Can a mock medication-taking learning activity enable pharmacy students to experience the range of barriers and facilitators to medication adherence? An analysis informed by the Theoretical Domains Framework and COM-B model.

Background: Pharmacy professionals are well-placed to provide medication adherence support to patients. The Capability, Opportunity, Motivation-Behaviour (COM-B) and Theoretical Domains Framework (TDF) are two complementary models previously applied to medication-taking behaviour. Understanding the patient-specific barriers and facilitators to adherence using psychological frameworks from the early stages of pharmacy education enables the design and delivery of effective interventions. Objectives: To examine whether a novel 'mock medicine' learning activity enabled students to experience the range of barriers and facilitators to medication adherence using the COM-B and TDF. Methods: A mock medicine activity was conducted with students at pharmacy schools in three universities in the UK, Norway, and Australia over one week. Percentage adherence was calculated for five dosing regimens; theoretical framework analysis was applied to map reflective statements from student logs to COM-B and TDF. Results: A total of 349 students (52.6%) returned completed logs, with high overall mean adherence (83.5%, range 0-100%). Analysis of the 277 (79.4%) students who provided reflective statements included barriers and facilitators that mapped onto one (9%), two (29%) or all three (62%) of the COM-B components and all fourteen TDF domains (overall mean = 4.04; Uni 1 = 3.72; Uni 2 = 4.50; Uni 3 = 4.38; range 1-8). Most frequently mapped domains were 'Environmental context and resources' (n = 199; 72%), 'Skills' (n = 186; 67%), 'Memory, attention and decision-making' (184; 66%) and 'Beliefs about capabilities' (n = 175; 63%). Conclusions: This is the first study to utilise both COM-B and TDF to analyse a proxy measure of medication adherence in pharmacy education. Data mapping demonstrated that students experienced similar issues to patients when prescribed a short course of medication. Importantly, all the factors influencing medication-taking reported by students were captured by these two psychological frameworks. Future educational strategies will involve students in the mapping exercise to gain hands-on experience of using these psychological constructs in practice.

Effects of reseeding with perennial ryegrass, chicory or tyfon in pure stands or mixtures on lamb performance postweaning.

Many producers reseed old pasture (OP) with the objective of improving lamb performance. Old pasture was reseeded with perennial ryegrass plus white clover (RGC), RGC plus chicory (CG), RGC plus tyfon (TG), chicory (C), or tyfon (T) to generate five treatments. In addition, a contiguous block of OP was included in the study for comparative purposes. Lambs (n = 286) were assigned to one of the five treatments plus the section of OP from just after weaning until slaughter. Grazing commenced 45 days after the desiccation of OP for reseeding. All lambs were managed by rotational grazing and drafted for slaughter at a specified target BW. No concentrate supplement was offered during the study. For the RGC, CG, TG, C and T treatments and the OP section, the average herbage NDF concentrations were 524, 473, 402, 352, 256 and 565 (SE 15.4) g/kg DM; total grazing days were 5 213, 4 005, 4 466, 2 262, 3 496 and 3 677 (SE 629.3); BW gain to slaughter was 211, 175, 205, 211, 199 and 203 (SE 9.5) g/day; days to slaughter were 91, 100, 84, 86, 78 and 88 (SE 4.8). Compared to RGC, the C, T, CG and TG treatments had lower herbage NDF concentrations (P < 0.05), and there was no difference (P > 0.05) for BW gain or days to slaughter. Chicory yielded significantly fewer (P < 0.05) grazing days, but tended to give a higher (P < 0.06) carcass weight than RGC. It is concluded that neither reseeding OP with RGC, including either C or T with RGC as a multispecies sward, nor growing pure stands of C or T increased (P < 0.05) lamb BW gain from weaning to slaughter or reduced days to slaughter.

Historical and future trends in emergency pituitary referrals: a machine learning analysis.

PURPOSE: Acute pituitary referrals to neurosurgical services frequently necessitate emergency care. Yet, a detailed characterisation of pituitary emergency referral patterns, including how they may change prospectively is lacking. This study aims to evaluate historical and current pituitary referral patterns and utilise state-of-the-art machine learning tools to predict future service use. METHODS: A data-driven analysis was performed using all available electronic neurosurgical referrals (2014-2021) to the busiest U.K. pituitary centre. Pituitary referrals were characterised and volumes were predicted using an auto-regressive moving average model with a preceding seasonal and trend decomposition using Loess step (STL-ARIMA), compared against a Convolutional Neural Network-Long Short-Term Memory (CNN-LSTM) algorithm, Prophet and two standard baseline forecasting models. Median absolute, and median percentage error scoring metrics with cross-validation were employed to evaluate algorithm performance. RESULTS: 462 of 36,224 emergency referrals were included (referring centres = 48; mean patient age = 56.7 years, female:male = 0.49:0.51). Emergency medicine and endocrinology accounted for the majority of referrals (67%). The most common presentations were headache (47%) and visual field deficits (32%). Lesions mainly comprised tumours or haemorrhage (85%) and involved the pituitary gland or fossa (70%). The STL-ARIMA pipeline outperformed CNN-LSTM, Prophet and baseline algorithms across scoring metrics, with standard accuracy being achieved for yearly predictions. Referral volumes significantly increased from the start of data collection with future projected increases (p < 0.001) and did not significantly reduce during the COVID-19 pandemic. CONCLUSION: This work is the first to employ large-scale data and machine learning to describe and predict acute pituitary referral volumes, estimate future service demands, explore the impact of system stressors (e.g. COVID pandemic), and highlight areas for service improvement.

Effects of joining at 7 months, and ewe genotype, on the performance of ewes to 19 months of age and that of their progeny to slaughter.

Two ways of reducing the cost of replacements are increasing litter size and number of litters produced; thus, the total weight of lamb carcass output per ewe lifetime. The effects of ewe genotype on the performance of ewes lambing at 1 year and of their progeny to slaughter, and the effect of age at first joining (7 or 19 months) on BW at ~19 months and survival to joining at 19 months were evaluated over two consecutive years, using 460 ewe lambs from three genotypes: Belclare (Bel), Suffolk × Belclare (Suf × Bel) and ≥ 75% Suffolk ancestry (Suf75). Lambs from the three genotypes were at a similar proportion of mature BW and half of the lambs, within genotype, were allocated to be joined for the first time at 7 or 19 months. The ewe lambs were managed in a grass-based rotational-grazing system, except when housed from December to March on a grass silage-based diet. Belclare ewes had larger litters (P < 0.001), reared more lambs per ewe joined (P < 0.01), were lighter at lambing and at 19 months (P < 0.01), were of smaller body size at 19 months (P < 0.001) and their progeny were lighter at weaning (P < 0.05) relative to Suf75 genotype; the Suf × Bel ewes were intermediate for most traits but had a significantly lower litter size (P < 0.05) than Bel ewes. Progeny from Suf × Bel ewes were 17 days younger at slaughter (P < 0.01) relative to those from Bel ewes. Ewe genotype had no effect (P > 0.05) on lamb mortality (born dead, total mortality to weaning), lambing assistance, number of ewes that failed to lamb, or on ewe survival to 19 months of age. Increasing ewe BW at joining increased the probability (P < 0.001) of rearing at least one lamb and this effect was consistent across genotypes. There were significant relationships (P < 0.001) between ewe BW at lambing and lamb BW at birth and at weaning of 0.053 (SE 0.0089) kg and 0.29 (SE 0.049) kg, respectively. Ewes that lambed at 1 year were 2 kg lighter (P < 0.001) at 19 months of age and had a smaller body size (P < 0.01) relative to those not joined. It is concluded that ewe genotype had a significant effect on number of lambs reared, and thus lamb carcass output. Whilst lambing at 1 year reduced BW by 2 kg when joined at 19 months, it did not affect ewe survival to that stage.

Comparative Study of Dissolution for Cannabidiol in EU and US Hemp Oil Products by HPLC.

For the first time, a simple and robust HPLC method has been developed for dissolution studies for cannabidiol (CBD) in hemp oil products. An isocratic elution of samples performed on SOLAS™ C18 150 mm x 4.6 mm, 5 µm column with a mobile phase consisting of 75:25 acetonitrile-water v/v, delivered at a flow rate 1.5 mL/minutes to variable wavelength detector using 214 nm. An in-house validated assay test was executed for calculating the purity of hemp oil products and also for considering the dissolution medium to be used. For dissolution studies, equivalent of 5 mg and/or 10 mg of the active was introduced into 500 mL of simulated gastric and intestinal fluids separately, and dissolution was performed at 50 rpm using paddles for 180 min. Dissolution profiles for hemp oil products purchased from the United States and Europe were compared. Additionally, dissolution testing was conducted to study the effect of percentage CBD release on increased agitation speed of 75 and 100 rpm and also, on extended dissolution runtime of 240 min.

Effects of grass and maize silage feed value, offering soybean meal with maize silage, and concentrate feed level in late pregnancy, on ewe and lamb performance.

In many countries, daily herbage accumulation on pasture declines towards zero during the winter period; thus, many pregnant ewes are housed and offered conserved forages supplemented with concentrate prior to parturition. The effects of forage type and feed value (FV), offering soybean meal with maize silage during mid and late pregnancy, and concentrate feed level in late pregnancy on the performance of ewes and their progeny (to slaughter) were evaluated. Ewes (n = 151) were assigned to one of nine treatments from mid-pregnancy until lambing. Medium FV and high FV grass silages (metabolisable energy concentrations of 10.7 and 12.0 MJ/kg DM) were offered ad libitum supplemented with either 15 or 25 kg concentrate/ewe during late pregnancy. Low and high DM maize silages (starch concentrations of 80 and 315 g/kg DM) were offered ad libitum either alone or with soybean meal (200 g/d) and supplemented with 15 kg concentrate during late pregnancy. A final treatment consisted of high FV grass silage supplemented with 5 kg soybean/ewe over the final 4 weeks of pregnancy. Ewes and lambs were put to pasture in a rotational-grazing system within 3 days of lambing. There were no interactions (P > 0.05) between grass silage FV and concentrate feed level for ewe or lamb traits. Increasing grass silage FV increased food intake (P < 0.001) during late pregnancy, ewe BW and body condition score (BCS) at lambing (P < 0.001), lamb BW at birth (P < 0.001) and weaning (P < 0.05), and reduced age at slaughter (P = 0.06). Increasing concentrate feed level increased metabolisable energy (P < 0.05) intake during late pregnancy but had no effect (P > 0.05) on ewe or lamb performance. Increasing maize DM at harvest and offering soybean meal with maize silage increased food intake (P < 0.001) and ewe BW and BCS at lambing (P < 0.05 or P < 0.01). Offering soybean meal with maize silage increased lamb BW at birth (P < 0.01) and reduced age at slaughter (P < 0.05). Reducing supplementation of high FV grass silage to 5 kg of soybean meal had no effect (P > 0.05) on animal performance. Replacing grass silage with maize silage did not affect (P > 0.05) BW gain of lambs. It is concluded that increasing the FV of the grass silage offered during pregnancy had the greatest positive impact on ewe and lamb performance.

The effect of fresh gas flow during induction of anaesthesia on sevoflurane usage: a quality improvement study.

Reducing fresh gas flow during inhalational anaesthesia results in cost savings and decreases environmental impact. We are interested in the influence of fresh gas flow on the early (induction) phase of overall fresh gas flow and vapour consumption. This stage is often excluded in studies of fresh gas flow. Data were collected from 3199 sevoflurane anaesthetics over an 11-month period in four operating theatres. We determined fresh gas flow at different stages of anaesthesia, and developed an explanatory model for the influence of the 'induction' period. Following a three-month collection of baseline data we emphasised the importance of the early phase to our department repeatedly over a two-week period. We explored the relationship between fresh gas flow and total vapour usage, and used a simple mathematical model to explore the effect of changes in the fresh gas flow and duration of the 'induction' phase. Mean fresh gas flow was 1.15 l.min-1 in the baseline period and 0.91 l.min-1 in the two months following our educational effort (p = 0.0005). In the following six months, mean fresh gas flow was 1.17 l.min-1 (p = 0.7726 compared with baseline). These results were driven by changes in both fresh gas flow and duration of the initial high-flow period. We found some correlation (R2  = 0.85) between overall fresh gas flow and vapour consumption; a 1 l.min-1 increase in fresh gas flow consumes an additional 18 ml.hr-1 of liquid sevoflurane. This preliminary study demonstrates that an episode of high fresh gas flow at the start of anaesthesia has a large and modifiable effect on overall fresh gas flow and vapour consumption. We also confirmed the linear relationship between fresh gas flow and vapour usage.

Cervical mucus sialic acid content determines the ability of frozen-thawed ram sperm to migrate through the cervix.

The aim of this study was to investigate the properties and to functionally characterize the cervical mucus that modulates sperm transport through the cervix by using ewe breeds with a divergent pregnancy rate (Belclare and Suffolk; high and low, respectively) following cervical insemination using frozen-thawed semen. Sperm number, as well as sialic acid and fucose content in both the channels and in the lumen of different regions of the cervix were quantified in inseminated Belclare and Suffolk ewes. Expression of glycosyltransferase and MUC genes, glycosidase activity and sialic acid speciation in follicular phase cervical tissue and mucus were assessed. More spermatozoa were found in the cervical channels in the region closest to the cervical os in Belclare than Suffolk ewes (P < 0.05) and Suffolk ewes had a higher sialic acid content in the cervical channels than Belclare ewes (P < 0.05) in all regions of cervix. Suffolk ewes had significantly higher expression of FUT1, ST6GAL1 and MUC5AC than Belclare ewes. There was no difference between the breeds in glycosidase activity (P > 0.05). Levels of Neu5Ac were higher in Belclare than Suffolk ewes (P < 0.05) and levels of Neu5Gc was higher in Suffolk than Belclare ewes (P < 0.05). Competitive sperm penetration assays demonstrated that frozen-thawed sperm progression increased when cervical mucus was incubated with sialyllactose prior to a sperm penetration test (P < 0.05). These results suggest that the difference between Belclare and Suffolk ewes in sperm transport with frozen-thawed semen is due to the higher concentration of sialic acid within channels, which binds to spermatozoa and reduces their ability to traverse the cervix.

Preoperative gastric residual volumes in fasted patients measured by bedside ultrasound: a prospective observational study.

The purpose of this prospective observational study was to measure gastric volumes in fasted patients using bedside gastric ultrasound. Patients presenting for non-emergency surgery underwent a gastric antrum assessment, using the two-diameter and free-trace methods to determine antral cross-sectional area (CSA). Gastric residual volume (GRV) was calculated using a validated formula. Univariate and multivariable analyses were performed to examine any potential relationships between 'at risk' GRVs (>100 ml) and patient factors. Two hundred and twenty-two successful scans were performed; of these 110 patients (49.5%) had an empty stomach, nine patients (4.1%) had a GRV >100 ml, and a further six patients (2.7%) had a GRV >1.5 ml/kg. There was no significant relationship between at risk GRV and obesity, diabetes mellitus, gastro-oesophageal reflux disease or opioid use, although our study had insufficient power to exclude an influence of one or more of these factors. Our results indicate that despite compliance with fasting guidelines, a small percentage of patients still have GRVs that pose a pulmonary aspiration risk. Anaesthetists should consider this background incidence when choosing anaesthesia techniques for their patients. While future observational studies are required to determine the role of preoperative bedside gastric ultrasound, it is possible that this technique may assist anaesthetists in identifying patients with 'at risk' GRVs.

An independent validation study of loci associated with nematode resistance in sheep.

Gastrointestinal nematode infection is a constraint on sheep production worldwide. Selective breeding programmes to enhance resistance to nematode infection are currently being implemented in a number of countries. Identification of loci associated with resistance to infection or causative mutations for resistance would enable more effective selection. Loci associated with indicator traits for nematode resistance has been identified in previous studies. In this study, Scottish Blackface, Texel and Suffolk lambs were used to validate the effects at eight genomic regions previously associated with nematode resistance (OAR3, 4, 5, 7, 12, 13, 14, 21). No SNP was significantly associated with nematode resistance at the region-wide level but seven SNPs in three of the regions (OAR4, 12, 14) were nominally associated with trichostrongyle egg count in this study and six of these were also significant when fitted as single SNP effects. Nematodirus egg count was nominally associated with SNPs on OAR3, 4, 7 and 12.

Plane of nutrition during the rearing phase for replacement ewes of four genotypes: II - effects on performance during first pregnancy and to weaning, and of their progeny.

Plane of nutrition (PN) offered to ewe replacements during the rearing phase (8 to 17 months) and first pregnancy may affect reproductive and progeny performance when replacements are joined at ~19 months. The effects of PN offered to spring-born ewe replacements during their first winter (winter_1), second summer (summer_2) and subsequent winter (mid and late pregnancy; winter_2) were evaluated, over 3 consecutive years, using 287 ewe lambs of four genotypes: Charmoise×Scottish Blackface (C×SBF), Belclare×SBF (Bel×SBF), Belclare (Bel) and Belclare×SBF (heterozygous for a gene, either FecGH or FecXG mutations, that increases ovulation rate: BelMG×SBF). Ewe lambs were offered, daily, a deferred-grazed herbage allowance (HA) of either 0.75 or 1.75 kg dry matter during winter_1. During summer_2 the replacements were set-stocked to maintain sward heights of 4 or 6 cm. Ewes were housed during mid and late pregnancy (winter_2) and offered either medium or high-feed-value grass silage, thus yielding a 2×2×2 factorial design. Increasing HA during winter_1 and residual sward height during summer_2 increased ewe BW post joining (P0.05) on number of lambs reared. Increasing HA during winter_1 increased lamb BW at birth (P<0.05) and reduced carcass fat score (P<0.05). There was a significant interaction between PN offered during winter_1 and summer_2 for average daily gain (ADG) from birth to 5 weeks (P<0.01) and from 5 to 10 weeks (P<0.05): progeny of replacements that experienced either a high or low PN throughout the rearing phase had lower ADG than lambs born to the replacements that experienced a low PN only during winter_1 or summer_2. Increasing PN during winter_2 increased ewe BW (P<0.001), lamb BW at birth (P<0.001) and weaning (P<0.05) and reduced the proportion of ewes that failed to lamb (P<0.01). Ewe genotype had a significant (P<0.001) effect on litter size (1.48 to 2.45), number of lambs reared, lamb BW at birth and weaning, lamb ADG and age at slaughter. It is concluded that, among the nutrition treatments examined, increasing PN during mid and late pregnancy had the greatest effect on ewe and progeny performance. Although increasing PN offered during winter_1 increased lamb birth BW, PN offered during summer_2 had no effect on lamb performance. There were no significant interactions between PN and ewe genotype.

Plane of nutrition during the rearing phase for replacement ewes of four genotypes: I - effects on growth and development, and on ovulation rate at first joining.

Plane of nutrition (PN) offered to ewe replacements during the rearing phase (8 to 17 months) affects BW at joining and may affect reproductive performance when joined at ~19 months. The effects of PN offered to ewe replacements during their first winter (winter_1) and second summer (summer_2) were evaluated over 3 consecutive years, using 287 spring-born ewe lambs, representing four genotypes (Belclare (Bel), Charmoise×Scottish Blackface (C×SBF), Bel×SBF (Bel×SBF), Bel×SBF that were heterozygous either FecGH or FecXG mutations that increase ovulation rate (OR) (BelMG×SBF)). During extended (deferred) grazing in winter_1 (November to March) the lambs were offered herbage DM allowances (HA) of 0.75 (L) or 1.75 (H) kg/day. During summer_2 (March to August) the replacements were set stocked to maintain sward heights of 4 (L) or 6 (H) cm, thus yielding a 2×2 factorial design for the nutrition treatments (71 to 73 animals/treatment). Increasing HA during winter_1, and residual sward height during summer_2, increased (P0.05) on OR. Mean BW at joining was 53.3, 56.8, 56.6 and 61.7 (SEM 0.74) kg for ewes offered the LL, LH, HL and HH treatments, respectively. Mean OR of C×SBF and BelMG×SBF differed (P<0.001) from that of Bel×SBF by -0.58 and +0.47, respectively. Correlations between linear measurements (LMs) of body size and BW at 7, 12 and 17 months were all positive and significant (P<0.001). The precision of the set of LMs as a predictor of BW at joining improved with age (R 2 0.46, 0.54 and 0.74 at 7, 12 and 17 months) but BW at a given age was a better predictor at all age time points. Chest girth was the best predictor, among the LMs, of BW at joining and its explanatory power increased with age. Equations developed using LMs at 7, 12 or 17 months, to predict BW at joining exhibited biases of -2.1, -1.5 and +0.9 kg, respectively. It is concluded that whilst altering PN during the rearing phase changed BW by 16% it had no effect on OR. There was no interaction (P>0.05) between the PN offered during winter_1 and summer_2, or PN and ewe genotype for BW at joining or OR. LMs, either at 7, 12 or 17 months of age, are useful predictors of BW at joining.

Cobalt supplementation, alone or in combination with vitamin B and selenium: Effects on lamb performance and mineral status.

The effects of supplementing lambs with Co, either alone or in combination with vitamin B and Se, were evaluated over the period from weaning to selection for slaughter. Two hundred and five recently weaned (at 14 wk [SD 0.44] of age), spring-born lambs of various crossbred genotypes were stratified, within genotype, by BW and gender and allocated to 3 groups, which were then allocated at random to one of the following treatments: no supplement (Control), a supplement containing Co only (Co-only), or a supplement containing Co, vitamin B, and Se (VitMin). The concentration of Co in the Co-only supplement was 2.1 mg/mL (as cobalt sulfate); the VitMin supplement contained Co (10 mg/mL, as cobalt acetate), vitamin B (200 µg/mL), and Se (0.25 mg/mL, as sodium selenite). Lambs on the Co-only and VitMin treatments received, every 14 d, 10 and 2.1 mL, respectively, of oral drench. Lambs were managed in a rotational-grazing system from 12 July (swards predominantly ) and selected for slaughter, on the basis of BW, after 45, 73, or 115 d; all remaining lambs were slaughtered on d 157. Supplementation, with Co-only or VitMin, increased BW at slaughter ( < 0.01), ADG ( < 0.001), carcass weight ( < 0.001), dressing percentage ( < 0.001), carcass fat score ( < 0.05), and the proportion selected for slaughter by d 73 ( < 0.05) and 115 ( < 0.01). Relative to the Control, the benefit to ADG from supplementation increased as the season advanced. Therefore, supplementation did not alter ( = 0.82) ADG between d 1 and 45 but increased ADG over the intervals d 45 to 73 ( < 0.01), d 73 to 115 ( < 0.001), and d 115 to 157 ( < 0.001). There was no difference ( > 0.05) between the Co-only and the VitMin treatments for ADG, carcass weight, fat score, or dressing percentage. Supplementation with Co increased liver Co concentration ( < 0.001), and lambs supplemented with VitMin had greater liver Co concentration than lambs supplemented with Co-only ( < 0.05); the concentrations were 0.18, 0.85, and 1.18 µmol/L for the Control, Co-only, and VitMin treatments, respectively. Treatment had no effect ( = 0.65) on the concentration of Se in kidney tissue. It is concluded that supplementation with Co increased lamb performance after weaning, that response to supplementation increased as the grazing season progressed, and that no extra benefit in performance resulted from augmenting supplementary Co with B and Se.

Immunogenomics of gastrointestinal nematode infection in ruminants - breeding for resistance to produce food sustainably and safely.

Gastrointestinal nematode (GIN) infection of ruminants represents a major health and welfare challenge for livestock producers worldwide. The emergence of anthelmintic resistance in important GIN species and the associated animal welfare concerns have stimulated interest in the development of alternative and more sustainable strategies aimed at the effective management of the impact of GINs. These integrative strategies include selective breeding using genetic/genomic tools, grazing management, biological control, nutritional supplementation, vaccination and targeted selective treatment. In this review, the logic of selecting for "resistance" to GIN infection as opposed to "resilience" or "tolerance" is discussed. This is followed by a review of the potential application of immunogenomics to genetic selection for animals that have the capacity to withstand the impact of GIN infection. Advances in relevant genomic technologies are highlighted together with how these tools can be advanced to support the integration of immunogenomic information into ruminant breeding programmes.

A pharmacological characterization of GABA, THIP and DS2 at binary α4ß3 and ß3δ receptors: GABA activates ß3δ receptors via the ß3(+)δ(-) interface.

There is growing evidence that GABA (γ-aminobutyric acid) can activate GABAA receptors (GABAARs) in the absence of an α subunit. In this study, we compared the pharmacology of homomeric and binary α4, ß3 or δ subunits with ternary α4ß3δ to identify subunit interfaces that contribute to the pharmacology of GABA, THIP, and DS2, and the antagonists, Zn(2+), gabazine and bicuculline. ß3δ receptors form functional GABA-gated channels when expressed in Xenopus oocytes with a pharmacology that differs to homomeric ß3, binary α4ß3 and ternary α4ß3δ receptors. GABA had similar potency at α4ß3 and ß3δ receptors (25µM and 26µM, respectively) but differed at α4ß3δ receptors where GABA exhibited a biphasic concentration-response (EC50 (1)=12.6nM; EC50 (2)=6.3µM). THIP activated ß3δ receptors (EC50=456µM) but was a more potent activator of α4ß3 (EC50=27µM) and α4ß3δ receptors (EC50 (1)=27.5nM; EC50 (2)=29.5µΜ), indicating that the α4 subunit significantly contribute to its potency. The δ-preferring modulator, DS2 had marginal or no effect at ß3δ and α4ß3 receptors, indicating a role for both the α4 and δ subunits for its potency. Gabazine inhibited GABA-elicited currents at ß3δ receptors whereas bicuculline activated these receptors. Mutational analysis verified that GABA binds to the ß3(+)δ(-) interface formed by the ß3 and δ subunits. In conclusion, evaluating agents against binary GABAARs such as ß3δ and α4ß3 receptors enables identification of interfaces that may contribute to the pharmacology of the more complex ternary α4ß3δ receptors.

Efficacy and safety of inhaled calcium lactate PUR118 in the ozone challenge model--a clinical trial.

BACKGROUND: The ozone challenge model can be used to assess the efficacy of anti-inflammatory compounds in early phases of clinical drug development. PUR118, a calcium salt based formulation engineered in the iSPERSE(TM) dry powder delivery technology, is a novel anti-inflammatory drug for COPD. Here we evaluated the efficacy and safety of three doses of PUR118 in attenuating ozone-induced airway inflammation in healthy volunteers. METHODS: In a single-blind, phase 1B proof of concept study, 24 subjects were enrolled to sequentially receive three doses of PUR118 (5.5 mg, n = 18; 11.0 mg, n = 18; 2.8 mg, n = 16). Each dose was inhaled 3 times (1, 13, 25 h, preceded by 2 puffs salbutamol) before the ozone exposure (250 ppb, 3 h intermittent exercise). Sputum was induced 3 h after the end of exposure. RESULTS: Sputum neutrophils, sputum CD14+ cells, as well as concentrations of IL1B, IL6, IL8, MMP9, and TNFA in sputum supernatant significantly increased after ozone exposure (n = 24). The percentage of sputum neutrophils (n = 12 who completed all treatments) did not change following treatment with different doses of PUR118. The high dose treatment group (n = 16) showed a decrease in the percentage and number of sputum macrophages (p ≤ 0.05) as well as a decrease in blood neutrophils (p = 0.04), and an increase in blood CD14 + cells (p = 0.04) compared to baseline. All dosages of PUR118 were safe and well tolerated. CONCLUSION: Ozone challenge resulted in the expected and significant increase of sputum inflammatory parameters. Treatment with multiple rising doses of PUR118 was safe and three applications within 25 h prior to the ozone challenge had small effects on ozone-induced airway inflammation. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01690949. Registered 12 September 2012.

Effects of shearing, forage type and feed value, concentrate feed level, and protein concentration on the performance of housed finishing lambs.

The effects of high and medium feed value grass silage or maize silage (MS), each offered with a range of concentrate feed levels, and ad libitum concentrate on the performance of finishing lambs, which were either shorn or unshorn, were evaluated. Three silages were used: 1 medium feed value (MFV) and 1 high feed value (HFV) grass silage (DM digestibility [DMD] of 71.3% and 74.5%, respectively) and 1 MS (DM and starch concentrations of 30.9% and 35.3% DM, respectively). The 3 silages were offered ad libitum with daily allowances of 0.4, 0.8, or 1.2 kg concentrate per lamb. Two additional treatments were: 1) MS supplemented with 0.4 kg of a low CP (LP) concentrate and 2) concentrate offered ad libitum with 0.5 kg/d of HFV grass silage. These 11 dietary treatments were offered to 264 crossbred Suffolk lambs (initial BW = 39.0 kg), half of which were unshorn and half of which were shorn, for a 54-d finishing period, resulting in 22 treatments. Shearing increased forage DMI (P < 0.001) but did not alter (P > 0.05) carcass weight, carcass gain, or ADG, and there was no interaction with dietary treatment. Reducing CP concentration of the concentrate offered with MS did not alter (P > 0.05) feed intake or lamb performance. Increasing concentrate feed level increased feed DMI and lamb performance (P < 0.001). The linear response in ADG to increased concentrate supplementation was greater (P = 0.012) for MFV than HFV grass silage, and a corresponding difference in carcass gain approached significance (P = 0.075). The linear response was greater for grass silage than for MS for ADG (P < 0.01) and carcass gain (P = 0.019). The response in lamb performance from increased concentrate supplementation was linear for HFV grass silage and MS but quadratic (P < 0.05) for the MFV grass silage, reflecting the large response for this silage when concentrate supplement was increased from 0.4 to 0.8 kg. Relative to the MFV grass silage supplemented with 0.8 kg concentrate, the potential concentrate-sparing effects of the HFV grass silage and MS were 0.16 and 0.32 kg concentrate per lamb daily, respectively. It is concluded that shearing lambs that are housed during finishing reduces the efficiency of the conversion of ME to carcass gain. Maize silage can replace HFV grass silage in the diet of finishing lambs. The total dietary protein concentration for finishing lambs can be reduced to 9% DM without adversely affecting carcass gain.

A hydrophobic area of the GABA ρ1 receptor containing phenylalanine 124 influences both receptor activation and deactivation.

Experimental evidence suggests that GABA ρ1 receptors are potential therapeutic targets for the treatment of a range of neurological conditions, including anxiety and sleep disorders. Homology modelling of the GABA ρ1 extracellular N-terminal domain has revealed a novel hydrophobic area that extends beyond, but not including the GABA-binding site. Phenylalanine 124 (F124) is predicted to be involved in maintaining the structural integrity of the orthosteric-binding site. We have assessed the activity of a series of GABA ρ1 receptors that incorporate a mutation at F124. Wild-type and mutant human GABA ρ1 subunits were expressed in Xenopus laevis oocytes and AD293 cells, and the pharmacology and kinetic properties of the receptors were measured using electrophysiological analysis. Mutation of F124 had minimal effect on receptor pharmacology. However, the rate of deactivation was significantly increased compared to wild type. This study provides further information about the role of residues within a novel hydrophobic area of the GABA ρ1 receptor. This knowledge can help future studies into the design of potent and subtype-selective ligands with therapeutic value.

The effects of an epithelial barrier protective cationic aerosol on allergen-induced airway inflammation in asthma: a randomized, placebo-controlled clinical trial.

Inhaled cationic airway lining modulator (iCALM) is a cationic aerosol therapy comprised of 1.29% calcium chloride dissolved in 0.9% isotonic saline that enhances the biophysical barrier function of the airway lining fluid and primes the host defense response. It's ability to attenuate bronchitis caused by inhaled particles was investigated using an allergen-inhalation model in a proof-of-concept study. In a randomized, double-blind, placebo-controlled cross-over trial of 6 mild atopic steroid-naïve asthmatic subjects, 3 doses of iCALM were well tolerated and they attenuated allergen-induced increase in sputum eosinophils, and levels of IL-5, MCP-1 and eotaxin. This study provides an opportunity to investigate the role of enhancing epithelial barrier to decrease airway inflammation provoked by inhaled particles in a variety of airway diseases.