with disorders of impulse conduction and muscle contraction in heart. In order to determine the extent to which receptor function changed in response to infection, infected neonatal rat cardiac myocytes in culture were compared with matched controls with regard to chronotropic response and Ca2+ mobilization following the application of adrenergic agonists. At 7-9 days in culture (5-7 days postinfection), spontaneous beat rates of control myocytes were four times as rapid as those in infected cells. Control cells responded to 10(-5) M isoproterenol (ISO) and 10(-6) M norepinephrine (NE) with increases in beat rate of 34 and 40%, respectively. Effects of ISO on infected cells were similar, and adenylate cyclase activity was similar in control and infected cells when measured in the presence of ISO alone or in combination with Gpp(NH)p. NE produced a more marked chronotropic response in infected cultures and altered Ca2+ mobilization. NE treatment increased Ca2+ levels in control cardiac myocytes from 51.8 +/- 4.4 to 113 +/- 16 nM (in 0 Ca2+ medium) and from 85.2 +/- 6.8 to 131.3 +/- 24.5 nM (1 mM external Ca2+). In infected cardiac myocytes, NE increased Ca2+ from 116.8 +/- 17 to 164.7 +/- 9.6 nM (in 0 Ca2+ medium) and from 132.2 +/- 13.2 to 162.5 +/- 0.3 nM (1 mM Ca2+ medium). Thus, basal and alpha-adrenergic-stimulated Ca2+ levels were higher in infected than uninfected myocytes regardless of the extracellular Ca2+ levels, although the fractional increase in infected myocytes was significantly lower than that in controls (1.4- and 1.2-fold vs 2.2- and 1.5-fold). Therefore, both chronotropic and Ca(2+)-mobilization responses to the alpha-adrenergic agonist NE are altered in T. cruzi-infected cardiac myocytes; the chronotropic response of similarly infected cells to the beta-adrenergic agonist ISO was not affected. These data indicating that T. cruzi infection may be associated with a dissociation in responses to these agonists suggest a possible mechanism to explain, in part, the cardiac dysfunction characteristic of Chagas' disease. MIME: Animals,-Newborn; Cells,-Cultured; Chagas-Cardiomyopathy-physiopathology; Guanylyl-Imidodiphosphate-pharmacology; Heart-physiology; Hydrogen-Ion-Concentration; Image-Processing,-Computer-Assisted; Isoproterenol-pharmacology; Myocardial-Contraction; Myocardium-metabolism; Norepinephrine-pharmacology; Prazosin-pharmacology; Rats-; Receptors,-Adrenergic,-alpha-physiology; Receptors,-Adrenergic,-beta-physiology; Signal-Transduction; Temperature-; Time-Factors MJME: *Calcium-metabolism; *Heart-parasitology; *Heart-Rate; *Myocardium-cytology; *Trypanosoma-cruzi-physiology TG: Animal; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI12770AINIAID; HL38449HLNHLBI; NS07512NSNINDS RN: 0; 0; 19216-56-9; 34273-04-6; 51-41-2; 7440-70-2; 7683-59-2 NM: Receptors,-Adrenergic,-alpha; Receptors,-Adrenergic,-beta; Prazosin; Guanylyl-Imidodiphosphate; Norepinephrine; Calcium; Isoproterenol AN: 94164265 UD: 9406 Record 119 of 131 - MEDLINE (R) 1994 TI: Quantitation of the interaction of the immunosuppressant deoxyspergualin and analogs with Hsc70 and Hsp90. AU: Nadeau-K; Nadler-SG; Saulnier-M; Tepper-MA; Walsh-CT AD: Department of Biological Chemistry, Harvard Medical School, Boston, Massachusetts 02138. SO: Biochemistry. 1994 Mar 8; 33(9): 2561-7 ISSN: 0006-2960 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Deoxyspergualin (DSG), a spermidinyl, alpha-hydroxyglycyl, 7-guanidinoheptanoyl peptidomimetic, shows immunosuppressive activity. In confirmation of a recent report that immobilized methoxyDSG selectively retains the heat shock protein Hsc70, we report here quantitative binding of DSG and analogs to both Hsc70 and the 90-kDa heat shock protein Hsp90. We have utilized affinity capillary electrophoresis to obtain Kd values for DSG and analogs, and stimulation of the ATPase activity of Hsc70 to obtain Km values for DSG, that are comparable and corroborative. Kd values are 4 microM for DSG binding to Hsc70 and 5 microM for DSG binding to Hsp90. Two active analogs, methoxy- and glycylDSG, bind with similar affinities. Glyoxylylspermidine and des(aminopropyl)DSG, two inactive metabolites, have much reduced affinity for Hsc70 and Hsp90. These data validate binding of these novel immunosuppressant agents to these molecular chaperones, at concentrations in the range of pharmacologically active doses, and indicate that further characterization of Hsc70 and/or Hsp90 as potential targets for DSG is warranted. MIME: Adenosinetriphosphatase-metabolism; Amino-Acid-Sequence; Binding,-Competitive; Cattle-; Mice-; Molecular-Sequence-Data; Peptides-chemistry; Peptides-metabolism; Protein-Binding; Structure-Activity-Relationship; Trypanosoma-cruzi MJME: *Carrier-Proteins-chemistry; *Guanidines-chemistry; *Heat-Shock-Proteins-chemistry; *Immunosuppressive-Agents-chemistry TG: Animal; Human; In-Vitro PT: JOURNAL-ARTICLE RN: EC; 0; 0; 0; 0; 0; 0; 84937-45-1 NM: Adenosinetriphosphatase; uncoating-protein; Carrier-Proteins; Guanidines; Heat-Shock-Proteins; Immunosuppressive-Agents; Peptides; 15-deoxyspergualin AN: 94162272 UD: 9406 Record 120 of 131 - MEDLINE (R) 1994 TI: Role in host cell invasion of Trypanosoma cruzi-induced cytosolic-free Ca2+ transients. AU: Tardieux-I; Nathanson-MH; Andrews-NW AD: Infectious Diseases Section, Yale University School of Medicine, New Haven, Connecticut 06510. SO: J-Exp-Med. 1994 Mar 1; 179(3): 1017-22 ISSN: 0022-1007 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Trypanosoma cruzi enters cells by a unique mechanism, distinct from phagocytosis. Invasion is facilitated by disruption of host cell actin microfilaments, and involves recruitment and fusion of host lysosomes at the site of parasite entry. These findings implied the existence of transmembrane signaling mechanisms triggered by the parasites in the host cells before invasion. Here we show that infective trypomastigotes or their isolated membranes, but not the noninfective epimastigotes, induce repetitive cytosolic-free Ca2+ transients in individual normal rat kidney fibroblasts, in a pertussis toxin-sensitive manner. Parasite entry is inhibited by buffering or depleting host cell cytosolic-free Ca2+, or by pretreatment with Ca2+ channel blockers or pertussis toxin. In contrast, invasion is enhanced by brief exposure of the host cells to cytochalasin D. These results indicate that a trypomastigote membrane factor triggers cytosolic-free Ca2+ transients in host cells through a G-protein-coupled pathway. This signaling event may promote invasion through modulation of the host cell actin cytoskeleton. MIME: Calcimycin-pharmacology; Cell-Line; Cell-Membrane-drug-effects; Cell-Membrane-physiology; Cytosol-metabolism; Host-Parasite-Relations-drug-effects; Kidney-parasitology; Kinetics-; Pertussis-Toxins-pharmacology; Rats-; Time-Factors MJME: *Calcium-metabolism; *Host-Parasite-Relations; *Trypanosoma-cruzi-physiology; *Trypanosoma-cruzi-pathogenicity TG: Animal; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: R29AI27260AINIAID; RO1AI32056AINIAID; P30DK34989DKNIDDK RN: 52665-69-7; 70323-44-3; 7440-70-2 NM: Calcimycin; Pertussis-Toxins; Calcium AN: 94157388 UD: 9406 Record 121 of 131 - MEDLINE (R) 1994 TI: Molecular and biochemical comparison of the 70-kDa heat shock proteins of Trypanosoma cruzi. AU: Olson-CL; Nadeau-KC; Sullivan-MA; Winquist-AG; Donelson-JE; Walsh-CT; Engman-DM AD: Department of Pathology, Northwestern University Medical School, Chicago, Illinois 60611. SO: J-Biol-Chem. 1994 Feb 4; 269(5): 3868-74 ISSN: 0021-9258 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: An analysis of the genetic organization, regulated expression and biochemical properties of the cytoplasmic/nuclear (hsp70) and mitochondrial (mtp70) 70-kDa heat shock proteins of Trypanosoma cruzi is presented. The two proteins are encoded by tandemly arranged gene families that are located on different chromosomes. Both are mildly heat-inducible but have different optimal temperatures for expression. During the switch from proliferation to differentiation that occurs during the growth of T. cruzi in culture, the hsp70 level decreases dramatically while the mtp70 level falls only slightly. The subcellular locations of the two proteins differ during heat shock. While mtp70 remains associated with the kinetoplast at all temperatures, hsp70 becomes more concentrated in the nucleus at higher temperatures. Biochemical analysis of hsp70 and mtp70 revealed both to be potent ATPases. Each protein binds ATP with a Km of about 70 microM and hydrolyzes ATP with a kcat of about 100 min-1, 100 times greater than the kcat of human hsp70. The high ATPase activities of hsp70 and mtp70 are further stimulated by incubation with peptides, suggesting that these trypanosome heat shock proteins have protein chaperone activity. Finally, mtp70, but not hsp70, was found to possess autophosphorylation activity in vitro, a property that it shares with prokaryotic hsp70. These findings demonstrate unique cellular and biochemical characteristics of T. cruzi mtp70 and hsp70 that suggest that they play distinct physiologic roles in the biology of the cell. MIME: Adenosinetriphosphatase-genetics; Adenosinetriphosphatase-metabolism; Amino-Acid-Sequence; Blotting,-Southern; Chromosome-Mapping; DNA,-Protozoan-genetics; DNA,-Protozoan-isolation-and-purification; Electrophoresis,-Polyacrylamide-Gel; Fluorescent-Antibody-Technique; Gene-Expression-Regulation-physiology; Glutathione-Transferases-biosynthesis; Glutathione-Transferases-isolation-and-purification; Heat-; Heat-Shock-Proteins-genetics; Heat-Shock-Proteins-metabolism; Kinetics-; Molecular-Sequence-Data; Oligopeptides-pharmacology; Phosphorylation-; Recombinant-Fusion-Proteins-biosynthesis; Recombinant-Fusion-Proteins-isolation-and-purification; Recombinant-Fusion-Proteins-metabolism; Trypanosoma-cruzi-growth-and-development; Trypanosoma-cruzi-genetics MJME: *Adenosinetriphosphatase-biosynthesis; *Heat-Shock-Proteins-biosynthesis; *Trypanosoma-cruzi-metabolism TG: Animal; Comparative-Study; Human; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: GM20011GMNIGMS RN: EC; EC; 0; 0; 0; 0 NM: Glutathione-Transferases; Adenosinetriphosphatase; DNA,-Protozoan; Heat-Shock-Proteins; Oligopeptides; Recombinant-Fusion-Proteins AN: 94148899 UD: 9405 Record 122 of 131 - MEDLINE (R) 1994 TI: Peroxynitrite inactivates thiol-containing enzymes of Trypanosoma cruzi energetic metabolism and inhibits cell respiration. AU: Rubbo-H; Denicola-A; Radi-R AD: Departamento de Bioquimica, Facultad de Medicina, Universidad de la Republica, Montevideo, Uruguay. SO: Arch-Biochem-Biophys. 1994 Jan; 308(1): 96-102 ISSN: 0003-9861 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Activated macrophages release peroxynitrite anion (ONOO-), which has been recently shown to be highly cytotoxic against Trypanosoma cruzi epimastigotes. In this work, we report that two critical enzymes for the energetic metabolism of the parasite, succinate dehydrogenase and fumarate reductase, are inactivated by biologically relevant concentrations of peroxynitrite. Enzyme inactivation was accompanied by a significant inhibition of succinate-dependent respiration in intact cells as well as in the membrane-rich fraction. Peroxynitrite also inhibited NADH-dependent oxygen consumption which depends almost exclusively on fumarate reductase activity in T. cruzi epimastigotes. Direct reactions of peroxynitrite anion with critical sulfhydryl residues of the two enzymes were responsible for most of the observed inactivation as indicated by the protection afforded by peroxynitrite scavengers and the reactivation of the enzymes by dithiothreitol. We propose that peroxynitrite-mediated inactivation of succinate dehydrogenase and fumarate reductase may be a key mechanism of macrophage-mediated cytotoxicity to T. cruzi, through inhibition of the energetic metabolism of the parasite. MIME: alpha-1-Antitrypsin-metabolism; Adenosinetriphosphatase-antagonists-and-inhibitors; Chelating-Agents-pharmacology; Ethylmaleimide-pharmacology; Kinetics-; Lung-metabolism; Mitochondria,-Heart-enzymology; NADH-Dehydrogenase-antagonists-and-inhibitors; Succinate-Dehydrogenase-metabolism; Trypanosoma-cruzi-drug-effects; Trypanosoma-cruzi-enzymology MJME: *Energy-Metabolism-drug-effects; *Nitrates-toxicity; *Succinate-Dehydrogenase-antagonists-and-inhibitors; *Trypanosoma-cruzi-metabolism TG: Animal; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC; EC; EC; 0; 0; 0; 128-53-0; 26404-66-0 NM: Succinate-Dehydrogenase; NADH-Dehydrogenase; Adenosinetriphosphatase; alpha-1-Antitrypsin; Chelating-Agents; Nitrates; Ethylmaleimide; peroxynitric-acid AN: 94145111 UD: 9405 Record 123 of 131 - MEDLINE (R) 1994 TI: Cardiac antigen-specific autoantibody production is associated with cardiomyopathy in Trypanosoma cruzi-infected mice. AU: Tibbetts-RS; McCormick-TS; Rowland-EC; Miller-SD; Engman-DM AD: Department of Pathology, Northwestern University, Chicago, IL 60611. SO: J-Immunol. 1994 Feb 1; 152(3): 1493-9 ISSN: 0022-1767 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: An inflammatory cardiomyopathy may develop in humans and experimental animals with chronic Trypanosoma cruzi infection (Chagas' disease). Among the possible mechanisms involved in the pathogenesis of Chagas' cardiomyopathy, induction of heart-specific autoimmune responses has recently received substantial experimental support. The goal of the current study was to determine whether cardiac Ag-specific antibodies are produced in T. cruzi-infected mice with heart disease and, if so, to determine their Ag specificities. Upon infection with the Brazil strain of T. cruzi, C57BL/6 mice develop a cardiomyopathy that is histologically similar to that observed in chronically infected humans. Antisera from these mice were found to react with three cardiac Ag, having relative molecular masses of 200, 150, and 53 kDa. p200 and p150 are specifically found in heart muscle, although p53 is found in skeletal muscle as well. C57BL/6 mice infected with the Guayas strain of T. cruzi, which do not develop cardiomyopathy, did not produce antibodies to p200, p150, or p53, indicating that these antibodies may be specific markers of cardiomyopathy. Finally, p200 and p53 were identified as the contractile protein myosin and the intermediate filament protein desmin, respectively. This last finding is of special interest, because antibodies specific for myosin or desmin have been detected in humans and experimental animals with other natural and experimental cardiomyopathies. This suggests that infection with particular strains of T. cruzi may lead to the development of a cardiac Ag-specific autoimmune disease, possibly involving one or more of the Ag identified in this study. MIME: Autoantibodies-biosynthesis; Chagas-Cardiomyopathy-pathology; Desmin-immunology; Mice-; Mice,-Inbred-C3H; Mice,-Inbred-C57BL; Molecular-Weight; Muscles-immunology; Myocardial-Diseases-pathology; Myocardium-pathology; Myosin-immunology; Trypanosoma-cruzi MJME: *Autoantibodies-immunology; *Autoantigens-immunology; *Chagas-Cardiomyopathy-immunology; *Myocardial-Diseases-immunology; *Myocardium-immunology TG: Animal; Female; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AR30692ARNIAMS; GM08061GMNIGMS; NS26543NSNINDS RN: 0; 0; 0; 0 NM: Autoantibodies; Autoantigens; Desmin; Myosin AN: 94132648 UD: 9405 SB: AIM Record 124 of 131 - MEDLINE (R) 1994 TI: Interleukin-6 (IL-6) production in mice infected with Trypanosoma cruzi: effect of its paradoxical increase by anti-IL-6 monoclonal antibody treatment on infection and acute-phase and humoral immune responses. AU: Truyens-C; Angelo-Barrios-A; Torrico-F; Van-Damme-J; Heremans-H; Carlier-Y AD: Laboratory of Parasitology, Faculty of Medicine, University of Brussels, Belgium. SO: Infect-Immun. 1994 Feb; 62(2): 692-6 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Trypanosoma cruzi infection of mice triggered endogenous production of interleukin-6 (IL-6) during the ascending phase of parasitemia. Injections of anti-IL-6 monoclonal antibody in infected mice at the time of the serum IL-6 peak paradoxically increased IL-6 levels to 60- to 80-fold those in infected mice receiving unrelated immunoglobulins. This early and transient increase in circulating IL-6 levels modified neither the immunoglobulin nor T. cruzi-specific antibody levels of immunoglobulin G1 (IgG1), IgG2a, IgG3, IgM, IgA, and IgE isotypes or the final outcome of infection nor the blood or tissular parasite levels. However, it tended to delay mortality of mice and to increase the levels of the acute-phase protein serum amyloid P component. MIME: Acute-Phase-Reaction-blood; Acute-Phase-Reaction-immunology; Amyloid-P-Component-metabolism; Antibodies,-Monoclonal-pharmacology; Antibodies,-Protozoan-blood; Chagas-Disease-blood; Chagas-Disease-parasitology; Immunoglobulin-Isotypes-blood; Interleukin-6-antagonists-and-inhibitors; Mice-; Mice,-Inbred-BALB-C; Time-Factors; Trypanosoma-cruzi-isolation-and-purification MJME: *Chagas-Disease-immunology; *Interleukin-6-biosynthesis TG: Animal; Male; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 0; 0; 0 NM: Amyloid-P-Component; Antibodies,-Monoclonal; Antibodies,-Protozoan; Immunoglobulin-Isotypes; Interleukin-6 AN: 94131608 UD: 9405 Record 125 of 131 - MEDLINE (R) 1994 TI: Isozyme variability of Trypanosoma brucei s.l.: genetic, taxonomic, and epidemiological significance. AU: Mathieu-Daude-F; Tibayrenc-M AD: UMR ORSTOM/CNRS 9926, Genetique Moleculaire des Parasites et des Vecteurs, Montpellier, France. SO: Exp-Parasitol. 1994 Feb; 78(1): 1-19 ISSN: 0014-4894 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Seventy-eight Trypanosoma brucei s.l. stocks from different hosts, representing the three Trypanosoma brucei subspecies and three Trypanosoma evansi stocks, were studied for variation at 18 polymorphic isozyme loci. The results were used to determine the genetic variability among stocks and to estimate gene flow among populations. Total genetic variability in T. brucei s.l. was less than that in Trypanosoma cruzi, the agent of Chagas' disease. Results support a clonal population structure in T. brucei, but do not preclude a hypothesis of occasional mating. However, some natural clones of T. brucei s.l. appear as genetically stable and should be considered as useful taxonomic units in applied studies. Greater genotypic diversity was observed in trypanosomes isolated from wild mammals. Numerical taxonomy methods identified a group of clones representing most of the human stocks from Central and West Africa. This group probably corresponds to Trypanosoma brucei gambiense "group I" (Gibson, Parasitology Today 2, 255-257, 1986). As reported elsewhere, genetic evidence of the subspecies Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense was ambiguous, suggesting that these taxa represent more "nosodemes" rather than actual genetic clades. MIME: Alleles-; Cloning,-Molecular; Cluster-Analysis; Gene-Frequency; Genetics,-Population; Genotype-; Isoenzymes-analysis; Linkage-Disequilibrium; Mammals-; Phylogeny-; Recombination,-Genetic; Trypanosoma-brucei-brucei-classification; Trypanosoma-brucei-brucei-genetics; Trypanosomiasis,-African-parasitology; Tsetse-Flies MJME: *Isoenzymes-genetics; *Trypanosoma-brucei-brucei-enzymology; *Trypanosomiasis,-African-epidemiology; *Variation-Genetics TG: Animal; Human; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0 NM: Isoenzymes AN: 94131048 UD: 9405 Record 126 of 131 - MEDLINE (R) 1994 TI: Trypanosoma cruzi epimastigote forms possess a Ca(2+)-calmodulin dependent protein kinase. AU: Ogueta-SB; Solari-A; Tellez-Inon-MT AD: Instituto de Investigaciones en Ingenieria Genetica y Biologia Molecular (INGEBI) UBA, Argentina. SO: FEBS-Lett. 1994 Jan 17; 337(3): 293-7 ISSN: 0014-5793 PY: 1994 LA: ENGLISH CP: NETHERLANDS AB: Trypanosoma cruzi epimastigote forms showed a tightly bound Ca(2+)-calmodulin-dependent protein kinase activity, which could be partially extracted from membranes and axonemes. The enzyme is constituted by subunits which were autophosphorylated in the absence of exogenous substrates. An antibody against CaM kinase II recognized a Ca(2+)- or Ca(2+)-CaM-dependent conformational epitope in these fractions. The detected bands were of molecular weights similar to the alpha and beta subunits of the corresponding bovine brain enzyme (60 and 50 kDa). Studies using [125I]CaM revealed the presence of a CaM-binding domain. These experiments confirm that the parasite possesses a particulate CaM kinase with characteristics similar to the bovine brain enzyme. MIME: Binding-Sites; Brain-enzymology; Calmodulin-metabolism; Calmodulin-Dependent-Protein-Kinases-chemistry; Calmodulin-Dependent-Protein-Kinases-metabolism; Cattle-; Cell-Fractionation; Cell-Membrane-enzymology; Cytoskeleton-enzymology; Flagella-enzymology; Microscopy,-Electron; Trypanosoma-cruzi-ultrastructure MJME: *Calmodulin-Dependent-Protein-Kinases-analysis; *Trypanosoma-cruzi-enzymology TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: EC 2.7.10.-; 0 NM: Calmodulin-Dependent-Protein-Kinases; Calmodulin AN: 94123773 UD: 9405 Record 127 of 131 - MEDLINE (R) 1994 TI: Endogenous IL-10 and IFN-gamma production controls thymic cell proliferation in mice acutely infected by Trypanosoma cruzi. AU: Leite-de-Moraes-MD; Minoprio-P; Dy-M; Dardenne-M; Savino-W; Hontebeyrie-Joskowicz-M AD: Unite d'Immunoparasitologie, Institut Pasteur, Paris, France. SO: Scand-J-Immunol. 1994 Jan; 39(1): 51-8 ISSN: 0300-9475 PY: 1994 LA: ENGLISH CP: ENGLAND AB: Thymocytes from mice with experimental Trypanosoma cruzi infection respond poorly to Con-A stimulation. However, the proliferative capacity of these cells is not impaired, as demonstrated by the fact that at high doses, exogenous rIL-2 restores thymidine uptake. This finding could be explained either by insufficient IL-2 production or by the appearance of inhibitory factors during T. cruzi infection. This paper shows that in response to Con A, IL-2 production is decreased in the model. Furthermore, the whole profile of cytokine production is modified, with a striking increase in IL-10, IFN-gamma, IL-4, IL-5 and IL-6 production. The results indicate that IL-10 plus IFN-gamma are responsible for the decrease in the Con A-induced proliferation since a normal proliferative response as well as normal IL-2 production can be restored if both cytokines are neutralized by adding their monoclonal antibodies (MoAbs). Evidence is provided also for an enhanced non-specific cytotoxicity of thymic cells from infected mice that might involve IL-4, IL-5 and IL-6. This is the first study demonstrating an alteration of thymic cell function by T. cruzi infection which results from overstimulation of IL-10 and IFN-gamma production. MIME: Antibodies,-Monoclonal; Antigens,-CD3-immunology; Cells,-Cultured; Concanavalin-A-immunology; Cytotoxicity,-Immunologic-immunology; Disease-Models,-Animal; Mice-; Mice,-Inbred-C57BL; T-Lymphocytes,-Cytotoxic-immunology; Thymus-Gland-cytology; Thymus-Gland-immunology MJME: *Chagas-Disease-immunology; *Interferon-Type-II-biosynthesis; *Interleukin-10-biosynthesis; *Lymphocyte-Transformation-immunology; *T-Lymphocytes-immunology TG: Animal; Support,-Non-U.S.-Gov't PT: JOURNAL-ARTICLE RN: 0; 0; 11028-71-0; 130068-27-8; 82115-62-6 NM: Antibodies,-Monoclonal; Antigens,-CD3; Concanavalin-A; Interleukin-10; Interferon-Type-II AN: 94120340 UD: 9404 Record 128 of 131 - MEDLINE (R) 1994 TI: Transcription and editing of cytochrome oxidase II RNAs in Trypanosoma cruzi. AU: Kim-KS; Teixeira-SM; Kirchhoff-LV; Donelson-JE AD: Department of Biochemistry, University of Iowa, Iowa City 52242. SO: J-Biol-Chem. 1994 Jan 14; 269(2): 1206-11 ISSN: 0021-9258 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The cytochrome oxidase subunit II (COII) gene is one of the maxicircle cryptogenes of kinetoplastids whose primary transcripts are sometimes modified by RNA editing to produce mature mRNAs. We determined the sequence of the COII gene in three strains of Trypanosoma cruzi (Y, Corpus Christi, and Tulahuen) and examined its developmental expression. Comparison of the RNA and DNA sequences encoding COII indicated that in the three strains of T. cruzi, four uridines are inserted in the pre-mRNA at the same positions as they are in the COII pre-mRNAs of Trypanosoma brucei, Leishmania tarentolae, and Crithidia fasciculata. The putative guide RNA (gRNA) sequence that serves as a template for the four uridine insertions is located in the 3'-untranslated region of the T. cruzi COII mRNA. Analysis of editing intermediates demonstrates that the COII gRNA remains attached to the pre-mRNA while participating in the formation of chimeric RNAs. Northern blots used to investigate stage-specific expression of the COII gene revealed RNAs of 800 and 900 nucleotides, similar in size to those present in T. brucei. In contrast to the differential expression observed in T. brucei, no difference occurs between the COII mRNA levels of insect and mammalian stages of T. cruzi. MIME: Amino-Acid-Sequence; Base-Sequence; DNA,-Complementary-genetics; Gene-Expression; Molecular-Sequence-Data; Nucleic-Acid-Hybridization; RNA,-Guide-genetics; RNA,-Messenger-genetics; RNA,-Protozoan-genetics; Sequence-Alignment; Sequence-Homology,-Nucleic-Acid; Transcription,-Genetic; Trypanosoma-cruzi-enzymology MJME: *Cytochrome-c-Oxidase-genetics; *RNA-Editing; *Trypanosoma-cruzi-genetics TG: Animal; Comparative-Study; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-Non-P.H.S.; Support,-U.S.-Gov't,-P.H.S. GS: COII PT: JOURNAL-ARTICLE CN: AI24711AINIAID; DK25295DKNIDDK RN: EC; 0; 0; 0; 0 NM: Cytochrome-c-Oxidase; DNA,-Complementary; RNA,-Guide; RNA,-Messenger; RNA,-Protozoan AN: 94117429 UD: 9404 SI: GENBANK/L22643 Record 129 of 131 - MEDLINE (R) 1994 TI: Treatment of chronic Chagas' disease with benznidazole: clinical and serologic evolution of patients with long-term follow-up. AU: Viotti-R; Vigliano-C; Armenti-H; Segura-E AD: Seccion Enfermedad de Chagas, Hospital Interzonal de Agudos Eva Peron, San Martin, Provincia de Buenos Aires, Argentina. SO: Am-Heart-J. 1994 Jan; 127(1): 151-62 ISSN: 0002-8703 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: Prescribing etiologic treatment for chronic Chagas' disease is highly controversial because of the difficulties involved in assessing its therapeutic efficacy--the low degree of parasitemia, the persistence of positive immunologic reactions, the lack of clinical findings to support each type of treatment, and the necessarily prolonged follow-up of the patient. An 8-year average follow-up was performed on 131 patients treated with benznidazole (5 mg/kg/day for 30 days) (TP) and 70 untreated patients (UTP) by serial electrocardiograms and analysis of the cardiomyopathic progress of the clinical groups, and by immunologic tests at both the beginning and end of the study. TPs presented less electrocardiographic changes during the follow-up period (4.2% vs 30%) and a lower frequency of deterioration in their clinical condition (2.1% vs 17%). The percentage of TPs who were serologically negative was 19.1% whereas 6% of the UTPs became serologically negative, a result that correlated with a lack of progress in the cardiomyopathy. Benznidazole treatment significantly decreased serologic titers, signifying parasitologic cure in two patients. MIME: Adult-; Aged-; Antibodies,-Protozoan-blood; Chagas-Disease-immunology; Chagas-Disease-physiopathology; Electrocardiography-; Follow-Up-Studies; Middle-Age; Serodiagnosis-; Treatment-Outcome; Trypanosoma-cruzi-immunology MJME: *Chagas-Disease-drug-therapy; *Nitroimidazoles-therapeutic-use; *Trypanocidal-Agents-therapeutic-use TG: Animal; Comparative-Study; Female; Human; Male PT: JOURNAL-ARTICLE RN: 0; 0; 0; 22994-85-0 NM: Antibodies,-Protozoan; Nitroimidazoles; Trypanocidal-Agents; benzonidazole AN: 94099293 UD: 9404 SB: AIM Record 130 of 131 - MEDLINE (R) 1994 TI: Biochemical analysis of the membrane and soluble forms of the complement regulatory protein of Trypanosoma cruzi. AU: Norris-KA; Schrimpf-JE AD: Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pennsylvania 15261. SO: Infect-Immun. 1994 Jan; 62(1): 236-43 ISSN: 0019-9567 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: A developmentally regulated, 160-kDa trypomastigote surface glycoprotein was previously shown to bind the third component of complement and to inhibit activation of the alternative complement pathway, thus providing the parasites a means of avoiding the lytic effects of complement. We now show that this complement regulatory protein (CRP) binds human C4b, a component of the classical pathway C3 convertase, and may therefore also act to restrict classical complement activation. Characterization of the extent of carbohydrate modification of the protein revealed extensive N-linked glycosylation and no apparent O-linked sugars. The CRP purified from parasites treated with an inhibitor of N-linked glycosylation exhibited a decreased binding affinity for C3b compared with that of the fully glycosylated protein. We have previously shown that the protein was anchored to the membrane via a glycosyl phosphatidylinositol linkage and was spontaneously shed from the parasite surface. The spontaneous release of CRP from the parasite surface may augment the protection of the parasites from complement-mediated lysis by the removal of complement-CRP complexes. The majority of the shed CRP had an apparent molecular mass of 160 kDa and lacked the glycolipid anchor, whereas the membrane form was recovered with the glycolipid anchor attached and had an apparent molecular mass of 185 kDa. Both the membrane form (185 kDa) and the soluble form (160 kDa) retained binding affinity for C3b. Evidence is presented to indicate that the conversion of the 185-kDa membrane form to the 160-kDa form is the result of cleavage by an endogenous phospholipase C. MIME: Complement-Inactivators-immunology; Complement-Inactivators-metabolism; Complement-3b-metabolism; Complement-4b-metabolism; Glycosylphosphatidylinositols-chemistry; Membrane-Glycoproteins-chemistry; Molecular-Weight; Protozoan-Proteins-chemistry; Solubility-; Structure-Activity-Relationship; Trypanosoma-cruzi-chemistry MJME: *Complement-Inactivators-chemistry; *Protozoan-Proteins-immunology; *Trypanosoma-cruzi-immunology TG: Animal; Support,-Non-U.S.-Gov't; Support,-U.S.-Gov't,-P.H.S. PT: JOURNAL-ARTICLE CN: AI32719AINIAID RN: 0; 0; 0; 0; 80295-43-8; 80295-50-7 NM: Complement-Inactivators; Glycosylphosphatidylinositols; Membrane-Glycoproteins; Protozoan-Proteins; Complement-3b; Complement-4b AN: 94086107 UD: 9403 Record 131 of 131 - MEDLINE (R) 1994 TI: Macrophages in experimental Chagas' disease. AU: Kuhn-RE AD: Wake Forest University, Winston-Salem, North Carolina. SO: Immunol-Ser. 1994; 60: 495-502 ISSN: 0092-6019 PY: 1994 LA: ENGLISH CP: UNITED-STATES AB: The foregoing provides a basis for considering that macrophages are important in basically every aspect of the host-parasite relationship in experimental Chagas' disease. The myriad of activities of macrophages and the diverse responses of these cells to T. cruzi and various stimulatory and inhibitory cytokines as measured both in vitro and in vivo are suggestive of the complexity of the cell-cell interactions of the host during the course of infection and the evasive activities of the parasite. MIME: Antibodies,-Protozoan-metabolism; Chagas-Disease-parasitology; Complement-metabolism; Cytokines-immunology; Cytotoxicity,-Immunologic; Disease-Models,-Animal; Immune-Tolerance; Macrophages-metabolism; Macrophages-parasitology; Mice-; Nitric-Oxide-metabolism; Phagocytosis-; Trypanosoma-cruzi-growth-and-development; Trypanosoma-cruzi-immunology; Trypanosoma-cruzi-pathogenicity MJME: *Chagas-Disease-immunology; *Macrophages-immunology TG: Animal; Human PT: JOURNAL-ARTICLE; REVIEW; REVIEW,-TUTORIAL RN: 0; 0; 10102-43-9; 9007-36-7 NM: Antibodies,-Protozoan; Cytokines; Nitric-Oxide; Complement AN: 94072678 UD: 9403