Single cell sequencing data identify distinct B cell and fibroblast populations in stricturing Crohn's disease.

Single cell RNA sequencing of human full thickness Crohn's disease (CD) small bowel resection specimens was used to identify potential therapeutic targets for stricturing (S) CD. Using an unbiased approach, 16 cell lineages were assigned within 14,539 sequenced cells from patient-matched SCD and non-stricturing (NSCD) preparations. SCD and NSCD contained identical cell types. Amongst immune cells, B cells and plasma cells were selectively increased in SCD samples. B cell subsets suggested formation of tertiary lymphoid tissue in SCD and compared with NSCD there was an increase in IgG, and a decrease in IgA plasma cells, consistent with their potential role in CD fibrosis. Two Lumican-positive fibroblast subtypes were identified and subclassified based on expression of selectively enriched genes as fibroblast clusters (C) 12 and C9. Cells within these clusters expressed the profibrotic genes Decorin (C12) and JUN (C9). C9 cells expressed ACTA2; ECM genes COL4A1, COL4A2, COL15A1, COL6A3, COL18A1 and ADAMDEC1; LAMB1 and GREM1. GO and KEGG Biological terms showed extracellular matrix and stricture organization associated with C12 and C9, and regulation of WNT pathway genes with C9. Trajectory and differential gene analysis of C12 and C9 identified four sub-clusters. Intra sub-cluster gene analysis detected 13 co-regulated gene modules that aligned along predicted pseudotime trajectories. CXCL14 and ADAMDEC1 were key markers in module 1. Our findings support further investigation of fibroblast heterogeneity and interactions with local and circulating immune cells at earlier time points in fibrosis progression. Breaking these interactions by targeting one or other population may improve therapeutic management for SCD.

Matching-Adjusted Indirect Comparisons of Filgotinib vs Vedolizumab, Tofacitinib, and Ustekinumab for Moderately to Severely Active Ulcerative Colitis.

BACKGROUND: Where head-to-head trials are lacking, indirect comparative effectiveness can aid treatment decisions. We conducted matching-adjusted indirect comparisons of clinical outcomes with filgotinib vs recently approved comparators (vedolizumab, tofacitinib, ustekinumab) in patients with moderately to severely active ulcerative colitis (UC). METHODS: Individual patient data from the SELECTION trial (NCT02914522) for filgotinib 200 mg were weighted to match average baseline characteristics of active treatment and placebo arms in comparator trials. Efficacy outcomes were compared for biologic-naive and biologic-experienced subgroups in induction and maintenance populations, if data were available. Safety and health-related quality of life outcomes were compared in the overall maintenance population. RESULTS: Filgotinib had a similar effect on efficacy outcomes compared with tofacitinib, ustekinumab, and subcutaneous vedolizumab in both the induction and maintenance populations. Filgotinib showed improved clinical response vs intravenous (IV) vedolizumab (odds ratio, 2.4; 95% confidence interval [CI], 1.0 to 5.5; P < .05) among the biologic-experienced induction population, and improved corticosteroid-free clinical remission (odds ratio, 15.2; 95% CI, 1.6 to 139.9; P < .05) among the biologic-naive maintenance population. Improved efficacy outcomes were reported with filgotinib compared with ustekinumab among the maintenance population. Higher estimates of serious adverse events were reported for filgotinib compared with vedolizumab IV 300 mg and tofacitinib 5 mg; however, imbalances were noted in their placebo groups. Health-related quality of life outcomes were similar between filgotinib and comparators. CONCLUSIONS: Matching-adjusted indirect comparison results suggest superiority of filgotinib 200 mg over vedolizumab IV in terms of clinical response and corticosteroid-free clinical remission in certain patient populations, noting small sample sizes and wide CIs, which may aid the selection of advanced therapies for moderately to severely active UC. A potential increased risk of serious adverse events was reported for filgotinib 200 mg vs vedolizumab IV and tofacitinib 5 mg, but findings should be interpreted with caution owing to underlying imbalances observed between the placebo groups of SELECTION and comparator trials.

Matching-adjusted indirect comparisons between filgotinib and subcutaneous vedolizumab, tofacitinib, and ustekinumab demonstrated similar effects on efficacy, safety, and health-related quality of life in patients with ulcerative colitis. Clinical response and corticosteroid-free remission were improved with filgotinib compared with intravenous vedolizumab.

Small-molecule Wnt inhibitors are a potential novel therapy for intestinal fibrosis in Crohns disease.

Intestinal fibrosis and stricture formation is an aggressive complication of Crohns disease (CD), linked to increased morbidity and costs. The present study investigates the contribution of Wingless-Int-1 (Wnt) signalling to intestinal fibrogenesis, considers potential cross-talk between Wnt and transforming growth factor ß1 (TGFß) signalling pathways, and assesses the therapeutic potential of small-molecule Wnt inhibitors. ß-catenin expression was explored by immunohistochemistry (IHC) in formalin-fixed paraffin embedded (FFPE) tissue from patient-matched nonstrictured (NSCD) and strictured (SCD) intestine (n=6 pairs). Functional interactions between Wnt activation, TGFß signalling, and type I collagen (Collagen-I) expression were explored in CCD-18Co cells and primary CD myofibroblast cultures established from surgical resection specimens (n=16) using small-molecule Wnt inhibitors and molecular techniques, including siRNA-mediated gene knockdown, immunofluorescence (IF), Wnt gene expression arrays, and western blotting. Fibrotic SCD tissue was marked by an increase in ß-catenin-positive cells. In vitro, activation of Wnt-ß-catenin signalling increased Collagen-I expression in CCD-18Co cells. Conversely, ICG-001, an inhibitor of ß-catenin signalling, reduced Collagen-I expression in cell lines and primary CD myofibroblasts. TGFß increased ß-catenin protein levels but did not activate canonical Wnt signalling. Rather, TGFß up-regulated WNT5B, a noncanonical Wnt ligand, and the Wnt receptor FZD8, which contributed directly to the up-regulation of Collagen-I through a ß-catenin-independent mechanism. Treatment of CCD-18Co fibroblasts and patient-derived myofibroblasts with the FZD8 inhibitor 3235-0367 reduced extracellular matrix (ECM) expression. Our data highlight small-molecule Wnt inhibitors of both canonical and noncanonical Wnt signalling, as potential antifibrotic drugs to treat SCD intestinal fibrosis. They also highlight the importance of the cross-talk between Wnt and TGFß signalling pathways in CD intestinal fibrosis.

In silico design and analyses of a multi-epitope vaccine against Crimean-Congo hemorrhagic fever virus through reverse vaccinology and immunoinformatics approaches.

Crimean Congo Hemorrhagic Fever virus (CCHFV) is a deadly human pathogen that causes an emerging zoonotic disease with a broad geographic spread, especially in Africa, Asia, and Europe, and the second most common viral hemorrhagic fever and widely transmitted tick-borne viral disease. Following infection, the patients are presented with a variety of clinical manifestations and a fatality rate of 40%. Despite the high fatality rate, there are unmet clinical interventions, as no antiviral drugs or vaccines for CCHF have been approved. Immunoinformatics pipeline and reverse vaccinology were used in this study to design a multi-epitope vaccine that may elicit a protective humoral and cellular immune response against Crimean-Congo hemorrhagic fever virus infection. Three essential virulent and antigenic proteins (S, M, and L) were used to predict seven CTL and 18 HTL epitopes that were non-allergenic, antigenic, IFN-γ inducing, and non-toxic. The epitopes were connected using linkers and 50S ribosomal protein L7/L12 was used as an adjuvant and raised a multi-epitope vaccine (MEV) that is 567 amino acids long. Molecular docking and simulation of the predicted 3D structure of the MEV with the toll-like (TLR2, TLR3, and TLR4) receptors and major histocompatibility complex (MCH-I and MCH-II) indicate high interactions and stability of the complexes, MM-GBSA free binding energy calculation revealed a favourable protein-protein complex. Maximum MEV expression was achieved with a CAI value of 0.98 through in silico cloning in the Drosophila melanogaster host. According to the immune simulation, IgG1, T-helper cells, T-cytotoxic cells, INF-γ, and IL-2 were predicted to be significantly elevated. These robust computational analyses demonstrated that the proposed MEV is effective in preventing CCHFV infections. However, it is still necessary to conduct both in vitro and in vivo experiments to validate the potential of the vaccine.

Acceptability, safety and tolerability of antidepressant repetitive transcranial magnetic stimulation for adolescents: A mixed-methods investigation.

OBJECTIVE: Major depressive disorder (MDD) is relatively common in adolescence, with far-reaching impacts. Current treatments frequently fail to alleviate depression severity for a substantial portion of adolescents. Repetitive transcranial magnetic stimulation (rTMS) may assist with this unmet clinical need. However, little is known about adverse events (AEs) experienced by adolescents receiving rTMS, subjective treatment experiences of adolescents and their parents, or treatment acceptability. METHODS: Fourteen adolescents (16.5 years ± 1.2; 71.4% female) with MDD received 20 sessions of either high-frequency (10 Hz; n = 7) left dorsolateral prefrontal cortex (DLPFC) or low-frequency (1 Hz; n = 7) right DLPFC rTMS. AEs were monitored at baseline and at weekly intervals via New York State Psychiatric Institute Side Effects Form for Children and Adolescents. Eight adolescents and nine parents participated in interviews regarding subjective treatment experience, analysed via content analysis. RESULTS: Drowsiness and lethargy were common AEs, reported by 92.3% of participants in week one. Number of AEs decreased throughout treatment course (after 5 treatments: M = 11.23, SD = 5.00; after 20 treatments: M = 8.92, SD = 5.95). Thirty-five AEs emerged during treatment, most commonly transient dizziness. Frequency, severity, and number of AEs reported were equivalent between treatment groups. Treatment adherence and satisfaction were high. Qualitative findings identified three themes relating to rTMS experience: Preparation and connection; Physical experience of treatment; and Perceived role of treatment. LIMITATIONS: Sample size was small, therefore findings are preliminary. CONCLUSIONS: rTMS was an acceptable treatment for adolescent MDD, from both adolescents' and parents' perspectives, and was safe and well-tolerated, as AE frequency and type did not differ from rTMS treatment courses in adults.

Adoption and continued use of mobile contact tracing technology: multilevel explanations from a three-wave panel survey and linked data.

OBJECTIVE: To identify the key individual-level (demographics, attitudes, mobility) and contextual (COVID-19 case numbers, tiers of mobility restrictions, urban districts) determinants of adopting the NHS COVID-19 contact tracing app and continued use overtime. DESIGN AND SETTING: A three-wave panel survey conducted in England in July 2020 (background survey), November 2020 (first measure of app adoption) and March 2021 (continued use of app and new adopters) linked with official data. PARTICIPANTS: N=2500 adults living in England, representative of England's population in terms of regional distribution, age and gender (2011 census). PRIMARY OUTCOME: Repeated measures of self-reported app usage. ANALYTICAL APPROACH: Multilevel logistic regression linking a range of individual level (from survey) and contextual (from linked data) determinants to app usage. RESULTS: We observe initial app uptake at 41%, 95% CI (0.39% to 0.43%), and a 12% drop-out rate by March 2021, 95% CI (0.10% to 0.14%). We also found that 7% of nonusers as of wave 2 became new adopters by wave 3, 95% CI (0.05% to 0.08%). Initial uptake (or failure to use) of the app associated with social norms, privacy concerns and misinformation about third-party data access, with those living in postal districts with restrictions on mobility less likely to use the app. Perceived lack of transparent evidence of effectiveness was associated with drop-out of use. In addition, those who trusted the government were more likely to adopt in wave 3 as new adopters. CONCLUSIONS: Successful uptake of the contact tracing app should be evaluated within the wider context of the UK Government's response to the crisis. Trust in government is key to adoption of the app in wave 3 while continued use is linked to perceptions of transparent evidence. Providing clear information to address privacy concerns could increase uptake, however, the disparities in continued use among ethnic minority participants needs further investigation.

The Representational Dynamics of Sequential Perceptual Averaging.

It is clear that humans can extract statistical information from streams of visual input, yet how our brain processes sequential images into the abstract representation of the mean feature value remains poorly explored. Using multivariate pattern analyses of electroencephalography recorded while human observers viewed 10 sequentially presented Gabors of different orientations to estimate their mean orientation at the end, we investigated sequential averaging mechanism by tracking the quality of individual and mean orientation as a function of sequential position. Critically, we varied the sequential variance of Gabor orientations to understand the neural basis of perceptual mean errors occurring during a sequential averaging task. We found that the mean-orientation representation emerged at specific delays from each sequential stimulus onset and became increasingly accurate as additional Gabors were viewed. Especially in frontocentral electrodes, the neural representation of mean orientation improved more rapidly and to a greater degree in less volatile environments, whereas individual orientation information was encoded precisely regardless of environmental volatility. The computational analysis of behavioral data also showed that perceptual mean errors arise from the cumulative construction of the mean orientation rather than the low-level encoding of individual stimulus orientation. Thus, our findings provide neural mechanisms to differentially accumulate increasingly abstract features from a concrete piece of information across the cortical hierarchy depending on environmental volatility.SIGNIFICANCE STATEMENT The visual system extracts behaviorally relevant summary statistical representation by exploiting statistical regularity of the visual stream over time. However, how the neural representation of the abstract mean feature value develops in a temporally changing environment remains poorly identified. Here, we directly recover the mean orientation information of sequentially delivered Gabor stimuli with different orientations as a function of their positions in time. The mean orientation representation, which is regularly updated, becomes increasingly accurate with increasing sequential position especially in the frontocentral region. Further, perceptual mean errors arise from the cumulative process rather than the low-level stimulus encoding. Overall, our study reveals a role of higher cortical areas in integrating stimulus-specific information into increasingly abstract task-oriented information.

CDK control pathways integrate cell size and ploidy information to control cell division.

Maintenance of cell size homeostasis is a property that is conserved throughout eukaryotes. Cell size homeostasis is brought about by the co-ordination of cell division with cell growth and requires restriction of smaller cells from undergoing mitosis and cell division, whilst allowing larger cells to do so. Cyclin-CDK is the fundamental driver of mitosis and therefore ultimately ensures size homeostasis. Here we dissect determinants of CDK activity in vivo to investigate how cell size information is processed by the cell cycle network in fission yeast. We develop a high-throughput single-cell assay system of CDK activity in vivo and show that inhibitory tyrosine phosphorylation of CDK encodes cell size information, with the phosphatase PP2A aiding to set a size threshold for division. CDK inhibitory phosphorylation works synergistically with PP2A to prevent mitosis in smaller cells. Finally, we find that diploid cells of equivalent size to haploid cells exhibit lower CDK activity in response to equal cyclin-CDK enzyme concentrations, suggesting that CDK activity is reduced by increased DNA levels. Therefore, scaling of cyclin-CDK levels with cell size, CDK inhibitory phosphorylation, PP2A, and DNA-dependent inhibition of CDK activity, all inform the cell cycle network of cell size, thus contributing to cell size homeostasis.

A long-term study of equine cheek teeth post-extraction complications: 428 cheek teeth (2004-2018).

BACKGROUND: Complications, especially delayed alveolar healing, are common following equine cheek teeth extraction, however, limited objective information is available on the prevalence and nature of these problems. OBJECTIVES: To document the type and prevalence of complications that occur following equine cheek tooth extraction and to identify possible risk factors for these complications that could be used to predict their occurrence and hopefully reduce their prevalence. STUDY DESIGN: Retrospective cohort study. METHODS: Clinical records of all cheek teeth extractions performed between February 2004 and September 2018 were examined and written questionnaires sent to owners. Details of post-extraction complications were analysed and logistic regression was used to evaluate potential associations between the likelihood of post-extraction nonhealing alveolus managed by the authors and the variables: age, breed, reason for extraction, Triadan position and extraction technique. RESULTS: Post-extraction complications were recorded following 58/428 extractions giving an overall complication rate of 13.6%, that caused a longer term clinical problem in 34/428 (7.9%) cases, with complications being asymptomatic or quickly self-resolving in the other 24 cases (5.6%). The most frequent complication was alveolar bone sequestration, including alveolar infection. Risk of developing a post-extraction alveolar disorder managed by the authors (n = 53) increased following extraction of the mandibular 06s, 07s or 08s compared with all other cheek teeth combined (P = .001); for cheek teeth with apical infections (P = .002) compared with those without; and following repulsion or minimally invasive transbuccal extraction (MTE) than following oral extraction (P = .01 and P = .02 respectively). MAIN LIMITATIONS: Length of time between exodontia and survey data collection for some cases, use of clinical records and survey data and biases associated with decision to treat. CONCLUSIONS: In agreement with previous studies, oral extraction had the lowest risk of complications. This study provides new information regarding the prevalence, types and risk of development of post extraction complications. Knowledge of these risk factors may help reduce these complications.

Ex Vivo Comparison of the Accuracy of a Clamped with a Hand-Held Drill Guide for Drilling Distal Third Metacarpals and Metatarsals in Equids.

OBJECTIVES: The aim of this study was to determine whether a clamped drill guide can be used effectively to drill across equine distal third metacarpals and metatarsals and to compare accuracy and speed of a drilling with a hand-held drill guide. STUDY DESIGN: Eighty equine cadaver limbs were divided between four surgeons with varying experience. The limbs were randomly allocated to clamped or hand-held drill guides and placed in a stand in an upright/standing position on a metal table. Drilling times from cis- to trans cortices were measured. Post-drilling computed tomography images were used to assess drilling deviations in the dorsopalmar and proximodistal directions. RESULTS: Mean proximodistal deviation of the drill hole was distad for both guides and significantly (p = 0.01) less for the clamped drill guide +0.35° (range: -5.42°, +6.61°, standard deviation [SD] ± 3.18°) than the hand-held drill guide +2.05° (range: -5.24°, +9.81°, SD ± 3.69°). Mean dorsopalmar/plantar deviation was non-significantly (p = 0.17) less for the clamped drill guide -0.27° (range: -9.72°, +6.58°, SD ± 3.57°) than the hand-held drill guide +0.60° (range: -9.74°, + 12.33°, SD ± 4.51°). Mean time taken to drill from cis to trans cortex was significantly (p < 0.001) shorter with the clamped drill guide (41.2s) than the hand-held guide (56.6s). CONCLUSION: The clamped drill guide could be used successfully on the equine distal third metacarpals and metatarsals and resulted in some improved accuracy and reduced drilling time compared with the hand-held guide. The use of the clamped drill guide in vivo could be supported. This may be of benefit for certain fracture repairs in equids.

Impact of sampling rate on statistical significance for single subject fMRI connectivity analysis.

A typical time series in functional magnetic resonance imaging (fMRI) exhibits autocorrelation, that is, the samples of the time series are dependent. In addition, temporal filtering, one of the crucial steps in preprocessing of functional magnetic resonance images, induces its own autocorrelation. While performing connectivity analysis in fMRI, the impact of the autocorrelation is largely ignored. Recently, autocorrelation has been addressed by variance correction approaches, which are sensitive to the sampling rate. In this article, we aim to investigate the impact of the sampling rate on the variance correction approaches. Toward this end, we first derived a generalized expression for the variance of the sample Pearson correlation coefficient (SPCC) in terms of the sampling rate and the filter cutoff frequency, in addition to the autocorrelation and cross-covariance functions of the time series. Through simulations, we illustrated the importance of the variance correction for a fixed sampling rate. Using the real resting state fMRI data sets, we demonstrated that the data sets with higher sampling rates were more prone to false positives, in agreement with the existing empirical reports. We further demonstrated with single subject results that for the data sets with higher sampling rates, the variance correction strategy restored the integrity of true connectivity.

Possible links between the lag structure in visual cortex and visual streams using fMRI.

Conventional functional connectivity analysis using functional magnetic resonance imaging (fMRI) measures the correlation of temporally synchronized brain activities between brain regions. Lag structure analysis relaxes the synchronicity constraint of fMRI signals, and thus, this approach might be better at explaining functional connectivity. However, the sources of the lag structure in fMRI are primarily unknown. Here, we applied lag structure analysis to the human visual cortex to identify the possible sources of lag structure. A total of 1,250 fMRI data from two independent databases were considered. We explored the temporal lag patterns between the central and peripheral visual fields in early visual cortex and those in two visual pathways of dorsal and ventral streams. We also compared the lag patterns with effective connectivity obtained with dynamic causal modeling. We found that the lag structure in early visual cortex flows from the central to peripheral visual fields and the order of the lag structure flow was consistent with the order of signal flows in visual pathways. The effective connectivity computed by dynamic causal modeling exhibited similar patterns with the lag structure results. This study suggests that signal flows in visual streams are possible sources of the lag structure in human visual cortex.

Epithelial down-regulation of the miR-200 family in fibrostenosing Crohn's disease is associated with features of epithelial to mesenchymal transition.

Intestinal mesenchymal cells deposit extracellular matrix in fibrotic Crohn's disease (CD). The contribution of epithelial to mesenchymal transition (EMT) to the mesenchymal cell pool in CD fibrosis remains obscure. The miR-200 family regulates fibrosis-related EMT in organs other than the gut. E-cadherin, cytokeratin-18 and vimentin expression was assessed using immunohistochemistry on paired strictured (SCD) and non-strictured (NSCD) ileal CD resections and correlated with fibrosis grade. MiR-200 expression was measured in paired SCD and NSCD tissue compartments using laser capture microdissection and RT-qPCR. Serum miR-200 expression was also measured in healthy controls and CD patients with stricturing and non-stricturing phenotypes. Extra-epithelial cytokeratin-18 staining and vimentin-positive epithelial staining were significantly greater in SCD samples (P = 0.04 and P = 0.03, respectively). Cytokeratin-18 staining correlated positively with subserosal fibrosis (P < 0.001). Four miR-200 family members were down-regulated in fresh SCD samples (miR-141, P = 0.002; miR-200a, P = 0.002; miR-200c, P = 0.001; miR-429; P = 0.004); miR-200 down-regulation in SCD tissue was localised to the epithelium (P = 0.001-0.015). The miR-200 target ZEB1 was up-regulated in SCD samples (P = 0.035). No difference in serum expression between patient groups was observed. Together, these observations suggest the presence of EMT in CD strictures and implicate the miR-200 family as regulators. Functional studies to prove this relationship are now warranted.

Label-free nanoscale optical metrology on myelinated axons in vivo.

In the mammalian nervous system, myelin provides electrical insulation for the neural circuit by forming a highly organized, multilayered thin film around the axon fibers. Here, we investigate the spectral reflectance from this subcellular nanostructure and devise a new label-free technique based on a spectroscopic analysis of reflected light, enabling nanoscale imaging of myelinated axons in their natural living state. Using this technique, we demonstrate three-dimensional mapping of the axon diameter and sensing of dynamic changes in the substructure of myelin at nanoscale. We further reveal the prevalence of axon bulging in the brain cortex in vivo after mild compressive trauma. Our novel tool opens new avenues of investigation by creating unprecedented access to the nanostructural dynamics of live myelinated axons in health and disease.

Autism Spectrum Disorder Related Functional Connectivity Changes in the Language Network in Children, Adolescents and Adults.

Autism spectrum disorder (ASD) is a neurodevelopmental disability with global implication. Altered brain connectivity in the language network has frequently been reported in ASD patients using task-based functional magnetic resonance imaging (fMRI) compared to typically developing (TD) participants. Most of these studies have focused on a specific age group or mixed age groups with ASD. In the current study, we investigated age-related changes in functional connectivity related measure, degree centrality (DC), in the language network across three age groups with ASD (113 children, 113 adolescents and 103 adults) using resting-state fMRI data collected from the autism brain imaging data exchange repository. We identified regions with significant group-wise differences between ASD and TD groups for three age cohorts using DC based on graph theory. We found that both children and adolescents with ASD showed decreased DC in Broca's area compared to age-matched TD groups. Adults with ASD showed decreased DC in Wernicke's area compared to TD adults. We also observed increased DC in the left inferior parietal lobule (IPL) and left middle temporal gyrus (MTG) for children with ASD compared to TD children and for adults with ASD compared to TD adults, respectively. Overall, functional differences occurred in key language processing regions such as the left inferior frontal gyrus (IFG) and superior temporal gyrus (STG) related to language production and comprehension across three age cohorts. We explored correlations between DC values of our findings with autism diagnostic observation schedule (ADOS) scores related to severity of ASD symptoms in the ASD group. We found that DC values of the left IFG demonstrated negative correlations with ADOS scores in children and adolescents with ASD. The left STG showed significant negative correlations with ADOS scores in adults with ASD. These results might shed light on the language network regions that should be further explored for prognosis, diagnosis, and monitoring of ASD in three age groups.

Arthroscopy of the Dorsal and Plantar Pouches of the Tarsocrural Joint for the Treatment of Osteochondritis Dissecans in the Horse: Clinical Features and Follow-Up.

OBJECTIVE: To describe the clinical details and pathology within the dorsal and plantar pouches of the tarsocrural joint of a population of horses that underwent arthroscopic surgery for tarsocrural osteochondritis dissecans (OCD). STUDY DESIGN: Retrospective cohort study. ANIMALS: Horses referred for arthroscopic treatment of tarsocrural OCD between 2005 and 2013 (102 horses; 144 joints). METHODS: Case records of all horses that had tarsocrural arthroscopy for OCD at Rossdales Equine Hospital, Newmarket, United Kingdom were included. Cases from 3 ECVS Diplomates were included, 1 of whom routinely examined 70 plantar pouches concurrently with routine dorsal pouch investigation. A structured questionnaire was used to obtain follow-up data alongside examination of racing records where appropriate. Descriptive data and 95% CI were calculated. RESULTS: Of the 70 joints that had both dorsal and plantar pouches examined, there was cartilage erosion/degeneration in 22 dorsal pouches (31.4%), cartilage wear lines in 32 plantar pouches (45.7%), and fragments were removed at the time of surgery from 7 plantar pouches (10.0%). Of the plantar pouches with wear lines, 18 (25.7%) had no evidence of cartilage abnormalities (separate from the OCD lesion) within the dorsal pouch. From the 102 horses with available follow-up, 34 horses (66.7%) achieved their intended use postsurgery. CONCLUSION: Routine plantar pouch investigation is warranted in cases of tarsocrural OCD to provide further information on the health of the joint and allows for removal of fragments from the plantar pouch that may not have been identified by routine diagnostic radiography.

TNFα induces co-trafficking of TRPV1/TRPA1 in VAMP1-containing vesicles to the plasmalemma via Munc18-1/syntaxin1/SNAP-25 mediated fusion.

Transient receptor potential (TRP) A1 and V1 channels relay sensory signals, yet little is known about their transport to the plasmalemma during inflammation. Herein, TRPA1 and TRPV1 were found on vesicles containing calcitonin gene-related peptide (CGRP), accumulated at sites of exo- and endo-cytosis, and co-localised on fibres and cell bodies of cultured sensory neurons expressing both. A proinflammatory cytokine, TNFα, elevated their surface content, and both resided in close proximity, indicating co-trafficking. Syntaxin 1-interacting protein, Munc18-1, proved necessary for the response to TNFα, and for TRPV1-triggered CGRP release. TNFα-induced surface trafficking of TRPV1 and TRPA1 required a synaptic vesicle membrane protein VAMP1 (but not 2/3), which is essential for CGRP exocytosis from large dense-core vesicles. Inactivation of two proteins on the presynaptic plasma membrane, syntaxin-1 or SNAP-25, by botulinum neurotoxin (BoNT)/C1 or /A inhibited the TNFα-elevated delivery. Accordingly, enhancement by TNFα of Ca(2+) influx through the upregulated surface-expressed TRPV1 and TRPA1 channels was abolished by BoNT/A. Thus, in addition, the neurotoxins' known inhibition of the release of pain transmitters, their therapeutic potential is augmented by lowering the exocytotic delivery of transducing channels and the resultant hyper-sensitisation in inflammation.

Histogenesis of the Oesophagus of Guinea Fowl (Numida meleagris) at Prehatch and Posthatch.

The histogenesis of the primordial oesophagus was studied to determine the period in which the tunics of the oesophagus developed and became functional in the helmeted guinea fowl (Numida meleagris). Eighteen embryos and nine keets were studied at prehatch and posthatch, respectively. Simple columnar epithelium surrounded by mesenchymal cells was obvious at the 8th day of embryonic development. By the 19th day of embryonic development, the four tunics, tunica mucosa, submucosa, tunica muscularis, and tunica adventitia/serosa, were beginning to differentiate from the mesenchymal cells and also the primordial oesophageal glands appeared as clusters of cells that invaginate from the epithelium. By the 27th day, the tunics were clearly differentiated and the primordial glands were fully developed as evident with positive reaction to Periodic Acid Schiff (PAS). The tunics of the muscularis were not well developed till at posthatch. This study therefore concludes that the primordial oesophagus is active at the late incubation due to mucin secretion by mucous glands but fully functional at posthatch since the tunica muscularis is completely developed at posthatch.