The spontaneous mouse mutant low set ears (Lse) is caused by tandem duplication of Fgf3 and Fgf4.

The external ear develops from an organized convergence of ventrally migrating neural crest cells into the first and second branchial arches. Defects in external ear position are often symptomatic of complex syndromes such as Apert, Treacher-Collins, and Crouzon Syndrome. The low set ears (Lse) spontaneous mouse mutant is characterized by the dominant inheritance of a ventrally shifted external ear position and an abnormal external auditory meatus (EAM). We identified the causative mutation as a 148 Kb tandem duplication on Chromosome 7, which includes the entire coding sequences of Fgf3 and Fgf4. Duplications of FGF3 and FGF4 occur in 11q duplication syndrome in humans and are associated with craniofacial anomalies, among other features. Intercrosses of Lse-affected mice revealed perinatal lethality in homozygotes, and Lse/Lse embryos display additional phenotypes including polydactyly, abnormal eye morphology, and cleft secondary palate. The duplication results in increased Fgf3 and Fgf4 expression in the branchial arches and additional discrete domains in the developing embryo. This ectopic overexpression resulted in functional FGF signaling, demonstrated by increased Spry2 and Etv5 expression in overlapping domains of the developing arches. Finally, a genetic interaction between Fgf3/4 overexpression and Twist1, a regulator of skull suture development, resulted in perinatal lethality, cleft palate, and polydactyly in compound heterozygotes. These data indicate a role for Fgf3 and Fgf4 in external ear and palate development and provide a novel mouse model for further interrogation of the biological consequences of human FGF3/4 duplication.

Identifying genetic determinants of inflammatory pain in mice using a large-scale gene-targeted screen.

ABSTRACT: Identifying the genetic determinants of pain is a scientific imperative given the magnitude of the global health burden that pain causes. Here, we report a genetic screen for nociception, performed under the auspices of the International Mouse Phenotyping Consortium. A biased set of 110 single-gene knockout mouse strains was screened for 1 or more nociception and hypersensitivity assays, including chemical nociception (formalin) and mechanical and thermal nociception (von Frey filaments and Hargreaves tests, respectively), with or without an inflammatory agent (complete Freund's adjuvant). We identified 13 single-gene knockout strains with altered nocifensive behavior in 1 or more assays. All these novel mouse models are openly available to the scientific community to study gene function. Two of the 13 genes (Gria1 and Htr3a) have been previously reported with nociception-related phenotypes in genetically engineered mouse strains and represent useful benchmarking standards. One of the 13 genes (Cnrip1) is known from human studies to play a role in pain modulation and the knockout mouse reported herein can be used to explore this function further. The remaining 10 genes (Abhd13, Alg6, BC048562, Cgnl1, Cp, Mmp16, Oxa1l, Tecpr2, Trim14, and Trim2) reveal novel pathways involved in nociception and may provide new knowledge to better understand genetic mechanisms of inflammatory pain and to serve as models for therapeutic target validation and drug development.

Large-scale discovery of mouse transgenic integration sites reveals frequent structural variation and insertional mutagenesis.

Transgenesis has been a mainstay of mouse genetics for over 30 yr, providing numerous models of human disease and critical genetic tools in widespread use today. Generated through the random integration of DNA fragments into the host genome, transgenesis can lead to insertional mutagenesis if a coding gene or an essential element is disrupted, and there is evidence that larger scale structural variation can accompany the integration. The insertion sites of only a tiny fraction of the thousands of transgenic lines in existence have been discovered and reported, due in part to limitations in the discovery tools. Targeted locus amplification (TLA) provides a robust and efficient means to identify both the insertion site and content of transgenes through deep sequencing of genomic loci linked to specific known transgene cassettes. Here, we report the first large-scale analysis of transgene insertion sites from 40 highly used transgenic mouse lines. We show that the transgenes disrupt the coding sequence of endogenous genes in half of the lines, frequently involving large deletions and/or structural variations at the insertion site. Furthermore, we identify a number of unexpected sequences in some of the transgenes, including undocumented cassettes and contaminating DNA fragments. We demonstrate that these transgene insertions can have phenotypic consequences, which could confound certain experiments, emphasizing the need for careful attention to control strategies. Together, these data show that transgenic alleles display a high rate of potentially confounding genetic events and highlight the need for careful characterization of each line to assure interpretable and reproducible experiments.

Custom and practice: a multi-center study of medicines reconciliation following admission in four acute hospitals in the UK.

BACKGROUND: Many studies have highlighted the problems associated with different aspects of medicines reconciliation (MR). These have been followed by numerous recommendations of good practice shown in published studies to decrease error; however, there is little to suggest that practice has significantly changed. The study reported here was conducted to review local medicines reconciliation practice and compare it to data within previously published evidence. OBJECTIVES: To determine current medicines reconciliation practice in four acute hospitals (A-D) in one region of the United Kingdom and compare it to published best practices. METHOD: Quantitative data on key indicators were collected prospectively from medical wards in the four hospitals using a proforma compiled from existing literature and previous, validated audits. Data were collected on: i) time between admission and MR being undertaken; ii) time to conduct MR; iii) number and type of sources used to ascertain current medication; and iv) number, type and potential severity of unintended discrepancies. The potential severity of the discrepancies was retrospectively dually rated in 10% of the sample using a professional panel. RESULTS: Of the 250 charts reviewed (54 Hospital A, 61 Hospital B, 69 Hospital C, 66 Hospital D), 37.6% (92/245) of patients experienced at least one discrepancy on their drug chart, with the majority of these being omissions (237/413, 57.1%). A total of 413 discrepancies were discovered, an overall mean of 1.69 (413/245) discrepancies per patient. The number of sources used to reconcile medicines varied with 36.8% (91/247) only using one source of information and the patient being used as a source in less than half of all medicines reconciliations (45.7%, 113/247). In three out of the four hospitals the discrepancies were most frequently categorized as potentially requiring increased monitoring or intervention. CONCLUSION: This study shows higher rates of unintended discrepancies per patient than those in previous studies, with omission being the most frequently occurring type of discrepancy. None of the four centers adhered to current UK guidance on medicines reconciliation. All four centers demonstrated a strong reliance on General Practitioner (GP)-based sources. A minority of discrepancies had the potential to cause injury to patients and to increase utilization of health care resources. There is a need to review current practice and procedures at transitions in care to improve the accuracy of medication history-taking at admission by doctors and to encourage pharmacy staff to use an increased number of sources to validate the medication history. Although early research indicates that safety can be improved through patient involvement, this study found that patients were not involved in the majority of reconciliation encounters.

Communicating medication changes to community pharmacy post-discharge: the good, the bad, and the improvements.

BACKGROUND: Communication between hospital and community pharmacists when a patient is discharged from hospital can improve the accuracy of medication reconciliation, thus preventing unintentional changes and ensuring continuity of supply. It allows problems to be resolved before a patient requires a further supply of medication post-discharge. Despite evidence demonstrating the benefits of sharing information, community pharmacists' willingness to receive information and advances in information technology (particularly electronic discharge medication summaries), there is little published evidence to indicate whether communication has improved over the last 15 years. This study aimed to explore community pharmacists' experience of information sharing by and with their local hospital and GP practices. OBJECTIVES: (1) To establish the extent to which community pharmacies currently receive discharge medication information, and for which patients.(2)To determine community pharmacy staff opinion on where and how current communication practice could be improved. SETTING: Community Pharmacies in one Primary Care Organisation (PCO) in England. METHOD: Semi-structured interviews conducted during visits to community pharmacies. MAIN OUTCOME MEASURE: Reported receipt of discharge medication information from hospitals and general practices. RESULTS: A total of 14 community pharmacies participated. Current provision of information to community pharmacies from hospitals regarding medication changes at discharge was reported to be inconsistent and lacking in quality. Where information was received it was predominantly for patients who receive their medicines in monitored dosage systems (MDS) rather than for the general population of patients. Some examples of "notable practice" were reported. CONCLUSION: Community pharmacists received post-discharge information rarely and mainly for patients where the hospital perceived the patient's medication issues as "complex". Practice was inconsistent overall. These findings suggest that the potential of community pharmacists to improve patient safety after discharge from hospital is not being utilised.