Avaliação do desempenho oral na alimentação de bebês após correção cirúrgica de patologias do trato gastrointestinal / Evaluation of oral performance in infant feeding after surgical correction of gastrointestinal tract pathologies

O processo de alimentação do recém-nascido pode ter interferência de vários fatores, sendo um deles a prematuridade, principalmente quando este bebê, além de pré-termo, apresenta alguma patologia do trato gastrointestinal. Em função disso, o objetivo desse artigo é relatar, através de uma série de casos, os achados na avaliação da alimentação por via oral de bebês submetidos a intervenções cirúrgicas em função de malformações congênitas ou outras patologias do trato gastrointestinal. Diante dos casos relatados foi possível identificar, por intermédio da avaliação fonoaudiológica, alterações significativas no desempenho alimentar, como fase oral desorganizada, escape extraoral do alimento e fadiga. Tais achados comprometem o progresso da alimentação por via oral e dificultam o desmame da dieta por via alternativa.(AU)

Aneuploidy and recombination in the human preimplantation embryo. Copy number variation analysis and genome-wide polymorphism genotyping.

RESEARCH QUESTION: What are the incidence and patterns of meiotic trisomies and recombination separately and in relation to each other at the blastocyst stage via single nucleotide polymorphism genotyping combined with array comparative genomic hybridization. DESIGN: Single nucleotide polymorphism microarrays were carried out on a total of 1442 blastocyst stage embryos derived from 268 fertile couples undergoing preimplantation genetic diagnosis for the purposes of avoiding transmittance of known single gene disorders to their offspring; 24-chromosome aneuploidy screening via array comparative genomic hybridization was carried out in parallel. RESULTS: One hundred per cent of meiotic trisomies identified in these embryos were of maternal origin and their incidence increased significantly with advancing maternal age (P < 0.0001). A total of 55.8% of meiotic trisomies were meiosis I-type and 44.2% were meiosis II-type. Certain chromosomes were affected more by meiosis I-type errors, whereas others experienced more meiosis II-type errors. A detailed recombination analysis was carried out for 11,476 chromosomes and 17,763 recombination events were recorded. The average number of recombination sites was 24.0 ± 0.3 for male meiosis and 41.2 ± 0.6 for female meiosis (autosomes only). Sex-specific differences were observed in the locations of recombination sites. Comparative analysis conducted between 190 euploid embryos and 69 embryos presenting maternal meiotic trisomies showed similar recombination rates (P = 0.425) and non-recombinant chromatid rates (P = 0.435) between the two categories; differences, however, were observed when analysing embryos affected with specific maternal meiotic trisomies. CONCLUSIONS: This study yielded unique data concerning recombination and the origin of aneuploidies observed during the first few days of life and provides a novel insight into these important biological processes.

Live births following Karyomapping of human blastocysts: experience from clinical application of the method.

The clinical application of a new, widely applicable method known as Karyomapping to carry out a total of 55 clinical cases of preimplantation genetic diagnosis (PGD) for single gene disorders is reported. Conventional polymerase chain reaction (PCR) testing was carried out in parallel to the new method for all cases. Clinical application of Karyomapping in this study resulted in three live births and nine clinical pregnancies out of 20 cases with a transfer. All in all, results presented in this study indicate that Karyomapping is a highly efficient, accurate and robust method for PGD of single gene disorders. Karyomapping can offer a more comprehensive assessment of the region of interest than conventional PCR analysis, allowing for more embryos to receive diagnosis (99.6% versus 96.8%), whereas its wide applicability reduces substantially the time that patients have to wait before starting their in vitro fertilization (IVF) cycle. Nonetheless, inclusion of elements of conventional PCR methodology, such as direct mutation detection, may be required in cases in which the gene of interest is in a region with reduced single nucleotide polymorphism (SNP) coverage (e.g. telomeric regions), when offering PGD for consanguineous couples, or in cases where no samples from additional family members are available.

Validation of a multiplex genotyping platform using a novel genomic database approach.

PURPOSE: Technological advances now allow for multiplex platforms to simultaneously test many genetic conditions. Typically, such platforms are validated by assaying samples with known genotypes and/or phenotypes and/or with synthetic plasmids; however, these methods have limitations and with the inclusion of rarer diseases and mutations, we can no longer rely solely on them. We used a novel genomic database to validate an expanded genetic carrier screening platform. METHODS: Our expanded carrier screening assay uses the Illumina Infinium iSelect HD Custom genotyping platform to test for 213 genetic diseases by assaying 1,663 pathogenic mutations. We leveraged two Coriell Institute biorepositories for validation: the Subcollection of Heritable Diseases and the 1000 Genomes Project. RESULTS: We measured 12,394 mutation observations in 206 samples, resulting in 246 true positives, 12,147 true negatives, 1 false positive, and no false negatives. Results demonstrated high sensitivity (99.99%) and specificity (99.99%). CONCLUSION: We successfully validated our platform with two biorepositories, demonstrating high sensitivity and specificity. The 1000 Genomes Project samples provided both positive and negative validation for mutations in genes not available through other biorepositories, expanding the depth of validated variants. We recommend including samples from the 1000 Genomes Project in the validation of future multiplex testing platforms.Genet Med advance online publication 30 July 2015Genetics in Medicine (2015); doi:10.1038/gim.2015.101.

Detection and phasing of single base de novo mutations in biopsies from human in vitro fertilized embryos by advanced whole-genome sequencing.

Currently, the methods available for preimplantation genetic diagnosis (PGD) of in vitro fertilized (IVF) embryos do not detect de novo single-nucleotide and short indel mutations, which have been shown to cause a large fraction of genetic diseases. Detection of all these types of mutations requires whole-genome sequencing (WGS). In this study, advanced massively parallel WGS was performed on three 5- to 10-cell biopsies from two blastocyst-stage embryos. Both parents and paternal grandparents were also analyzed to allow for accurate measurements of false-positive and false-negative error rates. Overall, >95% of each genome was called. In the embryos, experimentally derived haplotypes and barcoded read data were used to detect and phase up to 82% of de novo single base mutations with a false-positive rate of about one error per Gb, resulting in fewer than 10 such errors per embryo. This represents a ∼ 100-fold lower error rate than previously published from 10 cells, and it is the first demonstration that advanced WGS can be used to accurately identify these de novo mutations in spite of the thousands of false-positive errors introduced by the extensive DNA amplification required for deep sequencing. Using haplotype information, we also demonstrate how small de novo deletions could be detected. These results suggest that phased WGS using barcoded DNA could be used in the future as part of the PGD process to maximize comprehensiveness in detecting disease-causing mutations and to reduce the incidence of genetic diseases.

Genome-wide karyomapping accurately identifies the inheritance of single-gene defects in human preimplantation embryos in vitro.

PURPOSE: Our aim was to compare the accuracy of family- or disease-specific targeted haplotyping and direct mutation-detection strategies with the accuracy of genome-wide mapping of the parental origin of each chromosome, or karyomapping, by single-nucleotide polymorphism genotyping of the parents, a close relative of known disease status, and the embryo cell(s) used for preimplantation genetic diagnosis of single-gene defects in a single cell or small numbers of cells biopsied from human embryos following in vitro fertilization. METHODS: Genomic DNA and whole-genome amplification products from embryo samples, which were previously diagnosed by targeted haplotyping, were genotyped for single-nucleotide polymorphisms genome-wide detection and retrospectively analyzed blind by karyomapping. RESULTS: Single-nucleotide polymorphism genotyping and karyomapping were successful in 213/218 (97.7%) samples from 44 preimplantation genetic diagnosis cycles for 25 single-gene defects with various modes of inheritance distributed widely across the genome. Karyomapping was concordant with targeted haplotyping in 208 (97.7%) samples, and the five nonconcordant samples were all in consanguineous regions with limited or inconsistent haplotyping results. CONCLUSION: Genome-wide karyomapping is highly accurate and facilitates analysis of the inheritance of almost any single-gene defect, or any combination of loci, at the single-cell level, greatly expanding the range of conditions for which preimplantation genetic diagnosis can be offered clinically without the need for customized test development.

Parto normal ou cesariana? Fatores que influenciam na escolha da gestante / ¿Parto natural o cesárea? Factores que influyen en la elección de la mujer embarazada / Normal childbirth or cesarean? Factors affecting the choice of pregnancy

Objetivo: conhecer fatores que influenciam na decisão da via de parto de gestantes atendidas em uma Unidade de Saúde de Juazeiro-Bahia. Método: pesquisa qualitativa de caráter exploratório-descritivo, desenvolvido com gestantes acompanhadas em Unidade de Saúde entre março e junho de 2010. Participaram da entrevista semiestruturada 12 mulheres grávidas com idade igual ou superior a 18 anos, e para categorização do material utilizou-se análise de conteúdo. Resultados: a maior parte delas optaria pelo parto normal devido à rápida recuperação. A experiência anterior influiu na escolha da via de parto, seja esta da própria gestante ou outrem. Dentre as influências relatadas, destacaram-se: a mãe, profissionais de saúde e a mídia. Avaliaram que as informações perpassadas no pré-natal não eram satisfatórias. Considerações Finais: ressalta-se a relevância do acesso ao pré-natal de qualidade, com ações que proporcionem escolhas seguras, esclarecendo dúvidas e anseios da futura mãe, tranquilizando-a para o momento do parto.

Aim: to know the factors that influence the decision of the parturition mode of pregnant women attending Juazeiro-Bahia, Health Unit. Method: qualitative research if character exploratory-descriptive, developed with pregnant women followed in the Family Health Unit of Juazeiro-BA, conducted between March and June, 2010. Twelve pregnant women, with aged over 18 years, participated in the semi-structured interviews. For the categorization of the material, it was used content analysis. Results: most of them opted for normal parturition because has a rapid recovery. Previous experience influenced in parturition choice, whether herself or others. Among the influences reported, stood his mother, health professionals and the media. Noted that the information given in prenatal were unsatisfactory. Final Thoughts: it emphasizes the importance of access to a care quality prenatal, with actions that provide safe choices, clarifying doubts and reassures of the future mother, reassuring her to the time of parturition.

Objetivo: conocer los factores que influyen en la decisión del tipo de parto de las mujeres embarazadas en Juazeiro-BA. Método: investigación cualitativo con carácter exploratorio-descriptivo, desarrollado con mujeres embarazadas acompañadas en la Unidad de Salud de la Familia, entre marzo y junio de 2010. Participaron de las entrevistas semiestructuradas 12 mujeres embarazadas con edad igual o superior a 18 años, y para la clasificación de los materiales, utilizó el análisis de contenido. Resultados: la mayoría optaron por parto natural debido a la rápida recuperación. La experiencia anterior influye en la elección del parto. Entre las influencias reportadas, estaban la madre, profesionales de la salud y medios de comunicación. Tomó nota de que la información contenida en prenatal no era satisfactoria. Consideraciones finales: se destaca la importancia del acceso a un prenatal de calidad, con acciones que ofrecen opciones seguras, aclarando dudas de la madre y tranquilizándola para el parto.

Validation of microarray comparative genomic hybridization for comprehensive chromosome analysis of embryos.

OBJECTIVE: To validate and determine the best array-comparative genomic hybridization (aCGH; array-CGH) protocols for preimplantation genetic screening (PGS). DESIGN: Embryos had one cell removed as a biopsy specimen and analyzed by one of two array-CGH protocols. Abnormal embryos were reanalyzed by fluorescence in situ hybridization (FISH). SETTING: Reference laboratory. PATIENT(S): Patients donating embryos or undergoing PGS. INTERVENTION(S): Embryo biopsy, array-CGH, FISH reanalysis. MAIN OUTCOME MEASURE(S): Diagnosis, no result rate and error rate. RESULT(S): Method one produced 11.2% of embryos with no results and a 9.1% error rate compared with 3% and 1.9% for method two, respectively. Thereafter, only method two was used clinically. The aneuploidy rate for cleavage-stage embryos was 63.2%, significantly increasing with maternal age. The chromosomes most involved in aneuploidy were 16, 22, 21, and 15. We report the first live births after array-CGH combined with single blastomere biopsy. CONCLUSION(S): Array-CGH is proved to be highly robust (2.9% no results) and specific (1.9% error rate) when applied to rapid (24-hour) analysis of single cells biopsied from cleavage-stage embryos. This comprehensive chromosome analysis technique is the first to be validated by reanalyzing the same embryos with another technique (e.g., FISH). Unlike some alternative techniques for comprehensive chromosome screening, array-CGH does not require prior testing of parental DNA and thus advance planning and careful scheduling are unnecessary.

Comprehensive chromosome screening of polar bodies and blastocysts from couples experiencing repeated implantation failure.

OBJECTIVE: To identify and transfer cytogenetically normal embryos after screening all chromosomes of first and second polar bodies (PBs) or trophectoderm samples with the use of comparative genomic hybridization. DESIGN: Clinical research study. SETTING: In vitro fertilization clinic referring samples to a specialist preimplantation genetic diagnosis laboratory. PATIENT(S): Thirty-two couples with repeated implantation failure. INTERVENTION(S): Zygotes from patients with repeated implantation failure and poor response to ovarian stimulation underwent PB biopsy. Patients with repeated implantation failure who were candidates for blastocyst transfer received trophectoderm biopsy. Zygotes or blastocysts were vitrified while chromosome analysis took place. Euploid embryos were transferred during a subsequent cycle. MAIN OUTCOME MEASURE(S): Cytogenetic status and implantation and pregnancy rates. RESULT(S): The oocyte and blastocyst aneuploidy rates were 65.5% and 45.2%, respectively. Abnormalities affecting all chromosomes were detected. Implantation and pregnancy rates for the patients with PB biopsy were 11.5% and 21.4%, respectively, whereas for patients receiving blastocyst analysis they were 58.3% and 69.2%. CONCLUSION(S): Initial results for patients of advanced maternal age (39.8 years) with repeated implantation failure and poor ovarian response were encouraging. However, further study is required to confirm whether or not screening is beneficial. Blastocyst analysis was associated with high pregnancy rates, suggesting that comprehensive chromosome screening may assist patients with repeated implantation failure capable of producing blastocysts in achieving pregnancies.

Diabetes mellitus e câncer ginecológico / Diabetes mellitus and gynecological cancer

A ocidentalização dos hábitos de vida, como o sedentarismo, e uma dieta rica em açucares de rápida absorção vêm causando impactos notáveis em nossa sociedade, como o aumento da obesidade e dos casos de diabetes mellitus, uma vez que a relação entre estas duas afecções já está bem estabelecida. Conseqüentemente, as doenças relacionadas direta ou indiretamente ao diabetes e aos distúrbios metabólicos causados pela obesidade também têm se apresentado de maneira mais freqüente. Dentre os achados atuais de grandes estudos experimentais e epidemiológicos, merece destaque a associação observada entre o câncer ginecológico e o diabetes mellitus. Embora essa associação não se confirme em todos os tipos de câncer do sistema genital feminino, ela parece estar bem documentada e estabelecida para os cânceres de endométrio, de mama e do colo uterino