Establishment of two quantitative nested qPCR assays targeting the human EPO transgene.

For ethical and safety reasons it is critical to develop easily implemented assays with high sensitivity and specificity for gene doping surveillance. Two nested quantitative real-time PCR (qPCR) assays were developed that target the human EPO (hEPO) cDNA sequence in a circular form, representative of recombinant adeno-associated viral (rAAV) vector genomes found in vivo. Through an interlaboratory evaluation, the assays were validated and utilized in an in vitro blinded study. These assays are specific and extremely sensitive with a limit of detection (LOD) of 1 copy of circular plasmid DNA and a limit of quantification (LOQ) of 10 to 20 copies in the presence of 500 ng of human genomic DNA (hgDNA) extracted from WBCs. Additionally, using the two nested qPCR assays in a non-human primate study, where macaques were injected intramuscularly with a rAAV8 vector harboring a promoterless hEPO cDNA sequence, the viral vector was detected 8 to 14 weeks post-injection in macaque WBCs. The high sensitivity of the nested qPCR approach along with the capability of quantifying target DNA, make this approach a reliable tool for gene doping surveillance and the monitoring of exogenous DNA sequences.

Estudio comparativo del uso de la matriz ósea desmineralizada de origen humana y de la poliuretana derivada del aceite de mamona sobre el proceso de regeneración ósea. Estudio histométrico en calota de conejos / A comparative study on the use of demineralized bone matrix of human origin and polyurethane derived from castor oil in bone regeneration processes. A histometric study on rabbit calvaria

Objetivo. Analizar histométricamente el efecto de la matriz óseadesmineralizada de origen humana y de la resina poliuretana derivada delaceite de mamona, sobre el proceso de regeneración ósea. Diseño del estudio:fueron utilizados 24 conejos y en cada calota craniana fueron preparadasdos cavidades quirúrgicas, siendo una a cada lado de la sutura parietal.Los animales fueron divididos en dos grupos. En el grupo I la cavidadexperimental (lado derecho) fue rellenada con la poliuretana derivadadel aceite de mamona, mientras que en el grupo II la cavidad experimentalfue rellenada con la matriz ósea desmineralizada de origen humano. Lascavidades de control fueron rellenadas solo con la sangre del animal. Losanimales fueron sacrificados en los períodos postoperatorios de 04, 07 y15 semanas. Resultados. El análisis estadístico, realizado en base de losdatos de la histometría, mostró una mayor neoformación ósea de los gruposI y II en relación al grupo control, no existiendo diferencia estadísticacuando los grupos experimentales fueron comparados. Conclusiones. Eluso de poliuretana derivada del aceite de mamona y matriz ósea desmineralizadade origen humano mejoran el proceso de regeneración cuandolo comparamos a la regeneración ósea normal resultando que entre ambosmateriales tuvieron un comportamiento similar sin diferencias estadísticas significantes entre sí (AU)

Objective. The present study was aimed at carrying out a histological and histometric analysis on the effect of humande mineralized bone matrix and a polyurethane resin, derived of the castor bean oil, on the bone regeneration process. Design: For this,24 rabbits were used and in each calvarium two surgical bone defect were prepared, one on the right side and another on the left side of the parietal suture. The animals were divided into two groups. In group I the experimental defect (right side) was filled with polyurethane derived from oil of the castor oil plant, while in group II the experimental cavity was filled with demineralized bone matrix of human origin. The control cavities were just filled with the bloodof the animal. The animals were sacrificed in the postoperative period at 04, 07 and 15 weeks. Results. The statistical analysis, which was based on the data of the histometric study, showed larger bone neoformation in groups I and II in relation to the control groups, and there were no statistical differences when the experimental groups were compared. Conclusions. The use of polyurethane resinimplants and demineralized human bone matrix helped and optimized the regeneration process when compared with naturalbone regeneration, and similar behavior was obtained without statistical differences in both materials (AU)

A retrospective study of the potential complications during alveolar distraction osteogenesis in 55 patients.

The aim of this retrospective study was to analyse the outcome of alveolar distraction osteogenesis for the correction of vertical defects in a large series of 55 cases. The existing bone deficiencies were secondary to atrophy after periodontal disease or tooth extraction. The overall success rate of this technique was 89.1%. The complications presented during treatment were divided into minor (no effect on final result, but immediate intervention required) 14/55 patients (25.4%), and major (lead to technique failure) 6/55 patients (10.9%). The frequency of minor complications was 8/27 in the anterior maxillary region, 1/27 in the anterior mandibular region and 15/27 in the posterior mandibular region. The frequency of major complications was 5/6 in the posterior mandibular region and 1/6 in the anterior maxillary region. The mean alveolar height achieved was 6mm. The overall rate was 36.3%. On the basis of these results it was concluded that alveolar distraction osteogenesis is an effective technique to treat vertical alveolar ridge deficiencies.