Comparative PCR Analysis of Two Triplet Repeat Loci and Factors Influencing Their Mutability

Fourteen expanded trinucleotide loci have been found to cause neurological diseases including Huntington's Disease, myotonic dystrophy type 1 and spinocerebellar ataxia. The repeat sequence and allele sizes vary between each locus, and the flanking DNA is assumed to play a significant role in repeat instability. The aim of this project was to characterise the behaviour and mutability of two trinucleotide repeat loci with the same repeat sequence, but with different flanking DNA sequences. Comparing the mutation rates and average length changes of these loci in the male germline demonstrated a clear effect of the influence of the flanking sequence. The ERDAI locus has low %GC content in the flanking sequence and displays a low mutation rate, even for large length alleles, whereas CTG18.1 has a relatively high %GC content and demonstrates an increased mutation rate and average length change for similarly sized alleles. Utilising the sensitive SP-PCR technique gives a clear insight into how the CTG18.1 locus has responded dramatically to defects in mismatch repair, both in MMR deficient cell lines and HNPCC tissues, and possibly in an ovarian tumour. This investigation has also revealed a possible dominant negative mutation in the hMLH1 gene which maintains its influence on an expanded CTG18.1 allele in two generations of a HNPCC family. In conclusion, we have successfully characterised the behaviour of two trinucleotide repeat loci which highlights the significant effect of flanking DNA sequence on the stability of triplet repeats

The role of microbiology and pharmacy departments in the stewardship of antibiotic prescribing in European hospitals

This observational, cross-sectional study describes the role played by clinical microbiology and pharmacy departments in the stewardship of antibiotic prescribing in European hospitals. A total of 170 acute care hospitals from 32 European countries returned a questionnaire on antibiotic policies and practices implemented in 2001. Data on antibiotic use, expressed as De.ned Daily Doses per 100 occupied bed-days (DDD/100 BD) were provided by 139 hospitals from 30 countries. A total of 124 hospitals provided both datasets. 121 (71%) of Clinical Microbiology departments and 66 (41%) of Pharmacy departments provided out of hours clinical advice. 70 (41%) of microbiology/infectious disease specialists and 28 (16%) of pharmacists visited wards on a daily basis. The majority of laboratories provided monitoring of blood cultures more than once per day and summary data of antibiotic susceptibility testing (AST) for empiric prescribing (86% and 73% respectively). Most of the key laboratory and pharmacy-led initiatives examined did not vary signi.cantly by geographical location. Hospitals from the North and West of Europe were more likely to examine blood cultures more than once daily compared with other regions (p < 0.01). Hospitals in the North were least likely routinely to report susceptibility results for restricted antibiotics compared to those in the South-East and Central/Eastern Europe (p < 0.01). Hospital wards in the North were more likely to hold antibiotic stocks (100%) compared with hospitals in the South-East which were least likely (39%) (p < 0.001). Conversely, hospital pharmacies in the North were least likely to dispense antibiotics on an individual patient basis (16%) compared with hospital pharmacies from Southern Europe (60%) (p = 0.01). Hospitals that routinely reported susceptibility results for restricted antibiotics had signi.cantly lower median total antibiotic use in 2001 (p < 0.01). Hospitals that provided prescribing advice outside normal working hours had signi.cantly higher antibiotic use compared with institutions that did not provide this service (p = 0.01). A wide range of antibiotic stewardship measures was practised in the participating hospitals in 2001, although there remains great scope for expansion of those overseen by pharmacy departments. Most hospitals had active antibiotic stewardship programmes led by specialists in infection, although there is no evidence that these were associated with reduced antibiotic consumption. There was also no evidence that pharmacy services reduced the amount of antibiotics prescribed.The ARPAC study was funded by the European Commission (project QLK2-CT-2001-00915). F.M. MacKenzie was supported by the European Study Group on Antibiotic Policies to write this manuscript

Recommendations for validation testing of home pregnancy tests (HPTs) in Europe

Homepregnancy tests (HPTs) available in Europe include accuracy and other performance claims listed on their packaging. Due to the lack of guidance on the standardisation of such products, it is often difficult to replicate these claims when tested on a clinical sample, whether in a laboratory setting or by lay users. The In Vitro Diagnostic Regulation is a set of requirements that mandate comprehensive validation data on human pregnancy tests and other in vitro devices. It is due to replace the current European Directive (98/79/EC) and fully implemented in Europe by 2022. In June 2019, a panel of seven experts convened to discuss the validation studies required to provide the information needed to meet the new regulation for HPTs in Europe and proposed 15 recommendations for best practice. Defining best practice at all stages of validation of these important tests may ensure that tests marketed inEurope are fit for purpose, enabling lay users to be confident of the high quality of the HPT results they obtain. The panelists believe that the recommendations proposed here for the validation of HPTs may constructively contribute to improved standardisation of validation procedures in Europe.Peer reviewe

Patient involvement in the implementation of infection prevention and control guidelines and associated interventions: a scoping review

Objective: To explore patient involvement in the implementation of infection prevention and control (IPC) guidelines and associated interventions.Design: Scoping review. Methods: A methodological framework was followed to identify recent publications on patient involvement in the implementation of IPC guidelines and interventions. Initially, relevant databases were searched to identify pertinent publications (published 2013–2018). Reflecting the scarcity of included studies from these databases, a bidirectional citation chasing approach was used as a second search step. The reference list and citations of all identified papers from databases were searched to generate a full list of relevant references. A grey literature search of Google Scholar was also conducted.Results: From an identified 2078 papers, 14 papers were included in this review. Our findings provide insights into the need for a fundamental change to IPC, from being solely the healthcare professionals (HCPs) responsibility to one that involves a collaborative relationship between HCPs and patients. This change should be underpinned by a clear understanding of patient roles, potential levels of patient involvement in IPC and strategies to overcome barriers to patient involvement focusing on the professional–patient relationship (eg, patient encouragement through multimodal educational strategies and efforts to disperse professional’s power).Conclusions: There is limited evidence regarding the best strategies to promote patient involvement in the implementation of IPC interventions and guidelines. The findings of this review endorse the need for targeted strategies to overcome the lack of role clarity of patients in IPC and the power imbalances between patients and HCPs

Negative regulation of syntaxin4/SNAP-23/VAMP2-mediated membrane fusion by Munc18c <i>In Vitro</i>

Background: Translocation of the facilitative glucose transporter GLUT4 from an intracellular store to the plasma membrane is responsible for the increased rate of glucose transport into fat and muscle cells in response to insulin. This represents a specialised form of regulated membrane trafficking. Intracellular membrane traffic is subject to multiple levels of regulation by conserved families of proteins in all eukaryotic cells. Notably, all intracellular fusion events require SNARE proteins and Sec1p/Munc18 family members. Fusion of GLUT4-containing vesicles with the plasma membrane of insulin-sensitive cells involves the SM protein Munc18c, and is regulated by the formation of syntaxin 4/SNAP23/VAMP2 SNARE complexes. Methodology/Principal Findings Here we have used biochemical approaches to characterise the interaction(s) of Munc18c with its cognate SNARE proteins and to examine the role of Munc18c in regulating liposome fusion catalysed by syntaxin 4/SNAP23/VAMP2 SNARE complex formation. We demonstrate that Munc18c makes contacts with both t- and v-SNARE proteins of this complex, and directly inhibits bilayer fusion mediated by the syntaxin 4/SNAP23/VAMP2 SNARE complex. Conclusion/Significance Our reductionist approach has enabled us to ascertain a direct inhibitory role for Munc18c in regulating membrane fusion mediated by syntaxin 4/SNAP23/VAMP2 SNARE complex formation. It is important to note that two different SM proteins have recently been shown to stimulate liposome fusion mediated by their cognate SNARE complexes. Given the structural similarities between SM proteins, it seems unlikely that different members of this family perform opposing regulatory functions. Hence, our findings indicate that Munc18c requires a further level of regulation in order to stimulate SNARE-mediated membrane fusion

Antimicrobial Drug Use and Methicillin-resistant Staphylococcus aureus, Aberdeen, 1996–2000

Relationships between antimicrobial use and MRSA prevalence are analyzed in Aberdeen, Scotland

Evaluating the Quality of Research into a Single Prognostic Biomarker: A Systematic Review and Meta-analysis of 83 Studies of C-Reactive Protein in Stable Coronary Artery Disease

Background Systematic evaluations of the quality of research on a single prognostic biomarker are rare. We sought to evaluate the quality of prognostic research evidence for the association of C-reactive protein (CRP) with fatal and nonfatal events among patients with stable coronary disease. Methods and Findings We searched MEDLINE (1966 to 2009) and EMBASE (1980 to 2009) and selected prospective studies of patients with stable coronary disease, reporting a relative risk for the association of CRP with death and nonfatal cardiovascular events. We included 83 studies, reporting 61,684 patients and 6,485 outcome events. No study reported a prespecified statistical analysis protocol; only two studies reported the time elapsed (in months or years) between initial presentation of symptomatic coronary disease and inclusion in the study. Studies reported a median of seven items (of 17) from the REMARK reporting guidelines, with no evidence of change over time. The pooled relative risk for the top versus bottom third of CRP distribution was 1.97 (95% confidence interval [CI] 1.78–2.17), with substantial heterogeneity (I2 = 79.5). Only 13 studies adjusted for conventional risk factors (age, sex, smoking, obesity, diabetes, and low-density lipoprotein [LDL] cholesterol) and these had a relative risk of 1.65 (95% CI 1.39–1.96), I2 = 33.7. Studies reported ten different ways of comparing CRP values, with weaker relative risks for those based on continuous measures. Adjusting for publication bias (for which there was strong evidence, Egger's p<0.001) using a validated method reduced the relative risk to 1.19 (95% CI 1.13–1.25). Only two studies reported a measure of discrimination (c-statistic). In 20 studies the detection rate for subsequent events could be calculated and was 31% for a 10% false positive rate, and the calculated pooled c-statistic was 0.61 (0.57–0.66). Conclusion Multiple types of reporting bias, and publication bias, make the magnitude of any independent association between CRP and prognosis among patients with stable coronary disease sufficiently uncertain that no clinical practice recommendations can be made. Publication of prespecified statistical analytic protocols and prospective registration of studies, among other measures, might help improve the quality of prognostic biomarker research

Ethnographic study using Normalization Process Theory to understand the implementation process of infection prevention and control guidelines in Ireland.

OBJECTIVE: The aim of this study was to explore how infection prevention and control (IPC) guidelines are used and understood by healthcare professionals, patients and families. DESIGN: Ethnographic study with 59 hours of non-participant observation and 57 conversational interviews. Data analysis was underpinned by the Normalization Process Theory (NPT) as a theoretical framework. SETTING: Four hospitals in Ireland. PARTICIPANTS: Healthcare professionals, patient and families. RESULTS: Five themes emerged through the analysis. Four themes provided evidence of the NPT elements (coherence, cognitive participation, collective action and reflexive monitoring). Our findings revealed the existence of a 'dissonance between IPC guidelines and the reality of clinical practice' (theme 1) and 'Challenges to legitimatize guidelines' recommendations in practice' (theme 3). These elements contributed to 'Symbolic implementation of IPC guidelines' (theme 2), which was also determined by a 'Lack of shared reflection upon IPC practices' (theme 4) and a clinical context of 'Workforce fragmentation, time pressure and lack of prioritization of IPC' (theme 5). CONCLUSIONS: Our analysis identified themes that provide a comprehensive understanding of elements needed for the successful or unsuccessful implementation of IPC guidelines. Our findings suggest that implementation of IPC guidelines is regularly operationalised through the reproduction of IPC symbols, rather than through adherence to performance of the evidence-based recommendations. Our findings also provide insights into changes to make IPC guidelines that align with clinical work