Differential expression of PPP1R12A transcripts, including those harbouring alternatively spliced micro-exons, in placentae from complicated pregnancies.

INTRODUCTION: Placenta-associated pregnancy complications, including pre-eclampsia (PE) and intrauterine growth restriction (IUGR) are conditions postulated to originate from initial failure of placentation, leading to clinical sequelae indicative of endothelial dysfunction. Vascular smooth muscle aberrations have also been implicated in the pathogenesis of both disorders via smooth muscle contractility and relaxation mediated by Myosin Light Chain Phosphatase (MLCP) and the oppositional contractile action of Myosin Light Chain Kinase. PPP1R12A is a constituent part of the MLCP complex responsible for dephosphorylation of myosin fibrils. We hypothesize that alternative splicing of micro-exons result in isoforms lacking the functional leucine zipper (LZ) domain which may give those cells expressing these alternative transcripts a tendency towards contraction and vasoconstriction. METHODS: Expression was determined by qRT-PCR. Epigenetic profiling consisted of bisulphite-based DNA methylation analysis and ChIP for underlying histone modifications. RESULTS: We identified several novel transcripts with alternative micro-exon inclusion that would produce LZ- PPP1R12A protein. qRT-PCR revealed some isoforms, including the PPP1R12A canonical transcript, are differentially expressed in placenta biopsies from PE and IUGR samples compared to uncomplicated pregnancies. DISCUSSION: We propose that upregulation of PPP1R12A expression in complicated pregnancies may be due to enhanced promoter activity leading to increased transcription as a response to physiological stress in the placenta, which we show is independent of promoter DNA methylation.

Assessment of pet exposure by self-report in epidemiological studies of allergy and asthma: a systematic review.

OBJECTIVE: In epidemiological studies that aim to investigate the relationship between pet exposure and allergy/asthma, pet exposure is often ascertained by means of a questionnaire, but it is unclear which questionnaire items are used. The objective of this study was to systematically review self-reported pet exposure assessment in questionnaires used in epidemiological studies which explore the associations between pets and allergy/asthma. METHODS: A systematic literature search was conducted in PubMed and papers were selected according to pre-specified eligibility criteria. The pet exposure questions used were classified within a framework including direct pet contact, indirect pet contact (e.g. through carers or grandparents) and avoidance behaviour. Authors were contacted when the questions used were not reported in detail. RESULTS: Ninety-six full text papers were systematically reviewed. All studies assessed direct pet contact, but less than half (45%) explicitly assessed whether pets were allowed indoors. The vast majority of studies assessed both pet exposures during the first year of life and after the first year of life. The minority (13%) assessed whether pet(s) were kept at places regularly visited by the child and pet exposure in utero (15%). Even fewer studies assessed indirect contact to pets (n = 8) and avoidance behaviour (n = 10). CONCLUSIONS: In epidemiological studies, the ascertainment of pet exposure through questionnaires appears to vary greatly. This variation might partly explain the inconsistent and contradictory results of the effects of pet exposure on the development of allergy and asthma.