Erika H.E. Hoffmann

Child health and nutrition in the Western Brazilian Amazon: population-based surveys in two counties in Acre State

The article presents prevalence rates for malnutrition, intestinal parasitic infections, anemia, and iron deficiency in under-five children in a population-based cross-sectional survey performed in the urban area of two counties in the Western Brazilian Amazon, Assis Brasil (n = 200) and Acrelandia (n = 477). Available data included: (a) weight and height measurements, standardized as z-scores using the 1977 NCHS reference population, (b) diagnosis of current intestinal parasitic infection, (c) blood hemoglobin levels, and (d) plasma ferritin and soluble transferrin receptor levels. Overall prevalence rates of low weight-for-height, low weight-for-age, and low height-for-age were 3.7%, 8.7%, and 7.5%, respectively, with similar figures in the two towns. Intestinal parasites were detected in 32.5% children; helminths were uncommon. Anemia and iron deficiency were diagnosed in 30.6% and 43.5% of the children, respectively. Evidence of anemia was found in only 47.6% of the children with depleted iron reserves, indicating that hemoglobin measurement alone would severely underestimate the magnitude of iron deficiency in this population. In both towns, anemia and malnutrition were significantly more prevalent among children in the lowest socioeconomic stratum.

Geographical patterns of allelic diversity in the Plasmodium falciparum malaria-vaccine candidate, merozoite surface protein-2

The polymorphic merozoite surface protein-2 (MSP-2) of Plasmodium falciparum is a major malariavaccine candidate. In the present study, PCR and hybridization with allelic-specific probes were used to type the Msp-2 gene from isolates from hypo-endemic Brazil (N = 113), meso-endemic Vietnam (N = 208) and holo-endemic Tanzania (N = 67). The typing methods were designed to group isolates into the dimorphic allelic families FC27 and IC1 and to detect possible between-family recombination events. The analysis was complemented by a comparison of 156 Msp-2 sequences from the GenBank database with 12 additional sequences obtained during the present study. Statistically significant differences were detected in pair-wise comparisons of the distribution of Msp-2 allelic types in Brazil and Vietnam, and in Brazil and Tanzania, but not in Vietnam and Tanzania. The extent of allelic diversity in the Msp-2 gene, as estimated by the total number of different alleles found in a given parasite population and the mean multiplicity of infections, clearly paralleled the levels of malaria endemicity in the study areas. However, no correlation between age and multiplicity of infections was found in the subjects. The patterns of Msp-2 diversity in Brazil appeared to be temporally stable, since no significant difference was observed in the distribution of Msp-2 allelic types among isolates collected, 10–13 years apart, in the same area of Rondônia. Despite the extensive sequence diversity found in Msp-2 alleles, especially in the central repetitive region of the molecule, several instances of identical or nearly identical alleles were found among isolates from different countries and regions, possibly as a result of extensive homoplasy. No recombinant allele was detected by molecular typing in any of the study sites, and the GenBank database included only 12 recombinant sequences (representing 7% of all reported Msp-2 sequences), all of them with an IC1-type 5′ end and an FC27-type 3′ end. A single, putative, crossover site was characterised for all recombinant alleles. Most of the allelic diversity observed was therefore attributable to variation in the repetitive region of the gene, instead of recombination between alleles of dimorphic families (as commonly found, for example, in the Msp-1 gene). The implications of these findings for studies on the genetic and antigenic diversity of malarial parasites are discussed.

Antigenic Polymorphism and Naturally Acquired Antibodies to Plasmodium vivax Merozoite Surface Protein 1 in Rural Amazonians

ABSTRACT Merozoite surface protein 1 of Plasmodium vivax (PvMSP-1), a major target for malaria vaccine development, contains six highly polymorphic domains interspersed with conserved sequences. Although there is evidence that the sequence divergence in PvMSP-1 has been maintained over 5 million years by balanced selection exerted by the hosts acquired immunity, the variant specificity of naturally acquired antibodies to PvMSP-1 remains poorly investigated. Here, we show that 15 recombinant proteins corresponding to PvMSP-1 variants commonly found in local parasites were poorly recognized by 376 noninfected subjects aged 5 to 90 years exposed to malaria in rural Amazonia; less than one-third of them had detectable immunoglobulin G (IgG) antibodies to at least one variant of blocks 2, 6, and 10 that were expressed, although 54.3% recognized the invariant 19-kDa C-terminal domain PvMSP-119. Although the proportion of responders to PvMSP-1 variants increased substantially during subsequent acute P. vivax infections, the specificity of IgG antibodies did not necessarily match the PvMSP-1 variant(s) found in infecting parasites. We discuss the relative contribution of antigenic polymorphism, poor immunogenicity, and original antigenic sin (the skew in the specificity of antibodies elicited by exposure to new antigenic variants due to preexisting variant-specific responses) to the observed patterns of antibody recognition of PvMSP-1. We suggest that antibody responses to the repertoire of variable domains of PvMSP-1 to which subjects are continuously exposed are elicited only after several repeated infections and may require frequent boosting, with clear implications for the development of PvMSP-1-based subunit vaccines.

The South American Plasmodium falciparum var gene repertoire is limited, highly shared and possibly lacks several antigenic types

The Plasmodium falciparum var gene family encodes large variant antigens, which are important virulence factors, and also targets of the humoral host response. The frequently observed mild outcomes of falciparum malaria in many places of the Amazon area prompted us to ask whether a globally restricted variant (var) gene repertoire is present in currently circulating and older isolates of this area. By exhaustive analysis of var gene tags from 89 isolates and clones taken during many years from all over the Brazilian Amazon, we estimate that there are probably no more than 350-430 distinct sequence types, less than for any similar sized area studied so far. Detailed analysis of the var tags from genetically distinct clones obtained from single isolates revealed restricted and redundant repertoires suggesting either a low incidence of infective bites or restricted variant gene diversity in inoculated parasites. Additionally, we found a structuring of var gene repertoires observed as a higher pairwise typing sharing in isolates from the same microregion compared to isolates from different regions. Fine analysis of translated var tags revealed that certain Distinct Sequence Identifiers (DSIDs) were differently represented in Brazilian/South American isolates when compared to datasets from other continents. By global alignment of worldwide var DBLalpha sequences and sorting in groups with more than 76% identity, 125 clusters were formed and more than half of all genes were found in nine clusters with 50 or more sequences. While Brazilian/South American sequences were represented only in 64 groups, African sequences were found in the majority of clusters. DSID type 1 related sequences accumulated almost completely in one single cluster, indicating that limited recombination occurs in these specific var gene types. These data demonstrate the so far highest pairwise type sharing values for the var gene family in isolates from all over an entire subcontinent. The apparent lack of specific sequences types suggests that the P. falciparum transmission dynamics in the whole Amazon are probably different from any other endemic region studied and possibly interfere with the parasites ability to efficiently diversify its variant gene repertoires.

Genetic relatedness of Plasmodium falciparum isolates and the origin of allelic diversity at the merozoite surface protein-1 (MSP-1) locus in Brazil and Vietnam

BackgroundDespite the extensive polymorphism at the merozoite surface protein-1 (MSP-1) locus of Plasmodium falciparum, that encodes a major repetitive malaria vaccine candidate antigen, identical and nearly identical alleles frequently occur in sympatric parasites. Here we used microsatellite haplotyping to estimate the genetic distance between isolates carrying identical and nearly identical MSP-1 alleles.MethodsWe analyzed 28 isolates from hypoendemic areas in north-western Brazil, collected between 1985 and 1998, and 23 isolates obtained in mesoendemic southern Vietnam in 1996. MSP-1 alleles were characterized by combining PCR typing with allele-specific primers and partial DNA sequencing. The following single-copy microsatellite markers were typed : Polyα, TA42 (only for Brazilian samples), TA81, TA1, TA87, TA109 (only for Brazilian samples), 2490, ARAII, PfG377, PfPK2, and TA60.ResultsThe low pair-wise average genetic distance between microsatellite haplotypes of isolates sharing identical MSP-1 alleles indicates that epidemic propagation of discrete parasite clones originated most identical MSP-1 alleles in parasite populations from Brazil and Vietnam. At least one epidemic clone propagating in Brazil remained relatively unchanged over more than one decade. Moreover, we found no evidence that rearrangements of MSP-1 repeats, putatively created by mitotic recombination events, generated new alleles within clonal lineages of parasites in either country.ConclusionIdentical MSP-1 alleles originated from co-ancestry in both populations, whereas nearly identical MSP-1 alleles have probably appeared independently in unrelated parasite lineages.

Differential Recognition of Plasmodium falciparum Merozoite Surface Protein 2 Variants by Antibodies from Malaria Patients in Brazil

ABSTRACT Four variants of merozoite surface protein 2 (MSP-2) of Plasmodium falciparum were used in serology to examine whether changes in repeat units affect its recognition by antibodies during infection with parasites of known MSP-2 types. The results indicate that variation in MSP-2 repeats may represent a mechanism for parasite immune evasion.

Genetic diversity and differentiation in natural Plasmodium falciparum populations inferred by molecular typing of the merozoite surface proteins 1 and 2

Genetic diversity and differentiation, inferred by typing the polymorphic genes coding for the merozoite surface proteins 1 (Msp-1) and 2 (Msp-2), were compared for 345 isolates belonging to seven Plasmodium falciparum populations from three continents. Both loci yielded similar estimates of genetic diversity for each population, but rather different patterns of between-population differentiation, suggesting that natural selection on these loci, rather than the transmission dynamics of P. falciparum, determines the variation in allele frequencies among populations.

SU‐E‐I‐14: Evaluation of Image Quality Parameters of Small and Full Field of View Cone Beam Computed Tomography Dental Imaging Systems

PURPOSE CBCT systems are being used in dental preoperative planning, which rely on 3D surface model representations of the jaw, using a segmentation algorithm for extracting the bony tissues. However, CBCT systems have differences of imaging parameters, which affect the image quality and segmentation process, influencing the accuracy of the 3D surface models used in preoperative implant planning. The purpose of this study was to evaluate image quality parameters of different types of CBCT systems, showing their potentialities and limitations for preoperative planning. METHODS Images were acquired using the systems: i-CAT (Imaging Sciences International Inc., USA); ProMax 3Ds (Planmeca Oy, Finland); Kodak 9500 Cone Beam 3D and Kodak 9000C Cone Beam 3D (Kodak Dental Systems, USA). CBCT systems were divided into categories, related to their field of view size: Small Field of View (SFOV) and Full Field of View (FFOV) systems. Image quality parameters evaluated were: CT number accuracy, uniformity, noise level, and artifacts. Measurements were performed in slices around the center of the phantom. RESULTS CT numbers are highly affected by the amount of mass outside the reconstructed volume, confirming the relation between density variability and imaging volume. The mean CT number shows little correspondence to that of the traditional CT protocols. The variability is especially high in the case for SFOV systems. Uniformity artifacts occur at the top or bottom of the FOV, and Result in voxel values that are unsuitable for CT number accuracy analysis. SFOV systems showed a higher noise level (11,9%) comparing to FFOV systems (4.4%). CONCLUSION The experiments showed that the size and position of the FOV affect the image quality for all CBCT systems. The study was limited to the CBCT systems that were accessible when this study was conducted. Whether new CBCT systems models would perform differently remains to be investigated.