Papers by Elizeu Carvalho
Research in Microbiology, 1990
Protein antigens of Mycobacterium leprae have been identified by screening the),gtl 1, pYA626 and... more Protein antigens of Mycobacterium leprae have been identified by screening the),gtl 1, pYA626 and pHC79::M, leprae genomic libraries with pooled sera from leprosy patients and with antiserum to M. leprae cell wall protein (CWP) aggregate. Immunological screening of the),gtl I library with pooled sera from 21 lepromatous (LL) leprosy patients resulted in the identification of 19 antigens that are apparently different from previously identified M. leprae antigens. Five additional antigens were identified by screening the),gt I 1 library with pooled sera from 30 borderline tuberculoid or tuberculoid patients. Four other antigens were identified by screening the),gtl 1 library with anti-CWP. Two groups of recombinant cosmids were identified by screening the pHC79 library with LL patients' sera: one group specified proteins that reacted with monoclonal antibodies (mAb) against the 65-kDa protein and against the 18-kDa protein; the other group specified a 15-kDa protein that did not react with any of the mAb that were tested. One pYA626 clone also specified a 15-kDa protein that reacted with LL patients' sera, but did not react with any mAb. Genes specifying several of these antigens have been subcloned into the Asd + plasmid vector pYA292 and have been introduced into a Acya Acrp Aasd Salmonella typhimurium strain to evaluate the ability of individual M. leprae proteins to elicit immune responses against M. leprae infection.
Memórias do Instituto Oswaldo Cruz, 1988
Food and Chemical Toxicology, 1996
Stannous chloride (SnCI,) has been widely used in nuclear medicine as a reducing agent of pharmac... more Stannous chloride (SnCI,) has been widely used in nuclear medicine as a reducing agent of pharmaceutical products radiolabelled with technetium-99m. To verify whether the lethality induced by this salt coulcl be mediated by reactive oxygen species (ROS), Escherichiu coli cultures were treated with SnCl, in the presence of catalase, ROS scavengers or metal-ion chelators. The inactivation effect, as measured by survival determination, was abolished by thiourea, sodium benzoate, dipyridyl or catalase. The results suggest the participation of ROS, generated by a Fenton-like reaction, in the lethal effect induced by SKI?.
FEBS Letters, 1989
The cluster of alternated r,. and fl-tubulin genes in the genome of Trypanosoma cruzi was shown t... more The cluster of alternated r,. and fl-tubulin genes in the genome of Trypanosoma cruzi was shown to be transcribed into a single RNA molecule which upon processing gives rise to the mature ~-and fl-tubulin mRNAs. This conclusion was based on: (i) nuclear RNA species with the same molecular mass hybridize to both a-and fl-tubulin eDNA probes; (ii) $1 nuclease assay of the clustered tubulin genes has shown protected DNA fragments of the same size and of greater molecular mass than that corresponding to the mRNAs, hybridizable to both ~t-and fl-tubulin eDNA probes; (iii) fl-tubulin hybrid selected RNA is still able to hybridize to ~t-tubulin probe.
Trypanosoma cruzi: An in vitro cycle of cell differentiation in axenic culture
Experimental Parasitology, 1988
The operation of an in vitro cycle of cell differentiation of Trypanosoma cruzi in axenic culture... more The operation of an in vitro cycle of cell differentiation of Trypanosoma cruzi in axenic culture was obtained. When epimastigote forms, grown in LIT medium, were transferred to a modified LIT medium (E. Chiari, 1981, "Diferenciação do Trypanosoma cruzi em cultura." Ph.D. dissertation, Universidade Federal de Minas Gerais, Brazil), metacyclic trypomastigotes were generated. The latter, upon treatment with fresh human serum, and subsequent incubation in LIT medium gave origin to clusters of spheromastigote cells. The spheromastigotes were resistent to lysis mediated by the complement system and possess a morphology shown by optical and electron microscopy to be very similar to spheromastigotes derived from tissues of infected vertebrates. Blood-like trypomastigotes, or epimastigotes, could be obtained from spheromastigotes depending on the incubation conditions: at high serum concentration (55%) at 37 C, blood-like trypomastigotes were generated; by aging or heating (37 C), at low serum concentration (10%), epimastigotes were formed, closing the whole sequence of cell differentiation of T. cruzi. The molecular characterization of the different cell forms by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of metabolic pulse labeled proteins showed that the in vitro differentiated cells were distinct, not only by morphological criteria, but by differential gene expression as well. All the forms described could be obtained in large amounts (6 x 10(7) to 1 x 10(8)/ml), making it possible to perform preparative biochemical, molecular biological, and immunological experiments.
Testing the Ion AmpliSeq™ HID Y-SNP Research Panel v1 for performance and resolution in admixed South Americans of haplogroup Q
Forensic Science International: Genetics
Y haplogroups, defined by Y-SNPs, allow the reconstruction of the human Y chromosome genealogy, w... more Y haplogroups, defined by Y-SNPs, allow the reconstruction of the human Y chromosome genealogy, which is important for population, evolutionary and forensic genetics. In this study, Y-SNPs were typed and haplogroups inferred with the MPS Ion AmpliSeq™ HID Y-SNP Research Panel v1, as a high-throughput approach. Firstly, the performance of the panel was evaluated with different DNA input amounts, reagent volumes and cycle numbers. DNA-inputs from 0.5 to 1 ng generated the most balanced read depth. Combined with full reagent and 19 cycles, this offered the highest number of amplicons with a sequencing read depth of at least 20 reads. Secondly, the sub-haplogroups of 182 admixed South Americans and Greenlanders belonging to haplogroup Q were inferred and tested for potential improvement in resolution. Most samples were assigned to lineage Q-M3 with some samples assigned to lineages upstream (Q-M346, L56, L57; Q-L331, L53; Q-L54; Q-CTS11969, CTS11970) or parallel (Q-L330, L334; Q-Z780/M971) to Q-M3. Only one sample was assigned to a downstream lineage (Q-Z35615, Z35616). Most individuals of haplogroup Q with NAM ancestry could neither be distinguished from each other, nor from half of the Greenlandic samples. Typing additional, known SNPs within lineage Q-M3, Z19483 and SA05, increased the resolution of predicted haplogroups. The search for novel variants in the sequenced regions allowed the detection of 42 variants and the subdivision of lineage Q-M3 into new subclades. The variants found in six of these subclades were exclusive to certain South American countries. In light of the limited differentiation of haplogroup Q samples, the additional information on known or novel SNPs disclosed in this study when using MPS Ion AmpliSeq™ HID Y-SNP Research Panel v1 should be included in the Yleaf software, to increase the differentiation of lineage Q-M3.
Scientific Reports, 2020
San Basilio de Palenque is an Afro-descendant community near Cartagena, Colombia, founded in the ... more San Basilio de Palenque is an Afro-descendant community near Cartagena, Colombia, founded in the sixteenth century. The recognition of the historical and cultural importance of Palenque has promoted several studies, namely concerning the African roots of its first inhabitants. To deepen the knowledge of the origin and diversity of the Palenque parental lineages, we analysed a sample of 81 individuals for the entire mtDNA Control Region as well as 92 individuals for 27 Y-STRs and 95 for 51 Y-SNPs. The results confirmed the strong isolation of the Palenque, with some degree of influx of Native American maternal lineages, and a European admixture exclusively mediated by men. Due to the high genetic drift observed, a pairwise FST analysis with available data on African populations proved to be inadequate for determining population affinities. In contrast, when a phylogenetic approach was used, it was possible to infer the phylogeographic origin of some lineages in Palenque. Contradictin...
International Journal of Legal Medicine, 2020
Although many genes have been shown to be associated with human pigmentary traits and forensic pr... more Although many genes have been shown to be associated with human pigmentary traits and forensic prediction assays exist (e.g. HIrisPlex-S), the genetic knowledge about skin colour remains incomplete. The highly admixed Brazilian population is an interesting study population for investigation of the complex genotype-phenotype architecture of human skin colour because of its large variation. Here, we compared variants in 22 pigmentary genes with quantitative skin pigmentation levels on the buttock, arm, and forehead areas of 266 genetically admixed Brazilian individuals. The genetic ancestry of each individual was estimated by typing 46 AIM-InDels. The mean proportion of genetic ancestry was 68.8% European, 20.8% Sub-Saharan African, and 10.4% Native American. A high correlation (adjusted R 2 = 0.65, p < 0.05) was observed between nine SNPs and quantitative skin pigmentation using multiple linear regression analysis. The correlations were notably smaller between skin pigmentation and biogeographic ancestry (adjusted R 2 = 0.45, p < 0.05), or markers in the leading forensic skin colour prediction system, the HIrisPlex-S (adjusted R 2 = 0.54, p < 0.05). Four of the nine SNPs, OCA2 rs1448484 (rank 2), APBA2 rs4424881 (rank 4), MFSD12 rs10424065 (rank 8), and TYRP1 1408799 (rank 9) were not investigated as part of the HIrisPlex-S selection process, and therefore not included in the HIrisPlex-S model. Our results indicate that these SNPs account for a substantial part of the skin colour variation in individuals of admixed ancestry. Hence, we suggest that these SNPs are considered when developing future skin colour prediction models.
Forensic Science International: Genetics Supplement Series, 2019
The Blue Shark (Prionace glauca, Linnaeus, 1758) is one of the most abundant sharks in the epipel... more The Blue Shark (Prionace glauca, Linnaeus, 1758) is one of the most abundant sharks in the epipelagic zone (0-200 m) and its population is distributed in oceans of tropical and temperate climate. Currently, these animals have been considered predatory fishing target and are classified as low-risk or near-endangered, according to the Red List of International Union for the Conservation Nature (IUCN). Aiming study conservation aspects of the specie, the purpose of this study is verifying the occurrence of population under-structuring in Prionace glauca. This study was started by analysis of control region of mitochondrial DNA sequences of 254 Prionace glauca individuals, obtained from the Genbank database, as well as new sequences obtained from laboratory. The second step, sequencing of another 16 individuals of the same specie was made and added to the database for a new comparison. From the current results, we can verify that there are haplotypic difference in individuals of the same species, indicating that they belong to distinct lineages. The difference observed between haplotypes indicates that there is a great migratory potential of this specie. This study will be extended with a larger number of samples and the inclusion of nuclear markers, aiming at a better understanding of the processes that act at the population level.
PLOS ONE, 2019
Amphibians are the focus of a recent debate and public attention owing to the global decline in t... more Amphibians are the focus of a recent debate and public attention owing to the global decline in their populations worldwide. Amphibians are one of the most threatened and poorly known groups of vertebrates in several geographic areas, even though they play a central role in their own ecosystems. At different levels, amphibians make their contribution to proper ecosystem functioning. They act as regulators of the food web and nutrient cycling, and they also provide several valuable ecosystem services, e.g., as a food source and as animal models for lab research. In this sense, it seems clear that the maintenance of amphibian diversity should be one of the major goals for the several countries where their population decline is observed. However, we are still struggling with the very first step of this process, i.e., the correct identification of the amphibian species diversity. Over the past few decades, research on molecular identification of amphibians using DNA barcoding has encountered some difficulties related to high variability in the mitochondrial genome of amphibians, and a research gap is noticeable in the literature. We herein evaluated both COI and 16S rRNA mitochondrial genes for the molecular identification of frogs and tadpoles in a large fragment of the South American Atlantic Rainforest in Rio de Janeiro, Brazil. Our results suggest that both COI and 16S rRNA are informative markers for the molecular identification of the amphibian specimens with all specimens unambiguously identified at the species level. We also made publicly available 12 new sequences of Atlantic Rainforest amphibian species for the first time, and we discussed some conservation issues related to amphibians within the Atlantic Rainforest domains in the state of Rio de Janeiro, Brazil.
Forensic Science International: Genetics Supplement Series, 2017
One of the taxa suffering from intense poaching activities along its entire occurrence area, Dasy... more One of the taxa suffering from intense poaching activities along its entire occurrence area, Dasypus represents a genus of armadillos that comprises seven living species: D. hybridus, D. kappleri, D. novemcinctus, D. pilosus, D. sabanicola, D. septemcinctus, and D. yepesi. The aim of this study is to analyze the genetic diversity of D. novemcinctus in order to build a haplotype database for molecular identification purposes. Molecular analyses were performed using the database that includes 99 sequences with 762 bp of the mitochondrial gene Cytochrome b of D. novemcinctus from several localities including North, Central and South America. We performed a maximum likelihood analysis and constructed a median-joining network using the Geneious and PopART softwares respectively. Sequences of the remaining Dasypus species and other armadillos were also included for the molecular analyses. The obtained results suggest four distinct lineages (groups I, II, III and IV) among D. novemcinctus haplotypes. Group I was the largest, with 53 haplotypes and spread over Brazil, Paraguay and Peru. The results reinforced the use of the genetic marker Cytochrome b as an effective tool in the identification of D. novemcinctus.
Forensic Science International: Genetics Supplement Series, 2013
Forensic Science International: Genetics Supplement Series, 2015
The South American country of Peru is composed by a highly admixed population, with Native Americ... more The South American country of Peru is composed by a highly admixed population, with Native American, European and African genetic contributions. Some Native American groups in Peru underwent low admixture with Europeans or Africans and they have kept much of their culture and their original language. In this work we have studied one of these groups called Ashaninka, for the 27 Y chromosome specific STR loci that were included in the recently released YFiler Plus kit. The samples have been collected from 58 unrelated males belonging to 41 different communities located in the margins of the Amazonian rivers Pichis and Palcazú, in the district of Puerto Bermúdez, Pasco region, Peru. A high Y-STR haplotype diversity was found (1.0000±0.0022) with all haplotypes being unique in the studied sample. Two markers that usually present a high diversity in European populations showed very low values of diversity in the Ashaninka Native Americans, namely the DYS635 (Het=0.2263) and DYS437 (Het=0.1325). On the other hand, the DYS438 showed a much higher diversity in Ashaninka (Het=0.6582) group than that usually found in European populations. Apart from the multi-loci markers DYS385 and DYF387S1, more than one allele was also observed in one sample for DYS518 locus. This study represents the first report of haplotype frequencies for the YFiler Plus markers' set in a Native American population, showing a high diversity of haplotypes and, therefore, demonstrating their usefulness in forensic identification cases.
Heart failure and endothelial nitric oxide synthase G894T gene polymorphism frequency variations within ancestries
Nitric oxide : biology and chemistry, Jan 26, 2017
The G894T polymorphism in endothelial nitric oxide synthase enzyme gene plays an important role i... more The G894T polymorphism in endothelial nitric oxide synthase enzyme gene plays an important role in heart failure (HF) and its frequency varies among populations. We investigated this association in highly admixed samples in terms of ancestry. The cohort included 210 HF patients and 106 healthy individuals. Self-reported race and NYHA class were analyzed for HF patients. G894T polymorphism was analyzed by polymerase chain reaction (PCR) and by restriction fragment length polymorphism technique. Ancestry was estimated using a PCR reaction containing 46 autosomal ancestry informative markers and an analysis by capillary electrophoresis. The GG homozygous genotype had a higher frequency in HF patients (63.8%) than in healthy individuals (48.1%), showing an increased chance (odds ratio 1.90, 95% confidence interval 1.18-3.05). The ancestry profiles in patients and controls were similar, with a major European contribution (57.1% and 63.2%), followed by African (30.2% and 24.0%) and Native...
Y Chromosome STR haplotypes in different ethnic groups of Vietnam
Forensic Science International: Genetics, 2016
Forensic Science International: Genetics Supplement Series, 2015
The identification of species and the assignment of unidentified samples to a voucher reference d... more The identification of species and the assignment of unidentified samples to a voucher reference database are among the most requested forensic analyses involving crimes against wildlife. Genomic DNA from forensic or museum samples, however, are frequently degraded, hampering the analysis of DNA fragments longer than 500 bp. Among studies targeting non-human species identification, one of the most frequently used fragments is the mitochondrial Cytochrome C Oxidase subunit I (COI), in particular a 'barcode' region containing approximately 650 base pairs (bp). The main objective of this work was to develop a set of primers to amplify five shorter overlapping COI fragments for the neotropical marmosets (Callithrix spp.) to be used in samples from museum specimens. Taxidermized skins, bones, and hair tissues were sampled out of 9 museum specimens with a range archival from 81 to 2 years; PCR were performed with five mini-amplicons primers pairs and with the primer pair for the whole barcode COI fragment, followed by Sanger sequencing. As results, no amplification was observed using the primer pair for the longer COI sequence; however, using the new set of mini-amplicon primers, all types of samples were amplified. We established optimal PCR conditions for the employed primers and conclude that the mini-amplicon strategy for COI region typing is best suited for hardly degraded samples in forensic research, and in particular for museum samples. 2015 Elsevier Ireland Ltd. All rights reserved.
Forensic Science International: Genetics Supplement Series, 2013
Aquatic Invasions, 2015
The results of our study confirm the occurrence of the cryptic invader Ferrissia fragilis (Tryon,... more The results of our study confirm the occurrence of the cryptic invader Ferrissia fragilis (Tryon, 1863) in Brazil, a species of worldwide geographical distribution and with poorly known morphology that is pervasive in several countries. Specimens were collected in the states of Rio de Janeiro and Minas Gerais in southeastern Brazil. We describe their morphology, and analyze the similarity of haplotypes generated from these samples with those previously obtained for F. fragilis. Shell morphology was compared by light and scanning microscopy. Soft parts of stained dissected specimens were studied under the stereomicroscope. Molecular analysis was performed on three specimens from each sample using the mitochondrial cytochrome c oxidase I gene. Based on a comprehensive analysis, including both morphological and molecular methodologies, we were able to identify the examined specimens as F. fragilis.
38 Human Mutation, Vol. 35, No. 8 (2014), 1021-1032
Towards male individualization with rapidly mutating Y-chromosomal STRs Source: Human mutation (1098-1004)(2014)
ABSTRACT Relevant for various areas of human genetics, Y-chromosomal STRs (Y-STRs) are commonly u... more ABSTRACT Relevant for various areas of human genetics, Y-chromosomal STRs (Y-STRs) are commonly used for testing close paternal relationships amongst individuals and populations, and for male lineage identification. However, even the widely used 17-loci Yfiler set cannot resolve individuals and populations completely. Here, 52 centers generated quality-controlled data of 13 rapidly-mutating (RM) Y-STRs in 14,644 related and unrelated males from 111 worldwide populations. Strikingly, &amp;gt;99% of the 12,272 unrelated males were completely individualized. Haplotype diversity was extremely high (global: 0.9999985, regional: 0.99919-0.9999988). Haplotype sharing between populations was almost absent except for six (0.05%) of the 12,156 haplotypes. Haplotype sharing within populations was generally rare (0.8% non-unique haplotypes), significantly lower in urban (0.9%) than rural (2.1%) and highest in endogamous groups (14.3%). AMOVA revealed 99.98% of variation within populations, 0.018% among populations within groups, and 0.002% among groups. Of the 2,372 newly and 156 previously typed male relative pairs, 29% were differentiated including 27% of the 2,378 fathers/son pairs. Relative to Yfiler, haplotype diversity was increased in 86% of the populations tested and overall male relative differentiation was raised by 23.5%. Our study demonstrates the value of RM Y-STRs in identifying and separating unrelated and related males and provides a reference database. This article is protected by copyright. All rights reserved.
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Papers by Elizeu Carvalho