Expression of membrane-associated proteins within single emulsion cell facsimiles

MreB is a structural membrane-associated protein which is one of the key components of the bacterial cytoskeleton. Although it plays an important role in shape maintenance of rod-like bacteria, the understanding of its mechanism of action is still not fully understood. This study shows how segmented flow and microdroplet technology can be used as a new tool for biological in vitro investigation of this protein. In this paper, we demonstrate cell-free expression in a single emulsion system to express red fluorescence protein (RFP) and MreB linked RFP (MreB–RFP). We follow the aggregation and localisation of the fusion protein MreB–RFP in this artificial cell-like environment. The expression of MreB–RFP in single emulsion droplets leads to the formation of micrometer-scale protein patches distributed at the water/oil interface

Anger, Quality of Life and Mood in Multiple Sclerosis

This research was funded by The Multiple Sclerosis Society (UK).Peer reviewedPublisher PD

Vasopeptidase inhibition attenuates the progression of renal injury in subtotal nephrectomized rats

Vasopeptidase inhibition attenuates the progression of renal injury in subtotal nephrectomized rats.BackgroundVasopeptidase inhibitors are a new class of cardiovascular compounds that inhibit both angiotensin-converting enzyme (ACE) and neutral endopeptidase (NEP). The aim of the present study was to explore the effects of omapatrilat, a vasopeptidase inhibitor, on renal function and pathology in subtotally nephrectomized (STNx) rats.MethodsSTNx rats were randomized to four groups and treated for 12 weeks: no treatment (N = 14); omapatrilat at a low dose of 10 mg/kg (L, N = 12) and at a high dose of 40 mg/kg (H, N = 10); or an ACE inhibitor, fosinopril, at a dose of 10 mg/kg (N = 12). Sham-operated rats were used as control animals (N = 12).ResultsElevated blood pressure in STNx rats (174 ± 9mm Hg) was reduced by omapatrilat in a dose-dependent manner (L, 121 ± 3mm Hg; H, 110 ± 3mm Hg) and by fosinopril (149 ± 5mm Hg). Proteinuria in STNx rats (246 ± 73 mg/day) was reduced by treatment with fosinopril (88 ± 21 mg/day) and was normalized by treatment with omapatrilat (L, 30 ± 4 mg/day; H, 20 ± 2 mg/day vs. control 25 ± 1 mg/day). Decreased glomerular filtration rates, elevated plasma urea and creatinine and glomerulosclerosis, and tubulointerstitial fibrosis were ameliorated by omapatrilat and fosinopril to a similar degree. Compared with fosinopril, omapatrilat treatment was associated with increased plasma renin activity and decreased renal ACE and NEP binding in a dose-dependent manner.ConclusionThese findings suggest that vasopeptidase inhibition may provide a useful strategy for the treatment of progressive renal disease

The clinical effectiveness and cost-effectiveness of cetuximab (mono- or combination chemotherapy), bevacizumab (combination with non-oxaliplatin chemotherapy) and panitumumab (monotherapy) for the treatment of metastatic colorectal cancer after first-line chemotherapy (review of technology appraisal No.150 and part review of technology appraisal No. 118): a systematic review and economic model

Colorectal cancer is the third most commonly diagnosed cancer in the UK after breast and lung cancer. People with metastatic disease who are sufficiently fit are usually treated with active chemotherapy as first- or second-line therapy. Recently, targeted agents have become available including anti-epidermal growth factor receptor (EGFR) agents, for example cetuximab and panitumumab, and anti-vascular endothelial growth factor (VEGF) receptor agents, for example bevacizumab

Learning to prescribe - pharmacists' experiences of supplementary prescribing training in England

Background: The introduction of non-medical prescribing for professions such as pharmacy and nursing in recent years offers additional responsibilities and opportunities but attendant training issues. In the UK and in contrast to some international models, becoming a non-medical prescriber involves the completion of an accredited training course offered by many higher education institutions, where the skills and knowledge necessary for prescribing are learnt. Aims: to explore pharmacists' perceptions and experiences of learning to prescribe on supplementary prescribing (SP) courses, particularly in relation to inter-professional learning, course content and subsequent use of prescribing in practice. Methods: A postal questionnaire survey was sent to all 808 SP registered pharmacists in England in April 2007, exploring demographic, training, prescribing, safety culture and general perceptions of SP. Results: After one follow-up, 411 (51%) of pharmacists responded. 82% agreed SP training was useful, 58% agreed courses provided appropriate knowledge and 62% agreed that the necessary prescribing skills were gained. Clinical examination, consultation skills training and practical experience with doctors were valued highly; pharmacology training and some aspects of course delivery were criticised. Mixed views on inter-professional learning were reported – insights into other professions being valued but knowledge and skills differences considered problematic. 67% believed SP and recent independent prescribing (IP) should be taught together, with more diagnostic training wanted; few pharmacists trained in IP, but many were training or intending to train. There was no association between pharmacists' attitudes towards prescribing training and when they undertook training between 2004 and 2007 but earlier cohorts were more likely to be using supplementary prescribing in practice. Conclusion: Pharmacists appeared to value their SP training and suggested improvements that could inform future courses. The benefits of inter-professional learning, however, may conflict with providing professionspecific training. SP training may be perceived to be an instrumental 'stepping stone' in pharmacists' professional project of gaining full IP status

Antidepressant Prevention of Postnatal Depression

A systematic review sought to discover whether antidepressant drugs are effective in preventing postnatal depression. Only two small trials were found and it was not possible to draw firm conclusions

Piloting Eyes on the Baby: A Multiagency Training and Implementation Intervention Linking Sudden Unexpected Infant Death Prevention and Safeguarding

We describe the coproduction, pilot implementation, and user evaluation of an evidence-based training intervention addressing prevention of Sudden Unexpected Deaths in Infancy (SUDI) for the multiagency workforce supporting vulnerable families with babies in a northern English county. We aimed in this pilot study to improve knowledge, skills, and engagement of professionals and support staff providing services for vulnerable families with increased risk of SUDI. The training intervention was co-produced by the academic team and the project Steering Committee which comprised senior leaders from the local authority, health and care sectors, and third-sector organisations, and rolled out to multiagency teams between November 2022 and March 2023. Evaluation data were collected using a post-training questionnaire, followed up by the Normalisation Process Theory (NPT) NoMAD survey issued at two time-points post-training, and interviews with stakeholders. The evaluation, conducted from January to May 2023, aimed to assess how well the multiagency workforce accepted SUDI prevention as part of their remit and incorporated SUDI prevention activities into their everyday work. Most multiagency professionals and support staff were enthusiastic about the training and their role in SUDI prevention. Fewer health professionals completed the training than expected. Forty percent (397/993) of invited staff completed the training. Our results revealed initial lack of knowledge and confidence around SUDI prevention and targeted provision for vulnerable families which improved following the Eyes on the Baby training. The proportion of nonhealth professionals rating their knowledge of SUDI prevention as good or excellent increased significantly from 28% before training to 57% afterwards. Self-rated confidence in discussing SUDI prevention with families increased significantly from 71% to 97%. Health professionals’ ratings increased significantly for knowledge from 62% to 96%, and confidence from 85% to 100%. Use of NPT allowed us to identify that by the time of evaluation, the earliest adopters were cognitively involved with the programme and engaged in collective action, while later adopters had not yet reached this stage. We conclude that effective implementation of multiagency working for SUDI prevention can be accomplished by providing clear training and guidance for all staff who have regular or opportunistic contact with vulnerable families. Our next step is to evaluate the sustainability of MAW SUDI prevention over the medium to long term and assess the responses of recipient families to this approach

Race and sex associations with tacrolimus pharmacokinetics in stable kidney transplant recipients

Study Objective: This study investigated race and sex differences in tacrolimus pharmacokinetics and pharmacodynamics in stable kidney transplant recipients. Design and Setting: A cross-sectional, open-label, single center, 12-h pharmacokinetic-pharmacodynamic study was conducted. Tacrolimus pharmacokinetic parameters included area under the concentration-time curve (AUC0–12), AUC0–4, 12-h troughs (C12 h), maximum concentrations (Cmax), oral clearance (Cl), with dose-normalized AUC0–12, troughs, and Cmax with standardized adverse effect scores. Statistical models were used to analyze end points with individual covariate-adjustment including clinical factors, genotypic variants CYP3A5*3, CYP3A5*6, CYP3A5*7(CYP3A5*3*6*7) metabolic composite, and ATP binding cassette gene subfamily B member 1 (ABCB1) polymorphisms. Patients: 65 stable, female and male, Black and White kidney transplant recipients receiving tacrolimus and mycophenolic acid ≥6 months post-transplant were evaluated. Measurements and Main Results: Black recipients exhibited higher tacrolimus AUC0–12 (Race: p = 0.005), lower AUC* (Race: p \u3c 0.001; Race × Sex: p = 0.068), and higher Cl (Race: p \u3c 0.001; Sex: p = 0.066). Greater cumulative (Sex: p \u3c 0.001; Race × Sex: p = 0.014), neurologic (Sex: p = 0.021; Race × Sex: p = 0.005), and aesthetic (Sex: p = 0.002) adverse effects were found in females, with highest scores in Black women. In 84.8% of Black and 68.8% of White patients, the target AUC0–12 was achieved (p = 0.027). In 31.3% of White and 9.1% of Black recipients, AUC0–12 was \u3c100 \u3eng‧h/ml despite tacrolimus troughs in the target range (p = 0.027). The novel CYP3A5*3*6*7 metabolic composite was the significant covariate accounting for 15%–19% of tacrolimus variability in dose (p = 0.002); AUC0–12 h* (p \u3c 0.001), and Cl (p \u3c 0.001). Conclusions: Tacrolimus pharmacokinetics and adverse effects were different among stable kidney transplant recipient groups based upon race and sex with interpatient variability associated with the CYP3A5*3*6*7 metabolic composite. More cumulative, neurologic, and aesthetic adverse effects were noted among females. Tacrolimus regimens that consider race and sex may reduce adverse effects and enhance allograft outcomes by facilitating more patients to achieve the targeted AUC0–12 h

Beyond Single Nucleotide Polymorphisms: CYP3A5∗3 ∗6 ∗7 Composite and ABCB1 Haplotype Associations to Tacrolimus Pharmacokinetics in Black and White Renal Transplant Recipients

Interpatient variability in tacrolimus pharmacokinetics is attributed to metabolism by cytochrome P-450 3A5 (CYP3A5) isoenzymes and membrane transport by P-glycoprotein. Interpatient pharmacokinetic variability has been associated with genotypic variants for both CYP3A5 or ABCB1. Tacrolimus pharmacokinetics was investigated in 65 stable Black and Caucasian post-renal transplant patients by assessing the effects of multiple alleles in both CYP3A5 and ABCB1. A metabolic composite based upon the CYP3A5 polymorphisms: ∗3(rs776746), ∗6(10264272), and ∗7(41303343), each independently responsible for loss of protein expression was used to classify patients as extensive, intermediate and poor metabolizers. In addition, the role of ABCB1 on tacrolimus pharmacokinetics was assessed using haplotype analysis encompassing the single nucleotide polymorphisms: 1236C \u3e T (rs1128503), 2677G \u3e T/A(rs2032582), and 3435C \u3e T(rs1045642). Finally, a combined analysis using both CYP3A5 and ABCB1 polymorphisms was developed to assess their inter-related influence on tacrolimus pharmacokinetics. Extensive metabolizers identified as homozygous wild type at all three CYP3A5 loci were found in 7 Blacks and required twice the tacrolimus dose (5.6 ± 1.6 mg) compared to Poor metabolizers [2.5 ± 1.1 mg (P \u3c 0.001)]; who were primarily Whites. These extensive metabolizers had 2-fold faster clearance (P \u3c 0.001) with 50% lower AUC∗ (P \u3c 0.001) than Poor metabolizers. No differences in C12 h were found due to therapeutic drug monitoring. The majority of blacks (81%) were classified as either Extensive or Intermediate Metabolizers requiring higher tacrolimus doses to accommodate the more rapid clearance. Blacks who were homozygous for one or more loss of function SNPS were associated with lower tacrolimus doses and slower clearance. These values are comparable to Whites, 82% of who were in the Poor metabolic composite group. The ABCB1 haplotype analysis detected significant associations of the wildtype 1236T-2677T-3435T haplotype to tacrolimus dose (P = 0.03), CL (P = 0.023), CL/LBW (P = 0.022), and AUC∗ (P = 0.078). Finally, analysis combining CYP3A5 and ABCB1 genotypes indicated that the presence of the ABCB1 3435 T allele significantly reduced tacrolimus clearance for all three CPY3A5 metabolic composite groups. Genotypic associations of tacrolimus pharmacokinetics can be improved by using the novel composite CYP3A5∗3∗4∗5 and ABCB1 haplotypes. Consideration of multiple alleles using CYP3A5 metabolic composites and drug transporter ABCB1 haplotypes provides a more comprehensive appraisal of genetic factors contributing to interpatient variability in tacrolimus pharmacokinetics among Whites and Blacks

Multiagency approaches to preventing sudden unexpected death in infancy (SUDI): a review and analysis of UK policies

Background Recent reviews of sudden unexpected deaths in infancy (SUDI) in England recommend a multiagency working (MAW) approach to prevention but lack clear guidance around how this might be implemented.Aims In England, local authorities commission and oversee public health services. This review examines how local authority policies address implementation of MAW for SUDI prevention to understand local variations and identify strengths and weaknesses.Methods Using a comprehensive list of all metropolitan, county, unitary councils and London boroughs in England, we systematically searched local authority websites for relevant published documents and submitted freedom of information (FOI) requests where policies or guidance for SUDI prevention had not been sourced online. We extracted data from documents using a standardised form to summarise policy contents which were then collated, described and appraised.Findings We searched the websites of 152 council and London boroughs, identifying 36 relevant policies and guidelines for staff. We submitted 116 FOI requests which yielded 64 responses including six valid documents: 45% (52/116) of local authorities did not respond. Seventeen councils shared the same guidance under safeguarding partnerships; removal of duplicates resulted in 26 unique documents. Only 15% (4/26) of the documents included a detailed plan for how MAW approaches were to be implemented despite 73% (19/26) of the documents mentioning the importance of engaging the MAW in raising awareness of safe sleep for babies with vulnerable families. Five areas of variation were identified across policies: (1) scope, (2) responsibilities, (3) training, (4) implementation and (5) evaluation.Conclusions There are discrepancies between local authorities in England in whether and how MAW for SUDI prevention is carried out. Strengths and weaknesses of approaches are identified to inform future development of MAW for SUDI prevention