Sex determination of porcine embryos using a new developed duplex polymerase chain reaction procedure based on the amplification of repetitive sequences

Polymerase chain reaction (PCR)-based assays have become increasingly prevalent for sexing embryos. The aim of the present study was to develop a suitable duplex PCR procedure based on the amplification of porcine repetitive sequences for sexing porcine tissues, embryos and single cells. Primers were designed targeting the X12696 Y chromosome-specific repeat sequence (SUSYa and SUSYb; sex-related primer sets), the multicopy porcine-specific mitochondrial 12S rRNA gene (SUS12S; control primer set) and the X51555 1 chromosome repeat sequence (SUS1; control primer set). The specificity of the primer sets was established and the technique was optimised by testing combinations of two specific primer sets (SUSYa/SUS12S; SUSYb/SUS12S), different primer concentrations, two sources of DNA polymerase, different melting temperatures and different numbers of amplification cycles using genomic DNA from porcine ovarian and testicular tissue. The optimised SUSYa/SUS12S-and SUSYb/SUS12S-based duplex PCR procedures were applied to porcine in vitro-produced (IVP) blastocysts, cell-stage embryos and oocytes. The SUSYb/SUS12S primer-based procedure successfully sexed porcine single cells and IVP cell-stage embryos (100% efficiency), as well as blastocysts (96.6% accuracy; 96.7% efficiency). This is the first report to demonstrate the applicability of these repetitive sequences for this purpose. In conclusion, the SUSYb/SUS12S primer-based duplex PCR procedure is highly reliable and sensitive for sexing porcine IVP embryos. © 2013 CSIRO

Erratum to "Evaluation of boar sperm maturation after co-incubation with caput, corpus and cauda epididymal cultures"

International audienc

Evaluation of boar sperm maturation after co-incubation with caput, corpus and cauda epididymal cultures - Evaluation of boar sperm maturation in vitro

International audienc

Cluster of Type IV Secretion Genes in Helicobacter pylori's Plasticity Zone

Some genes present in only certain strains of the genetically diverse gastric pathogen Helicobacter pylori may affect its phenotype and/or evolutionary potential. Here we describe a new 16.3-kb segment, 7 of whose 16 open reading frames are homologs of type IV secretion genes (virB4, virB7 to virB11, and virD4), the third such putative secretion gene cluster found in H. pylori. This segment, to be called tfs3, was discovered by subtractive hybridization and chromosome walking. Full-length and truncated tfs3 elements were found in 20 and 19%, respectively, of 94 strains tested, which were from Spain, Peru, India, and Japan. A tfs3 remnant (6 kb) was found in an archived stock of reference strain J99, although it was not included in this strain's published genome sequence. PCR and DNA sequence analyses indicated the following. (i) tfs3's ends are conserved. (ii) Right-end insertion occurred at one specific site in a chromosomal region that is varied in gene content and arrangement, the “plasticity zone.” (iii) Left-end insertion occurred at different sites in each of nine strains studied. (iv) Sequences next to the right-end target in tfs3-free strains were absent from most strains carrying full-length tfs3 elements. These patterns suggested insertion by a transposition-like event, but one in which targets are chosen with little or no specificity at the left end and high specificity at the right end, thereby deleting the intervening DNA

The urban burden of disease estimation for policy-making in 1000 European cities

In Europe, over 70% of the population lives in urban areas. Cities promote innovation and wealth creation; however, they are also a main source of pollution, disease and mortality, to some extent linked with suboptimal urban and transport planning practices. The aim of the project is to improve the estimation of health impacts and socioeconomic costs of environmental stressors related to urban and transport planning, advance modelling approaches and strengthen evidence-based policy-making. The analyses focus on air pollution, noise, temperature/heat and lack of green spaces for nearly 1000 European cities. We are (1) developing an overall urban burden of disease impact assessment framework; (2) identifying health and wellbeing indicators; (3) obtaining exposure-response relationships for environmental stressors and health outcomes of interest; (4) developing tools and guidelines for cost-benefit analyses; (5) obtaining health and environmental stressors data for cities for 2015, 2018, 2021 and 2024 and (6) calculating health and socioeconomic impacts and monitor trends. Specific urban and transport planning scenarios will be developed for case-studies cities (Barcelona, Brussels, Manchester, Munich, Utrecht, Sofia, Warsaw and Zagreb) and include additional aspects such as mobility, physical activity and inequities. The science-to-policy translation is a strong component of this research project where we are conducting consultation with and providing knowledge translation to cities and stakeholders, and sharing good practices, for greater science-policy impact. We strive to make the change on city-level policy and action plans, as well as on national and EU level policy making. By improving health impact and socioeconomic costs estimations and engaging with stakeholders we expect to generate impact and promote healthier urban and transport planning practices in European cities

The urban burden of disease estimation for policy-making in 1000 European cities

In Europe, over 70% of the population lives in urban areas. Cities promote innovation and wealth creation; however, they are also a main source of pollution, disease and mortality, to some extent linked with suboptimal urban and transport planning practices. The aim of the project is to improve the estimation of health impacts and socioeconomic costs of environmental stressors related to urban and transport planning, advance modelling approaches and strengthen evidence-based policy-making. The analyses focus on air pollution, noise, temperature/heat and lack of green spaces for nearly 1000 European cities. We are (1) developing an overall urban burden of disease impact assessment framework; (2) identifying health and wellbeing indicators; (3) obtaining exposure-response relationships for environmental stressors and health outcomes of interest; (4) developing tools and guidelines for cost-benefit analyses; (5) obtaining health and environmental stressors data for cities for 2015, 2018, 2021 and 2024 and (6) calculating health and socioeconomic impacts and monitor trends. Specific urban and transport planning scenarios will be developed for case-studies cities (Barcelona, Brussels, Manchester, Munich, Utrecht, Sofia, Warsaw and Zagreb) and include additional aspects such as mobility, physical activity and inequities. The science-to-policy translation is a strong component of this research project where we are conducting consultation with and providing knowledge translation to cities and stakeholders, and sharing good practices, for greater science-policy impact. We strive to make the change on city-level policy and action plans, as well as on national and EU level policy making. By improving health impact and socioeconomic costs estimations and engaging with stakeholders we expect to generate impact and promote healthier urban and transport planning practices in European cities