Three-dimensional (3D) food printing is a rapidly emerging technology offering unprecedented potential for customised food design and personalised nutrition. Here, we evaluate the technological advances in extrusion-based 3D food printing and its possibilities to promote healthy and sustainable eating. We consider the challenges in implementing the technology in real-world applications. We propose viable applications for 3D food printing in health care, health promotion and food waste upcycling. Finally, we outline future work on 3D food printing in food safety, acceptability and economics, ethics and regulations.

OBJECTIVES/GOALS: The goal of this proposal is to develop a technology that combines calcium imaging via confocal microscopy, and force measurement via monolayer stress microscopy to perform simultaneous quantitative measurements of agonist-induced Ca2+ and mechanical signals in HASMCs. METHODS/STUDY POPULATION: The methods by which second messenger signals and changes in mechanical forces determine specific physiological responses are complex. Recent studies point to the importance of temporal and spatial encoding in determining signal specificity. Hence, approaches that probe both chemical and mechanical signals are needed. We combine hyperspectral imaging for second messenger signal measurements, monolayer stress microscopy for mechanical force measurements, and S8 analysis software for quantifying localized signals. Imaging was performed using an excitation-scanning hyperspectral microscope. Hyperspectral images were unmixed to identify signals from fluorescent labels and microparticles. Images were analyzed to quantify localized force dynamics through monolayer stress microscopy. RESULTS/ANTICIPATED RESULTS: Results indicate that localized and transient cellular signals can be quantified and mapped within cell populations. Importantly, these results establish a method for simultaneous interrogation of cellular signals and mechanical forces that may play synergistic roles in regulating downstream cellular physiology in confluent monolayers. DISCUSSION/SIGNIFICANCE: We will measure the distribution of chemical and mechanical signals within cells, providing insight into the dynamics of cell signaling. Studies will have implication in the understanding of infections, drug delivery in which non-uniform distributions of drugs are a certainty, and in understanding coordinated responses in cellular systems.

Hydrodynamic clogging in planar channels is studied via direct numerical simulation for the first time, utilising a novel numerical test cell and stochastic methodology with special focus on the influence of electrostatic forces. Electrostatic physics is incorporated into an existing coupled lattice Boltzmann-discrete element method framework, which is verified rigorously. First, the dynamics of the problem is governed by the Stokes number, $St$. At low $St$, the clogging probability, $P$, increases with $St$ due to increasing collision frequency. At high $St$, however, $P$ decreases with $St$ due to quadratic scaling of hydrodynamic force acting on arches. Under electrostatic forces, clogging is well represented by the wall adhesion number, $Ad_w$. For $Ad_w \lesssim 4$, the mechanical dependence on $St$ is exhibited, while for $4 < Ad_w < 20$, there is a transition to high $P$ as sliding along, and attachment to, the channel surface occurs increasingly. For $Ad_w \gtrsim 20$, clogging occurs with $P > 0.95$. Particle agglomeration, however, can also decrease $P$ due to diminished interaction with channel walls. Distinct parametric regions of clogging are also observed in relation to the channel width, while a critical width $w/d^*=2.6$ is reported, which increases to $w/d^*=4$ with strong electrostatic surface attachment. The number of particles that form stable arches across a planar channel is determined to be $n=\left \lceil {w/d}\right \rceil + 1$. Finally, sensitivity to the Coulomb friction coefficient is determined in favour of calibrating numerical parameters to bulk system behaviour. The greatest sensitivities occur in situations where the arch stability is lowest, while clogging becomes independent of friction for strong wall adhesion.

Plenoptic particle image velocimetry and surface pressure measurements were used to analyse the early development of leading-edge vortices (LEVs) created by a flat-plate wing of aspect ratio 2 rolling in a uniform flow parallel to the roll axis. Four cases were constructed by considering two advance coefficients, $J=0.54$ and 1.36, and two wing radii of gyration, $R_g/c=2.5$ and 3.25. In each case, the wing pitch angle was articulated such as to achieve an angle of attack of $33^{\circ }$ at the radius of gyration of the wing. The sources and sinks of vorticity were quantified for a chordwise rectangular control region, using a vorticity transport framework in a non-inertial coordinate system attached to the wing. Within this framework, terms associated with Coriolis acceleration provide a correction to tilting and spanwise convective fluxes measured in the rotating frame and, for the present case, have insignificant values. For the baseline case ($J=0.54, R_g/c=3.25$), three distinct spanwise regions were observed within the LEV, with distinct patterns of vortex evolution and vorticity transport mechanisms in each region. Reducing the radius of gyration to $R_g/c=2.5$ resulted in a more stable vortex with the inboard region extending over a broader spanwise range. Increasing advance ratio eliminated the conical vortex, resulting in transport processes resembling the mid-span region of the baseline case. Although the circulation of the LEV system was generally stronger at the larger advance coefficient, the shear-layer contribution was diminished.

The cornerstone of obesity treatment is behavioural weight management, resulting in significant improvements in cardio-metabolic and psychosocial health. However, there is ongoing concern that dietary interventions used for weight management may precipitate the development of eating disorders. Systematic reviews demonstrate that, while for most participants medically supervised obesity treatment improves risk scores related to eating disorders, a subset of people who undergo obesity treatment may have poor outcomes for eating disorders. This review summarises the background and rationale for the formation of the Eating Disorders In weight-related Therapy (EDIT) Collaboration. The EDIT Collaboration will explore the complex risk factor interactions that precede changes to eating disorder risk following weight management. In this review, we also outline the programme of work and design of studies for the EDIT Collaboration, including expected knowledge gains. The EDIT studies explore risk factors and the interactions between them using individual-level data from international weight management trials. Combining all available data on eating disorder risk from weight management trials will allow sufficient sample size to interrogate our hypothesis: that individuals undertaking weight management interventions will vary in their eating disorder risk profile, on the basis of personal characteristics and intervention strategies available to them. The collaboration includes the integration of health consumers in project development and translation. An important knowledge gain from this project is a comprehensive understanding of the impact of weight management interventions on eating disorder risk.

The purpose of this experiment was to investigate the welfare of pregnant mares kept in straight stalls and given only limited exercise, conditions that are similar to those encountered in the pregnant mare urine industry. Sixteen pregnant mares (eight in each of two years) were randomly assigned to two groups: Ex (exercised in a paddock for 30 min per day) or NoEx (exercised for one 30 min period every 14 days). The horses were housed in straight (or ‘tie’) stalls for six months and had ad libitum access to grass hay. Each horse's behaviour was recorded on videotape once per week for 24 h. The major behaviours were eating hay, standing, and stand-resting (head down and one hind limb flexed). There was no difference between the behaviours or the number of foot lifts per min of the Ex and NoEx groups in their stalls. Nine of 16 mares were not observed in recumbency throughout the whole of the six-month observation period, suggesting that horses with no previous experience in straight stalls may be reluctant to lie down. Thirteen of 16 mares dropped to their knees at least once, probably when they were REM sleeping while standing. There were no significant differences between the Ex and the NoEx mares in baseline plasma Cortisol levels or in Cortisol response to ACTH. Following 30 min of exercise, NoEx mares showed an increase in Cortisol from 5.0 to 5.4 μg dL−1, whereas Ex mares showed a decrease from 4.6 to 3.6 µg dL−1. The NoEx horses that had been confined for two weeks trotted more (NoEx = 22 [6-38; median and range]% of time; Ex = 2.4 [0-8.7]%) and galloped more (NoEx = 6 [2-8]%; Ex = 0 [0-4]%) than the Ex that were released daily, but walked less (NoEx = 17 [10-26]%; Ex = 35 [20-40]%) and grazed less (NoEx = 0%; Ex = 3 [0-12]%). Confined horses show rebound locomotion — that is, a compensatory increase — when released from confinement, indicating a response to exercise deprivation.

Captive bolts or firearms are unsuitable for euthanasia of livestock when an intact brain is required for diagnostics. Injectable barbiturates can be used, but this method carries risk of poisoning animals eating the carcase. Intravenous saturated salt solutions have been used to euthanase heavily sedated ruminants and are cheap, readily available and not a risk to scavenging animals. However, there is concern that they may be painful or cause distress to animals that are not unconscious. This study aimed to determine the suitability of saturated salt solutions, in combination with xylazine, as a method of euthanasia of ruminants using a sheep model. Thirty-two sheep were sedated with xylazine (0.4 mg kg-1 IM) and euthanased with an intravenous overdose of pentobarbitone (PENT; n = 10), saturated potassium chloride (KCL; n = 11) or saturated magnesium sulphate (MGS; n = 10). Time until end of rhythmic breathing and cardiac arrest, and movement events were recorded. Conscious perception of pain was evaluated by measuring cortical brain activity by electroencephalography (EEG). There was no evidence of perceived pain or unpleasant sensory experience for any treatment as indicated by P50, P95 and Ptot and so all methods were deemed humane. Time until transient EEG was comparable for all treatments. Time until onset of isoelectric EEG was prolonged for KCL. Animals euthanased with KCL consistently exhibited severe reflex movements during infusion (eg kicking, convulsion). No severe movement events were observed in animals euthanased with MGS, hence, physiological and movement data support the preferential use of MGS over KCL.

Humane slaughter implies that an animal experiences minimal pain and distress before it is killed. Stunning is commonly used to induce insensibility but can lead to variable results or be considered unsatisfactory by some religious groups. Microwave energy can induce insensibility in rats, and high power equipment has recently been developed for sheep and cattle. We examined the effectiveness of different settings for microwave energy delivery, power and duration, to induce insensibility based on electroencephalography (EEG) of anaesthetised cows, using the minimal anaesthesia model. All applications resulted in the appearance of seizure-like complexes in the EEG, a pattern considered incompatible with awareness. Shorter duration of application resulted in more rapid EEG changes, as quickly as 3 s. Higher power resulted in a longer duration of EEG suppression, at least 37 s and up to 162 s. Microwave energy can induce insensibility in cattle based on seizure-like complexes in the EEG.

Timely euthanasia on swine farms can help to reduce the incidence of poor welfare outcomes for compromised pigs (Sus scrofa) when recovery is prolonged or impossible. Timely euthanasia relies upon caretakers’ abilities to identify compromised pigs and administer euthanasia in various environments. To determine appropriate timelines and most common reasons for on-farm euthanasia, an online survey was conducted with members of the United States National Pork Board. Additionally, two focus groups were conducted to investigate barriers and possible solutions associated with timely euthanasia. Clinical signs related to poor locomotion (57.6%), prolapses (47.2%), and hernias (43.5%) were identified by the greatest percentage of respondents who believed immediate euthanasia was warranted, while a greater percentage of respondents believed euthanasia was not warranted for clinical signs related to the integumentary (90.3%), reproductive (75.8%), and respiratory (67.5%) systems. The most common reason for euthanasia was poor body condition in pre-weaned piglets and non-ambulatory or severely weak for both breeding and non-breeding pigs. In the focus groups, two themes were identified when evaluating barriers to euthanasia on-farm, and participants agreed that making timely decisions relies upon several dimensions of risk analysis. An unsupportive farm culture was identified as a critical barrier to timely euthanasia decision-making, suggesting that caretaker characteristics may play a role in the success of any timely euthanasia programme. This present study has highlighted areas for future research and demonstrated the need to extend educational efforts both to swine industry leaders and producers to improve overall animal welfare by ensuring timely euthanasia in swine.

As part of surveillance of snail-borne trematodiasis in Knowsley Safari (KS), Prescot, United Kingdom, a collection was made in July 2021 of various planorbid (n = 173) and lymnaeid (n = 218) snails. These were taken from 15 purposely selected freshwater habitats. In the laboratory emergent trematode cercariae, often from single snails, were identified by morphology with a sub-set, of those most accessible, later characterized by cytochrome oxidase subunit 1 (cox1) DNA barcoding. Two schistosomatid cercariae were of special note in the context of human cercarial dermatitis (HCD), Bilharziella polonica emergent from Planorbarius corneus and Trichobilharzia spp. emergent from Ampullacaena balthica. The former schistosomatid was last reported in the United Kingdom over 50 years ago. From cox1 analyses, the latter likely consisted of two taxa, Trichobilharzia anseri, a first report in the United Kingdom, and a hitherto unnamed genetic lineage having some affiliation with Trichobilharzia longicauda. The chronobiology of emergent cercariae from P. corneus was assessed, with the vertical swimming rate of B. polonica measured. We provide a brief risk appraisal of HCD for public activities typically undertaken within KS educational and recreational programmes.

Humpback whales (Megaptera novaeangliae) exhibit maternally driven fidelity to feeding grounds, and yet occasionally occupy new areas. Humpback whale sightings and mortalities in the New York Bight apex (NYBA) have been increasing over the last decade, providing an opportunity to study this phenomenon in an urban habitat. Whales in this area overlap with human activities, including busy shipping traffic leading into the Port of New York and New Jersey. The site fidelity, population composition and demographics of individual whales were analysed to better inform management in this high-risk area. Whale watching and other opportunistic data collections were used to identify 101 individual humpback whales in the NYBA from spring through autumn, 2012–2018. Although mean occurrence was low (2.5 days), mean occupancy was 37.6 days, and 31.3% of whales returned from one year to the next. Individuals compared with other regional and ocean-basin-wide photo-identification catalogues (N = 52) were primarily resighted at other sites along the US East Coast, including the Gulf of Maine feeding ground. Sightings of mother-calf pairs were rare in the NYBA, suggesting that maternally directed fidelity may not be responsible for the presence of young whales in this area. Other factors including shifts in prey species distribution or changes in population structure more broadly should be investigated.

Cesario claims that all bias research tells us is that people “end up using the information they have come to learn as being probabilistically accurate in their daily lives” (sect. 5, para. 4). We expose Cesario's flawed assumptions about the relationship between accuracy and bias. Through statistical simulations and empirical work, we show that even probabilistically accurate responses are regularly accompanied by bias.

The shear-induced migration of dense suspensions with continuously distributed (polydisperse) particle sizes is investigated in planar channel flows for the first time. A coupled lattice Boltzmann–discrete element method numerical framework is employed and validated against benchmark experimental results of bulk shear-induced migration and segregation by particle size. Distinct dependence on the particle size distribution is shown in the flowing (non-plugged) regime (where the bulk solid volume fraction, $\bar{\phi}$, $\leq 0.3$) resulting from a dual dependence on the particle self-diffusivity and local rheology imposed by the particle pressure gradient. Close agreement between statistically equivalent bidisperse and polydisperse suspensions suggests that the bulk migration, and by extension the shear-induced diffusivity, is completely characterised by the first three statistical moments of the particle size distribution. For both bidisperse and polydisperse suspensions in the plugging regime, $\bar {\phi }\geq 0.4$, the smallest particles preferentially form the plugs, causing the largest particles to segregate to the channel walls. This effect is accentuated as $\bar {\phi }$ increases and has not been reported in the literature hitherto. It is proposed that smaller particles preferentially form the plugs due to their higher shear-rate fluctuations, which completely dominate particle motion near the plug where the mean shear rate vanishes. Finally, increasing inertia causes a greater bulk migration towards the channel walls, but increased mid-plane migration for the largest particles due to the dependence of the particle self-diffusivity on the particle Reynolds number. As $\bar {\phi }$ increases shear-induced migration dominates and these inertial effects disappear, as does dependence on the particle size distribution.

Important advances in biomedical and behavioral research ethics have occurred over the past few decades, many of them centered on identifying and eliminating significant harms to human subjects of research. Comprehensive attention has not been paid to the totality of harms experienced by animal subjects, although scientific and moral progress require explicit appraisal of these harms. Science is a public good and the prioritizing within, conduct of, generation of, and application of research must soundly address questions about which research is morally defensible and valuable enough to support through funding, publication, tenure, and promotion. Likewise, educational pathways of re-imagined science are critical.

Background: Children with pathogenic variations in SCN8A can present with early infantile epileptic encephalopathy-13, benign familial infantile seizures-5 or intellectual disability alone without epilepsy. In this case series, we discuss six children with variants in SCN8A managed at BC Children’s Hospital. Methods: We describe clinical and genetic results on six individuals with SCN8A variants identified via clinical or research next-generation sequencing. Functional consequences of two SCN8A variants were assessed using electrophysiological analyses in transfected cells. Results: Clinical findings ranged from normal development with well-controlled epilepsy to significant developmental delay with treatment-resistant epilepsy. Phenotypes and genotypes in our cohort are described in the table below. Functional analysis supported gain-of-function in P2 and loss-of-function in P4. Conclusions: Our cohort expands the clinical and genotypic spectrum of SCN8A-related disorders. We establish functional evidence for two missense variants in SCN8A, including LoF variant in a patient with intellectual disability, and autism spectrum disorder without seizures.

Table for P.120