EMT and mesendoderm markers upregulated in neurospheres after 48 h induction with serum and Lif.

<p>(<b>A</b>): Immunostaining for Slug, E-cadherins and N-cadherins (<b>B</b>): Immunostaining for mesendoderm markers Sox17 and Brachyury (T). (<b>C</b>): Expression profile of Slug, N-cadherins, E-cadherins, Sox17 and Brachyury of 48 h serum and Lif induced neurospheres measured by QPCR relative to neurospheres cultured in standard neural stem cell media. Abbreviations: Ncad, N-cadherins; Ecad, E-cadherins. Scale bars: 50 µm.</p

Schematic diagram depicting our interpretation of the results.

<p>Neural stem cells derived from later stages of development have been shown to reprogram to iPS cells via overexpression of <i>Oct4</i> transcription factor (Kim et al., 2009). Signalling alone (BMPs and bFGF), however, can induce dedifferentiation of neural stem cells into a neural crest phenotype (Sailer et al., 2005). TGFβ and Jak/Stat pathways can induce a further dedifferentiation to mesendoderm-like phenotype providing evidence for extracellular signaling regulated cell plasticity of neural stem cells (this work, red arrow).</p

Effect of signalling pathway inhibitors on upregulation of mesendoderm markers during 48 h induction of neurospheres.

<p>Expression levels: ++++ very strong; +++ strong; ++good; + weak; − no expression.</p

<i>In vivo</i> differentiation of 48 h induced neural stem cells.

<p>Staining of chick embryo paraffin sections for mesenchymal marker N-cadherin, tight junction marker ZO-1, endoderm marker Sox17, and epithelial marker E-cadherin. Although some of the labelled cells express E-cadherin, the staining pattern suggest the cells do not achieve a complete integration to ectoderm lineage. However, high integration towards mesoderm and endoderm lineages is confirmed by N-cadherin, ZO-1 and Sox17 stainings. Although induced (green) cells show higher efficiency of integration, both induced (green) and non-induced (red) cells once incorporated into these tissues express respective lineage markers and acquire similar morphological characteristics to their neighbouring host cells.</p

Expression of pluripotency associated proteins in serum-free cultured neurospheres and 48 h serum and Lif induced neurospheres.

<p>(<b>A</b>): Morphology in serum-free (upper panel) and 48 h serum/Lif (lower panel) conditions. (<b>B</b>): Transcriptional profile of pluripotency associated factors Oct4, Sox2, c-Myc, Klf4 and Nanog of serum-free cultured neurospheres and 48 h treated neurospheres. Data is expressed as ΔΔCT and normalized to ES cells. (<b>C</b>): Immunostaining for Oct4, Nanog, Sox2, SSEA1 and alkaline phosphatase. Abbreviations: AP, alkaline phosphatase; SSEA1, stage specific embryonic antigen 1. Scale bars: 50 µm.</p

<i>In vitro</i> differentiation of neurospheres in serum and Lif conditions.

<p>(<b>A</b>): Immunostaining for Brachyury (T), Sox17, Nanog and Oct4 at 5 days after induction in serum and Lif conditions (<b>B</b>): At 10 days post induction with serum and Lif, cells exhibit very heterogeneous morphologies, indicating the presence of different cell types. (<b>C</b>): Immunostaining for GFAP and αSMA in 10 day induced cultures. Abbreviations: GFAP, glial fibrillary acidic protein; αSMA, alpha smooth muscle actin. Scale bars: 50 µm.</p

Supplementary Table 1 from Shared stressful experiences affect social proximity in Merino sheep

Mean (and standard error) of weather parameters of different cohorts across four study day

Supplementary Figure 1 from Shared stressful experiences affect social proximity in Merino sheep

RTK rovers fitted to the backs of experimental shee

Towards a more practical attention bias test to assess affective state in sheep

<div><p>Tests for attention bias potentially offer more rapid assessment of affective state in animals than existing cognitive methods. An attention bias test has previously been developed for sheep and validated as a measure of anxious states. The 3 minute test assessed behavioural responses of sheep in an enclosed arena after brief exposure to the threat of a dog. Experiment 1 of the current study aimed to refine the previously developed method, removing the need for a habituation period and shortening the test duration. Sheep were given either an anxiolytic drug, an anxiogenic drug or a control treatment prior to testing to induce contrasting affective states. Differences in behaviour were found between the treatment groups within the first 45s of the test, indicating the original test duration could be shortened from 180 s. During testing, 36 of 40 animals in the control and anxiolytic groups ate the novel feed offered in the test, indicating it is not necessary to habituate animals to a feed container. Experiment 2 aimed to confirm the responses measured in the test were primarily towards the dog rather than other aspects of the test environment. Sheep exposed to an empty window at the beginning of the test behaved differently to those which were exposed to a dog, indicating sheep behaviour in the test is at least partially a response to the dog. A third group of sheep were also tested with the dog immediately after having small data loggers attached to their necks. Behaviour of these sheep did not differ from the sheep tested without loggers, indicating data logger attachment did not impact their behaviour in the test. In both experiments, treatments did not appear to modify activity (zones crossed), which we propose indicates the test was primarily detecting valence of the affective state rather than arousal.</p></div

Machine Learning and Taguchi DOE Combined Approach for Modeling Dynamic Ultrasound-Assisted Fresh-Cut Leafy Green Sanitation

Chlorine-based fresh produce sanitation is a dynamic process, and sanitation efficiency is limited due to chlorine degradation. Here, ultrasound was coupled with a benchtop sanitation system to enhance chlorine sanitizer efficiency in fresh-cut leafy green sanitation. Taguchi design of experiments (DOE) and machine learning (ML) were combined to model the relationship between sanitation condition parameters and sanitation outcomes. Multiple ML algorithms were fitted, tuned, and compared for performance using 127 experimental trials (training-to-validation ratio = 3:1). Gaussian process regression (GPR) models showed the best performance in predicting sanitation outcomes of chemical oxygen demand (COD, R2 = 0.73), remaining Escherichia coli O157:H7 on the leaf surface (“Surface Microbe”, R2 = 0.88), and E. coli O157:H7 concentration in sanitation water (“Water Microbe”, R2 = 1.00). Cut size and agitation speed were identified as the most critical input parameters. An initial free chlorine concentration over 20 mg/L was recommended to minimize the E. coli O157:H7 concentration in sanitation water. This work showcases the combined approach of ML and DOE in optimizing fresh-cut produce sanitation. Moreover, it provides a solution for overcoming the difficulties of modeling multiple controllable and uncontrollable factors with reduced experimental runs