RDDSVM: accurate prediction of A-to-I RNA editing sites from sequence using support vector machines.

Adenosine to inosine (A-to-I) editing in RNA is involved in various biological processes like gene expression, alternative splicing, and mRNA degradation associated with carcinogenesis and various human diseases. Therefore, accurate identification of RNA editing sites in transcriptome is valuable for research and medicine. RNA-seq is very useful for the detection of RNA editing events in condition-specific cells. However, computational analysis methods of RNA-seq data have considerable false-positive risks due to mapping errors. In this study, we developed a simple machine learning method using support vector machines to train sequence and structure information derived from flanking sequences of experimentally verified A-to-I editing sites to predict new A-to-I editing sites in RNA. The highest performance results were obtained by the model that utilizes the composition of the triplet sequence elements in the flanking regions of the in A-to-I editing sites. Using this model, the SVM classifier also showed high performance on experimentally verified data providing a sensitivity of 92.8%, specificity of 77.1%, and accuracy of 90.2%. To compare the predictive capacity of our method with other classifiers that use sequence information, we have used validated human A-to-I RNA editing sites by Sanger sequencing. Out of 58 validated editing sites, our method recognized 53 of them correctly with an accuracy of 91.4% outperforming other classifiers. As to our knowledge, this is the first case of utilization of the composition of the triplet sequence elements neighboring A-to-I editing sites for the prediction of new A-to-I editing sites in RNA. The methodology is very easy to perform and computationally low demanding making it a convenient and valuable choice for facilities with low sources. To facilitate the usage of the method publicly, we developed an open-source program called RDDSVM to perform prediction on candidate A-to-I RNA editing sites using support vector machines.

Successful haematopoietic stem cell transplantation in 44 children from healthy siblings conceived after preimplantation HLA matching.

Haematopoietic stem cell transplantation (HSCT) remains the best therapeutic option for many acquired and inherited paediatric haematological disorders. Unfortunately, the probability of finding an HLA matched donor is limited. An alternative technique is PGD combined with HLA matching, which offers the possibility of selecting unaffected embryos that are HLA compatible with the sick child, with the aim of possible use of stem cells from the resulting baby in future. Since the first successful report for Fanconi anaemia a decade ago, the therapeutic success of this technique was reported in a few cases and for a limited number of disorders. Here, we report full recovery of 44 sick children who received HSCT from healthy infants conceived after pre-implantation HLA matching for the following 10 indications; beta-thalassaemia, Wiskott-Aldrich syndrome, Fanconi anaemia, sickle cell anaemia, acute myeloid leukaemia, acute lymphoblastic leukaemia, Glanzmann's thrombasthaenia, Diamond-Blackfan anaemia, X-linked adrenoleukodystrophy and mucopolysaccharidosis type I. No serious complications were observed among recipients and donors. Graft failure occurred in four children with beta-thalassaemia where a second HSCT was planned. Preimplantation HLA matching is a reliable technique and provides a realistic option for couples seeking treatment for an affected child when no HLA-matched donor is available.

Birth of a healthy infant after preimplantation genetic diagnosis by sequential blastomere and trophectoderm biopsy for ß-thalassemia and HLA genotyping.

BACKGROUND: Preimplantation genetic diagnosis (PGD) is a widely used technique for couples at genetic risk and involves the diagnosis and transfer of unaffected embryos generated through in vitro fertilization (IVF) techniques. STUDY DESIGN: For those couples who are at risk of transmitting a genetic disease to their offspring, preimplantation embryos can be selected according to their genetic status as well as human leukocyte antigen (HLA) compatibility with the affected child. Stem cells from the resulting baby's umbilical cord blood can be used for transplantation to the affected sibling without graft rejection. RESULTS: Here we report successful hematopoietic stem cell transplantation (HSCT) after the birth of a healthy infant, who was born after successful PGD testing with both cleavage stage and blastocyst stage biopsy for the purpose of diagnosis of ß-thalassemia and HLA compatibility. CONCLUSION: The specific feature of this work is not only to have the first successful HSCT achieved in Bulgaria after using preimplantation HLA typing technique, it also demonstrates how to accomplish this success via cross-border collaboration of different units, which makes the application of these sophisticated methods possible in hospitals not having the necessary equipments and expertise.