The fork protection complex recruits FACT to reorganize nucleosomes during replication

Chromosome replication depends on efficient removal of nucleosomes by accessory factors to ensure rapid access to genomic information. Here, we show this process requires recruitment of the nucleosome reorganization activity of the histone chaperone FACT. Using single-molecule FRET, we demonstrate that reorganization of nucleosomal DNA by FACT requires coordinated engagement by the middle and C-terminal domains of Spt16 and Pob3 but does not require the N-terminus of Spt16. Using structure-guided pulldowns, we demonstrate instead that the N-terminal region is critical for recruitment by the fork protection complex subunit Tof1. Using in vitro chromatin replication assays, we confirm the importance of these interactions for robust replication. Our findings support a mechanism in which nucleosomes are removed through the coordinated engagement of multiple FACT domains positioned at the replication fork by the fork protection complex

Investigation of Laurus Tamala leaves extract as an environmentally acceptable corrosion inhibitor for soft steel in 1M HCl: Electrochemical, DFT, and surface characterization techniques

Laurus Tamala leaves extract (LTLE) has been employed as a soft steel corrosion inhibitor in a 1M Hydrochloric acid media. Chemical (weight loss) and electrochemical investigations were carried out to assess the corrosion rate and percentage inhibition efficiency of the extract. The electrochemical polarization results have demonstrated that plant leaves extract functions as a mixed type inhibitor. The stability of the inhibitor is tested at elevated temperatures by weight loss method. The corrosion inhibition mechanism is interpreted through adsorption mechanism, and the LTLE components has obeyed the Langmuir adsorption isotherm for soft steel. The interaction of the components of the extract is assessed through FT-IR technique. The surface morphology, roughness and hydrophobicity in presence and absence of the extract have been characterized through SEM, AFM and water contact angle techniques respectively. The highest inhibitory efficiency is 96.21% for 24 h as recorded by weight loss method. Additionally, the DFT computations has revealed the inhibitor’s adsorption through electron donor-acceptor interactions

Training primary care providers in opioid deprescribing and chronic pain management based on local guidance: a pre–post study of attitude change

Background. Local chronic non-cancer pain guidance recommends that general practitioners should consider opioid deprescribing and referral to multidisciplinary healthcare providers for behaviourally based treatments. We designed a training package called AIMM (Assess, Inform, Manage and Monitor) to reinforce this stewardship.Aim. To identify whether participation in AIMM training effectively aligned clinicians’ attitudes with local guidance for treating chronic non-cancer pain.Design and setting. In 2014–15, the AIMM training was tested using a pre–post-test non-randomised design at two sites in NSW, Australia. The primary outcome measure was an 11-item, study-specific, pain attitude questionnaire (PAQ).Method. Step one of AIMM training involved online completion of the PAQ and review of a specialist pain website. Step two involved attendance at two face-to-face, two-hour interactive workshops led by pain experts who addressed opioid deprescribing and switching to broader care. A repeat PAQ survey was completed at the conclusion of the second workshop.Results. Nineteen participants attended the workshops, including general practitioners (n = 7), nurses (n = 5), exercise physiologists (n = 2), a dietitian (n = 1), community pharmacists (n = 2) and psychologists (n = 2). Significant shifts in six attitudes occurred, including prescribing less pain medication, greater emphasis on social reconnection, increasing planned activity and adopting anti-inflammatory nutrition (p  .05). Responses to the item regarding expectations of a positive recovery was not aligned with local guidance and no significant attitudinal change was found. Four other attitudes were aligned with local guidance at baseline and did not change during the study.Conclusions. Online information and face-to-face training can achieve a change in healthcare provider attitudes towards non-pharmacological treatment of chronic non-cancer pain. Further work is needed to assess whether attitudinal changes are maintained and translate into behavioural change

Basic science232. Certolizumab pegol prevents pro-inflammatory alterations in endothelial cell function

Background: Cardiovascular disease is a major comorbidity of rheumatoid arthritis (RA) and a leading cause of death. Chronic systemic inflammation involving tumour necrosis factor alpha (TNF) could contribute to endothelial activation and atherogenesis. A number of anti-TNF therapies are in current use for the treatment of RA, including certolizumab pegol (CZP), (Cimzia ®; UCB, Belgium). Anti-TNF therapy has been associated with reduced clinical cardiovascular disease risk and ameliorated vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to investigate the mechanisms underpinning CZP effects on TNF-activated human endothelial cells. Methods: Human aortic endothelial cells (HAoECs) were cultured in vitro and exposed to a) TNF alone, b) TNF plus CZP, or c) neither agent. Microarray analysis was used to examine the transcriptional profile of cells treated for 6 hrs and quantitative polymerase chain reaction (qPCR) analysed gene expression at 1, 3, 6 and 24 hrs. NF-κB localization and IκB degradation were investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was conducted to detect microparticle release from HAoECs. Results: Transcriptional profiling revealed that while TNF alone had strong effects on endothelial gene expression, TNF and CZP in combination produced a global gene expression pattern similar to untreated control. The two most highly up-regulated genes in response to TNF treatment were adhesion molecules E-selectin and VCAM-1 (q 0.2 compared to control; p > 0.05 compared to TNF alone). The NF-κB pathway was confirmed as a downstream target of TNF-induced HAoEC activation, via nuclear translocation of NF-κB and degradation of IκB, effects which were abolished by treatment with CZP. In addition, flow cytometry detected an increased production of endothelial microparticles in TNF-activated HAoECs, which was prevented by treatment with CZP. Conclusions: We have found at a cellular level that a clinically available TNF inhibitor, CZP reduces the expression of adhesion molecule expression, and prevents TNF-induced activation of the NF-κB pathway. Furthermore, CZP prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions and measurement of microparticles has potential as a novel prognostic marker for future cardiovascular events in this patient group. Disclosure statement: Y.A. received a research grant from UCB. I.B. received a research grant from UCB. S.H. received a research grant from UCB. All other authors have declared no conflicts of interes

Investigation of Laurus Tamala leaves extract as an environmentally acceptable corrosion inhibitor for soft steel in 1M HCl: Electrochemical, DFT, and surface characterization techniques

492-505Laurus Tamala leaves extract (LTLE) has been employed as a soft steel corrosion inhibitor in a 1M Hydrochloric acid media. Chemical (weight loss) and electrochemical investigations were carried out to assess the corrosion rate and percentage inhibition efficiency of the extract. The electrochemical polarization results have demonstrated that plant leaves extract functions as a mixed type inhibitor. The stability of the inhibitor is tested at elevated temperatures by weight loss method. The corrosion inhibition mechanism is interpreted through adsorption mechanism, and the LTLE components has obeyed the Langmuir adsorption isotherm for soft steel. The interaction of the components of the extract is assessed through FT-IR technique. The surface morphology, roughness and hydrophobicity in presence and absence of the extract have been characterized through SEM, AFM and water contact angle techniques respectively. The highest inhibitory efficiency is 96.21% for 24 h as recorded by weight loss method. Additionally, the DFT computations has revealed the inhibitor’s adsorption through electron donor-acceptor interactions