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Frontal plane ankle motion is important for balance in walking but is seldom controlled in robotic prostheses. This article describes the design, control and performance of a semi-active two-degree-of-freedom robotic prosthetic ankle. The mechanism uses a non-backdrivable wedge cam system based on rotating inclined planes, allowing actuation only during swing phases for low power, light weight and compactness. We present details of the mechanism and its kinematic and mechatronic control, and a benchtop investigation of the system’s speed and accuracy in ankle angle control. The two-axis ankle achieves angular reorientation movements spanning ±10 deg in any direction in less than 0.9 s. It achieves a plantarflexion/dorsiflexion error of 0.35 ± 0.27 deg and an inversion/eversion error of 0.29 ± 0.25 deg. Backdriven motion during walking tests is negligible. Strengths of the design include self-locking behavior for low power and simple kinematic control. Two-axis ankle angle control could enable applications such as balance augmentation, turning assistance, and wearable perturbation training.

Abstract Background Gastrointestinal bleed (GIB) has high incidence in traumatic spinal cord injured (tSCI) patients and can frequently be life-threatening, especially early post-injury. Several risk factors often compound bleeding risk, some are unique to this patient population. Normally, clinical suspicion for GIB arises from symptoms like coffee-ground emesis, hematemesis, melena or even hematochezia. A hemoglobin drop may be a late sign. Due to tSCI, however, patients often experience neurogenic bowels and dysautonomia, which may delay symptom presentation and complicate timely diagnosis of GIB. We report a case of an almost clinically silent GI bleed in the context of acute cervical tSCI. Case presentation A 21-year-old female presented with cervical cord transection at C-7 in the setting of motor vehicle rollover, for which surgical decompression was performed. During the acute injury phase, she also received a 10-day course of dexamethasone for symptomatic COVID-19 pneumonia. Two weeks after injury, she underwent percutaneous endoscopic gastrostomy (PEG) placement which demonstrated normal gastric and duodenal anatomy. One week later, a large spike (10x) in blood urea nitrogen: creatinine (BUN: Cr) ratio raised concern for GIB, but hemoglobin remained stable, and stool color remained unchanged. The following day, a gastroenterology consult was requested under increased suspicion of GIB from a sudden 3.5 g/dL hemoglobin drop. The patient received blood transfusion and pantoprazole. An upper endoscopy was performed, revealing three small duodenal ulcers. Melanotic stool ensued afterwards. Conclusions Due to dysautonomia, clinical presentation of GIB can be significantly delayed in the tSCI patient population, leaving them vulnerable to succumb to illness. This case illustrates the possibility of an interval in which the patient was bleeding, with the sole indicator being an elevated BUN. Our case calls for closer monitoring of and vigilance for tSCI patients, and possibly employment of different strategies to reduce the incidence and enhance early detection of GIB in tSCI patients to subsequently decrease the morbidity and mortality associated with it.

BRD4 is a bromodomain extraterminal domain family member and functions primarily as a chromatin reader regulating genes involved in cell-fate decisions. Here, we bred Brd4fl/fl Ox40-Cre mice in which Brd4 was conditionally deleted in OX40-expressing cells to examine the role of BRD4 in regulating immune responses. We found that the Brd4fl/fl Ox40-Cre mice developed profound alopecia and dermatitis, while other organs and tissues were not affected. Surprisingly, lineage-tracing experiments using the Rosa26fl/fl-Yfp mice identified a subset of hair follicle stem cells (HFSCs) that constitutively express OX40, and deletion of Brd4 specifically in such HFSCs resulted in cell death and a complete loss of skin hair growth. We also found that death of HFSCs triggered massive activation of the intradermal γδ T cells, which induced epidermal hyperplasia and dermatitis by producing the inflammatory cytokine IL-17. Interestingly, deletion of Brd4 in Foxp3+ Tregs, which also constitutively express OX40, compromised their suppressive functions, and this, in turn, contributed to the enhanced activation of γδ T cells, as well as the severity of dermatitis and hair follicle destruction. Thus, our data demonstrate an unexpected role of BRD4 in regulating skin follicle stem cells and skin inflammation.

Objectives Damage control laparotomy (DCL) remains an important tool in the trauma surgeon’s armamentarium. Inconsistency in reporting standards have hindered careful scrutiny of DCL outcomes. We sought to develop a core outcome set (COS) for DCL clinical studies to facilitate future pooling of data via meta-analysis and Bayesian statistics while minimizing reporting bias.Methods A modified Delphi study was performed using DCL content experts identified through Eastern Association for the Surgery of Trauma (EAST) ‘landmark’ DCL papers and EAST ad hoc COS task force consensus.Results Of 28 content experts identified, 20 (71%) participated in round 1, 20/20 (100%) in round 2, and 19/20 (95%) in round 3. Round 1 identified 36 potential COS. Round 2 achieved consensus on 10 core outcomes: mortality, 30-day mortality, fascial closure, days to fascial closure, abdominal complications, major complications requiring reoperation or unplanned re-exploration following closure, gastrointestinal anastomotic leak, secondary intra-abdominal sepsis (including anastomotic leak), enterocutaneous fistula, and 12-month functional outcome. Despite feedback provided between rounds, round 3 achieved no further consensus.Conclusions Through an electronic survey-based consensus method, content experts agreed on a core outcome set for damage control laparotomy, which is recommended for future trials in DCL clinical research. Further work is necessary to delineate specific tools and methods for measuring specific outcomes.Level of evidence V, criteria

FOXP3 is the lineage-defining transcription factor for Tregs, a cell type critical to immune tolerance, but the mechanisms that control FOXP3 expression in Tregs remain incompletely defined, particularly as it relates to signals downstream of TCR and CD28 signaling. Herein, we studied the role of IRF4 and BATF3, two transcription factors upregulated upon T cell activation, to the conversion of conventional CD4+ T cells to FOXP3+ T cells (iTregs) in vitro. We found that IRF4 must partner with BATF3 to bind to a regulatory region in the Foxp3 locus where they cooperatively repress FOXP3 expression and iTreg induction. In addition, we found that interactions of these transcription factors are necessary for glycolytic reprogramming of activated T cells that is antagonistic to FOXP3 expression and stability. As a result, Irf4 KO iTregs show increased demethylation of the critical CNS2 region in the Foxp3 locus. Together, our findings provide important insights how BATF3 and IRF4 interactions integrate activating signals to control CD4+ cell fate decisions and govern Foxp3 expression.

Cerebral palsy (CP) is caused by a variety of factors attributed to early brain damage, resulting in permanently impaired motor control, marked by weakness and muscle stiffness. To find out if altered physiology of spinal motoneurons (MNs) could contribute to movement deficits, we performed whole-cell patch-clamp in neonatal rabbit spinal cord slices after developmental injury at 79% gestation. After preterm hypoxia-ischemia (HI), rabbits are born with motor deficits consistent with a spastic phenotype including hypertonia and hyperreflexia. There is a range in severity, thus kits are classified as severely affected, mildly affected, or unaffected based on modified Ashworth scores and other behavioral tests. At postnatal day (P)0–5, we recorded electrophysiological parameters of 40 MNs in transverse spinal cord slices using whole-cell patch-clamp. We found significant differences between groups (severe, mild, unaffected and sham control MNs). Severe HI MNs showed more sustained firing patterns, depolarized resting membrane potential, and fired action potentials at a higher frequency. These properties could contribute to muscle stiffness, a hallmark of spastic CP. Interestingly altered persistent inward currents (PICs) and morphology in severe HI MNs would dampen excitability (depolarized PIC onset and increased dendritic length). In summary, changes we observed in spinal MN physiology likely contribute to the severity of the phenotype, and therapeutic strategies for CP could target the excitability of spinal MNs.

Enhancers are cis-regulatory elements in the genome that cooperate with promoters to control target gene transcription. Unlike promoters, enhancers are not necessarily adjacent to target genes and can exert their functions regardless of enhancer orientations, positions and spatial segregations from target genes. Thus, for a long time, the question as to how enhancers act in a temporal and spatial manner attracted considerable attention. The recent discovery that enhancers are also abundantly transcribed raises interesting questions about the exact roles of enhancer RNA (eRNA) in gene regulation. In this review, we highlight the process of enhancer transcription and the diverse features of eRNA. We review eRNA functions, which include enhancer-promoter looping, chromatin modifying, and transcription regulating. As eRNA are transcribed from active enhancers, they exhibit tissue and lineage specificity, and serve as markers of cell state and function. Finally, we discuss the unique relationship between eRNA and super enhancers in phase separation wherein eRNA may contribute significantly to cell fate decisions.

Background Exercise and heat trigger dehydration and an increase in extracellular fluid osmolality, leading to deficits in exercise performance and thermoregulation. Evidence from previous studies supports the potential for deep-ocean mineral water to improve recovery of exercise performance post-exercise. We therefore wished to determine whether acute rehydration and muscle strength recovery was enhanced by deep-ocean mineral water following a dehydrating exercise, compared to a sports drink or mountain spring water. We hypothesized that muscle strength would decrease as a result of dehydrating exercise, and that recovery of muscle strength and hydration would depend on the type of rehydrating fluid. Methods Using a counterbalanced, crossover study design, female (n = 8) and male (n = 9) participants performed a dehydrating exercise protocol under heat stress until achieving 3% body mass loss. Participants rehydrated with either deep-ocean mineral water (Deep), mountain spring water (Spring), or a carbohydrate-based sports drink (Sports) at a volume equal to the volume of fluid loss. We measured relative hydration using salivary osmolality (Sosm) and muscle strength using peak torque from a leg extension maneuver. Results Sosm significantly increased (p

Feline obesity elicits a plethora of metabolic responses leading to comorbidities, with potential reversal during weight loss. The specific metabolic alterations and biomarkers of organ dysfunction are not entirely understood. Untargeted, high-throughput metabolomic technologies may allow the identification of biological components that change with weight status in cats, increasing our understanding of feline metabolism. The objective of this study was to utilize untargeted metabolomic techniques to identify biomarkers and gain mechanistic insight into the serum metabolite changes associated with reduced food intake and weight loss in overweight cats. During a four-wk baseline period, cats were fed to maintain body weight. For 18 wk following baseline, cats were fed to lose weight at a rate of ~1.5% body weight/wk. Blood serum metabolites were measured at wk 0, 1, 2, 4, 8, 12, and 16. A total of 535 named metabolites were identified, with up to 269 of them being altered (p- and q-values < 0.05) at any time point. A principal component analysis showed a continual shift in metabolite profile as weight loss progressed, with early changes being distinct from those over the long term. The majority of lipid metabolites decreased with weight loss; however, ketone bodies and small lipid particles increased with weight loss. The majority of carbohydrate metabolites decreased with weight loss. Protein metabolites had a variable result, with some increasing, but others decreasing with weight loss. Metabolic mediators of inflammation, oxidative stress, xenobiotics, and insulin resistance decreased with weight loss. In conclusion, global metabolomics identified biomarkers of reduced food intake and weight loss in cats, including decreased markers of inflammation and/or altered macronutrient metabolism.

The language of gene expression displays topological symmetry. An important step during gene expression is the binding of transcriptional proteins to DNA promoters adjacent to a gene. Some proteins bind to many promoters in a genome, defining a regulon of genes wherein each promoter might vary in DNA sequence relative to the average consensus. Here we examine the linguistic organization of gene promoter networks, wherein each node in the network represents a promoter and links between nodes represent the extent of base pair-sharing. Prior work revealed a fractal nucleus in several σ-factor regulons from Escherichia coli. We extend these findings to show fractal nuclei in gene promoter networks from three bacterial species, E. coli, Bacillus subtilis, and Pseudomonas aeruginosa. We surveyed several non-σ transcription factors from these species and found that many contain a nucleus that is both visually and numerically fractal. Promoter footprint size scaled as a negative power-law with both information entropy and fractal dimension, while the latter two parameters scaled positively and linearly. The fractal dimension of the diffuse networks (dB = ~1.7) was close to that expected of a diffusion limited aggregation process, confirming prior predictions as to a possible mechanism for development of this structure.

Gene promoter networks (GPNs) are systems-level representations of the base pair-sharing relationships (graph edges) among promoters (graph nodes). It has been shown in the bacterium E. coli that these networks can contain a fractal nucleus of strong associations suggesting a self-organizing complexity. Here I report results of twenty seven in silico simulations for a diffusion limited aggregation model which accounts for much of the fractal structure previously observed in GPNs. Parameters varied in the model included (a) the frequency of gene duplication events, and the extent of (b) attraction and (c) repulsion presented by the DNAprotein binding chemistry. Both duplication and attraction had significant effects on fractal topology of the GPN nucleus, whereas repulsion due to DNA-protein binding chemistry did not, at least for the levels explored in these simulations. Since repulsion is thought to be a key feature of fractal networks, it is likely that the repulsion in GPNs arises from the sparseness of the promoter space. The generation of a finite random set of promoters leads to sparse occupancy of promoter space which itself presents a considerable repulsion away from the consensus motif, working against the DNA-binding protein's efforts to organize the system of promoters over evolutionary time. This interplay between attractive and repulsive forces in a GPN is sufficient to generate a fractal topology.

Dogs participating in endurance exercise, including herding, hunting and racing have a greater energy requirement and may be more susceptible to nutrient depletion, electrolyte imbalance and metabolic stress. The objective of the present study was to investigate the acute response to unstructured mixed exercise in American Foxhounds fed a nutrient-fortified endurance diet. Thirty-nine adult Foxhound dogs (median age: 5·0, range: 2–10 years and median body weight (BW): 36·4, range: 24·9–49·5 kg) were allotted to a standard performance diet (Control) or nutrient-fortified endurance diet for adult dogs (Test). Dogs were balanced by sex, age, BW and athletic performance between diets. All male dogs were intact, whereas all the female dogs were spayed. After 80 d on diet, blood samples were collected via jugular puncture at baseline (0 h), and at 3 and 25 h post-exercise (mean: 17·7 (sem 0·92) km run over 2–3 h). Plasma taurine concentration and complete amino acid (AA) profile, serum chemistry and creatine kinase were measured. Serum chemistry profile remained within normal ranges throughout the study. A significant (P

In response to global economic duress and heightened consumer awareness of nutrition and health, sustainable and natural ingredients are in demand. Identification of alternative sources of nitrogen and amino acids, including taurine, may help meet dietary requirements while fostering sustainability and natural feeding approaches. Twenty plants, eighteen marine algae and five insect species were analysed. All samples were freeze-dried, hydrolysed and filtered prior to amino acid analysis. Samples for amino acids were analysed in duplicate and averaged. Nitrogen was analysed and crude protein (CP) determined by calculation. With the exception of taurine concentration in soldier fly larvae, all insects exceeded both the National Research Council's canine and feline minimal requirements (MR) for growth of all essential amino acids (EAA) and CP. Although some plants and marine algal species exceeded the canine and feline MR for growth for EAA and CP, only very low concentrations of taurine were found in plants. Taurine concentration in insects was variable but high, with the greatest concentration found in ants (6·42 mg/g DM) and adult flesh flies (3·33 mg/g DM). Taurine was also high in some macroalgae, especially the red algal species: Mazaella spp. (4·11 mg/g DM), Porphyra spp. (1·22 mg/g DM) and Chondracanthus spp. (6·28 mg/g DM). Preliminary results suggest that insects and some marine algal species may be practical alternatives to traditional protein and supplemental taurine sources in pet foods. Safety, bioavailability, palatability and source variability of alternative items as food ingredients should be investigated prior to incorporation into canine and feline diets.

The benefits of whole grain consumption have been studied in human subjects, but little research exists on their effects in dogs. The objective of the present study was to test the effects of resistant starch (RS) in the diet of healthy adult dogs. Twelve adult Miniature Schnauzer dogs (eight males, four females; mean age: 3·3 (1·6) years; mean body weight: 8·4 (1·2) kg; mean body condition score: D/ideal) were randomly allotted to one of three treatment groups, which consisted of different amounts of RS supplied in a biscuit format. Dogs received either 0, 10 or 20 g biscuits per d (estimated to be 0, 2·5 or 5 g RS per d) that were fed within their daily energetic allowance. A balanced Latin square design was used, with each treatment period lasting 21 d (days 0–17 adaptation; days 18–21 fresh and total faecal collection). All dogs were fed the same diet to maintain body weight throughout the study. Dogs fed 5 g RS per d had lower (P = 0·03) fat digestibility than dogs fed 0 gRS per d, but DM, organic matter and crude protein digestibilities were not affected. Faecal fermentative end-products, including SCFA and branched-chain fatty acids, ammonia, phenols and indoles, and microbial populations were not affected. The minor changes observed in the present study suggest the RS doses provided to the dogs were too low. Further work is required to assess the dose of RS required to affect gut health.

The impostor phenomenon was introduced over four decades ago and has become an increasingly popular research topic. Nonetheless, few empirical investigations offer a quantitative analysis of the patterns and structures within the impostor phenomenon&apos;s literature base. This limits researchers to an incomplete and reactive understanding of the scope and trajectory of research about the impostor phenomenon. The current investigation utilized bibliometric analyses (e.g., science mapping and performance analysis) to illuminate prominent contributors and the latent themes within the impostor phenomenon literature. The findings identified important characteristics of the impostor phenomenon literature, such as the most dominant or published authors, frequently cited articles, its global existence across countries, and its rapidly increasing rate of published articles. The findings also illuminated research streams within the impostor phenomenon literature and identified the emerging trend of examining the impostor phenomenon within the STEM context. Implications for future impostor phenomenon research and scholarship are discussed., Author(s): Steven Stone-Sabali [sup.1] , Donte L. Bernard [sup.2] , Kristen J. Mills [sup.1] , Preston R. Osborn [sup.3] Author Affiliations: (1) https://ror.org/00rs6vg23, grid.261331.4, 0000 0001 2285 7943, Department of [...]

Metal transfer to femoral heads may result from impingement against the metallic acetabular shell following subluxation/dislocation, or when metallic debris enters the articulation zone. Such transfers roughen the head surface, increasing polyethylene wear in total hip replacements. Presently, we examined the surface roughness of retrieved femoral heads with metallic transfer. Profilometry revealed roughness averages in regions of metal transfer averaging 0.380 𝜇m for CoCr and 0.294 𝜇m for ZrO2 which were one order of magnitude higher than those from non-implanted controls. Scanning electron microscopy (SEM) revealed adherent transfers on these retrievals, with titanium presence confirmed by electron dispersive spectroscopy. Due to the concern for increased wear, metal transfer was induced on non-implanted heads, which were then articulated against flat polyethylene discs in multidirectional sliding wear tests. Increased polyethylene wear was associated with these specimens as compared to unaltered controls. SEM imaging provided visual evidence that the transfers remained adherent following the wear tests. Pre- and post-test roughness averages exceeded 1 𝜇m for both the CoCr and ZrO2 heads. Overall, these results suggest that metal transfer increases the surface roughness of CoCr and ZrO2 femoral heads and that the transfers may remain adherent following articulation against polyethylene, leading to increased polyethylene wear.

ObjectivesDamage control laparotomy (DCL) remains an important tool in the trauma surgeon's armamentarium. Inconsistency in reporting standards have hindered careful scrutiny of DCL outcomes. We sought to develop a core outcome set (COS) for DCL clinical studies to facilitate future pooling of data via meta-analysis and Bayesian statistics while minimizing reporting bias.MethodsA modified Delphi study was performed using DCL content experts identified through Eastern Association for the Surgery of Trauma (EAST) 'landmark' DCL papers and EAST ad hoc COS task force consensus.ResultsOf 28 content experts identified, 20 (71%) participated in round 1, 20/20 (100%) in round 2, and 19/20 (95%) in round 3. Round 1 identified 36 potential COS. Round 2 achieved consensus on 10 core outcomes: mortality, 30-day mortality, fascial closure, days to fascial closure, abdominal complications, major complications requiring reoperation or unplanned re-exploration following closure, gastrointestinal anastomotic leak, secondary intra-abdominal sepsis (including anastomotic leak), enterocutaneous fistula, and 12-month functional outcome. Despite feedback provided between rounds, round 3 achieved no further consensus.ConclusionsThrough an electronic survey-based consensus method, content experts agreed on a core outcome set for damage control laparotomy, which is recommended for future trials in DCL clinical research. Further work is necessary to delineate specific tools and methods for measuring specific outcomes.Level of evidenceV, criteria.

We are developing thin tile scintillator detectors with silicon photomultiplier (SiPM) readout for use in a multi-layer fast-neutron tracker. The tracker is based on interleaved Timepix and plastic scintillator layers. The thin 15 × 15 × 2 mm plastic scintillators require suitable optical readout in order to detect and measure the energy lost by energetic protons that have been recoiled by fast neutrons. Our first prototype used dual SiPMs, coupled to opposite edges of the scintillator tile using light-guides. An alternative readout geometry was designed in an effort to increase the fraction of scintillation light detected by the SiPMs. The new prototype uses a larger SiPM array to cover the entire top face of the tile. This paper details the comparative performance of the two prototype designs. A deuterium-tritium (DT) fast-neutron source was used to compare the relative light collection efficiency of the two designs. A collimated UV light source was scanned across the detector face to map the uniformity. The new prototype was found to have 9.5 times better light collection efficiency over the original design. Both prototypes exhibit spatial non-uniformity in their response. Methods of correcting this non-uniformity are discussed.

The objective of the present study was to evaluate the changes in blood metabolites, AA profile, and oxidative stress markers in American Foxhound dogs fed a nutrient-fortified endurance diet while undergoing unstructured endurance exercise over several months. Thirty-six adult American Foxhound dogs (mean age: 4.5, range 2 to 10 yr and mean BW: 34.7, range: 23.1 to 46.9 kg) were selected to participate in the study. Prior to the study, all dogs consumed a commercial diet for 16 wk. After collecting baseline blood samples, dogs were assigned to a standard commercial performance diet (control) or a nutrient-fortified dog food (test). Dogs were balanced by gender, age, body weight, and athletic performance between diets. During the study, dogs underwent 78 bouts of exercise, with approximately 22 km/bout. Blood samples were collected after 40, 75, 138, and 201 d on study (October 2012 to March 2013). All blood metabolites were similar at baseline and serum chemistry profile remained within normal ranges throughout the study. Over time, plasma taurine and vitamin E concentrations decreased (P < 0.05) in dogs fed the control diet but were maintained or increased (P < 0.05) in dogs fed the treatment diet. Also, plasma creatinine and triglycerides were lower (P < 0.05) and blood phosphorus and alkaline phosphatase were higher (P < 0.05) in dogs fed the treatment diet. Vitamin E and taurine status of dogs appear to be affected by extended endurance exercise. These data suggest dogs undergoing endurance exercise may benefit from supplementation of vitamin E and taurine to minimize oxidation and maintain taurine status.

BackgroundRecent studies have suggested the length of treatment of intra-abdominal infections (IAIs) can be shortened without detrimental effects on patient outcomes. However, data from high-risk patient populations are lacking. We hypothesized that patients at high risk for treatment failure will benefit from a longer course of antimicrobial therapy.MethodsPatients enrolled in the Study to Optimize Peritoneal Infection Therapy (STOP-IT) trial were evaluated retrospectively to identify risk factors associated with treatment failure, which was defined as the composite outcome of recurrent IAI, surgical site infection, or death. Variables were considered risk factors if there was a positive statistical association with treatment failure. Patients were then stratified according to the presence and number of these risk factors. Univariable analyses were performed using the Kruskal-Wallis, χ2, and Fisher exact tests. Logistic regression controlling for risk factors and original randomization group, either a fixed four-day antimicrobial regimen (experimental) or a longer course based on clinical response (control), also was performed.ResultsWe identified corticosteroid use, Acute Physiology and Chronic Health Evaluation II score ≥5, hospital-acquired infection, or a colonic source of IAI as risk factors associated with treatment failure. Of the 517 patients enrolled, 263 (50.9%) had one or two risk factors and 16 (3.1%) had three or four risk factors. The rate of treatment failure rose as the number of risk factors increased. When controlling for randomization group, the presence and number of risk factors were independently associated with treatment failure, but the duration of antimicrobial therapy was not.ConclusionsWe were able to identify patients at high risk for treatment failure in the STOP-IT trial. Such patients did not benefit from a longer course of antibiotic administration. Further study is needed to determine the optimum duration of antimicrobial therapy in high-risk patients.

Three extruded dog diets were created for a nutritional study with different primary protein sources (BAS: lamb meal (LM) and deboned lamb (DL); CHK: chicken meal, LM, and DL; PEA: dried ground pea, LM, and DL). All diets were processed using the same single‐screw extruder, shipped from the processing facility on the same day, and transported under the same conditions in January 2021. After 8 months of storage in a temperature and humidity‐controlled room in September 2021, only the PEA diet was molded upon inspection. Mold and mycotoxin analysis of all diets was conducted in both September 2021 and at expiry in January 2022, which confirmed mold and mycotoxin contamination to some degree in all diets and most pronounced in the PEA diet across both timepoints. Nutrient analysis of all diets was conducted at production and 2 months post‐expiry in March 2022. As expected, fatty acid and vitamin contents of all diets decreased between sampling timepoints, and amino acid contents generally remained stable. Methionine decreased by 14% in CHK, cystine decreased by 15% and 20% in CHK and PEA, respectively, tyrosine decreased by 30%, 25%, and 27% across BAS, CHK, and PEA, respectively, and taurine decreased by 50%, 42%, and 55% across BAS, CHK, and PEA, respectively. Inaccurate measurement of the PEA diet moisture content post‐production likely led to mold development which may also negatively impact the availability of nutrients and could put dogs at risk for mycotoxicosis and nutrient deficiencies if not closely monitored, but controlled studies are required. [ABSTRACT FROM AUTHOR]