A novel isolator-based system promotes viability of human embryos during laboratory processing

In vitro fertilisation (IVF) and related technologies are arguably the most challenging of all cell culture applications. The starting material is a single cell from which one aims to produce an embryo capable of establishing a pregnancy eventually leading to a live birth. Laboratory processing during IVF treatment requires open manipulations of gametes and embryos, which typically involves exposure to ambient conditions. To reduce the risk of cellular stress, we have developed a totally enclosed system of interlinked isolator-based workstations designed to maintain oocytes and embryos in a physiological environment throughout the IVF process. Comparison of clinical and laboratory data before and after the introduction of the new system revealed that significantly more embryos developed to the blastocyst stage in the enclosed isolator-based system compared with conventional open-fronted laminar flow hoods. Moreover, blastocysts produced in the isolator-based system contained significantly more cells and their development was accelerated. Consistent with this, the introduction of the enclosed system was accompanied by a significant increase in the clinical pregnancy rate and in the proportion of embryos implanting following transfer to the uterus. The data indicate that protection from ambient conditions promotes improved development of human embryos. Importantly, we found that it was entirely feasible to conduct all IVF-related procedures in the isolator-based workstations

Identification of candidate transmission-blocking antigen genes in Theileria annulata and related vector-borne apicomplexan parasites

Background: Vector-borne apicomplexan parasites are a major cause of mortality and morbidity to humans and livestock globally. The most important disease syndromes caused by these parasites are malaria, babesiosis and theileriosis. Strategies for control often target parasite stages in the mammalian host that cause disease, but this can result in reservoir infections that promote pathogen transmission and generate economic loss. Optimal control strategies should protect against clinical disease, block transmission and be applicable across related genera of parasites. We have used bioinformatics and transcriptomics to screen for transmission-blocking candidate antigens in the tick-borne apicomplexan parasite, Theileria annulata. Results: A number of candidate antigen genes were identified which encoded amino acid domains that are conserved across vector-borne Apicomplexa (Babesia, Plasmodium and Theileria), including the Pfs48/45 6-cys domain and a novel cysteine-rich domain. Expression profiling confirmed that selected candidate genes are expressed by life cycle stages within infected ticks. Additionally, putative B cell epitopes were identified in the T. annulata gene sequences encoding the 6-cys and cysteine rich domains, in a gene encoding a putative papain-family cysteine peptidase, with similarity to the Plasmodium SERA family, and the gene encoding the T. annulata major merozoite/piroplasm surface antigen, Tams1. Conclusions: Candidate genes were identified that encode proteins with similarity to known transmission blocking candidates in related parasites, while one is a novel candidate conserved across vector-borne apicomplexans and has a potential role in the sexual phase of the life cycle. The results indicate that a ‘One Health’ approach could be utilised to develop a transmission-blocking strategy effective against vector-borne apicomplexan parasites of animals and humans

Transfer of mouse blastocysts exposed to ambient oxygen levels can lead to impaired lung development and redox balance

DOGAN, GOKSEL/0000-0002-4583-3140; CINAR KUL, Bengi/0000-0002-8955-0097WOS: 000510017700006PubMed: 31504752In vitro culture under atmospheric oxygen puts embryos under oxidative stress and impairs preimplantation development. However, to what extent this process alters the redox balance in the perinatal period remains largely unknown. the aim of the present study was to examine if the redox balance is altered in the lung tissue of fetuses generated through transfer of mouse embryos exposed to atmospheric oxygen at different stages of development and to determine if this has any effect on lung morphogenesis and gene expression. Two experimental groups (EGs) were generated by transferring in vitro- and in vivo-derived blastocysts to pseudo-pregnant females. in vivo-developed fetuses served as control. Enzymatic/nonenzymatic antioxidants, malondialdehyde (MDA) levels, total antioxidant capacity, stage of lung development and gene expression were evaluated on day 18 of pregnancy. Weight of fetuses was significantly less in both experimental cohorts (ANOVA, P < 0.001 versus control), associated with delayed lung development, higher amounts of MDA (ANOVA, P < 0.001 versus control) and altered expression of several genes in oxidative stress/damage pathways. Evidence gathered in the present study indicates that pre-implantation stress caused by culture under atmospheric oxygen, even for a short period of time, leads to fetal growth restriction, impaired lung development and redox balance along with dysregulation of several genes in oxidative stress response. Absence of an EG in which in vitro embryo culture was performed at 5% oxygen and the use of genetically heterogeneous F2 fetuses are the limitations of the study. in any case, the long-term impact of such dramatic changes in the developmental programming of resulting fetuses warrants further investigations.Scientific and Technological Research Council of Turkey (TUB.ITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [112O259]This work was funded by the Scientific and Technological Research Council of Turkey (TUB.ITAK, Project no: 112O259)

Genetic exchange and sub-structuring in Theileria annulata populations

Tropical theileriosis, caused by the tick-transmitted protozoan Theileria annulata, is a major disease of cattle in many regions of the developing world. Development and deployment of current and novel control strategies should take into account the potential diversity of parasite populations and therefore it is important that the nature and basis for this genetic variation is investigated and quantified. The recently completed genome sequence of T. annulata provided an opportunity to develop a panel of genetic markers for population studies. Bioinformatic screening of the genome sequence identified micro- and mini-satellite loci, which were PCR amplified from a series of diverse parasite stocks in order to characterise their polymorphism and determine their species-specificity. A panel of 10 polymorphic markers was then selected for population genetic studies and used to genotype both cloned and uncloned isolates maintained as cell lines of T. annulata isolated from different countries. Uncloned isolates comprised a multiplicity of genotypes, while cloned isolates showed evidence of a single haploid genome. Population genetic analysis revealed a high level of genotypic diversity both between and within countries, as measured by the number of distinct multilocus genotypes. Based on measures of genetic differentiation (F&lt;sub&gt;ST&lt;/sub&gt; and Nei's genetic distance), the parasite populations are geographically sub-structured and these measurements positively correlate with geographical distance. To investigate whether genetic exchange occurs at random and the populations are panmictic, the level of linkage disequilibrium was measured using the index of association across all loci. Linkage disequilibrium was observed when isolates from all countries were treated as a single population but when the isolates from Tunisia were analysed separately, linkage equilibrium was observed