Methods for blood group antigens detection: cost-effectiveness analysis of phenotyping and genotyping

Abstract Background Alloimmunization is a major problem in transfusion practice due to the clinical complications of the patients and the difficulty of choosing a unit of compatible blood product. Serological methods are widely used in blood banks, but they not always determine the phenotype. Thus, genotyping is an important complement to the serology tool as it allows one to predict the phenotype from deoxyribonucleic acid (DNA) with high accuracy. Objective To compare the centrifugation gel, microarray, Restriction Fragment Length Polymorphismone PCR (PCR-RFLP) and Sequence-Specific Primer PCR (PCR-SSP) techniques, in terms of cost, reaction time and reliability of the results. Methods The RHCE, Kidd, Kell and Duffy blood group systems were chosen to determine the approximate cost of each technique, considering the reagents used in both methods and considering only one sample. The time required for the development of each reaction was obtained at the Maringa Regional Blood Center and Immunogenetics Laboratory at the State University of Maringa. Data from Microarray reactions were obtained at the Campinas Blood Center. The results of phenotyping and genotyping of the 16 samples were compiled in a spreadsheet and compared. Results The PCR-SSP was more economical compared to other methods, and the serological method was faster than the molecular methods. However, all methods proved to be effective and safe in the detection of erythrocyte antigens. Conclusion Analyzing the advantages and limitations of the molecular and serological methods tested in this study, we note that both are important and complementary. However, the choice of a methodology depends on the reality and needs of each health service.

Frequências fenotípicas dos grupos sanguíneos Kell, Duffy e Kidd em doadores de sangue do Hemonúcleo de Apucarana, sul do Brasil / Phenotypic frequencies of the Kell, Duffy and Kidd blood groups in blood donors from Apucarana Blood Center, southern Brazil

Objetivo: O objetivo desse estudo foi determinar as frequências fenotípicas dos grupos sanguíneos Kell, Duffy e Kidd em uma população paranaense. Métodos: Foram avaliadas as frequências desses grupos sanguíneos em 1.759 doadores de sangue fenotipados no Hemonúcleo de Apucarana, sul do Brasil. A fenotipagem foi realizada pela aglutinação em gel-teste e os dados foram obtidos pelo sistema Report Smith-Access, da rede Hemepar. Resultados: Essa população apresentou uma distribuição das frequências fenotípicas de Kell, Kidd e Duffy compatível com populações caucasianas. Para averiguar esse fato, nós comparamos nossos dados com aqueles de uma população da mesma região do Paraná, composta principalmente por caucasianos. O fenótipo Fy(a+b-) foi mais frequente na população de Apucarana do que na população de Maringá (22,68 vs. 12,50%, P<0,001), enquanto que o fenótipo Fy (a+b+) foi menos frequente (37,24 vs. 48,0%, P<0,001). Conclusão: As frequências fenotípicas de três grupos sanguíneos foram determinadas e poderão ser utilizadas pelos Serviços de Hematologia e Hemoterapia na busca de unidades de concentrados de hemácias com fenótipos desejados e no cálculo da incidência de doadores compatíveis, em casos de receptores com múltiplos aloanticorpos, além de poderem ser utilizadas para comparações antropológicas e em estudos de associação com doenças.

Infertilidade masculina decorrente de microdeleções no cromossomo Y / Male infertility caused by microdeletions on Y chromosome

Objetivo: Abordar a infertilidade masculina causada pela microdeleção no cromossomo Y e apresentar possíveis tratamentos por meio das técnicas de reprodução humana assistida. Métodos: Levantamento de dados da literatura científica na área da medicina reprodutiva. Resultados: Quando comparadas com outras causas de infertilidade, as microdeleções do cromossomo Y são relativamente frequentes. O cromossomo Y é essencial para a determinação sexual masculina e no seu braço longo há regiões responsáveis pela espermatogênese. São elas AZFa, AZFb e AZFc. Essas regiões podem ser deletadas e por conter múltiplos genes essenciais para a espermatogênese podem causar infertilidade masculina. Graças aos avanços da medicina, hoje vários casos de infertilidade são tratáveis por meio das técnicas de reprodução assistida. Dentre as técnicas, a MSOME se destaca por ser uma metodologia que seleciona apenas espermatozoides morfologicamente normais para serem usados na inseminação e aumentar as chances de gestação. Conclusões: A infertilidade masculina tem aumentado consideravelmente nos últimos anos e as causas genéticas são uma das grandes consequências disso. As microdeleções do cromossomo Y podem causar desde uma oligozoospermia leve a uma azoospermia, a depender da região AZF afetada. Para as causas mais leves, o casal pode recorrer a algumas técnicas de reprodução assistida e para as causas mais graves a solução para o casal pode ser usar gametas doados.(AU)

Objective: Address male infertility caused by microdeletions on Y chromosome and present possible treatment through assisted human reproduction techniques. Methods: Survey data from the scientific literature in the field of reproductive medicine. Results: When compared with other causes of infertility, the microdeletions on Y chromosome are the relatively frequent. The Y chromosome is essential for male sex determination and there are in his long arm regions responsible for spermatogenesi, it's they AZFa, AZFb and AZFc. Such regions may be deleted causing male infertility by contain multiple genes essential for spermatogenesis. Thanks to advances in medicine, now several cases of infertility are treatable through assisted reproduction techniques. Among the techniques, the MSOME stands out as a methodology that selects only morphologically normal sperm to be used in insemination increasing the chances of pregnancy. Conclusions: Male infertility has increased considerably in recent years and the genetic causes are one ofthe major consequences ofthis. Y chromosome microdeletions can cause mild oligozoospermia or azoospermia depending on the AZF region affected. For lighter causes, the couple may use some assisted reproductive techniques and to the most serious causes the solution to the couple is using donated gametes.(AU)

Importance of immune response genes in hemophilia A

Hemophilia A is a disease caused by a deficiency of coagulation factor VIII resulting from genetic inheritance linked to chromosome X. One treatment option is the administration of plasma or recombinant FVIII. However, some patients develop inhibitors or antibodies against this factor. Inhibitors are alloantibodies that bind to the epitope of factor VIII causing it to be recognized by the immune system as a foreign peptide. This is the most serious complication in hemophilia patients in respect to replacement therapy. Some studies have suggested that genetic factors influence the development of factor VIII inhibitors such as ethnicity, family history, mutations in the factor VIII gene and in genes of the immune system. The aim of this study was to conduct a literature review to assess the influence of genetic factors of immune response genes, especially genes of the major histocompatibility complex and cytokines, which may be related to the development of factor VIII inhibitors in hemophilia A patients. Understanding these risk factors will help to determine future differential treatment in the control and prevention of the development of inhibitors.

Study of the myenteric and submucous plexuses after BAC treatment in the intestine of rats

A morphological and quantitative study in the ileal and colonic myenteric and submucous plexuses of rats after BAC denervation was performed. Four groups were employed: SI--ileum control; CBI--denervated ileum; SC--colon control; and CBC--denervated colon. We used the Myosin-V immunohistochemistry technique to study the myenteric and submucous plexuses. In the submucous plexus of the ileum and colon there was not a significant decrease in the number of neurons/mm2 and of ganglia/mm2. The denervation of the myenteric plexus in the group CBI was 44.7

and in the group CBC, 68.3

. In the myenteric plexus there was also a significant decrease in the number of ganglia/mm2 (13.8

in group CBI and 52.14

in group CBC) and in the number of neurons/ganglion (33.9

in group CBI and 39.6

in group CBC). The morphological analyses showed that there was an alteration in the shape of the ganglia of the ileal and colonic myenteric plexus. The area of the cell bodies had a significant increase both in the myenteric and the submucous plexus in groups CBI and CBC. These data demonstrate that the BAC treatment causes morphologic and quantitative changes in the myenteric plexus and quantitative changes in the cell body area of the submucous plexus.