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M A Miles

Publications and source records attributed to M A Miles.

At least 109 records · Page 6Linked to original sources
PubMed1984

Anion-exchange separation for neotropical trypanosomes: a preliminary trial and a description of Trypanosoma devei from the tamarin Saguinus midas niger.

Anion-exchange separation trials using DEAE-cellulose columns were performed with blood from two single species of marsupial and edentate, three species of rodent and single species of carnivore, primate, cayman and lizard. Trypanosoma cruzi was isolated from Didelphis marsupialis, Dasypus novemcinctus and Coendou sp. T. (Megatrypanum) devei was isolated from the tamarin Saguinus midas niger and the mensural characters of the organism were redescribed. Anion-exchange separation was considered to be a valuable procedure for the taxonomist searching for new or little-known trypanosomes.

Animal Population Groups↗
PubMed1984

Isozyme heterogeneity and numerical taxonomy of Trypanosoma cruzi stocks from Chile.

Fifty-three stocks of Trypanosoma cruzi were isolated in Chile, 13 from patients, 32 from the domestic triatomine vector Triatoma infestans, and 8 from the silvatic and peridomestic vector T. spinolai. The majority of isolates from triatomine bugs were made by the direct culture of infected faeces. The 53 stocks and a single clone were characterized by a combination of starch-gel and cellulose acetate enzyme electrophoresis. Three groups of T. cruzi stocks were apparent from either simple visual comparisons of isozyme profiles or numerical taxonomy. The groups were designated Chilean zymodeme (Z) 1, which was similar to Brazilian Z1, Chilean Z2a, similar to Brazilian Z2 and Chilean Z2b, similar to Bolivian Z2 and with prominent heterozygous isozyme profiles. Chilean Z1 was isolated only from T. spinolai colonizing farm walls inhabited by the rodent Octodon degus. Chilean Z2a and Z2b were both isolated from domestic T. infestans T. infestans and man, in some cases within the same household. Hardy-Weinberg equilibria were not found amongst a group of 22 stocks from a single locality and deviations from theoretical Hardy-Weinberg distributions were compatible with the absence of genetic exchange in the sampled population of T. cruzi.

Alanine Transaminase↗
PubMed1984

Zymodeme and species specificities of monoclonal antibodies raised against Trypanosoma cruzi.

A series of hybrid cell lines was generated by the fusion of Sp2/0 myeloma cells with spleen cells from BALB/c mice that had been immunized or infected with Trypanosoma cruzi zymodemes (Z1, Z2, Z3 ). Four immunoglobulin isotypes, IgM, IgG1, IgG2a and IgG3 were represented amongst the monoclonal antibodies secreted by 22 hybridoma clones. On indirect immunofluorescence (IFAT) antibodies bound to flagellum, cytoplasm, cell membrane or stained the whole organism. Two antibodies were epimastigote-specific. Enzyme-linked immunosorbent assays (ELISA) and a dot immunobinding test were used to evaluate the zymodeme and species specificities of 13 antibodies: four reacted with all T. cruzi zymodemes tested, two reacted strongly with all except Z1, two predominantly with Z1, two predominantly with Z2, and three predominantly with Z3 . Two IgM antibodies cross reacted with Trypanosoma rangeli, T. brucei, Leishmania mexicana, L. braziliensis and L. donovani. Five antibodies were used in a preliminary immunobinding test, performed blindly, to compare monoclonal reactivities and zymodeme groups. The results suggested a correlation between the two methods of characterization. Anti-T. cruzi monoclonal antibodies are considered to have important applications to epidemiological studies and the improved diagnosis and prognosis of Chagas' disease.

Animals↗
PubMed1984

Trypanosoma cruzi from the Paraguayan Chaco: isoenzyme profiles of strains isolated at Makthlawaiya.

At Makthlawaiya, in the Paraguayan Chaco, the prevalence of Trypanosoma (Schizotrypanum) cruzi infection among both domestic Triatoma infestans and domestic dogs was 38%, and IgG anti-T. cruzi antibody was detected by the quantitative enzyme-linked immunosorbent assay (ELISA) in 80% (105/133) of human sera. Ninety percent (25/28) of T. cruzi strains isolated from both T. infestans and dogs showed heterozygous isoenzyme profiles for glucose phosphate isomerase, phosphoglucomutase and 6-phosphogluconate dehydrogenase. These strains appeared to be closely related to Bolivian zymodeme 2. Three Paraguayan T. cruzi strains showed homozygous isoenzyme profiles, similar to those of major Brazilian zymodemes. It was concluded that T. cruzi strains with heterozygous isoenzyme profiles predominate in domestic transmission cycles in this highly endemic area of the Paraguayan Chaco.

Animals↗
PubMed1984

Quantitative differences in the expression of a 72,000 molecular weight cell surface glycoprotein (GP72) in Trypanosoma cruzi zymodemes.

A high affinity monoclonal antibody, 8G2 B9, was used to assess the expression of a 72,000 m.w. glycoprotein ( GP72 ) in isoenzyme-typed T. cruzi strains ( zymodemes ). Western blotting analysis of T. cruzi clones showed that 8G2 B9 bound strongly to GP72 and also suggested that this antigen was absent or weakly detectable in T. cruzi zymodeme 1 (Z1) strains. Purified 8G2 B9 was radiolabeled with 125I and used in an inhibition radioimmune binding assay to compare the quantities of GP72 in different zymodemes . Ninety-six T. cruzi strains were assayed, of which 36 were Z1, 36 were Z2, five were Z3 , and 19 were Z2 (heterozygous). Most (64%) Z1 strains lacked detectable GP72 , whereas this antigen was always detected Z2 and Z2 (heterozygous) strains. There was an 18-fold difference between geometric mean values for the quantities of GP72 (expressed as nanograms per milligram total cell protein) in Z1 and Z2 strains (Z1, 36 ng/mg; Z2, 639 ng/mg; p less than 0.001). There were also significant differences between the geometric mean values for Z2 and Z2 (heterozygous) strains, i.e., 639 ng/mg and 1648 ng/mg, respectively (p less than 0.001). GP72 was detected in four of the Z3 strains in quantities ranging from 740 to 3640 ng/mg. The absolute amounts of antigen in GP72 -positive strains were low, comprising less than 1% of the total cell protein. The specificities of two other anti- GP72 monoclonal antibodies, 7C6 D7 and WIC 29.26, were compared with 8G2 B9. Both antibodies completely inhibited the binding of 8G2 B9 to GP72 in solid phase immunoassays, suggesting that they reacted with the same antigenic determinants. The results show that monoclonal antibody-based assessments of the expression of GP72 correlate with zymodeme classification, and they also suggest that the monoclonal antibodies recognize major antigenic determinants on GP72 . It should be possible to use 8G2 B9 as an immunologic marker to additionally investigate the clinical significance of T. cruzi zymodemes and the biologic significance of GP72 .

Animals↗
PubMed1984

Chagas' disease in the Amazon basin IV. Host records of Trypanosoma cruzi zymodemes in the states of Amazonas and Rondonia, Brazil.

Of 151 Trypanosoma cruzi stocks from 117 different individual hosts collected in the states of Amazonas and Rondonia, 147 from 113 hosts were identified as zymodeme 1 (Z1). These included T. cruzi stocks from three marsupial species, two rodent species and three triatomine species although most were from the common opossum, Didelphis marsupialis. One T. cruzi stock from Rhodnius robustus was identified as Z1 with a Z3 PGM character, one from Sciurus sp. as Z3 and two from Monodelphis brevicaudata and Panstrongylus geniculatus as Z3 with a Z1 ASAT character. The ways in which stocks were isolated and grown up in vitro did not influence isozyme profile. These results support earlier evidence from Pará State that Z2 is absent from the Amazon basin and that the distribution of T. cruzi zymodemes in this region is quite different to that in endemic areas on the south of the continent.

Animals↗
PubMed1983

Purified Trypanosoma cruzi specific glycoprotein for discriminative serological diagnosis of South American trypanosomiasis (Chagas' disease).

Chagas' disease, leishmaniasis, and Trypanosoma rangeli infection are endemic and their distributions overlap in vast regions of South and Central America. Serological cross-reactivities can confuse epidemiological studies of these infections, and their differential diagnosis has been assigned a high priority by the World Health Organisation. A lectin-affinity-purified, 90,000 molecular weight glycoprotein (GP90) is present in the known principal strains (zymodemes) of Trypanosoma cruzi and absent from Leishmania and T rangeli. Patients with T cruzi infection have antibody to GP90, whereas patients with leishmaniasis do not and the two infections can be distinguished in an ELISA system using this antigen. In a mouse model, the same test can differentiate between T cruzi and T rangeli infections. Antigens purified by affinity chromatography clearly provide a practical basis for very precise, even strain-specific, diagnostic tests.

Antibodies↗
PubMed1983

A genetic comparison between Brazilian and Bolivian zymodemes of Trypanosoma cruzi.

The enzyme profiles of three major Brazilian Trypanosoma cruzi zymodemes (Brazilian Z1, Brazilian Z2, Brazilian Z3) and of two principal Bolivian zymodemes were compared. Relationships were assessed both intuitively and by calculating genetic distances. One of the Bolivian zymodemes, Bolivian Z1, was closely related to the Brazilian Z1. The second Bolivian zymodeme, Bolivian Z2, was related to brazilian Z2 but differed from all the Brazilian zymodemes in the occurrence of typical heterozygous isozyme patterns in five out of 12 enzyme loci. Parental stocks and clones of Bolivian Z2 had the same putative heterozygous patterns. The evidence from enzyme profiles on the ploidy of T. cruzi and the possibility of recombination was considered. The presence of putative heterozygous patterns in Bolivian Z2 supported the hypothesis that T. cruzi is diploid. The definition of T. cruzi as a single polytypic species or as a species complex was considered to be dependent on the presence or absence of genetic recombination between or within the zymodemes, which has not been demonstrated in the ecotopes so far examined.

Animals↗
PubMed1983

The polypeptide profiles of strains of the Trypanosoma subgenera Schizotrypanum and Trypanozoon: peptideme characterization.

Examination, by SDS-PAGE, of the polypeptide profiles of 36 Brazilian strains of Trypanosoma (Schizotrypanum) cruzi has revealed five peptidemes (subpopulations having similar polypeptide profiles) recognizing differences not apparent from zymodeme analysis. T. cruzi zymodeme (Z)1 has been subdivided into peptidemes 3, 4 and 5 (one example of Z2 was included in peptideme 5). Peptideme 1 consisted mostly of Z2 strains except for one example each of Z1 and Z3. One strain of Z2 (Tc 003) was sufficiently dissimilar to merit separation into peptideme 2. Peptideme characterization was not possible with the Trypanozoon trypanosomes since there were no significant differences in the polypeptide profiles of 56 strains from Africa and South America; their profiles could only be compared in the molecular weight ranges 94,000 to 70,000 and below 36,000 because of distortion resulting from the variant specific surface glycoproteins. It was possible to differentiate between eight VATs of T. (Trypanozoon) brucei (BruTat 2 to 9) by isoelectric focusing of their soluble proteins.

Animals↗
PubMed1983

Vertebrate hosts and vectors of Trypanosoma rangeli in the Amazon Basin of Brazil.

A total of 46 Trypanosoma rangeli stocks were isolated from naturally infected mammals and triatomine vectors. Twenty-two stocks were from the common opossum (Didelphis marsupialis), one from the brown "4-eyed" opossum (Metachirus nudicaudatus), one from the anteater (Tamandua tetradactyla), one from the coati (Nasua nasua), seven from Rhodnius pictipes and 14 from Rhodnius robustus. Two stocks were also isolated from recently fed sandflies (Lutzomyia sp., Shannoni group). The stocks were identified as T. rangeli on the basis of natural or experimental salivary gland infections in Rhodnius, inoculative (anterior station) transmission to mice, morphological parameters in parasitemic mice and comparisons of isozyme profiles with a known stock of T. rangeli isolated from man. Three other trypanosome stocks from D. marsupialis, T. tetradactyla and the three-toed sloth (Bradypus tridactylus) were morphologically similar to T. rangeli in culture but had quite different isozyme profiles and were not identified. It is concluded that T. rangeli is widely distributed in Amazonas, Pará and Rondonia States of Brazil, and probably extends into other regions where R. pictipes and R. robustus are known to occur. R. pictipes is light-attracted into houses and occasionally transmits Chagas' disease to man. It is likely that T. rangeli is also occasionally transmitted to man in the Amazon basin.

Animals↗
PubMed1983

A monoclonal antibody with specificity for Trypanosoma cruzi, central and peripheral neurones and glia.

Of 26 hybridomas secreting anti-Trypanosoma cruzi antibodies, two reacted with murine brain and spinal cord extracts but not other tissues. Both antibodies (5H7 and 3H3) had identical isotypes (IgM) and specificities as judged by western blotting. Antigens of molecular weight 58,000 and 37,000 in mouse brain and spinal cord, and 58,000 and 35,000 in T. cruzi were detected. Different tissue antigens were recognized by a previously reported T. cruzi mammalian neural tissue cross-reacting monoclonal antibody CE5; on immunofluorescence 5H7 stained some glia, and some central and peripheral neurones in rat tissue sections. Pooled sera from mice chronically infected with T. cruzi competed with binding of 5H7 to T. cruzi antigens.

Animals↗
PubMed1982

Detection of antibodies to Trypanosoma cruzi in the South American opossum (Didelphis marsupialis).

Stocks of Trypanosoma cruzi belonging to two different zymodemes, one usually associated with silvatic reservoir hosts and the other not normally found in wild reservoir hosts, were used as sources of diagnostic antigens in enzyme-linked immunosorbent assays for the detection of antibodies to T. cruzi in Didelphis marsupialis. Both antigen preparations reacted with antibodies in sera from animals found to be infected by conventional parasitological techniques and also in sera from a proportion of the remaining animals in which it was not possible to detect trypanosomes.

Animals↗
PubMed1982

Leishmaniasis in Brazil: XVII. Enzymic characterization of a Leishmania from the armadillo, Dasypus novemcinctus (Edentata), from Pará State.

A comparison of enzyme profiles, by starch-gel electrophoresis, has distinguished a Leishmania of armadillos (Dasypus novemcinctus), from Pará State, north Brazil, from Leishmania braziliensis braziliensis, L. braziliensis guyanensis, L. mexicana amazonensis, L. donovani sensu lato (from Bahia, Brazil), and L. hertigi deanei. The parasite was separated from L. b. braziliensis and L. b. guyanensis by 8 of the 14 enzymes used (ASAT, ALAT, PGM, GPI, G6PD, PEP, MPI and GD), although differences in the mobility of some of the enzymes were small. At least 9 of the enzymes separated the organism from L. m. amazonensis, L. donovani s.l., and L. h. deanei.

Animals↗
PubMed1982

Molecular size of enzymes in Trypanosoma cruzi considered in relationship to the genetic interpretation of isozyme patterns.

The molecular size of seven Trypanosoma cruzi enzymes, chosen because of their frequent use in studying trypanosome populations, has been found to be similar to that of their mammalian equivalents. Malic enzyme (NADP-dependent malate dehydrogenase, decarboxylating, EC 1.1.1.40) from T. cruzi has an apparent molecular size of only half that of the mammalian enzyme. The probable subunit structure of these T. cruzi enzymes has been deduced from the molecular weights by comparison with mammalian data. The results are compatible with recent interpretations of isozyme data implying the existence of genetic diploidy in trypanosomes.

Animals↗