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To conclude, in this study we validated our previous results on whole genome methylation analysis in independent cohort of SMA patients. The results of this study confirm that methylation changes in the regulatory regions of SLC23A2 and NCOR2 are associated with SMA severity. Taking into account the data from ENCODE visualization, we suspect that DNA methylation changes might lead to expression changes of these genes. However, we could not define a difference in the expression level of NCOR2 between severe and mild types of SMA patients. Thus, further studies are needed to access if DNA methylation changes of these genes are meaningful for their WZ8040 transcriptional regulation in patients with different SMA types. The methylation changes in ARHGAP22, CDK2AP1, CHML and RPL9 were not detected among SMA patients of different types. However, it is not excluded that the difference between SMA patients and controls, indicated in our previous study, is meaningful for SMA pathogenesis. Our findings are limited to DNA methylation analysis in leukocytes. It should be taking in account that DNA methylation in leukocytes might not reflect the methylation level of the same genes in the motor neurons. Therefore our findings might require further confirmation by investigating methylation and expression levels of SLC23A2 and NCOR2 genes in disease-related tissue. However, we believe that analysis of methylation status in blood leukocytes is valuable in case of SMA as it is the most accessible biological material and could potentially be used in clinical practice for precise SMA severity detection and as a biomarker during SMA pharmacotherapy. Acute kidney injury is a common problem implicated in a substantial proportion of hospital admissions and the incidence is increasing. It is associated with a marked increase in mortality and also leads to prolonged hospital stay, increased secondary care costs and possibly accelerated decline in longterm kidney function. AKI has many and often multifactorial aetiologies. However, an important cause is the use of ACE inhibitor and Angiotensin-II Receptor Antagonists drugs which are associated with AKI in a range of settings, particularly during acute hypovolaemic illness. The increased risk of AKI among patients taking these medications has been recognised by the UK National Institute for Health and Clinical Excellence and the international organisation Kidney Disease: Improving Global Outcomes, both of which recommend that patients with chronic kidney disease should stop taking them if they become acutely unwell. There are many evidence based indications for use of ACE inhibitors and ARAs and national guidelines recommend treatment with them for a number of chronic conditions including hypertension, chronic kidney disease with proteinuria, and heart failure with left ventricular dysfunction. The result is that these medicines are the second most commonly prescribed in English primary care, accounting for 6% of all prescriptions. Due to increasing prevalence of chronic comorbidities in older people they are commonly used in the elderly: in Belgium, 7.3% of the population were treated with long-term ACE inhibitors or ARAs and this rose to 36% for people aged 80 years or more. However, despite their frequent use, it is not known to what extent increasing use of these medications has contributed to the increasing incidence of AKI on a population level. This is in part because observational studies on this topic are confounded by indication. The conditions for which ACE inhibitors and ARAs are indicated are themselves associated with increased risk of AKI. Therefore increasing incidence of AKI may reflect increasing prevalence of comorbidities, independently of medications used. We hypothesised that if these medications were playing a causal role, changes in prescribing would be associated with changes in hospital admission with AKI within general practices. We therefore conducted a longitudinal ecological analysis using routinely-collected national hospital administrative data to determine whether hospital admission rates with AKI in England are associated with increased prescribing of ACE inhibitor and ARA therapy. All data used in this study relates to the period 1st April 2007 to 31st March 2011. We used prescribing data from the English National Health Service Prescription Services’ Prescribing Database. This provides data for each English general practice for the total number of prescriptions that were prescribed and subsequently dispensed, although information about the quantity of medication provided is not captured.

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