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A biological method for peptide synthesis provides increased production capacity of inexpensive peptide pharmaceuticals with environmentally safe procedures relative to current chemical peptide synthesis. Most precursor peptides are readily produced from yeast and bacterial systems using recombinant DNA technologies but require C-terminal amidation for maximum biological activity. Peptidylglycine α-amidating monooxygenase (PAM) is the only enzyme that catalyses the C-terminal amidation of peptides in vivo through its two catalytic cores, peptidylglycine α-hydroxylating monooxygenase (PHM) and peptidylglycine αamidating lyase (PAL). The cost and limited quantities of the commercial PAM variants available have necessitated research into low cost, scalable quantities of PAM and peptide amidation to enable inexpensive biological peptide production. In the present study, an assay for measuring the product of PAM activity, glyoxylate, was developed based on a 2-aminobenzaldehyde-glycine-glyoxylate (AGG) absorbance assay. The AGG chromophore synthesised was identified with ultra-performance liquid chromatography mass-spectroscopy (UPLC-MS). PAM activity was measurable with glyoxylate between 25 µM and 1600 µM with the AGG assay. Furthermore, the activity of PHM alone was measured by the inclusion of an alkaline hydrolysis step to lyse glyoxylate as a substitute for PAL catalytic activity. Multiple candidate proteins and DNA sequences for PAM were identified by genetic sequence searches and a novel fungal PHM modelled in silico. Fourteen PAM, PHM, PAL and truncated constructs were expressed in the non-conventional yeast host, Yarrowia lipolytica. The novel fungal PHM's nutrient, temperature, and pH conditions were optimised to maximise protein expression. Enzyme purification was optimised with scalable industrial appropriate methods to purify milligram amounts of fungal PHM. The AGG assay was validated with a commercially obtained PAM, demonstrating a simple medium-throughput method to measure PAM activity. The novel fungal PHM was characterised with a pH optimum of 4.0 and maximum enzymatic activity at 45°C. Deglycosylation of fungal PHM enhanced enzyme activity by 1.83 fold, but lowered the temperature optimum to 37°C. The novel PHM and alkaline hydrolysis catalysed the conversion of the peptide pharmaceutical precursor for exenatide into its final bioactive form.

The world has changed significantly since the day I was born. One could argue that its fantastic political, moral, philosophical, and environmental gyrations have led to an inevitable collision between what is absolute and that which is not. Added to this widening gyre is the fundamental role of data and information in seating emperors on their respective thrones. The shift of power away from the halls of legislature and the proletariat into the hands of the emerging data bourgeoisie has led to the need for a new type of leadership, focused on using the prevalence of data in adversarial ways to shape society (and indeed the world) in new ways.

Older workers who are confident about the changes accompanying retirement report higher well-being. We have developed an index to measure retirement confidence – the Retirement Confidence Index (RCI). A six-stage approach was used to develop the index items, including (i) a literature review to catalogue retirement confidence components; (ii) a consultation with a panel of experts to review the proposed indicators and combine components according to their meaning; (iii) normalisation of the selected components to make them comparable; (iv) weighting of the top-level dimensions using experts’ judgement; (v) linear aggregation of the dimension scores according to their corresponding relative weight; and (vi) correlation of the composite score with a self-report measure of retirement confidence. Based on the review of the literature, a list of nine sub-components (financial literacy, financial attitude and behaviour, financial control, financial anxiety, physical health, mental health, social connectedness, goal setting for retirement and future uncertainties) was compiled. Subsequently, these components were grouped into four broad dimensions. Correlations between these dimensions (social, financial awareness and skills, health and well-being, and retirement awareness and planning dimensions) and the corresponding self-reported measures were as high as r = 0.555, r = 0.603, r = 0.591 and r = 0.569, reflecting 30.8%, 36.3%, 34.9% and 32.3% shared variance with the corresponding self-reported indices, respectively. The Retirement Confidence Index provides the foundation for future research to measure retirement confidence, with the aim of identifying deficient RCI dimensions and directing efforts to targeted policies to ensure older workers are confident about retirement.

Background Perturbation-based training (PBT) is a balance training method that causes a trip like event requiring a rapid step response to regain balance. There are numerous examples in the literature demonstrating the effectiveness of PBT but the need to use an expensive treadmill in a scientific laboratory limits the general applicability of PBT as a community-based intervention. A possible solution is to rapidly stop the treadmill belt during walking using the e-brake as the perturbation event. Importantly this could be performed on a commercially available, lower cost treadmill. Therefore, the purpose of this study was to evaluate the effectiveness of a commercial treadmill during PBT. Methods Seventeen participants completed either 9 weeks of PBT or conventional balance training based on ACSM guidelines. During an initial and final testing session participants balance recovery performance was evaluated. Participants were released from a forward static lean angle and asked to recover with a single step, during this test their movement was recorded and subsequently used to determine the Margin or Stability pre- and post-training. Participants were tracked for 6 months following the intervention and falls were recorded on a weekly basis. Results and Summary: There was no difference in balance recovery performance between groups following the training intervention and there was no difference in fall rate between groups in the 6-month follow-up period. We conclude that overall using the e-brake of a commercial treadmill is ineffective as a PBT strategy as it elicits no greater benefit than conventional exercise training.

Perturbation-based training (PBT) is a balance training method presenting a high-challenge to balance which is extremely effective when compared to conventional training approaches. Common PBT methods use rapid treadmill belt translations with varying numbers of perturbations (20-1400 perturbations over 1-8wks). Importantly, the joint loads experienced during a stumble may be high enough to feasibly fracture bone (up to 12.7 Body Weights). However, the contact loads experienced during PBT are unknown. Because of increasing the prevalence of PBT it is necessary to specifically evaluate the range of joint loads experienced during PBT. Twelve participants completed a single PBT session of 24 perturbations. During both training and testing, participants movements were measured using a motion capture system which tracks body movements and records the forces under the feet. Hip joint contact loads were determined using Computational Musculoskeletal Modelling utilizing open-source software OpenSim. These techniques estimate the magnitude and pattern of force development of individual muscles and subsequently estimate the internal loads experienced by the hip joint.Hip joint contact loads were 4.90 ± 1.27 BW which is substantially lower that those previously reported by Graham et al. (2016) and Bergman et al. (2004) and is lower than the 5.5BW spontaneous fracture load boundary estimated by Schileo et al. (2014). Comparing the initial perturbation to the final perturbation revealed a 22% reduction in contact loads. We conclude that PBT performed using rapid translations on a treadmill are likely safe but suggest caution for individuals with poor bone mineral density or reduced neuromuscular function.

The Irish Johne's Control Programme (IJCP) provides a long-term approach to the voluntary control of Johne's disease (JD) in Ireland, strongly supported by Irish cattle industry leadership. It leverages the establishment of Animal Health Ireland for control of animal diseases not regulated by the European Union. The IJCP has four objectives: facilitate protection against spread of JD to uninfected farms; reduce the level of infection when present; assure markets of JD control in Ireland; and improve calf health and farm biosecurity. Key IJCP elements are an annual veterinary risk assessment and management plan (VRAMP), annual whole herd test (WHT) by ELISA on blood or milk samples with ancillary faecal PCR testing of ELISA reactors, and Targeted Advisory Service on Animal Health (TASAH) investigations of infected herds. There are pathways for assurance of herds with continuing negative tests and for management of test-positive herds. Herdowners are responsible for on-farm activities, and specifically-trained (approved) veterinary practitioners have a pivotal role as technical advisors and service providers. The programme is supported by training of veterinarians, performance of testing in designated laboratories, documentation of policies and procedures, innovative data management for herd and test activities and for programme administration, training, and broad communication and awareness activities. Tools and systems are refined to address emerging issues and enhance the value of the programme. An Implementation Group comprising industry, government and technical leaders sets strategic direction and policy, advised by a Technical Working Group. Shared funding responsibilities are agreed by key stakeholders until 2022 to support herds in the programme to complete requirements. Herd registrations have increased steadily to exceed 1,800. National bulk tank milk surveillance is also being deployed to identify and recruit test-positive herds with the expectation that they have a relatively high proportion of seropositive animals. The programme will continue to innovate and improve to meet farmer and industry needs.

© 2021 Elsevier Ltd Stellite 6 (Co-29.5%Cr-5%W-1.2%C in wt%) is traditionally used as a hardfacing material in the primary circuit of pressurised water reactors (PWRs) due to its good corrosion and wear resistance in water at up to 300 °C. In this study, pin-on-disc type sliding contact tribocorrosion testing was conducted on HIPed Stellite 6 at 20 °C and 250 °C using a bespoke tribometer to simulate a primary circuit environment. Transmission electron microscopy (TEM), scanning electron microscopy (SEM), and X-ray diffraction (XRD) were used to characterize, for the first time, the material affected by tribocorrosion. Whilst the material loss increases by 16–39 times when the test temperature is increased from 20 °C to 250 °C, the mechanisms of degradation and deformation remain largely unchanged. Furthest from the sliding contact, strain is principally accommodated by the deformation-induced transformation of the γ Co-based matrix to ε-martensite. Closer to the sliding contact, the ε-martensite phase accommodates further strain via twinning and dislocation slip. At the sliding contact the intense deformation generates a nanocrystalline structure. The tribologically affected material is resistant to plastic strain localisation; this confines wear to the nanoscale where the synergistic effects of chemical degradation and mechanical deformation permit the removal of nanoscale particulates (corrosion enhanced nanowear (tribocorrosion)). The increased wear rate at 250 °C is attributed to a temperature dependent increase in corrosion enhanced nanowear. The degradation mechanisms revealed are important for the design of future hardfacings.

The trace element composition of olivine is becoming increasingly important in petrological studies due to the ubiquity of olivine in the Earth's upper mantle and in primitive magmatic rocks. The LA-ICP-MS method allows for the routine analysis of trace elements in olivine to sub-ppm levels, but a major drawback of this method is the lack of knowledge about possible downhole fractionation effects when non matrix-matched calibration is used. In this contribution, we show that matrix-matched (i.e., olivine-based) calibration is preferable for small laser spot sizes ( 2 , and total laser shot counts of ≤250 (e.g., 5 Hz repetition rate for 50 s) are used in order to minimize fractionation effects between olivine and silicate glass calibration materials. We demonstrate the applicability of matrix-matched calibration on olivine from a suite of different mantle peridotite xenoliths sampled by kimberlites and alkali basalts from on-craton and off-craton localities.

Background and aim Acute gastrointestinal bleeding carries poor outcomes unless prompt endoscopic hemostasis is achieved. Mortality in these patients remains significant. Hemospray is a novel intervention that creates a mechanical barrier over bleeding sites. We report the largest dataset of patient outcomes after treatment with Hemospray from an international multicenter registry. Patients and methods Prospective data (Jan 2016-May 2018) from 12 centers across Europe were collected. Immediate hemostasis was defined as endoscopic cessation of bleeding within 5 min after application of Hemospray. Rebleeding was defined as subsequent drop in hemoglobin, hematemesis, persistent melena with hemodynamic compromise post-therapy. Results Three hundred and fourteen cases were recruited worldwide (231 males, 83 females). Median pretreatment Blatchford score was 11 (IQR: 8-14) and median complete Rockall score (RS) was 7 (IQR: 6-8) for all patients. Peptic ulcer disease (PUD) was the most common pathology (167/314 = 53%) and Forrest Ib the most common bleed type in PUD (100/167 = 60%). 281 patients (89.5%) achieved immediate hemostasis after successful endoscopic therapy with Hemospray. Rebleeding occurred in 29 (10.3%) of the 281 patients who achieved immediate hemostasis. Seven-day and 30-day all-cause mortality were 11.5% (36/314) and 20.1% (63/314), respectively (lower than the predicted rates as per the RS). Similar hemostasis rates were noted in the Hemospray monotherapy (92.4%), combination therapy (88.7%) and rescue therapy (85.5%) groups. Conclusions These data show high rates of immediate hemostasis overall and in all subgroups. Rebleeding and mortality rates were in keeping/lower than predicted rates.

The endoscopic detection of esophageal cancer is suboptimal in both patients referred with dyspeptic symptoms and those enrolled in Barrett’s surveillance programs. MCM5 expression in cells collected from gastric fluid may be correlated with the presence of dysplasia or adenocarcinoma. Analysis of this biomarker may improve the detection of cancer. Sixty-one patients were enrolled at a single UK referral center. From each patient, 5–10 ml of gastric fluid was aspirated endoscopically. Patients were categorized according to their histology, normal, non-dysplastic Barrett’s (NDBE), high-grade dysplastic Barrett’s (HGD), and esophageal adenocarcinoma (EAC). All histology was confirmed by Seattle protocol biopsies or endoscopic mucosal resection. Samples were centrifuged, and the cell pellet was lysed. MCM5 expression levels were quantified using a proprietary immunoassay. The mean MCM5 expression was compared between groups by Kruskal–Wallis test. ROC curves were also used to assess diagnostic utility. The mean expression of MCM5 increases as patients progress from a normal esophagus to NDBE, HGD, and EAC (14.4; 49.8; 112.3; and 154.1, respectively). There was a significant difference in the MCM5 expression of patients with a normal esophagus compared to those with EAC (p = 0.04). There was a trend toward higher MCM5 expression in patients with EAC compared to those with NDBE (p = 0.34). MCM5 expression was a fair discriminator (AUC 0.70 [95% CI 0.57–0.83]) between patients without neoplasia (normal and NDBE) and those with early neoplasia (HGD and EAC). MCM5 expression in gastric fluid samples can differentiate patients with a histologically normal esophagus compared to those with early adenocarcinoma. Larger, powered studies are needed to assess whether it can be used to differentiate those with HGD from NDBE.

peer-reviewed Background Quantitative genetic studies suggest the existence of variation at the genome level that affects the ability of cattle to resist to parasitic diseases. The objective of the current study was to identify regions of the bovine genome that are associated with resistance to endo-parasites. Methods Individual cattle records were available for Fasciola hepatica-damaged liver from 18 abattoirs. Deregressed estimated breeding values (EBV) for F. hepatica-damaged liver were generated for genotyped animals with a record for F. hepatica-damaged liver and for genotyped sires with a least one progeny record for F. hepatica-damaged liver; 3702 animals were available. In addition, individual cow records for antibody response to F. hepatica on 6388 genotyped dairy cows, antibody response to Ostertagia ostertagi on 8334 genotyped dairy cows and antibody response to Neospora caninum on 4597 genotyped dairy cows were adjusted for non-genetic effects. Genotypes were imputed to whole-sequence; after edits, 14,190,141 single nucleotide polymorphisms (SNPs) and 16,603,644 SNPs were available for cattle with deregressed EBV for F. hepatica-damaged liver and cows with an antibody response to a parasitic disease, respectively. Association analyses were undertaken using linear regression on one SNP at a time, in which a genomic relationship matrix accounted for the relationships between animals. Results Genomic regions for F. hepatica-damaged liver were located on Bos taurus autosomes (BTA) 1, 8, 11, 16, 17 and 18; each region included at least one SNP with a p value lower than 10−6. Five SNPs were identified as significant (q value

BACKGROUND: Gastric antral vascular ectasia (GAVE) is a rare cause of gastrointestinal bleeding, often causing iron deficiency anaemia. Previous studies have looked at the management of this with argon plasma coagulation, laser therapy and endoscopic band ligation. METHODS: This was a single-centre prospective study to evaluate the efficacy and safety of radiofrequency ablation (RFA) in patients with GAVE with persistent anaemia refractory to at least one session of first-line endoscopic therapy. Patients were treated with a through-the-scope (TTS) radiofrequency catheter at two endoscopic sessions six weeks apart. The primary outcome was change in haemoglobin at six months posttreatment. The secondary outcomes were reduction in blood or iron requirements, endoscopic surface area regression and complications. RESULTS: Twenty patients were treated. The mean change in haemoglobin at six months was +12.6 g/l (95% confidence interval 11.7–24.3 g/l), paired t test p

Background Esophageal adenocarcinoma carries a poor prognosis and therefore treatment of early neoplasia arising in the precursor condition Barrett’s esophagus (BE) is desirable. Visible lesions arising in BE need endoscopic mucosal resection for accurate staging and removal. Resection modalities include a cap-based system with snare and custom-made multiband mucosectomy (MBM) devices (Duette, Cook Medical Ltd). A new MBM device has recently become available (Captivator, Boston Scientific Ltd). Objectives A retrospective pilot study to compare the efficacy, safety, specimen size and histology of endoscopic mucosal resection (EMR) specimens resected with two MBM devices (Cook Duette and Boston Captivator) in treatment naive patients undergoing endoscopic therapy for BE neoplasia. Methods Consecutive EMR procedures carried out by a single experienced endoscopist were analysed. All visible lesions were marked and resected using one of the two MBM devices. All resected specimens were analysed by the same two experienced pathologists. The resected specimens in both groups were analysed for maximum diameter, minimum diameter, surface area and depth. Results Twenty consecutive patients were analysed (18M + 2F; mean age 74) in the Duette group and 20 (17M + 3F; mean age 72) in the Captivator group. A total of 58 specimens were resected in the Duette and 63 in the Captivator group. Min diameter, max diameter, surface area and depth of the ER specimens resected by the Captivator device were significantly larger than that by the Duette device [min diameter 9.89 mm vs 9.07 mm (p = 0.019); max diameter: 13.54 mm vs 12.38 mm (p = 0.024); surface area: 135.40 mm2 vs 113.89 mm2 (p = 0.005); depth 3.71 mm vs 2.89 (p = 0.001)]. Conclusions These two MBM devices showed equivalent efficacy and safety outcomes, but the EMR Captivator device resected specimens with a larger area in the esophagus when compared with the Duette device. A possible advantage of this is in situations where en bloc resections with fewer EMRs are desirable for larger lesions.

Background and Aim Barrett's esophagus is associated with increased risk of esophageal adenocarcinoma. The optimal management of low‐grade dysplasia arising in Barrett's esophagus remains controversial. We performed a retrospective study from a tertiary referral center for Barrett's esophagus neoplasia, to estimate time to progression to high‐grade dysplasia/esophageal adenocarcinoma in patients with confirmed low‐grade dysplasia compared with those with downstaged low‐grade dysplasia from index presentation and referral. We analyzed risk factors for progression. Methods We analyzed consecutive patients with low‐grade dysplasia in Barrett's esophagus referred to a single tertiary center (July 2006–October 2018). Biopsies were reviewed by at least two expert pathologists. Results One hundred and forty‐seven patients referred with suspected low‐grade dysplasia were included. Forty‐two of 133 (32%) of all external referrals had confirmed low‐grade dysplasia after expert histopathology review. Multivariable analysis showed nodularity at index endoscopy (P, Our large retrospective study from a tertiary Barrett's center shows clear variability in the diagnosis of low‐grade dysplasia. Nodularity at index endoscopy and location of low‐grade dysplasia were associated with risk of progression in true low‐grade dysplasia patients. Endoscopic therapy and resources should potentially be more focused in this sub‐cohort of patients.