An automated deep learning pipeline for EMVI classification and response prediction of rectal cancer using baseline MRI: a multi-centre study.

The classification of extramural vascular invasion status using baseline magnetic resonance imaging in rectal cancer has gained significant attention as it is an important prognostic marker. Also, the accurate prediction of patients achieving complete response with primary staging MRI assists clinicians in determining subsequent treatment plans. Most studies utilised radiomics-based methods, requiring manually annotated segmentation and handcrafted features, which tend to generalise poorly. We retrospectively collected 509 patients from 9 centres, and proposed a fully automated pipeline for EMVI status classification and CR prediction with diffusion weighted imaging and T2-weighted imaging. We applied nnUNet, a self-configuring deep learning model, for tumour segmentation and employed learned multiple-level image features to train classification models, named MLNet. This ensures a more comprehensive representation of the tumour features, in terms of both fine-grained detail and global context. On external validation, MLNet, yielding similar AUCs as internal validation, outperformed 3D ResNet10, a deep neural network with ten layers designed for analysing spatiotemporal data, in both CR and EMVI tasks. For CR prediction, MLNet showed better results than the current state-of-the-art model using imaging and clinical features in the same external cohort. Our study demonstrated that incorporating multi-level image representations learned by a deep learning based tumour segmentation model on primary MRI improves the results of EMVI classification and CR prediction with good generalisation to external data. We observed variations in the contributions of individual feature maps to different classification tasks. This pipeline has the potential to be applied in clinical settings, particularly for EMVI classification.

Effects of heat stress on development, quality and survival of Bos indicus and Bos taurus embryos produced in vitro.

Heat stress is an important cause of poor development and low survival rates in bovine embryos. Experiments were conducted to test the hypothesis that Bos indicus embryos are more resistant to heat stress than are Bos taurus embryos. In experiment 1, Nelore and Jersey embryos from oocyte pick-up-derived oocytes were submitted to heat stress (96 hours post-insemination, 41 °C, 6 hours), developmental ratios were assessed at Day 7 (Day 0 = day of fertilization), and blastocysts were frozen for RNA extraction. Experiment 2 evaluated expression of COX2, CDX2, HSF1, and PLAC8 in previously frozen blastocysts. In experiment 3, Nellore and Angus embryos from oocyte pick-up-derived oocytes were submitted to heat stress (96 hours post-insemination, 41 °C, 12 hours) and transferred to recipients on Day 7. In experiment 4, embryos developed as in experiment 3 were fixed for Terminal deoxynucleotidyl transferase dUTP nick end labeling labeling and total cell counting. In experiment 1, heat stress decreased the percentage of Jersey oocytes that became blastocysts, but had no effect on Nellore embryos (34.6%, 25.0%, 39.5%, and 33.0% for Jersey control, Jersey heat-stressed, Nellore control, and Nellore heat-stressed oocytes, respectively; P < 0.05). In experiment 2, heat stress decreased (P < 0.05) expression of CDX2 and PLAC8, with higher expression of these genes in Nellore embryos than in Jersey embryos. Heat stress also decreased (P < 0.05) expression of COX2 in Jersey embryos, but had no effect on Nellore embryos. Expression of HSF1 was decreased (P < 0.05) by heat stress in both breeds, with a greater effect in Nellore embryos. In experiment 3, heat stress tended (P = 0.1) to decrease the percentage of pregnancies among cows (Day 30 to 35) that received Angus embryos. In experiment 4, heat stress increased (P < 0.05) the percentage of apoptotic blastomeres, but had no breed-specific effects. In addition, Nellore embryos had fewer (P < 0.05) Terminal deoxynucleotidyl transferase dUTP nick end labeling- positive blastomeres than did Angus embryos. We concluded that the detrimental effects of heat stress were dependent upon embryo breed and were more evident in Bos taurus embryos than in Bos indicus embryos.

Sine ars scientia nihil est: Leonardo da Vinci and beyond.

The aim of this article is to reflect on the relationship between art and science so far as it concerns a symposium on neurosciences. We undertake a historical overview of that relationship, paying particular attention to the sui generis case of Leonardo da Vinci, who very often is regarded as the man who worked on art and science with equal ease. We then explain why his idea of merging these two forms of knowledge failed, considering the clear-cut distinction between art and science in his time. With this clarification, we explore the matter today. We look at Raphael's The Transfiguration, in which the representation of the possessed boy is seen by neuroscientists as indicative of an epileptic seizure. We also look at the ideas of neuroscientists Semir Zeki and Vilayanur Ramachandran, who study particular aspects of brain function and suggest a new merging of art and science.

Investigation of the substrate specificity of a beta-glycosidase from Spodoptera frugiperda using site-directed mutagenesis and bioenergetics analysis.

The specificity of the Spodoptera frugiperda digestive beta-glycosidase (Sfbetagly50) for fucosides, glucosides and galactosides is determined by noncovalent interactions of glycone 6-OH and glycone 4-OH with the active-site residues Q39 and E451. Site-directed mutagenesis and enzyme steady-state kinetics were described, showing that replacement of E451 with glutamine increased the preference of Sfbetagly50 for glucosides in comparison to galactosides, whereas replacing E451 with serine had the opposite effect. In contrast, the replacement of E451 with aspartate did not change Sfbetagly50 specificity. The energy of the interactions formed by these different residues with the axial and equatorial glycone 4-OH were also measured, showing that the increase in preference for galactosides resulted from a larger energy decrease in the interaction with equatorial 4-OH than with axial 4-OH (22.6 vs. 13.9 kJ x mol(-1)), whereas the increase in preference for glucosides was caused by an energy reduction in the interaction with the axial 4-OH (5.1 kJ x mol(-1)). The introduction of glutamine at position 451 or of asparagine at position 39 increased the preference of Sfbetagly50 for fucosides in comparison to galactosides, whereas the presence of aspartate or serine at position 451 had less effect on this preference. The hydrolysis of fucosides was favored because glutamine at position 451 increased a steric hindrance with 6-OH of 7.1 kJ x mol(-1) and asparagine at position 39 disrupted a favorable interaction with this same hydroxyl. In conclusion, it is proposed that the specificity of new beta-glycosidase mutants can be predicted by combining and adding energy of the enzyme-substrate interactions evaluated in the present study.

The role of residues R97 and Y331 in modulating the pH optimum of an insect beta-glycosidase of family 1.

The activity of the digestive beta-glycosidase from Spodoptera frugiperda (Sfbetagly50, pH optimum 6.2) depends on E399 (pKa = 4.9; catalytic nucleophile) and E187 (pKa = 7.5; catalytic proton donor). Homology modelling of the Sfbetagly50 active site confirms that R97 and Y331 form hydrogen bonds with E399. Site-directed mutagenesis showed that the substitution of R97 by methionine or lysine increased the E399 pKa by 0.6 or 0.8 units, respectively, shifting the pH optima of these mutants to 6.5. The substitution of Y331 by phenylalanine increased the pKa of E399 and E187 by 0.7 and 1.6 units, respectively, and displaced the pH optimum to 7.0. From the observed deltapKa it was calculated that R97 and Y331 contribute 3.4 and 4.0 kJ.mol(-1), respectively, to stabilization of the charged E399, thus enabling it to be the catalytic nucleophile. The substitution of E187 by D decreased the pKa of residue 187 by 0.5 units and shifted the pH optimum to 5.8, suggesting that an electrostatic repulsion between the deprotonated E399 and E187 may increase the pKa of E187, which then becomes the catalytic proton donor. In short the data showed that a network of noncovalent interactions among R97, Y331, E399 and E187 controls the Sfbetagly50 pH optimum. As those residues are conserved among the family 1 beta-glycosidases, it is proposed here that similar interactions modulate the pH optimum of all family 1 beta-glycosidases.