RHCE variant alleles and risk of alloimmunization in Brazilians.

Variant RHCE alleles are found mainly in Afro-descendant individuals, as well as in patients with sickle cell disease (SCD). The most common variants are related to the RHCE*ce allele, which can generate partial e and c antigens. Although RHCE variant alleles have been extensively studied, defining their clinical significance is a difficult task. We evaluated the risk of RhCE alloimmunization as a consequence of partial antigens in patients with a positive phenotype transfused with red blood cell (RBC) units with the corresponding antigen. A retrospective study was performed with Brazilian patients, evaluating the number of antigen-positive transfused RBC units (incompatible due to partial antigen) in 27 patients with SCD carrying RHCE variant alleles who did not develop antibodies as well as evaluating the variants present in 12 patients with partial phenotype and correlated antibody (one patient with SCD and 11 patients with other pathologies). Two patients showed variant alleles with molecular changes that had not yet been described. Variant RHCE alleles were identified in a previous study using molecular methods. RHCE*ceVS.01 was the most frequent allele found among the patients without antibodies. Six patients with partial c antigen had a mean of 3.8 c+ RBC units transfused, and 10 patients with partial e antigen were exposed for a mean of 7.2 e+ RBC units. Among the variant alleles found in alloimmunized patients, the most frequent was RHCE*ceAR, which was found in five patients; the antibodies developed were anti-hrS and/or anti-c. Our results showed that RHCE*ceVS.01 is indeed the most frequent variant allele in our cohort of patients with SCD, but the partial antigens that were identified have low risk of alloimmunization. RHCE*ceAR is the most impactful variant in the Brazilian population with high risk of alloimmunization and clinically significant anti-hrS formation.

Challenges in providing compatible blood with Rh genotype-matching in Brazilian patients with sickle cell disease.

OBJECTIVES: The aim of this study was to verify the possibility of performing prophylactic Rh genotype-matching in Brazilian patients with sickle cell disease (SCD) and identify the genotypes that are lacking or insufficient in our donor cohort. BACKGROUND: Rh alloimmunisation is still a challenge in transfused patients with SCD. Rh genotype-matching may mitigate Rh alloimmunisation. METHODS/MATERIALS: We examined the transfusion requests for antigen-matched donor units in SCD patients with Rh variants and compared the Rh altered alleles in the patients to the Rh allele frequency in a selected donor population. For each patient and donor, RBC antigen genotyping was performed using HEA, RHD and RHCE BeadChip arrays. Sequencing was used to clarify inconclusive results. Twenty-one patients and 956 Brazilian blood donors were genotyped. RESULTS: According to our matching strategies, 47·6% of patients filled most of their unit requests, but 52·4% of patients had insufficient donors to fill their annual transfusion needs. We found different combinations of RHCE variant alleles in patients and donors, but the most frequent genotypes that are lacking or insufficient in our donor cohort are those associated with the lack of hrB and hrS high prevalence antigens and those co-inherited with altered RHD alleles. CONCLUSION: Our study shows that the provision of compatible blood with Rh genotype-matching in Brazilian patients with SCD can be feasible but challenging and, efficient strategies of recruitment of African-Brazilian donors must be developed.

HLA polymorphisms and risk of red blood cell alloimmunisation in polytransfused patients with sickle cell anaemia.

BACKGROUND: Red blood cell (RBC) alloimmunisation is an event that may occur due to factors such as numerous blood transfusions, age, gender and genetic factors such as human leukocyte antigen (HLA). AIMS/OBJECTIVES: The aim of the present study was to investigate the possibility of alloimmunisation to red blood cell group antigens associated with the HLA of individuals and to relate alloimmunisation to risk factors. METHODS: A total of 172 polytransfused patients with sickle cell anaemia (SCA) (44 alloimmunised, 128 non-alloimmunised) participated in this study. Blood group genotyping was performed by the DNA microarray method and HLA genotyping by polymerase chain reaction - specific sequence of oligonucleotides. RESULTS: The number of transfusions received directly influenced the incidence of alloimmunisation, and the most common alloantibodies were against Rh (48·8%) and Kell (17%) systems. The HLA-C*06 and HLA-DQB1*03 variants were significantly higher in alloimmunised patients. The HLA-DRB1*04 and HLA-DRB1*11 were more often found in individuals who developed the alloantibodies anti-Fya and anti-K, respectively. CONCLUSION: This study suggests that polytransfused patients with SCA possessing the HLA-DQB1*03 and HLA-C*06 allele variants are more susceptible to alloimmunisation. In addition, HLA-DRB1*04 and HLA-DRB1*11 alleles were seen to be associated with the production of anti-Fya and anti-K antibodies, respectively.

Vacuna oral (OM-89) en la profilaxis de infección urinaria recurrente: una revisión sistemática realista con metaanálisis / Oral vaccine (OM-89) in the recurrent urinary tract infection prophylaxis: a realistic systematic review with meta-analysis

Objetivo: Evaluar la eficacia del extracto de Escherichia coli (OM-89) en la profilaxis de la infección del tracto urinario (ITU) no complicada recurrente a través de una revisión sistemática contemporánea y metaanálisis. Métodos: Los criterios de inclusión fueron ensayos aleatorios doble ciego usando 6 mg al día de OM-89 administrado por vía oral durante 3 meses, con un mínimo de 3 meses de seguimiento. Los resultados fueron la frecuencia de bacteriuria en 3 y 6 meses, disuria en 6 meses e ITU en 6 meses. Bases de datos: PubMed, MEDLINE, Cochrane Collaboration y sus referencias clave. Tras el análisis por 3 revisores independientes se identificaron 15 ensayos aleatorizados doble ciego, 10 documentos fueron excluidos debido a defectos de los métodos y 5 utilizados para el análisis de datos debido al doble cegamiento y la presentación de informes de abandonos. Resultados: Entre 5 estudios seleccionados la fecha de publicación varió desde 1985 hasta 2005, totalizando 396 pacientes en el grupo OM-89 y 392 en el grupo control. En total, hubo 61 abandonos en el grupo control y 76 en el grupo OM-89. Como una importante limitación no hubo descripción apropiada de sus metodologías, y ninguno de los estudios describió conflicto de intereses o compromiso con la industria farmacéutica. Todos los estudios fueron multicéntricos, a excepción de 2, que no mostraron claridad sobre ocultamiento de la asignación. Todos los estudios muestran beneficios a favor de la vacuna. Conclusiones: La literatura actual sobre ensayos controlados aleatorizados prospectivos que evalúan el uso de la vacuna oral de OM-89 en la profilaxis recurrente de ITU es de baja calidad, limitada únicamente a los primeros 6 meses y con definición variable de bacteriuria e ITU. Aunque todos los estudios muestran beneficio a favor de la vacuna, no se identificó ningún ensayo robusto, resultando en una alta heterogeneidad en los datos analizados. Además, el sesgo de publicación no pudo excluirse, y futuros estudios de mayor calidad están garantizados añadiendo seguimiento a medio (> 12 meses) y largo plazo

Objective: To evaluate the efficacy of Escherichia Coli extract (OM-89) in the prophylaxis of recurrent uncomplicated urinary tract infection (UTI) through a contemporary systematic review and meta-analysis. Methods: Inclusion criteria were double-blind randomized trials using orally administrated OM-89, 6 mg daily, during three months with a minimum of three months of monitoring. Outcomes were the frequency of bacteriuria in 3 and 6 months, dysuria in 6 months and UTI in 6 months. Databases: PubMed, MEDLINE, Cochrane Collaboration and their key references. After analysis by three independent reviewers, 15 double-blind randomized trials were identified, 10 papers excluded due to methods flaws and 5 used for data analysis due to double blinding and reporting drop-outs. Results: Among 5 selected studies the date of publication ranged from 1985 to 2005, totalizing 396 patients in the OM-89 group and 392 in the control group. Overall, there were 61 dropouts in the control group and 76 in the OM-89 group. As a major limitation there was no appropriate description of their methodologies and none of the studies described conflict of interest or commitment to the pharmaceutical industry. All studies were multi-centric, except for two, which showed no clarity on allocation concealment. All studies show benefit in favor of vaccine. Conclusions: Current literature on prospective randomized controlled trials evaluating the use of oral OM-89 vaccine in the recurrent UTI prophylaxis is of low quality, limited to the first six months only and with variable definition of bacteriuria and UTI. Although all studies show benefit in favor of vaccine, no robust trial was identified, resulting in a high heterogeneity in the data analyzed. Also, publication bias could not be excluded and future higher quality studies are warranted adding intermediate (> 12 months) and long-term follow-up

RHCE variants inherited with altered RHD alleles in Brazilian blood donors.

BACKGROUND: The high homology and opposite orientation of RH genes promote rearrangements between them and generate a large number of RHD and RHCE variants which can be inherited together. Searching of RHD-CE genotypes predicting partial antigens in donors is of interest in order to find more closely matched donors for African descent patients. This study aimed to evaluate a molecular approach to search for RhCE variants in a cohort of individuals with altered expression of D antigen and determine the association of RH variant alleles in Brazilian blood donors. METHODS: From 80,961 blood samples tested, 421 with atypical D typing results were studied. The samples were phenotyped for C, c, E, e antigens. Rh variants were identified using molecular techniques. RESULTS: All 421 samples had altered RHD alleles, being 56·3% of them partial D. Among them, 94·9% presented variant RHCE*ce and the most common associations were: RHD*weak D type 4.2.2 with RHCE*ceAR; RHD*DAR linked to RHCE*ceVS.02; RHD*weak D type 4.0 linked to RHCE*ceVS.02 and RHCE*ce (c.48C, c.105T, c.733G, c.744C, c.1025T). Among the samples with RhCE variants, 10·6% predict partial c, partial e, hr(B) - and/or hr(S) - and 100% express low prevalence antigens. CONCLUSION: Targeting individuals with altered expression of D antigen can be a good strategy for finding donors with RhCE variants. In our study 94·9% of the partial D samples revealed altered RHCE variant alleles and 5·7% of the samples with altered RHD allele predicted partial c, partial e and the lack of the high prevalence hr(B) and hr(S) antigens.

International society of blood transfusion working party on red cell immunogenetics and terminology: report of the Seoul and London meetings.

The Working Party has met twice since the last report: in Seoul, South Korea 2014, and in London, UK 2015, both in association with the International Society of Blood Transfusion (ISBT) Congress. As in previous meetings, matters pertaining to blood group antigen nomenclature were discussed. Eleven new blood group antigens were added to seven blood group systems. This brings the current total of blood group antigens recognized by the ISBT to 346, of which 308 are clustered within 36 blood groups systems. The remaining 38 antigens are currently unassigned to a known blood group system.

Oral vaccine (OM-89) in the recurrent urinary tract infection prophylaxis: a realistic systematic review with meta-analysis. / Vacuna oral (OM-89) en la profilaxis de infección urinaria recurrente: una revisión sistemática realista con metaanálisis.

OBJECTIVE: To evaluate the efficacy of Escherichia Coli extract (OM-89) in the prophylaxis of recurrent uncomplicated urinary tract infection (UTI) through a contemporary systematic review and meta-analysis. METHODS: Inclusion criteria were double-blind randomized trials using orally administrated OM-89, 6mg daily, during three months with a minimum of three months of monitoring. Outcomes were the frequency of bacteriuria in 3 and 6 months, dysuria in 6 months and UTI in 6 months. DATABASES: PubMed, MEDLINE, Cochrane Collaboration and their key references. After analysis by three independent reviewers, 15 double-blind randomized trials were identified, 10 papers excluded due to methods flaws and 5 used for data analysis due to double blinding and reporting drop-outs. RESULTS: Among 5 selected studies the date of publication ranged from 1985 to 2005, totalizing 396 patients in the OM-89 group and 392 in the control group. Overall, there were 61 dropouts in the control group and 76 in the OM-89 group. As a major limitation there was no appropriate description of their methodologies and none of the studies described conflict of interest or commitment to the pharmaceutical industry. All studies were multi-centric, except for two, which showed no clarity on allocation concealment. All studies show benefit in favor of vaccine. CONCLUSIONS: Current literature on prospective randomized controlled trials evaluating the use of oral OM-89 vaccine in the recurrent UTI prophylaxis is of low quality, limited to the first six months only and with variable definition of bacteriuria and UTI. Although all studies show benefit in favor of vaccine, no robust trial was identified, resulting in a high heterogeneity in the data analyzed. Also, publication bias could not be excluded and future higher quality studies are warranted adding intermediate (>12 months) and long-term follow-up.

Abundance of Harpy and Crested Eagles from a reservoir-impact area in the Low- and Mid-Xingu River.

In the Brazilian Amazon, two monospecific genera, the Harpy Eagle and Crested Eagle have low densities and are classified by IUCN as Near Threatened due to habitat loss, deforestation, habitat degradation and hunting. In this study, we evaluate occurrence of these large raptors using the environmental surveys database from Belo Monte Hydroelectric Power Plant. Integrating the dataset from two methods, we plotted a distribution map along the Xingu River, including records over a 276-km stretch of river. Terrestrial surveys (RAPELD method) were more efficient for detecting large raptors than standardized aquatic surveys, although the latter were complementary in areas without modules. About 53% of the records were obtained during activities of wildlife rescue/flushing, vegetation suppression or in transit. Between 2012 and 2014, four Harpy Eagles were removed from the wild; two shooting victims, one injured by collision with power lines and one hit by a vehicle. Also, seven nests were mapped. The mean distance between Harpy Eagle records was 15 km along the river channel, with a mean of 20 km between nests near the channel, which allowed us to estimate 20 possible pairs using the alluvial forest, riverine forest and forest fragments. Territories of another ten pairs will probably be affected by inundation of the Volta Grande channel, which is far from the main river. The average distance between Crested Eagle records was 16 km along the river channel. The only nest found was 1.3 km away from a Harpy Eagle nest. The remnant forests are under threat of being replaced by cattle pastures, so we recommend that permanently protected riparian vegetation borders (APP) be guaranteed, and that forest fragments within 5 km of the river be conserved to maintain eagle populations.

Lesion Activity Assessment (LAA) in Conjunction With International Caries Detection and Assessment System (ICDAS) for Occlusal Caries Diagnosis in Permanent Teeth.

OBJECTIVE: The aim of this study was to investigate the clinical performance and to validate the Lesion Activity Assessment (LAA) in conjunction with the International Caries Detection and Assessment System (ICDAS) for occlusal caries diagnosis in permanent teeth. METHODS: Patients with erupted or partially erupted third molars were recruited from the surgery clinic of the School of Dentistry of the Universidade Federal de Minas Gerais, Brazil. A calibrated examiner evaluated 49 teeth using the ICDAS-LAA criteria. The histologic criterion proposed by Ekstrand and others was used to validate severity at the thresholds D1 (outer half of the enamel), D2 (inner half of the enamel and outer third of the dentin), and D3 (inner or middle third of the dentin). Lesion activity was validated using 0.1% methyl red solution. RESULTS: The method demonstrated good reliability (weighted kappa for severity=0.60; unweighted kappa for activity=0.61). The ICDAS presented a higher performance for lesion detection (area under the receiver operating characteristic curve [Az]=0.79) using the threshold D3. At the thresholds D1 and D2, the results for Az were 0.57 and 0.74, respectively. Regarding the ICDAS-LAA, Az = 0.59. CONCLUSIONS: Clinical protocols can use ICDAS for the severity diagnosis of occlusal caries, but the LAA performance was poor.

Abundance of Harpy and Crested Eagles from a reservoir-impact area in the Low- and Mid-Xingu River

Abstract In the Brazilian Amazon, two monospecific genera, the Harpy Eagle and Crested Eagle have low densities and are classified by IUCN as Near Threatened due to habitat loss, deforestation, habitat degradation and hunting. In this study, we evaluate occurrence of these large raptors using the environmental surveys database from Belo Monte Hydroelectric Power Plant. Integrating the dataset from two methods, we plotted a distribution map along the Xingu River, including records over a 276-km stretch of river. Terrestrial surveys (RAPELD method) were more efficient for detecting large raptors than standardized aquatic surveys, although the latter were complementary in areas without modules. About 53% of the records were obtained during activities of wildlife rescue/flushing, vegetation suppression or in transit. Between 2012 and 2014, four Harpy Eagles were removed from the wild; two shooting victims, one injured by collision with power lines and one hit by a vehicle. Also, seven nests were mapped. The mean distance between Harpy Eagle records was 15 km along the river channel, with a mean of 20 km between nests near the channel, which allowed us to estimate 20 possible pairs using the alluvial forest, riverine forest and forest fragments. Territories of another ten pairs will probably be affected by inundation of the Volta Grande channel, which is far from the main river. The average distance between Crested Eagle records was 16 km along the river channel. The only nest found was 1.3 km away from a Harpy Eagle nest. The remnant forests are under threat of being replaced by cattle pastures, so we recommend that permanently protected riparian vegetation borders (APP) be guaranteed, and that forest fragments within 5 km of the river be conserved to maintain eagle populations.

Resumo Na Amazônia brasileira dois gêneros mono-específicos, Harpia e Morphnus, caracterizam-se por baixa densidade e estão classificados pelo IUCN como Quase Ameaçados, porém ocorrem sobre grande parte do território nacional, suas principais ameaças são a fragmentação florestal, a degradação de hábitat e a caça. Neste estudo avaliamos a abundância destas duas grandes aves de rapina utilizando a base de dados dos programas ambientais da UHE Belo Monte, integrando-se dois métodos para construir um mapa de distribuição ao longo de 245 km do rio Xingu. Os levantamentos terrestres pelo método RAPELD mostraram-se mais eficientes para os registros de grandes águias quando comparado aos esforços padronizados aquáticos, entretanto estes foram complementares na ausência de módulos. Cinquenta e tres por cento foram registros ocasionais durante a supressão da vegetação, afugentamento ou deslocamento. Entre 2012-2014 quatro harpias foram removidas da natureza, dois indivíduos alvo de disparos, uma por colisão com rede elétrica, e outra por atropelamento. Sete ninhos mapeados, a distância média entre os registros de harpia na calha e margens do rio foi de 15 km, 20 km distância média entre ninhos, o que permitiu estimar um total de 20 casais utilizando as florestas aluviais em uma distancia de 270 km, incluindo matas ciliares e os fragmentos fora da margem do rio. Estimamos que territórios de outros 10 casais usando a Volta Grande longe do rio principal também serão afetados pela inundação. A distância média entre os registros de Morphnus foi 16 km ao longo do rio, o único ninho mapeado estava distante 1.3 km do ninho de Harpia. Estes fragmentos florestais estão sendo substituídos por pastagens, ressaltando-se a importância da manutenção das áreas de preservação permanentes (APP) e a proteção destes fragmentos em diversos formatos de áreas de proteção, para diminuir a degradação dos mesmos e garantir a manutenção das populações destes grandes predadores na região do rio Xingu.

Abundance of Harpy and Crested Eagles from a reservoir-impact area in the Low- and Mid-Xingu River / Abundância de Gavião-real e Gavião-real falso numa área sob impacto de reservatório no Baixo e Médio rio Xingu

In the Brazilian Amazon, two monospecific genera, the Harpy Eagle and Crested Eagle have low densities and are classified by IUCN as Near Threatened due to habitat loss, deforestation, habitat degradation and hunting. In this study, we evaluate occurrence of these large raptors using the environmental surveys database from Belo Monte Hydroelectric Power Plant. Integrating the dataset from two methods, we plotted a distribution map along the Xingu River, including records over a 276-km stretch of river. Terrestrial surveys (RAPELD method) were more efficient for detecting large raptors than standardized aquatic surveys, although the latter were complementary in areas without modules. About 53% of the records were obtained during activities of wildlife rescue/flushing, vegetation suppression or in transit. Between 2012 and 2014, four Harpy Eagles were removed from the wild; two shooting victims, one injured by collision with power lines and one hit by a vehicle. Also, seven nests were mapped. The mean distance between Harpy Eagle records was 15 km along the river channel, with a mean of 20 km between nests near the channel, which allowed us to estimate 20 possible pairs using the alluvial forest, riverine forest and forest fragments. Territories of another ten pairs will probably be affected by inundation of the Volta Grande channel, which is far from the main river. The average distance between Crested Eagle records was 16 km along the river channel. The only nest found was 1.3 km away from a Harpy Eagle nest. The remnant forests are under threat of being replaced by cattle pastures, so we recommend that permanently protected riparian vegetation borders (APP) be guaranteed, and that forest fragments within 5 km of the river be conserved to maintain eagle populations.

Na Amazônia brasileira dois gêneros mono-específicos, Harpia e Morphnus, caracterizam-se por baixa densidade e estão classificados pelo IUCN como Quase Ameaçados, porém ocorrem sobre grande parte do território nacional, suas principais ameaças são a fragmentação florestal, a degradação de hábitat e a caça. Neste estudo avaliamos a abundância destas duas grandes aves de rapina utilizando a base de dados dos programas ambientais da UHE Belo Monte, integrando-se dois métodos para construir um mapa de distribuição ao longo de 245 km do rio Xingu. Os levantamentos terrestres pelo método RAPELD mostraram-se mais eficientes para os registros de grandes águias quando comparado aos esforços padronizados aquáticos, entretanto estes foram complementares na ausência de módulos. Cinquenta e tres por cento foram registros ocasionais durante a supressão da vegetação, afugentamento ou deslocamento. Entre 2012-2014 quatro harpias foram removidas da natureza, dois indivíduos alvo de disparos, uma por colisão com rede elétrica, e outra por atropelamento. Sete ninhos mapeados, a distância média entre os registros de harpia na calha e margens do rio foi de 15 km, 20 km distância média entre ninhos, o que permitiu estimar um total de 20 casais utilizando as florestas aluviais em uma distancia de 270 km, incluindo matas ciliares e os fragmentos fora da margem do rio. Estimamos que territórios de outros 10 casais usando a Volta Grande longe do rio principal também serão afetados pela inundação. A distância média entre os registros de Morphnus foi 16 km ao longo do rio, o único ninho mapeado estava distante 1.3 km do ninho de Harpia. Estes fragmentos florestais estão sendo substituídos por pastagens, ressaltando-se a importância da manutenção das áreas de preservação permanentes (APP) e a proteção destes fragmentos em diversos formatos de áreas de proteção, para diminuir a degradação dos mesmos e garantir a manutenção das populações destes grandes predadores na região do rio Xingu.