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Indirect immunofluorescence is usually restricted to 3-5 markers per preparation, limiting analysis of coexistence. A solution containing 2-mercaptoethanol and sodium dodecyl sulfate (2-ME/SDS) can elute indirect immunofluorescence labelling (i.e. primary antisera followed by fluorophore-conjugated secondary antisera) and has been used for sequential staining of sections. The aim of this study was to test whether 2-ME/SDS is effective for eluting indirect immunofluorescent staining (with primary antisera visualised by fluorophore-coupled secondary antisera) in wholemount preparations. We also analysed how 2-ME/SDS may work and used this understanding to devise additional uses for immunofluorescence in the nervous system. 2-ME/SDS appears to denature unfixed proteins (including antisera used as reagents) but has much less effect on antigenicity of formaldehyde-fixed epitopes. Moieties linked by strong biotin-streptavidin bonds are highly resistant to elution by 2-ME/SDS. Two primary antisera raised in the same species can be applied without spurious cross-reactivity, if a specific order of labelling is followed. The first primary antiserum is followed by a biotinylated secondary, then a tertiary of fluorophore-conjugated streptavidin. The preparation is then exposed to 2-ME/SDS, which has minimal impact on labelling by the first primary/secondary/tertiary combination. However, when this is followed by a second primary antiserum (raised in the same species), followed by a fluorophore-conjugated secondary antiserum, the intervening 2-ME/SDS exposure prevents cross-reactivity between primary and secondary antisera of the two layers. A third property of 2-ME/SDS is that it reduces lipofuscin autofluorescence, although it also raises background fluorescence and strongly enhances autofluorescence of erythrocytes. In summary, 2-ME/SDS is easy to use, cost-effective and does not require modified primary antisera. It can be used as the basis of a multi-layer immunohistochemistry protocol and allows 2 primary antisera raised in the same species to be used together.

Purpose – This paper outlines a contemporary conceptual framework for the embedding of experiential learning into a business consultancy module. Experiential learning is a fundamental teaching approach that allows students to apply theory into a working business context. Design/methodology/approach – As a conceptual and not an empirical paper, the methodological approach was to draw upon the literature reviewed and to build a framework to support student learning through a business consultancy module. Findings – Exploration of the literature suggests that there are four elements critical to student learning in experiential learning environments: action, reflection, social and context. A framework has been developed utilising these elements with the interaction between the factors being key to developing learning. Research limitations/implications – So far, the framework is conceptual, and further research is needed to explore its use when staff members are developing these types of modules and to understand the interaction of the factors over the course of the student learning experience. Originality/value – The originality comes from the intersection and interaction between the core factors in experiential learning, which enables this framework to move thinking beyond more static models and hence work in a more fluid student learning environment.

Background and AimsViscerofugal neurons (VFNs) have cell bodies in the myenteric plexus and axons that project to sympathetic prevertebral ganglia. In animals they activate sympathetic motility reflexes and may modulate glucose metabolism and feeding. We used rapid retrograde tracing from colonic nerves to identify VFNs in human colon for the first time, using ex vivo preparations with multi-layer immunohistochemistry.MethodsColonic nerves were identified in isolated preparations of human colon and set up for axonal tracing with biotinamide. After fixation, labeled VFN cell bodies were subjected to multiplexed immunohistochemistry for 12 established nerve cell body markers.ResultsBiotinamide tracing filled 903 viscerofugal nerve cell bodies (n = 23), most of which (85%) had axons projecting orally before entering colonic nerves. Morphologically, 97% of VFNs were uni-axonal. Of 215 VFNs studied in detail, 89% expressed ChAT, 13% NOS, 13% calbindin, 9% enkephalin, 7% substance P and 0 of 123 VFNs expressed CART. Few VFNs contained calretinin, VIP, 5HT, CGRP, or NPY. VFNs were often surrounded by dense baskets of axonal varicosities, probably reflecting patterns of connectivity; VAChT+ (cholinergic), SP+ and ENK+ varicosities were most abundant around them. Human VFNs were diverse; showing 27 combinations of immunohistochemical markers, 4 morphological types and a wide range of cell body sizes. However, 69% showed chemical coding, axonal projections, soma-dendritic morphology and connectivity similar to enteric excitatory motor neurons.ConclusionViscerofugal neurons are present in human colon and show very diverse combinations of features. High proportions express ChAT, consistent with cholinergic synaptic outputs onto postganglionic sympathetic neurons in prevertebral ganglia.

Abstract Background Constipation severity is often defined by symptoms including feelings of complete evacuation, straining, stool frequency and consistency. These descriptors are mostly obtained in the absence of laxative use. For many constipated patients laxative usage is ubiquitous and long standing. Our aim was to determine the impact of laxative use upon the stereotypic constipation descriptors. Methods Patients with confirmed slow transit constipation completed 3-week stool diaries, detailing stool frequency and form, straining, laxative use and pain and bloating scores. Each diary day was classified as being under laxative affect (laxative affected days) or not (laxative unaffected days). Unconditional logistic regression was used to assess the affects of laxatives on constipation symptoms. Results Ninety four patients with scintigraphically confirmed slow transit constipation were enrolled in the study. These patients reported a stool frequency of 5.6 ± 4.3 bowel motions/week, only 21 patients reported P P Conclusions The reporting of frequent and loose stools with abdominal pain and/or bloating is common in patients with slow transit constipation. While laxative use is a significant contributor to altering stool frequency and form, laxatives have no apparent affect on pain or bloating or upon a patients feeling of complete evacuation. These factors need to be taken into account when using constipation symptoms to define this population.

Business Schools in Great Britain are currently facing a number of major challenge due to the conflicting objectives that need to be met. The movement towards a metric-driven, performative approach that has occurred in the last decade has meant that the need to meet set measures that focus on the outcomes of students’ study have become imperative. However, focusing solely on these outcomes neglects a second set of objectives that relate to the journey students take during their studies. The need to ensure that both the process and the outcome of students’ studies are stressed is particularly challenging in British Business Schools which are often characterised by a large and diverse student and staff population. This think piece posits that by using a loose/tight cultural approach alongside a Schrödinger's leadership style, leaders can mitigate some of the challenges that are currently faced in the sector.

Background Irritable bowel syndrome with diarrhoea is characterised by frequent, loose or watery stools with associated urgency, resulting in marked reduction of quality of life. Ondansetron, a 5-hydroxytryptamine-3 receptor antagonist, has been shown to benefit patients with irritable bowel syndrome with diarrhoea. Objective To evaluate the effect of ondansetron in irritable bowel syndrome with diarrhoea. Design Phase III, parallel-group, randomised, double-blind, multicentre, placebo-controlled trial in 400 patients, with embedded mechanistic studies. Setting Hospital, primary care and community. Participants Eighty participants meeting Rome IV criteria for irritable bowel syndrome with diarrhoea. Intervention Ondansetron 4 mg (dose titrated up to two tablets three times a day) or matched placebo for 12 weeks. Main outcome measures Clinical – Primary patient-reported end point was % ‘Food and Drug Administration-defined responders’ over 12 weeks. Secondary end points were worst abdominal pain intensity, worst urgency, stool consistency, stool frequency, anxiety, depression and dyspepsia at 12 and 16 weeks. Main outcome measures Mechanistic – Whole gut transit time, faecal water, protease (FP), bile acids and assessment of rectal sensitivity using a barostat. Results Clinical – The study closed early due to slow recruitment. Between 1 January 2018 and 11 May 2020, 80 patients were recruited and randomised (20% of target), 37 to ondansetron, 43 to placebo. Discontinuations (4 ondansetron; 2 placebo) meant 75 completed the 12-week trial treatment. There were four protocol violations. In the intention-to-treat analysis, 15 (40.5%) on ondansetron were primary end-point responders (95% CI 24.7% to 56.4%), and 12 (27.9%) on placebo (95% CI 14.5% to 41.3%), p = 0.19, adjusted OR 1.93 (0.73, 5.11). Pain intensity reduction occurred in 17 (46.0%) on ondansetron (95% CI 29.9% to 62.0%) and 16 (37.2%) on placebo (95% CI 22.8% to 51.7%), p = 0.32. Improvement in stool consistency occurred in 25 (67.6%) on ondansetron (95% CI 52.5% to 82.7%) and 22 (51.2%) on placebo (95% CI 36.2% to 66.1%), p = 0.07. Use of rescue medication, loperamide, was lower on ondansetron [7 (18.9%) vs. 17 (39.5%)]. Average stool consistency in the final month of treatment reduced significantly more on ondansetron, adjusted mean difference –0.5 [standard error (SE) 0.25, 95% CI (–1.0 to –0.02), p = 0.042]. Ondansetron improved dyspepsia score (SFLDQ), adjusted mean difference –3.2 points [SE 1.43, 95% CI (–6.1 to –0.4), p = 0.028]. There were no serious adverse events. Mechanistic – mean (SD). Ondansetron increased whole gut transit time between baseline and week 12 by 3.8 (9.1) hours on ondansetron, significantly more than on placebo –2.2 (10.3), p = 0.01. Mean volume to reach urgency threshold using the barostat increased on ondansetron by 84 (61) ml and 38 (48) ml on placebo, n = 8, p = 0.26. Ondansetron did not significantly alter protease, faecal water or bile acids. Changes in referral pathways substantially reduced referrals, impairing recruitment, which meant the study was underpowered. Conclusion Our results are consistent with previous studies and confirmed ondansetron improves stool consistency and urgency but showed minor effect on pain. We plan to undertake a simplified version of this trial overcoming the changed referral pathways by recruiting in primary care, using software linked to primary care records to identify and randomise patients with irritable bowel syndrome with diarrhoea to ondansetron or placebo and remotely follow their progress; thus minimising barriers to recruitment. Trial registration This trial is registered as ISRCTN17508514. Funding This project was funded by the National Institute for Health and Care Research (NIHR) Efficacy and Mechanism Evaluation programme and will be published in full in Efficacy and Mechanism Evaluation; Vol. 10, No. 9. See the NIHR Journals Library website for further project information. Plain language summary Background Irritable bowel syndrome with diarrhoea is characterised by frequent, loose, or watery bowel movements with marked reduction of quality of life. A previous small study suggested ondansetron benefits patients with irritable bowel syndrome with diarrhoea. Methods A clinical trial aiming to recruit 400 patients meeting established criteria for irritable bowel syndrome with diarrhoea from 18 centres throughout the UK. Patients received either ondansetron or placebo for 12 weeks but neither the investigator and nor patient could tell which they were receiving. They recorded their worst abdominal pain, stool frequency and consistency daily. The main end point was the proportion of patients meeting a standard recommended by the U.S. Food and Drug Administration (FDA). Being called a “FDA responder” meant they showed reductions to both pain and days with loose bowel movements. Other less important end points included pain intensity, stool consistency and frequency. We also measured the time for content to pass through the gut (whole gut transit time). Results The study closed early due to slow recruitment with 80 patients randomised. There were 40.5% of responders in the ondansetron group and 27.9% in the placebo group; however, due to low numbers these differences could be due to chance. Ondansetron produced a significant improvement in average stool consistency in the final month of treatment. Ondansetron slowed whole gut transit time which increased from baseline to 12 weeks by a mean of 3.8 hours, while it fell 2.2 hours on placebo, a difference unlikely to be due to chance. Conclusion These results are consistent with previous studies showing that ondansetron improves stool consistency and slows transit. However, because the numbers recruited were smaller than planned, the apparent improvement in “FDA responder” rate could have been due to chance. A further larger trial is needed to confirm the benefit of ondansetron which should be done in primary care where most patients are to be found. Scientific summary Background Irritable bowel syndrome (IBS), which affects around 10% of the population, accounts for 1.8 million consultations/year in primary care in England and Wales (0.6 million patients). Symptoms of IBS with diarrhoea (IBS-D) include frequent, loose or watery stools with associated urgency, which can severely limit socialising, travelling and eating out, with resulting marked reduction in quality of life and loss of work productivity. Around one-third of all IBS patients meet Rome criteria for IBS-D. When patients are asked to rank symptoms in order of importance, the erratic bowel habit is rated first, followed by abdominal pain and, for those with diarrhoea, urgency. Ondansetron, a 5-hydroxytryptamine-3 (5HT3) receptor antagonist, has an excellent safety record for over 20 years as an antiemetic, but is only exceptionally used in the treatment of IBS-D. It has, however, been shown to slow colonic transit and in a small randomised, placebo-controlled, crossover pilot study, benefited patients with IBS-D. While the current trial was ongoing, a separate trial in the USA using a fixed dose bimodal release formulation (3 mg ondansetron +9 mg delayed release formulation) also reported improvement in stool consistency but not pain. Objectives Our primary aim was to determine the efficacy of generic ondansetron compared to placebo in controlling the symptoms of IBS-D using the US FDA-recommended combined end point in which a responder is defined as a patient who met the response criteria for both pain and bowel habit for 6 out of 12 weeks of the trial. Secondary end points included its effect on the characteristic abnormalities of stool consistency, frequency and urgency as well as abdominal pain, satisfactory relief of IBS symptoms, mood and use of rescue medication and to determine the effect of 12 weeks ondansetron over the 1 month after discontinuation, as well as safety. The study also included mechanistic studies to examine the correlation of rectal sensitivity and compliance, faecal bile acids (FBAs) and proteases and postprandial sigmoid motility with the baseline symptoms of our IBS-D patients. We also attempted to determine whether ondansetron significantly altered these biomarkers compared to placebo. Methods Treatment of irritable bowel syndrome using titrated ondansetron trial (TRITON) was a multisite, parallel-group, randomised, double-blinded, placebo-controlled trial, with embedded mechanistic studies within selected sites. Our aim was to determine the superiority of ondansetron compared with placebo. We aimed to randomise 400 patients with IBS-D on a 1 : 1 basis to receive either ondansetron or placebo. Both treatments were administered for 12 weeks in oral doses ranging from 4 mg every third day to 24 mg daily. Dose titration was undertaken in the first 2 weeks of the study to avoid constipation, which at a standard dose occurs in one-quarter of patients. This was achieved by frequent consultation with the research nurse, starting with 1 × 4 mg tablet per day and increasing in increments every 2 days to a maximum of 2 tablets thrice daily. If constipation developed, the treatment was stopped to allow the return of bowel movements and then restarted at a lower dose, typically one every alternate day or one every second day. Rescue medication of loperamide was discouraged but allowed exceptionally for uncontrolled diarrhoea and was documented in the daily diary. The primary outcome of response for both reduction in pain intensity and improvement in stool consistency was assessed over the 12 weeks post randomisation. Secondary and safety outcomes were measured up to 16 weeks post randomisation. Symptoms that were recorded daily included (1) stool consistency and abdominal pain (measured by both paper diary and daily text message); (2) stool frequency, urgency of defaecation, use of rescue medication (defined as the use of loperamide) over 12 weeks of treatment and the answer to the question in the diary ‘Overall, have you had satisfactory relief from your IBS symptoms in the past week?’. Irritable bowel syndrome symptom severity [measured by the IBS Severity Scoring System (IBS-SSS)], dyspepsia [using the Short Form Leeds Dyspepsia Questionnaire (SFLDQ)], quality of life and mood [using the IBS Quality of Life (IBS-QOL) and Hospital Anxiety and Depression Scale (HADS) questionnaires], and somatic symptoms [using the Patient Health Questionnaire 12 Somatic Symptoms (PHQ-12) questionnaire] were assessed by patient-reported questionnaires at the baseline and 12 weeks post randomisation. The trial also assessed possible underlying mechanisms of any effect of ondansetron on changes in the primary and secondary end points. Whole gut transit was measured at baseline and 12 weeks using radio-opaque markers and an abdominal X-ray. High-resolution manometry was performed at baseline and after 8–11 weeks of treatment at two centres to assess whether ondansetron decreased the number of high-amplitude propagating contractions (HAPCs) or increased the percentage time occupied by cyclical retrograde propagated contractions. Barostat assessment was performed at baseline and after 8–11 weeks of treatment at two centres in order to assess if ondansetron increases rectal compliance or decreases sensitivity (manifested as increased pressure thresholds for pain and urgency). Stool samples were assessed for faecal water % (FW), faecal protease (FP) and FBAs. Clinical results The study closed early due to slow recruitment with just 80 patients randomised; 37 to ondansetron and 43 to placebo. Four patients discontinued ondansetron and one placebo during 12-week randomised treatment. Four were excluded from the per-protocol population due to major protocol violations. In the intention to treat (ITT) analysis, 15 patients (40.5%) on ondansetron achieved the primary end point response [95% confidence interval (CI) 24.7% to 56.4%], compared to 12 (27.9%) patients on placebo (95% CI 14.5% to 41.3%), p = 0.19, adjusted OR 1.93 (0.73, 5.11). Response for pain intensity reduction was achieved by 17 (46.0%) on ondansetron (95% CI 29.9% to 62.0%) and 16 (37.2%) on placebo (95% CI 22.8% to 51.7%), p = 0.32, adjusted OR 1.61 (0.63 to 4.12). Response for stool consistency improvement was reported by 25 (67.6%) on ondansetron (95% CI 52.5% to 82.7%) and 22 (51.2%) on placebo (95% CI 36.2% to 66.1%), p = 0.07, adjusted OR 2.45 (0.92, 6.52). Overall use of the rescue medication, loperamide, was 39.5% (n = 17) on placebo compared with 18.9% (n = 7) on ondansetron. However, by week 12, loperamide use fell to 13.5% on ondansetron versus 25.6% on placebo. Average stool consistency in the final month of treatment fell significantly more on ondansetron than placebo, adjusted mean difference –0.5 [standard error (SE) 0.25, 95% CI (–1.0 to –0.02), p = 0.042]. Ondansetron improved the dyspepsia score (SFLDQ) significantly more than placebo; the largest reduction being in symptoms of indigestion and nausea. The adjusted mean difference in the total score compared to placebo was –3.2 points [SE 1.43, 95% CI (–6.1, to –0.4), p = 0.028]. Ondansetron was well tolerated with most adverse reactions being mild or moderate and not significantly greater than on placebo. The commonest was constipation, reported in 32% on ondansetron and 23% on placebo, of which 75% and 80%, respectively, were rated as mild. Just two patients withdrew citing constipation as the cause. Mechanistic results Results are expressed as mean (SD). Comparing baseline and week 12 showed ondansetron increased average whole gut transit 3.78 (9.1) hours on ondansetron significantly more than placebo –2.2 (10.3), p = 0.01. Mean volume to reach urgency threshold using the barostat increased on ondansetron by 84 (61) ml and 38 (48) ml on placebo, n = 8; the difference was not significant, p = 0.26. Too few underwent manometry to allow meaningful assessment of the effect of ondansetron but anecdotally one patient who had a dramatic clinical improvement showed a loss of HAPCs and an increase in retrograde contractions, but this could have been due to chance. Ondansetron appeared not to significantly alter FP, though overall the increase in whole gut transit time from baseline to week 12 was correlated with a decrease in FP. There were no significant changes in FBAs and no evidence that ondansetron altered these though we did confirm that we had effectively excluded those with bile acid diarrhoea (BAD). The ratio of secondary to primary bile acids, a measure of bacterial metabolism of bile acids, increased substantially on ondansetron from 9.7 (7.08) to 21.4 (32.9) and less so on placebo from 22.84 (58.23) to 28.61 (31.42). However, owing to small numbers and wide variability these differences were not significant. Limitations Two previous studies in Nottingham had recruited 120 and 136 IBS-D patients within 2 years so we did not anticipate problems with recruitment. However, changes in referral pathways from primary to secondary care substantially reduced referrals to our coinvestigators who were all in secondary care, thus impairing recruitment. The power calculations required 400 to achieve 90% power to detect a 15% difference in primary end point, so the study is substantially underpowered. Use of loperamide did somewhat complicate interpretation since those on placebo used more rescue medication reducing the size of the effect on transit and stool consistency. Conclusion Despite being underpowered for our primary end point, our results are consistent with previous studies and confirmed ondansetron improves stool consistency but showed little effect on pain. Ondansetron significantly slowed whole gut transit time. Ondansetron reduced sensitivity to rectal distension more than placebo without altering compliance, but numbers were too small to achieve statistical significance. This could plausibly contribute to the reduction in urgency and stool frequency but needs repeating with larger numbers to be sure it was not due to chance. We found no evidence that rectal sensitivity was related to either faecal protease or bile acids. The manometry studies were underpowered but anecdotally ondansetron appeared to alter rectosigmoid motor patterns in a way that could reduce inflow of stool to the rectum. Future work We plan to do a simplified version of this trial, using an efficient and remote process, to overcome the changed referral pathways by recruiting in primary care. We will search for patients who have had a diagnosis of chronic diarrhoea and the recommended screening including a normal full blood count, a negative tissue transglutaminase (excluding coeliac disease) and a normal faecal calprotectin using software linked to primary care records. This will allow rapid screening of large numbers of patients to identify and approach patients with IBS-D who meet criteria to take part in a randomised trial of ondansetron or placebo, thus minimising barriers to recruitment. We would remove the pain threshold, which would increase the number of eligible patients and facilitate recruitment. Not allowing loperamide as rescue medication would simplify interpretation and dropouts would be treated as treatment failures. Further streamlining by removing all additional tests that were included in the current trial, as well as efficient trial processes, including e-consent, remote blood and stool samples (if required), and online questionnaires would also optimise recruitment. Study registration This trial is registered as ISRCTN17508514. Funding This project was funded by the National Institute for Health and Care Research (NIHR) Efficacy and Mechanism Evaluation programme and will be published in full in Efficacy and Mechanism Evaluation; Vol. 10, No. 9. See the NIHR Journals Library website for further project information.

BackgroundThis manuscript summarizes consensus reached by the International Anorectal Physiology Working Group (IAPWG) for the performance, terminology used, and interpretation of anorectal function testing including anorectal manometry (focused on high-resolution manometry), the rectal sensory test, and the balloon expulsion test. Based on these measurements, a classification system for disorders of anorectal function is proposed.MethodsTwenty-nine working group members (clinicians/academics in the field of gastroenterology, coloproctology, and gastrointestinal physiology) were invited to six face-to-face and three remote meetings to derive consensus between 2014 and 2018.Key recommendationsThe IAPWG protocol for the performance of anorectal function testing recommends a standardized sequence of maneuvers to test rectoanal reflexes, anal tone and contractility, rectoanal coordination, and rectal sensation. Major findings not seen in healthy controls defined by the classification are as follows: rectoanal areflexia, anal hypotension and hypocontractility, rectal hyposensitivity, and hypersensitivity. Minor and inconclusive findings that can be present in health and require additional information prior to diagnosis include anal hypertension and dyssynergia.Conclusions and inferencesThis framework introduces the IAPWG protocol and the London classification for disorders of anorectal function based on objective physiological measurement. The use of a common language to describe results of diagnostic tests, standard operating procedures, and a consensus classification system is designed to bring much-needed standardization to these techniques.

Background and Aims: Gut functions including motility, secretion, and blood flow are largely controlled by the enteric nervous system. Characterizing the different classes of enteric neurons in the human gut is an important step to understand how its circuitry is organized and how it is affected by disease. Methods: Using multiplexed immunohistochemistry, 12 discriminating antisera were applied to distinguish different classes of myenteric neurons in the human colon (2596 neurons, 12 patients) according to their chemical coding. All antisera were applied to every neuron, in multiple layers, separated by elutions. Results: A total of 164 combinations of immunohistochemical markers were present among the 2596 neurons, which could be divided into 20 classes, with statistical validation. Putative functions were ascribed for 4 classes of putative excitatory motor neurons (EMN1–4), 4 inhibitory motor neurons (IMN1–4), 3 ascending interneurons (AIN1–3), 6 descending interneurons (DIN1–6), 2 classes of multiaxonal sensory neurons (SN1–2), and a small, miscellaneous group (1.8% of total). Soma-dendritic morphology was analyzed, revealing 5 common shapes distributed differentially between the 20 classes. Distinctive baskets of axonal varicosities surrounded 45% of myenteric nerve cell bodies and were associated with close appositions, suggesting possible connectivity. Baskets of cholinergic terminals and several other types of baskets selectively targeted ascending interneurons and excitatory motor neurons but were significantly sparser around inhibitory motor neurons. Conclusions: Using a simple immunohistochemical method, human myenteric neurons were shown to comprise multiple classes based on chemical coding and morphology and dense clusters of axonal varicosities were selectively associated with some classes.

A rhythmic expression of clock genes occurs within the cells of multiple organs and tissues throughout the body, termed “peripheral clocks.” Peripheral clocks are subject to entrainment by a multitude of factors, many of which are directly or indirectly controlled by the light-entrainable clock located in the suprachiasmatic nucleus of the hypothalamus. Peripheral clocks occur in the gastrointestinal tract, notably the epithelia whose functions include regulation of absorption, permeability, and secretion of hormones; and in the myenteric plexus, which is the intrinsic neural network principally responsible for the coordination of muscular activity in the gut. This review focuses on the physiological circadian variation of major colonic functions and their entraining mechanisms, including colonic motility, absorption, hormone secretion, permeability, and pain signalling. Pathophysiological states such as irritable bowel syndrome and ulcerative colitis and their interactions with circadian rhythmicity are also described. Finally, the classic circadian hormone melatonin is discussed, which is expressed in the gut in greater quantities than the pineal gland, and whose exogenous use has been of therapeutic interest in treating colonic pathophysiological states, including those exacerbated by chronic circadian disruption.

The causes of a pyrexia of unknown origin (PUO) are broadly spread across infectious, malignant, or inflammatory conditions. The authors report a case of a 43-year-old female who initially presented with hypoxia and fevers secondary to a presumed acute viral or bacterial infectious pulmonary insult. However, fevers persisted despite pulmonary imaging remaining normal. The case subsequently evolved into a PUO with multiple lines of investigation proving inconclusive. No definitive diagnosis was made at the time of discharge; however, cessation of hydralazine was noted to coincide with the last of the patient’s fevers. Following the discharge, her anti-histone antibodies returned a positive result, and the diagnosis of drug-induced lupus was made with hydralazine as the presumed trigger.

Abstract The speed of pellet propulsion through the isolated guinea pig distal colon in vitro significantly exceeds in vivo measurements, suggesting a role for inhibitory mechanisms from sources outside the gut. The aim of this study was to investigate the effects of sympathetic nerve stimulation on three different neurogenic motor behaviors of the distal colon: transient neural events (TNEs), colonic motor complexes (CMCs), and pellet propulsion. To do this, segments of guinea pig distal colon with intact connections to the inferior mesenteric ganglion (IMG) were set up in organ baths allowing for simultaneous extracellular suction electrode recordings from smooth muscle, video recordings for diameter mapping, and intraluminal manometry. Electrical stimulation (1–20 Hz) of colonic nerves surrounding the inferior mesenteric artery caused a statistically significant, frequency‐dependent inhibition of TNEs, as well as single pellet propulsion, from frequencies of 5 Hz and greater. Significant inhibition of CMCs required stimulation frequencies of 10 Hz and greater. Phentolamine (3.6 μM) abolished effects of colonic nerve stimulation, consistent with a sympathetic noradrenergic mechanism. Sympathetic inhibition was constrained to regions with intact extrinsic nerve pathways, allowing normal motor behaviors to continue without modulation in adjacent extrinsically denervated regions of the same colonic segments. The results demonstrate differential sensitivities to sympathetic input among distinct neurogenic motor behaviors of the colon. Together with findings indicating CMCs activate colo‐colonic sympathetic reflexes through the IMG, these results raise the possibility that CMCs may paradoxically facilitate suppression of pellet movement in vivo, through peripheral sympathetic reflex circuits.

BackgroundIn the human large bowel, sacral parasympathetic nerves arise from S2 to S4, project to the pelvic plexus (“hypogastric plexus”) and have post-ganglionic axons entering the large bowel near the rectosigmoid junction. They then run long distances orally or aborally within the bowel wall forming “ascending nerves” or “shunt fascicles” running in the plane of the myenteric plexus. They form bundles of nerve fibres that can be distinguished from the myenteric plexus by their straight orientation, tendency not to merge with myenteric ganglia and greater width.AimTo identify reliable marker(s) to distinguish these bundles of ascending nerves from other extrinsic and intrinsic nerves in human colon.MethodsHuman colonic segments were obtained with informed consent, from adult patients undergoing elective surgery (n = 21). Multi-layer immunohistochemical labelling with neurofilament-H (NF200), myelin basic protein (MBP), von Willebrand factor (vWF), and glucose transporter 1 (GLUT1), and rapid anterograde tracing with biotinamide, were used to compare ascending nerves and lumbar colonic nerves.ResultsThe rectosigmoid and rectal specimens had 6–11 ascending nerves spaced around their circumference. Distal colon specimens typically had 1–3 ascending nerves, with one located near the mesenteric taenia coli. No ascending nerves were observed in ascending colon specimens. GLUT1 antisera labelled both sympathetic lumbar colonic nerves and ascending nerves in the gut wall. Lumbar colonic nerves joined the myenteric plexus and quickly lost GLUT1 labelling, whereas GLUT1 staining labelled parasympathetic ascending nerves over many centimetres.ConclusionAscending nerves can be distinguished in the colorectum of humans using GLUT1 labelling combined with NF200.

Purpose: The purpose of this paper is to explore the level enterprise language is articulated within programme level documentation, and then, using EntreComp (Bacigalupo et al., 2016) as a framework, investigate the extent by which curricula is underpinned by enterprise competencies. Design/methodology/approach: This study adopted a qualitative approach firstly through a desk analysis of undergraduate programme documentation across one UK University (n=60), followed by face-to-face interviews with programme leaders (n=25). Findings: Findings revealed a lack of reference to enterprise in programme documentation as well as a confusion of the language associated with enterprise and entrepreneurship. However, all participants in the study were able to articulate opportunities afforded to students within the programme to practise entrepreneurial competencies from the EntreComp framework. Originality/value: Whilst the limitation of this study is that it has been carried out in just one university, the practical implication of these findings provide supporting evidence that the EntreComp framework can be used to build a whole programme approach to embedding enterprise. This remains to be tested in future research. As the EntreComp framework is relatively new, then this piece of research is original as it is amongst the first to report on the framework.

Purpose: To confirm feasibility of recording pressure along the length of the urethra using a multi-sensor fibre-optic pressure catheter; to identify the spatial and temporal features of changes in pressure along the urethra at sites related to specific striated pelvic floor muscles; and to investigate the relationship between urethral pressures and activation of individual pelvic floor muscles estimated from ultrasound imaging. Materials and Methods: Proof-of-concept study including one male (47 years old) and one female (33 years old). A multi-sensor fibre optic pressure catheter (10 mm sensor separation) was inserted into the urethra. Pressure data were recorded simultaneously with trans-perineal ultrasound imaging measures of pelvic floor muscle activity during sub-maximal and maximal voluntary contractions and evoked coughs. Results: Pressure changes along the urethra were recorded in all tasks in both participants. Face validity of interpretation of pressure measures with respect to individual muscles was supported by correlation with ultrasound-measured displacements induced by the relevant muscles. Onset of pressure increase occurred in a distal to proximal sequence in the urethra of the male but not the female during voluntary contraction. Peak urethral pressures varied in location, timing and amplitude between tasks. Evoked cough induced in the greatest urethral pressure increase across all tasks for both participants. Conclusions: The high spatial resolution pressure catheter provide viable and valid recordings of urethral pressure in a male and female. Data provide preliminary evidence of sex differences in spatial and temporal distribution of urethral pressure changes.

This study sets out to investigate physical education (PE) teachers' perceptions of the use of an online professional development resource to support making reasonable adjustment for learners with disability. The Equality Act (2010) called on all UK schools to ensure access to PE was equitable for all learners. This means schools have to adopt strategies for making reasonable adjustments so as not to disadvantage learners with any special educational need and disability. Teachers report that they are working towards an inclusive classroom; however, parents of learners with disabilities report that there remains a lack of opportunities for their children to engage with activity. Using purposive sampling, participants were selected for the study after which they were asked to complete a continuing professional development online training course on the subject of making reasonable adjustments for learning with disabilities. Through one-to-one interviews, qualitative data were collected and analysed through thematic analysis. The findings indicated that the online resource was greeted positively by staff and not only supported the PE teachers but also increased their awareness of different approaches to making reasonable adjustments. The findings also indicated that the issue of making reasonable adjustments and the use of the online development tool need to be driven from the school senior leadership team if it is to have value for the whole school community.

While there is a robust literature examining the patterns and causes of teacher turnover, few articles to date have critically examined the measures of turnover used in these studies. Yet, an assessment of the way turnover is measured is important, as the measures become the means by which the "problem" of turnover becomes defined and its varying dimensions understood. In this conceptual essay, we outline a typology of teacher turnover measures, discussing both measures used in existing teacher turnover literature as well as new measures that we have developed. We illustrate each of the measures using 10 years of administrative data from Texas. We discuss how the measures can help illuminate different ways in which staff instability can affect schools and identify schools that suffer from particularly severe staffing issues. We conclude with implications for policymakers and researchers who may seek to apply these measures to future empirical studies.

Fisheries involve complex problems not easily addressed by a single discipline, methodology, or set of stakeholders. In 2010, the Canadian Fisheries Research Network (CFRN) was initiated to increase fisheries research capacity in Canada through interdisciplinary and inclusive research collaborations. As post-graduate students in the network, we reflected on the type of training necessary to tackle fisheries problems and reviewed opportunities available at Canadian universities to receive such training. This paper presents an overview of fisheries education currently available in Canada, reflects on our training within the CFRN, and proposes improvements to fisheries education and research. Our review of the subject revealed few dedicated fisheries programs, limited interdisciplinary programs, few specialized fisheries training programs, and a heavy reliance on academic supervisors to secure research opportunities in fisheries. In contrast, the CFRN enhanced our training by deliberately focusing on tools and techniques to address fisheries issues, providing venues to foster interdisciplinary and inclusive research collaborations, and exposing the realities of stakeholder collaborations. We call for post-graduate-level fisheries education and research that is interdisciplinary, collaborative, and inclusive to produce well-rounded scientists and managers, and we suggest ways that universities, researchers, and funding agencies can incorporate these themes into fisheries education and research.

Manual analysis of human high-resolution colonic manometry data is time consuming, non-standardized and subject to laboratory bias. In this article we present a technique for spectral analysis and statistical inference of quasiperiodic spatiotemporal signals recorded during colonic manometry procedures. Spectral analysis is achieved by computing the continuous wavelet transform and cross-wavelet transform of these signals. Statistical inference is achieved by modeling the resulting time-averaged amplitudes in the frequency and frequency-phase domains as Gaussian processes over a regular grid, under the influence of categorical and numerical predictors specified by the experimental design as a functional mixed-effects model. Parameters of the model are inferred with Hamiltonian Monte Carlo. Using this method, we re-analyzed our previously published colonic manometry data, comparing healthy controls and patients with slow transit constipation. The output from our automated method, supports and adds to our previous manual analysis. To obtain these results took less than two days. In comparison the manual analysis took 5 weeks. The proposed mixed-effects model approach described here can also be used to gain an appreciation of cyclical activity in individual subjects during control periods and in response to any form of intervention.

The Equality Act called on British schools to "avoid as far as possible by reasonable means, the disadvantage which a disabled pupil experiences". Teachers, therefore, must be creative and flexible in order to meet the needs and optimise the capabilities of all pupils. Using focus group interviews, this article explores the influence of an online resource on pre-service teachers' perceptions of making reasonable adjustments for children with special educational needs and disabilities. Pre-service teachers appeared committed to making reasonable adjustments, with reports of the online resource being particularly influential on their planning and assessing progress. The influence of the resource was less significant on those pre-service teachers with previous experience of making reasonable adjustments.

Purpose: The purpose of this paper is to explore from an employer's perspective the skills, attributes and capabilities required of a sports graduate, whilst also discovering how a student is expected to demonstrate these skills in the context of a sports organisation. Design/methodology/approach: This study adopted a qualitative interpretive case study approach through the use of face-to-face interviews with six employers from sports organisations involved in the delivery of sport. Interviews lasted between 30 and 42 minutes and were recorded. This was followed by a thematic review to allow for common themes to be selected and represented. The results were then reviewed and evaluated by a further 15 industry professionals and sports educators. Findings: The results suggest employers seek skills that are characteristically enterprise/entrepreneurship skills, together with an articulation of a "sports graduate" mindset. This mindset is described by the employers as being something which is demonstrated through a combination of behaviours and attributes. The paper concluded that in the classroom, the use of case studies and challenges, where students have to respond to and solve problems by the very nature of the activity, is optimal. Originality/value: The originality of this paper lies in both the context of study and the integration of enterprise and entrepreneurship skills that are needed for the changing world of work in sport. The value of the papers is in both the employer's description of the graduate mindset and also examples of how the skills can be applied in the context of sport.

Students spend 12 to 14 years in school settings learning in what could be considered a carefully controlled and structured environment. Higher education may not offer the same landscape to students and it appears that many enter with unrealistic conceptions of what is expected of them and are faced with different approaches to aspects of teaching, learning and assessment. This qualitative study explores the perceptions of second-year and final-year students in relation to their expectations whilst studying at university. Focus groups were used across two programmes in one university faculty to ascertain student expectations and what they perceived as important. From the thematic analysis, four areas were highlighted by the students as key to the transition into university these were directed time, non-directed time, support and relationships. Overall these students where positive about the university experience and the levels of support offered to them, particularly noting that working in peer learning groups (PLGs) was beneficial. Issues were raised around the timetabling of face-to-face contact time and the value of the experience and this is an area that needs further research as is understanding the complexity of the students' lives outside of the institution.

With graduate employability being high on many universities' agendas, it becomes vital that a curriculum delivers not only subject knowledge but also the opportunity for students to develop and enhance industry-specific skills. This paper is concerned with how a work-based project can support skill development and considers the views of students and employers as to the application of these skills in the context of a sports undergraduate programme in a UK university. Using a mixed-method approach to data collection, 30 students and 5 project hosts took part in the study. The research findings reported a mismatch between the students' and hosts' viewpoints on whether skills were developed as a result of the project. Students appeared to have a lack of understanding how to apply the skills in the context of a sports organisation and were viewed by the employers as having an inflated opinion of their ability.

Objective To quantify the implementation of the third Joint British Societies’ Consensus Recommendations for the Prevention of Cardiovascular Disease (JBS3) after coronary event.Methods Using a cross-sectional survey design, patients were consecutively identified in 36 specialist and district general hospitals between 6 months and 2 years, after acute coronary syndrome or revascularisation procedure and invited to a research interview. Outcomes included JBS3 lifestyle, risk factor and therapeutic management goals. Data were collected using standardised methods and instruments by trained study nurses. Blood was analysed in a central laboratory and a glucose tolerance test was performed.Results 3926 eligible patients were invited to participate and 1177 (23.3% women) were interviewed (30% response). 12.5% were from black and minority ethnic groups. 45% were persistent smokers, 36% obese, 52.9% centrally obese, 52% inactive; 30% had a blood pressure >140/90 mm Hg, 54% non-high-density lipoprotein ≥2.5 mmol/L and 44.3% had new dysglycaemia. Prescribing was highest for antiplatelets (94%) and statins (85%). 81% were advised to attend cardiac rehabilitation (86%

This study sets out to investigate physical education (PE) teachers’ perceptions of the use of an online professional development resource to support making reasonable adjustment for learners with disability. The Equality Act (2010) called on all UK schools to ensure access to PE was equitable for all learners. This means schools have to adopt strategies for making reasonable adjustments so as not to disadvantage learners with any special educational need and disability. Teachers report that they are working towards an inclusive classroom; however, parents of learners with disabilities report that there remains a lack of opportunities for their children to engage with activity. Using purposive sampling, participants were selected for the study after which they were asked to complete a continuing professional development online training course on the subject of making reasonable adjustments for learning with disabilities. Through one-to-one interviews, qualitative data were collected and analysed through thematic analysis. The findings indicated that the online resource was greeted positively by staff and not only supported the PE teachers but also increased their awareness of different approaches to making reasonable adjustments. The findings also indicated that the issue of making reasonable adjustments and the use of the online development tool need to be driven from the school senior leadership team if it is to have value for the whole school community.

The majority of patients with an anorectal malformation (ARM) have associated congenital anomalies. It is well established that all patients diagnosed with an ARM should undergo systematic screening, including renal, spinal, and cardiac imaging. This study aimed to evaluate the findings and completeness of screening, following local implementation of standardized protocols. A retrospective cohort study was performed assessing all patients with an ARM managed at our tertiary pediatric surgical center, following a standardized protocol implementation for VACTERL screening (January 2016–December 2021). Cohort demographics, medical characteristics, and screening investigations were analyzed. Findings were compared with our previously published data (2000–2015), conducted prior to protocol implementation. One hundred twenty-seven (64 male, 50.4%) children were eligible for inclusion. Complete screening was performed in 107/127 (84.3%) children. Of these, one or more associated anomalies were diagnosed in 85/107 (79.4%), whilst the VACTERL association was demonstrated in 57/107 (53.3%). The proportion of children that underwent complete screening increased significantly in comparison with those assessed prior to protocol implementation (RR 0.43 [CI 0.27–0.66]; p < 0.001). Children with less complex ARM types were significantly less likely to receive complete screening (p = 0.028). Neither presence of an associated anomaly, nor prevalence of the VACTERL association, differed significantly by ARM type complexity. Screening for associated VACTERL anomalies in children with ARM was significantly improved following standardized protocol implementation. The prevalence of associated anomalies in our cohort supports the value of routine VACTERL screening in all children with ARM, regardless of malformation type. II. • The majority of patients with an anorectal malformation (ARM) have associated congenital anomalies and should undergo systematic screening. • Despite this, screening is often performed incompletely. • Screening significantly improved following standardized protocol implementation. • The prevalence of associated anomalies supports VACTERL screening in all children with ARM, regardless of malformation type. [ABSTRACT FROM AUTHOR]

Students spend 12 to 14 years in school settings learning in what could be considered a carefully controlled and structured environment. Higher education may not offer the same landscape to students and it appears that many enter with unrealistic conceptions of what is expected of them and are faced with different approaches to aspects of teaching, learning and assessment. This qualitative study explores the perceptions of second-year and final-year students in relation to their expectations whilst studying at university. Focus groups were used across two programmes in one university faculty to ascertain student expectations and what they perceived as important. From the thematic analysis, four areas were highlighted by the students as key to the transition into university these were directed time, non-directed time, support and relationships. Overall these students where positive about the university experience and the levels of support offered to them, particularly noting that working in peer learning groups (PLGs) was beneficial. Issues were raised around the timetabling of face-to-face contact time and the value of the experience and this is an area that needs further research as is understanding the complexity of the students’ lives outside of the institution.

With graduate employability being high on many universities’ agendas, it becomes vital that a curriculum delivers not only subject knowledge but also the opportunity for students to develop and enhance industry-specific skills. This paper is concerned with how a work-based project can support skill development and considers the views of students and employers as to the application of these skills in the context of a sports undergraduate programme in a UK university. Using a mixed-method approach to data collection, 30 students and 5 project hosts took part in the study. The research findings reported a mismatch between the students’ and hosts’ viewpoints on whether skills were developed as a result of the project. Students appeared to have a lack of understanding how to apply the skills in the context of a sports organisation and were viewed by the employers as having an inflated opinion of their ability.

Objectives: Exertional-heat stress generates a thermoregulatory strain that exacerbates splanchnic hypoperfusion and sympathetic drive, but the effects on gastrointestinal function are poorly defined. The study aimed to determine the effects of exertional-heat stress on gastric myoelectrical activity, orocecal transit time (OCTT), and gastrointestinal symptoms (GIS).Design: Randomised cross-over study.Methods: Endurance runners (n = 16) completed 2 h of running at 60 % V̇O2max in 35 °C (HOT) and 22 °C (TEMP) ambient conditions. Surface electrogastrography (cEGG) was recorded pre- and post-exercise to determine gastric myoelectrical activity, a lactulose challenge was used to determine OCTT, and GIS were recorded using a modified visual analogue scale tool.Results: Post-exercise Tre [HOT:38.8(38.5 to 39.0)°C and TEMP:38.1(37.8 to 38.4)°C] and Δ Tre [HOT:2.2(2.0 to 2.4)°C and TEMP:1.5(1.2 to 1.8)°C] was higher on HOT compared to TEMP (p < 0.001). Normal gastric myoelectrical cycle frequency reduced (p = 0.010) on HOT [-11.7(-20.8 to -2.6)%], but this decrease did not differ (p = 0.058) from TEMP [-2.7(-8.3 to 3.0)%]. Bradygastria increased post-exercise on both trials (HOT:11.3(2.3 to 20.4)%, p = 0.030; and TEMP:7.4(2.1 to 12.6)%, p = 0.009). OCTT did not differ between trials (p = 0.864) with transit response classified as very slow on both HOT (99(68 to 131)min) and TEMP (98(74 to 121)min). GIS incidence was higher on HOT (88 %) compared to TEMP (81 %), in accordance with greater total-GIS and upper-GIS severity (p = 0.005 and p = 0.033, respectively).Conclusions: Running for 2 h at 60 % V̇O2max in either hot or temperate ambient conditions instigates perturbations in myoelectrical activity and OCTT, with GIS incidence and severity greater in hot conditions. [ABSTRACT FROM AUTHOR]