First experimental study of carbon dioxide digital subtraction lymphangiography

Fourteen pigs with an average weight of 17 kg were used in this study. Under general anesthesia and magnification 1-3 ccm/kg of carbon dioxide were administered in the lymph vessels of the front and rear legs. Imaging of the peripheral lymph vessels, lymph nodes and the thoracic duct was achieved with digital subtraction angiography. The quality of lymphangiography was satisfactory and comparable with that of the standard non-ionic contrast agent. It is anticipated that further technical evolution will permit the application of CO2/DSA lymphangiography to man. Carbon dioxide is non-nephrotoxic and is non-allergic; it is inexpensive, can be administered in unlimited quantity and is quickly eliminated via the pulmonary system

Next generation sequencing of SNPs for non-invasive prenatal diagnosis: challenges and feasibility as illustrated by an application to beta-thalassaemia

beta-Thalassaemia is one of the most common autosomal recessive single-gene disorder worldwide, with a carrier frequency of 12% in Cyprus. Prenatal tests for at risk pregnancies use invasive methods and development of a non-invasive prenatal diagnostic (NIPD) method is of paramount importance to prevent unnecessary risks inherent to invasive methods. Here, we describe such a method by assessing a modified version of next generation sequencing (NGS) using the Illumina platform, called 'targeted sequencing', based on the detection of paternally inherited fetal alleles in maternal plasma. We selected four single-nucleotide polymorphisms (SNPs) located in the beta-globin locus with a high degree of heterozygosity in the Cypriot population. Spiked genomic samples were used to determine the specificity of the platform. We could detect the minor alleles in the expected ratio, showing the specificity of the platform. We then developed a multiplexed format for the selected SNPs and analysed ten maternal plasma samples from pregnancies at risk. The presence or absence of the paternal mutant allele was correctly determined in 27 out of 34 samples analysed. With haplotype analysis, NIPD was possible on eight out of ten families. This is the first study carried out for the NIPD of beta-thalassaemia using targeted NGS and haplotype analysis. Preliminary results show that NGS is effective in detecting paternally inherited alleles in the maternal plasma