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The aim of this study was to indentify the involvement of leptin receptors (LepR) in astrocytes in hippocampal synaptic transmission and plasticity and metabolism. To this end we used a genetic mouse model (GFAP-LepR -/- ) of specific LepR ablation in GFAP positive cells and recorded excitatory postsynaptic potentials (fEPSPs) within the CA1 area. Glutamate (Glu) uptake and the expression of Glu transporters (EEAT3, GLT-1 and GLAST) and enzymes involved in Glu metabolism (glutamine synthase, GABA decarboxylase 65 and 67) were quantified. Modifications in the expression of GFAP, the glucose transporter (GLUT)-1, and the monocarboxylate transporters MCT-2 and MCT-4, were also analyzed. The results show that depletion of LepR in GFAP positive cells reduced basal synaptic transmission within the CA1 area and impaired N-methyl-d-aspartate (NMDA)-evoked long-term depression (NMDA-LTD). Hippocampal slices from GFAP-LepR -/- mice displayed lower Glu uptake efficacy together with up-regulation of GLT-1, glutamine synthase, GFAP and GLUT-1. In conclusion, astrocyte LepRs are involved in the maintenance of Glu homeostasis and Glu neurotransmission within the hippocampus. Our findings support a role of hippocampal LepRs in synaptic plasticity, which could have an impact on memory and learning processes., (Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.)

Background and Objective: Prostate cancer is one of the most common diseases affecting men worldwide. The Gleason scoring system is the primary diagnostic and prognostic tool for prostate cancer. Furthermore, recent reports indicate that the presence of patterns of the Gleason scale such as the cribriform pattern may also correlate with a worse prognosis compared to other patterns belonging to the Gleason grade 4. Current clinical guidelines have indicated the convenience of highlight its presence during the analysis of biopsies. All these requirements suppose a great workload for the pathologist during the analysis of each sample, which is based on the pathologist's visual analysis of the morphology and organisation of the glands in the tissue, a time-consuming and subjective task. In recent years, with the development of digitisation devices, the use of computer vision techniques for the analysis of biopsies has increased. However, to the best of the authors' knowledge, the development of algorithms to automatically detect individual cribriform patterns belonging to Gleason grade 4 has not yet been studied in the literature. The objective of the work presented in this paper is to develop a deep-learning-based system able to support pathologists in the daily analysis of prostate biopsies. This analysis must include the Gleason grading of local structures, the detection of cribriform patterns, and the Gleason scoring of the whole biopsy., Methods: The methodological core of this work is a patch-wise predictive model based on convolutional neural networks able to determine the presence of cancerous patterns based on the Gleason grading system. In particular, we train from scratch a simple self-design architecture with three filters and a top model with global-max pooling. The cribriform pattern is detected by retraining the set of filters of the last convolutional layer in the network. Subsequently, a biopsy-level prediction map is reconstructed by bi-linear interpolation of the patch-level prediction of the Gleason grades. In addition, from the reconstructed prediction map, we compute the percentage of each Gleason grade in the tissue to feed a multi-layer perceptron which provides a biopsy-level score., Results: In our SICAPv2 database, composed of 182 annotated whole slide images, we obtained a Cohen's quadratic kappa of 0.77 in the test set for the patch-level Gleason grading with the proposed architecture trained from scratch. Our results outperform previous ones reported in the literature. Furthermore, this model reaches the level of fine-tuned state-of-the-art architectures in a patient-based four groups cross validation. In the cribriform pattern detection task, we obtained an area under ROC curve of 0.82. Regarding the biopsy Gleason scoring, we achieved a quadratic Cohen's Kappa of 0.81 in the test subset. Shallow CNN architectures trained from scratch outperform current state-of-the-art methods for Gleason grades classification. Our proposed model is capable of characterising the different Gleason grades in prostate tissue by extracting low-level features through three basic blocks (i.e. convolutional layer + max pooling). The use of global-max pooling to reduce each activation map has shown to be a key factor for reducing complexity in the model and avoiding overfitting. Regarding the Gleason scoring of biopsies, a multi-layer perceptron has shown to better model the decision-making of pathologists than previous simpler models used in the literature., Competing Interests: Declaration of Competing Interest The authors whose names are listed immediately below certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria, educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stoch ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or profesional relationships, affilliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Facial information is processed by our brain in such a way that we immediately make judgments about, for example, attractiveness or masculinity or interpret personality traits or moods of other people. The appearance of each facial feature has an effect on our perception of facial traits. This research addresses the problem of measuring the size of these effects for five facial features (eyes, eyebrows, nose, mouth, and jaw). Our proposal is a mixed feature-based and image-based approach that allows judgments to be made on complete real faces in the categorization tasks, more than on synthetic, noisy, or partial faces that can influence the assessment. Each facial feature of the faces is automatically classified considering their global appearance using principal component analysis. Using this procedure, we establish a reduced set of relevant specific attributes (each one describing a complete facial feature) to characterize faces. In this way, a more direct link can be established between perceived facial traits and what people intuitively consider an eye, an eyebrow, a nose, a mouth, or a jaw. A set of 92 male faces were classified using this procedure, and the results were related to their scores in 15 perceived facial traits. We show that the relevant features greatly depend on what we are trying to judge. Globally, the eyes have the greatest effect. However, other facial features are more relevant for some judgments like the mouth for happiness and femininity or the nose for dominance., (© The Author(s) 2020.)

Background and Objective: Early neonatal death is a worldwide challenge with 1 million newborn deaths every year. The primary cause of these deaths are complications during labour and birth asphyxia. The majority of these newborns could have been saved with adequate resuscitation at birth. Newborn resuscitation guidelines recommend immediate drying, stimulation, suctioning if indicated, and ventilation of non-breathing newborns. A system that will automatically detect and extract time periods where different resuscitation activities are performed, would be highly beneficial to evaluate what resuscitation activities that are improving the state of the newborn, and if current guidelines are good and if they are followed. The potential effects of especially stimulation are not very well documented as it has been difficult to investigate through observations. In this paper the main objective is to identify stimulation activities, regardless if the state of the newborn is changed or not, and produce timelines of the resuscitation episode with the identified stimulations., Methods: Data is collected by utilizing a new heart rate device, NeoBeat, with dry-electrode ECG and accelerometer sensors placed on the abdomen of the newborn. We propose a method, NBstim, based on time domain and frequency domain features from the accelerometer signals and ECG signals from NeoBeat, to detect time periods of stimulation. NBstim use causal features from a gliding window of the signals, thus it can potentially be used in future realtime systems. A high performing feature subset is found using feature selection. System performance is computed using a leave-one-out cross-validation and compared with manual annotations., Results: The system achieves an overall accuracy of 90.3% when identifying regions with stimulation activities., Conclusion: The performance indicates that the proposed NBstim, used with signals from the NeoBeat can be used to determine when stimulation is performed. The provided activity timelines, in combination with the status of the newborn, for example the heart rate, at different time points, can be studied further to investigate both the time spent and the effect of different newborn resuscitation parameters., (Copyright © 2020. Published by Elsevier B.V.)

Crude protein (CP) levels in commercial broiler ( Gallus gallus ) diets, optimized for maximum yield production vs. feed cost, have only begun to be assessed for impact on immune function. In order to study immune effects of dietary CP levels, different starter phase (day 1-14) diets were fed to 230 Ross 708 male broiler chicks randomly assigned at 1 day of age into two treatment groups. Group 1: Standard diet (STD) contained 3,000 kcal AMEn/kg energy and 23.78% CP; and Group 2: Reduced crude protein diet (RCP) contained 3,000 kcal AMEn/kg energy and 21.23% CP. From day 15-35 a common standard grower/finisher diet (3,150 kcal AMEn/kg energy and 22.18% CP) was allocated to both groups. Zymosan, a glycan derived from yeast cell walls that binds to TLR 2 and Dectin-1, was used for intra-abdominal challenge. Results demonstrated that a reduced crude protein starter diet (21.23 vs. 23.78% CP) between age 1-14, while maintaining the same levels of metabolizable energy and essential amino acids, did not affect broilers growth performance or lymphoid organ weights ( P > 0.05). Interestingly, basal leukocyte levels in the RCP group significantly ( P < 0.01) increased in the blood compartment at d35 in the unchallenged birds. Significant enhancements to leukocyte infiltration into the abdominal cavity were also detected post-immune challenge with zymosan (day 14 and day 35; P < 0.01). Post-challenge levels of TNF-α, IL-1β, and CXCL8 gene expression cells collected from the abdominal cavity were not affected by the diets ( P > 0.05). Moreover, dietary treatments did not influence percentage of ROS producing cells in the abdominal cavity ( P > 0.05). To our best knowledge, this is the first study that reports the impacts of reduced crude protein diet on the innate immune response of poultry to an acute inflammation model in the abdominal cavity. Overall, our results highlight that reduced crude protein diets can be used without negatively impacting broiler performance and may enhance the capacity of broilers to recruit leukocytes upon infection., (Copyright © 2020 Kamely, He, Wakaruk, Whelan, Naranjo and Barreda.)

Optical coherence tomography (OCT) is a well-established bedside imaging modality that allows analysis of skin structures in a non-invasive way. Automated OCT analysis of skin layers is of great relevance to study dermatological diseases. In this paper, an approach to detect the epidermal layer along with the follicular structures in healthy human OCT images is presented. To the best of the authors' knowledge, the approach presented in this paper is the only epidermis detection algorithm that segments the pilosebaceous unit, which is of importance in the progression of several skin disorders such as folliculitis, acne, lupus erythematosus, and basal cell carcinoma. The proposed approach is composed of two main stages. The first stage is a Convolutional Neural Network based on U-Net architecture. The second stage is a robust post-processing composed by a Savitzky-Golay filter and Fourier Domain Filtering to fully define the borders belonging to the hair follicles. After validation, an average Dice of 0.83 ± 0.06 and a thickness error of 10.25 μ m is obtained on 270 human skin OCT images. Based on these results, the proposed method outperforms other state-of-the-art methods for epidermis segmentation. It demonstrates that the proposed image segmentation method successfully detects the epidermal region in a fully automatic way in addition to defining the follicular skin structures as main novelty., (Copyright © 2020 del Amor, Morales, Colomer, Mogensen, Jensen, Israelsen, Bang and Naranjo.)

Generalized nucleus segmentation techniques can contribute greatly to reducing the time to develop and validate visual biomarkers for new digital pathology datasets. We summarize the results of MoNuSeg 2018 Challenge whose objective was to develop generalizable nuclei segmentation techniques in digital pathology. The challenge was an official satellite event of the MICCAI 2018 conference in which 32 teams with more than 80 participants from geographically diverse institutes participated. Contestants were given a training set with 30 images from seven organs with annotations of 21,623 individual nuclei. A test dataset with 14 images taken from seven organs, including two organs that did not appear in the training set was released without annotations. Entries were evaluated based on average aggregated Jaccard index (AJI) on the test set to prioritize accurate instance segmentation as opposed to mere semantic segmentation. More than half the teams that completed the challenge outperformed a previous baseline. Among the trends observed that contributed to increased accuracy were the use of color normalization as well as heavy data augmentation. Additionally, fully convolutional networks inspired by variants of U-Net, FCN, and Mask-RCNN were popularly used, typically based on ResNet or VGG base architectures. Watershed segmentation on predicted semantic segmentation maps was a popular post-processing strategy. Several of the top techniques compared favorably to an individual human annotator and can be used with confidence for nuclear morphometrics.

Estimated blind people in the world will exceed 40 million by 2025. To develop novel algorithms based on fundus image descriptors that allow the automatic classification of retinal tissue into healthy and pathological in early stages is necessary. In this paper, we focus on one of the most common pathologies in the current society: diabetic retinopathy. The proposed method avoids the necessity of lesion segmentation or candidate map generation before the classification stage. Local binary patterns and granulometric profiles are locally computed to extract texture and morphological information from retinal images. Different combinations of this information feed classification algorithms to optimally discriminate bright and dark lesions from healthy tissues. Through several experiments, the ability of the proposed system to identify diabetic retinopathy signs is validated using different public databases with a large degree of variability and without image exclusion.

Objectives: The objectives of the study were to compare the free serum concentrations after different fosphenytoin loading dose strategies in patients younger than 6 months old and to investigate the frequency of seizure cessation following a loading dose of fosphenytoin., Methods: This retrospective cohort study included neonates and infants admitted to a 150-bed children's hospital between August 1, 2014, and February 1, 2018. Patients were included if they were younger than 6 months old and had a postload free phenytoin serum concentration collected during the specified time frame. Patients were identified through a database query screening for the inclusion criteria. Patients were separated into 2 groups with the 15 mg/kg group as per protocol and the 20 mg/kg group as noted in common practice. Data collection included demographic information, fosphenytoin dose, time of administration of the fosphenytoin loading dose, time of sampling, free phenytoin serum concentration results, concomitant antiepileptic agents, albumin serum concentration, and total bilirubin serum concentration., Results: Forty-one patients were included for analysis, 12 in the 15 mg/kg group and 29 in the 20 mg/kg group. The average free phenytoin concentration after the loading dose was 2.45 ± 0.54 mg/L in the 15 mg/kg group and 2.52 ± 0.66 mg/L in the 20 mg/kg group. Seizure cessation after the fosphenytoin loading dose was achieved in 3 of 12 (25%) patients in the 15 mg/kg group and in 13 of 29 (45%) patients in the 20 mg/kg group (p = 0.305)., Conclusions: The study demonstrates that a traditional range of fosphenytoin loading dose (15-20 mg/kg) led to elevated postloading dose free phenytoin serum concentrations in the majority of patients with a seizure cessation rate of approximately 39%. The question remains as to what the optimal dose and target concentration should be in this patient population to achieve the best efficacy without risking associated toxicities., Competing Interests: Disclosure The authors declare no conflicts or financial interest in any product or service mentioned in the manuscript, including grants, equipment, medications, employment, gifts, and honoraria. The authors had full access to all data and take responsibility for the integrity and accuracy of the data analysis., (Copyright Pediatric Pharmacy Association. All rights reserved. For permissions, email: mhelms@pediatricpharmacy.org 2020.)

Glaucoma is one of the leading causes of irreversible but preventable blindness in working age populations. Color fundus photography (CFP) is the most cost-effective imaging modality to screen for retinal disorders. However, its application to glaucoma has been limited to the computation of a few related biomarkers such as the vertical cup-to-disc ratio. Deep learning approaches, although widely applied for medical image analysis, have not been extensively used for glaucoma assessment due to the limited size of the available data sets. Furthermore, the lack of a standardize benchmark strategy makes difficult to compare existing methods in a uniform way. In order to overcome these issues we set up the Retinal Fundus Glaucoma Challenge, REFUGE (https://refuge.grand-challenge.org), held in conjunction with MICCAI 2018. The challenge consisted of two primary tasks, namely optic disc/cup segmentation and glaucoma classification. As part of REFUGE, we have publicly released a data set of 1200 fundus images with ground truth segmentations and clinical glaucoma labels, currently the largest existing one. We have also built an evaluation framework to ease and ensure fairness in the comparison of different models, encouraging the development of novel techniques in the field. 12 teams qualified and participated in the online challenge. This paper summarizes their methods and analyzes their corresponding results. In particular, we observed that two of the top-ranked teams outperformed two human experts in the glaucoma classification task. Furthermore, the segmentation results were in general consistent with the ground truth annotations, with complementary outcomes that can be further exploited by ensembling the results., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Research has shown that methionine+ cysteine (M+C) requirements may be higher when chickens are infected with Eimeria app. In a 4 × 2 factorial design, broilers (11 to 21 D) were fed one of 4 corn-soybean meal-based diets containing either 0.6, 0.8, 0.9, or 1.0% standardized ileal digestible (SID) M+C; on day 14, broilers from each diet were gavaged with either phosphate-buffered saline (PBS) or a commercial coccidiosis vaccine (at 100 × vaccine dose) which provide a mixture of live Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts. Growth performance was recorded from day 11 to 21. Plasma and intestinal luminal samples were collected on days 14 and 21. Intestine lesion scores and fecal oocyst counts were conducted on day 21. Regardless of dietary SID M+C levels, compared to PBS gavaged broilers, the Eimeria-challenged broilers had (1) decreased (P < 0.05) body weight gain (BWG), feed intake (FI), and gain-to-feed ratio (G:F); (2) increased (P < 0.05) intestinal lesion scores and fecal oocyst counts; (3) increased (P < 0.05) plasma anti-Eimeria IgG, and intestinal luminal total IgA and anti-Eimeria IgA concentrations; and (4) increased (P < 0.05) levels of duodenum luminal gamma interferon (IFN-γ) and interleukin-10 (IL-10), as well as jejunum and cecum luminal IFN-γ concentrations. Regardless of Eimeria challenge, when compared to 0.6% SID M+C, broilers fed ≥0.8% SID M+C had (1) increased (P < 0.05) BWG, FI, and G:F and (2) increased (P < 0.05) levels of jejunum luminal total IgA. After Eimeria challenge, broilers fed 0.8% SID M+C had increased (P < 0.05) levels of jejunum luminal anti-Eimeria IgA compared to broilers fed diets containing 0.6 and 1.0% SID M+C. Collectively, in 11- to 21-D broilers, the growth suppression caused by Eimeria infection could not be mitigated by further increasing dietary M+C alone ≥0.8%. Further research should investigate interactions between dietary M+C and other nutrients for support of immune function and growth in pathogen-challenged broilers., (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background and Objective: Prostate cancer is one of the most common male tumors. The increasing use of whole slide digital scanners has led to an enormous interest in the application of machine learning techniques to histopathological image classification. Here we introduce a novel family of morphological descriptors which, extracted in the appropriate image space and combined with shallow and deep Gaussian process based classifiers, improves early prostate cancer diagnosis., Method: We decompose the acquired RGB image in its RGB and optical density hematoxylin and eosin components. Then, we define two novel granulometry-based descriptors which work in both, RGB and optical density, spaces but perform better when used on the latter. In this space they clearly encapsulate knowledge used by pathologists to identify cancer lesions. The obtained features become the inputs to shallow and deep Gaussian process classifiers which achieve an accurate prediction of cancer., Results: We have used a real and unique dataset. The dataset is composed of 60 Whole Slide Images. For a five fold cross validation, shallow and deep Gaussian Processes obtain area under ROC curve values higher than 0.98. They outperform current state of the art patch based shallow classifiers and are very competitive to the best performing deep learning method. Models were also compared on 17 Whole Slide test Images using the FROC curve. With the cost of one false positive, the best performing method, the one layer Gaussian process, identifies 83.87% (sensitivity) of all annotated cancer in the Whole Slide Image. This result corroborates the quality of the extracted features, no more than a layer is needed to achieve excellent generalization results., Conclusion: Two new descriptors to extract morphological features from histological images have been proposed. They collect very relevant information for cancer detection. From these descriptors, shallow and deep Gaussian Processes are capable of extracting the complex structure of prostate histological images. The new space/descriptor/classifier paradigm outperforms state-of-art shallow classifiers. Furthermore, despite being much simpler, it is competitive to state-of-art CNN architectures both on the proposed SICAPv1 database and on an external database., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Recent works show that generative adversarial networks (GANs) can be successfully applied to image synthesis and semi-supervised learning, where, given a small labeled database and a large unlabeled database, the goal is to train a powerful classifier. In this paper, we trained a retinal image synthesizer and a semi-supervised learning method for automatic glaucoma assessment using an adversarial model on a small glaucoma-labeled database and a large unlabeled database. Various studies have shown that glaucoma can be monitored by analyzing the optic disc and its surroundings, and for that reason, the images used in this paper were automatically cropped around the optic disc. The novelty of this paper is to propose a new retinal image synthesizer and a semi-supervised learning method for glaucoma assessment based on the deep convolutional GANs. In addition, and to the best of our knowledge, this system is trained on an unprecedented number of publicly available images (86926 images). This system, hence, is not only able to generate images synthetically but to provide labels automatically. Synthetic images were qualitatively evaluated using t-SNE plots of features associated with the images and their anatomical consistency was estimated by measuring the proportion of pixels corresponding to the anatomical structures around the optic disc. The resulting image synthesizer is able to generate realistic (cropped) retinal images, and subsequently, the glaucoma classifier is able to classify them into glaucomatous and normal with high accuracy (AUC = 0.9017). The obtained retinal image synthesizer and the glaucoma classifier could then be used to generate an unlimited number of cropped retinal images with glaucoma labels.

Background: Non-alcoholic fatty liver disease (NAFLD), a condition that leads to fibrosis, is caused by intake of very high-fat diets (HFDs). However, while the negative impact on the liver of these diets has been an issue of interest, systematic research on the effect of HFDs are lacking., Objective: To characterize the overall impact of HFDs on both molecular and morphological signs of liver remodeling., Methods: A study was conducted on male C57BL/6J mice to assess the effect of 4- and 8-week HFDs (60% kcal from fat) on (i) liver steatosis and fibrosis, and (ii) expression of factors involved in inflammation and angiogenesis., Results: After an 8-week HFD, vascular endothelial growth factor type-2 receptor (VEGF-R2) and fatty acid translocase/trombospondin-1 receptor (CD36) were overexpressed in liver tissue of mice given HFDs. These changes suggest impaired liver angiogenesis and occurred together with (i) increased GPR78-BiP and EIF2α phosphorylation, suggesting endoplasmic reticulum stress, (ii) induction of Col1a1 gene expression, a marker of fibrosis, and (iii) increased CD31 immunolabeling, consistent with active angiogenesis and fibrosis., Conclusion: Our data show that very HFDs promote a rapid inflammatory response, as well as deregulation of angiogenesis, both consistent with development of liver fibrosis., (Copyright © 2019 SEEN y SED. Publicado por Elsevier España, S.L.U. All rights reserved.)

Analysis of histopathological image supposes the most reliable procedure to identify prostate cancer. Most studies try to develop computer aid-systems to face the Gleason grading problem. On the contrary, we delve into the discrimination between healthy and cancerous tissues in its earliest stage, only focusing on the information contained in the automatically segmented gland candidates. We propose a hand-driven learning approach, in which we perform an exhaustive hand-crafted feature extraction stage combining in a novel way descriptors of morphology, texture, fractals and contextual information of the candidates under study. Then, we carry out an in-depth statistical analysis to select the most relevant features that constitute the inputs to the optimised machine-learning classifiers. Additionally, we apply for the first time on prostate segmented glands, deep-learning algorithms modifying the popular VGG19 neural network. We fine-tuned the last convolutional block of the architecture to provide the model specific knowledge about the gland images. The hand-driven learning approach, using a nonlinear Support Vector Machine, reports a slight outperforming over the rest of experiments with a final multi-class accuracy of 0.876 ± 0.026 in the discrimination between false glands (artefacts), benign glands and Gleason grade 3 glands.

Background: Most current algorithms for automatic glaucoma assessment using fundus images rely on handcrafted features based on segmentation, which are affected by the performance of the chosen segmentation method and the extracted features. Among other characteristics, convolutional neural networks (CNNs) are known because of their ability to learn highly discriminative features from raw pixel intensities., Methods: In this paper, we employed five different ImageNet-trained models (VGG16, VGG19, InceptionV3, ResNet50 and Xception) for automatic glaucoma assessment using fundus images. Results from an extensive validation using cross-validation and cross-testing strategies were compared with previous works in the literature., Results: Using five public databases (1707 images), an average AUC of 0.9605 with a 95% confidence interval of 95.92-97.07%, an average specificity of 0.8580 and an average sensitivity of 0.9346 were obtained after using the Xception architecture, significantly improving the performance of other state-of-the-art works. Moreover, a new clinical database, ACRIMA, has been made publicly available, containing 705 labelled images. It is composed of 396 glaucomatous images and 309 normal images, which means, the largest public database for glaucoma diagnosis. The high specificity and sensitivity obtained from the proposed approach are supported by an extensive validation using not only the cross-validation strategy but also the cross-testing validation on, to the best of the authors' knowledge, all publicly available glaucoma-labelled databases., Conclusions: These results suggest that using ImageNet-trained models is a robust alternative for automatic glaucoma screening system. All images, CNN weights and software used to fine-tune and test the five CNNs are publicly available, which could be used as a testbed for further comparisons.

Classification or typology systems used to categorize different human body parts have existed for many years. Nevertheless, there are very few taxonomies of facial features. Ergonomics, forensic anthropology, crime prevention or new human-machine interaction systems and online activities, like e-commerce, e-learning, games, dating or social networks, are fields in which classifications of facial features are useful, for example, to create digital interlocutors that optimize the interactions between human and machines. However, classifying isolated facial features is difficult for human observers. Previous works reported low inter-observer and intra-observer agreement in the evaluation of facial features. This work presents a computer-based procedure to automatically classify facial features based on their global appearance. This procedure deals with the difficulties associated with classifying features using judgements from human observers, and facilitates the development of taxonomies of facial features. Taxonomies obtained through this procedure are presented for eyes, mouths and noses., Competing Interests: The authors have declared that no competing interests exist.

Fourier transform infrared (FTIR) spectroscopy is a highly versatile tool for cell and tissue analysis. Modern commercial FTIR microspectroscopes allow the acquisition of good-quality hyperspectral images from cytopathological samples within relatively short times. This study aims at assessing the abilities of FTIR spectra to discriminate different types of cultured skin cell lines by different computer analysis technologies. In particular, 22700 single skin cells, belonging to two non-tumoral and two tumoral cell lines, were analysed. These cells were prepared in three different batches that included each cell type. Different spectral preprocessing and classification strategies were considered, including the current standard approaches to reduce Mie scattering artefacts. Special care was taken for the optimisation, training and evaluation of the learning models in order to avoid possible overfitting. Excellent classification performance (balanced accuracy between 0.85 and 0.95) was achieved when the algorithms were trained and tested with the cells from the same batch. When cells from different batches were used for training and testing the balanced accuracy reached values between 0.35 and 0.6, demonstrating the strong influence of sample preparation on the results and comparability of cell FTIR spectra. A deep study of the most optimistic results was performed in order to identify perturbations that influenced the final classification., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A study with a 2 × 2 factorial arrangement was conducted to determine the effects of 2 dietary crude protein levels, high (CPh) or low (CPl), supplemented with free amino acids (AA), and 2 ages at photo stimulation (PS)-early (21 wk; PSe) or late (23 wk; PSl)-on reproduction traits of broiler breeders and progeny performance. Diets were isocaloric, and calculated CP content of the CPl diets was 15 g/kg lower than the CPh diets during all phases. A total of 480 female and 64 male Ross 308 breeders of 20 wk of age were used. Total egg production was similar between CPl and CPh birds during phase 1 and 2 but was reduced by 2.8 eggs for CPl birds during phase 3. For the overall laying period, CPl birds tended (P = 0.075) to produce 4.7 fewer total eggs. Hatchability of set eggs was similar between CPl and CPh birds during phases 1 and 2 but tended (P = 0.064) to be lower for CPl birds in phase 3. PSe birds showed an advanced age at sexual maturity and age at peak production of 4.6 and 5.3 d, respectively, resulting in 2.5 more total eggs during phase 1. During phase 1, PSe birds showed an almost 5% increased fertility. Chick production in phase 1 was higher for PSe birds resulting in a tendency (P = 0.071) to higher overall chick production of almost 8 chicks. Progeny from early PS breeders showed an overall significant lower feed conversion ratio (FCR). It was concluded that egg and chick production during phases 1 and 2 were not affected by dietary CP level, but egg and chick production was reduced for CPl birds during phase 3. On the other hand, PSe birds showed an increased number of chicks. It is possible to decrease CP level of breeder diets with comparable reproduction from 22 to 46 wk; however, this is questionable for phase 3. For maximal chick production, early PS is recommended.

This paper addresses the functional localization of intra-patient images of the brain. Functional images of the brain (fMRI and PET) provide information about brain function and metabolism whereas anatomical images (MRI and CT) supply the localization of structures with high spatial resolution. The goal is to find the geometric correspondence between functional and anatomical images in order to complement and fuse the information provided by each imaging modality. The proposed approach is based on a variational formulation of the image registration problem in the frequency domain. It has been implemented as a C/C[Formula: see text] library which is invoked from a GUI. This interface is routinely used in the clinical setting by physicians for research purposes (Inscanner, Alicante, Spain), and may be used as well for diagnosis and surgical planning. The registration of anatomic and functional intra-patient images of the brain makes it possible to obtain a geometric correspondence which allows for the localization of the functional processes that occur in the brain. Through 18 clinical experiments, it has been demonstrated how the proposed approach outperforms popular state-of-the-art registration methods in terms of efficiency, information theory-based measures (such as mutual information) and actual registration error (distance in space of corresponding landmarks).

Background and Objective: This paper presents BRAIM, a computer-aided diagnosis (CAD) system to help clinicians in diagnosing and treatment monitoring of brain diseases from magnetic resonance image processing. BRAIM can be used for early diagnosis of neurodegenerative diseases such as Parkinson, Alzheimer or Multiple Sclerosis and also for brain lesion diagnosis and monitoring., Methods: The developed CAD system includes different user-friendly tools for segmenting and determining whole brain and brain structure volumes in an easy and accurate way. Specifically, three types of measurements can be performed: (1) total volume of white, gray matter and cerebrospinal fluid; (2) brain structure volumes (volume of putamen, thalamus, hippocampus and caudate nucleus); and (3) brain lesion volumes., Results: As a proof of concept, some study cases were analyzed with the presented system achieving promising results. In addition to be used to quantify treatment effectiveness in patients with brain lesions, it was demonstrated that BRAIM is able to classify a subject according to the brain volume measurements using as reference a healthy control database created for this purpose., Conclusions: The CAD system presented in this paper simplifies the daily work of clinicians and provides them with objective and quantitative volume data for prospective and retrospective analyses., (Copyright © 2017 Elsevier B.V. All rights reserved.)

This paper investigates discrimination capabilities in the texture of fundus images to differentiate between pathological and healthy images. For this purpose, the performance of local binary patterns (LBP) as a texture descriptor for retinal images has been explored and compared with other descriptors such as LBP filtering and local phase quantization. The goal is to distinguish between diabetic retinopathy (DR), age-related macular degeneration (AMD), and normal fundus images analyzing the texture of the retina background and avoiding a previous lesion segmentation stage. Five experiments (separating DR from normal, AMD from normal, pathological from normal, DR from AMD, and the three different classes) were designed and validated with the proposed procedure obtaining promising results. For each experiment, several classifiers were tested. An average sensitivity and specificity higher than 0.86 in all the cases and almost of 1 and 0.99, respectively, for AMD detection were achieved. These results suggest that the method presented in this paper is a robust algorithm for describing retina texture and can be useful in a diagnosis aid system for retinal disease screening.

When exposed to mixtures of glucose and fructose, as occurs during the fermentation of grape juice into wine, Saccharomyces cerevisiae uses these sugars at different rates. Moreover, glucose and fructose are transported by the same hexose transporters (HXT), which present a greater affinity for glucose, so that late in fermentation, fructose becomes the predominant sugar. Only a few commercial fermentation activators are available to optimally solve the problems this entails. The aim of this study was to investigate the relation between HXT3 gene expression and fructose/glucose discrepancy in two different media inoculated with a commercial wine strain of S. cerevisiae in the presence of three metabolic activators. Fermentation kinetics, vitality and major metabolites were also measured. Rehydration with ergosterol improved the area under the curve and the growth rate (µ max ) in both studied media. Also, the fructose/glucose discrepancy values were improved with all activator treatments, highlighting rehydration in the presence of ascorbic acid. The yeast rehydration process was demonstrated to influence HXT3 expression under the studied conditions. Tetrahydrofolic acid treatment greatly influenced HXT3 gene expression, especially on the 12th day of the fermentation process. To a lesser extent, ergosterol and ascorbic acid also improved this parameter.

Tudor staphylococcal nuclease (Tudor-SN) and Argonaute (Ago) are conserved components of the basic RNA interference (RNAi) machinery with a variety of functions including immune response and gene regulation. The RNAi machinery has been characterized in tick vectors of human and animal diseases but information is not available on the role of Tudor-SN in tick RNAi and other cellular processes. Our hypothesis is that tick Tudor-SN is part of the RNAi machinery and may be involved in innate immune response and other cellular processes. To address this hypothesis, Ixodes scapularis and I. ricinus ticks and/or cell lines were used to annotate and characterize the role of Tudor-SN in dsRNA-mediated RNAi, immune response to infection with the rickettsia Anaplasma phagocytophilum and the flaviviruses TBEV or LGTV and tick feeding. The results showed that Tudor-SN is conserved in ticks and involved in dsRNA-mediated RNAi and tick feeding but not in defense against infection with the examined viral and rickettsial pathogens. The effect of Tudor-SN gene knockdown on tick feeding could be due to down-regulation of genes that are required for protein processing and blood digestion through a mechanism that may involve selective degradation of dsRNAs enriched in G:U pairs that form as a result of adenosine-to-inosine RNA editing. These results demonstrated that Tudor-SN plays a role in tick RNAi pathway and feeding but no strong evidence for a role in innate immune responses to pathogen infection was found.

The risk assessment in terrestrial environments has been scarcely studied for mixtures of organic contaminants. To estimate toxicity due to these compounds, an ecotoxicological test may be done with the appropriate organism and biomarker. Photosynthesis is principally performed at photosystem II, and its efficiency is affected by any environmental stress. Consequently, the measure of this efficiency may be a good indicator of toxicity if different parameters are employed, e.g., the quantum efficiency of photosystem II and the photochemical quenching coefficient. We did a series of assays to determine the toxicity of two organic contaminants, ibuprofen and perfluorooctanoic acid, using a higher plant (Sorghum bicolor). The results showed more toxicity for the perfluorinated compound and greater sensibility for the quantum efficiency of photosystem II. Regarding the binary combination, three methods were applied to calculate EC50: combination index, concentration addition, and independent action. Synergistic behavior is the principal toxicological profile for this mix. Therefore, the combination index, which considers interactions among chemicals, gave the best estimation to determine risk indices. We conclude that the inhibition of photosynthesis efficiency can be a useful tool to determine the toxicity of the mixtures of organic pollutants and to estimate ecological risks in terrestrial environments.

Motion tracking systems are commonly used in virtual reality-based interventions to detect movements in the real world and transfer them to the virtual environment. There are different tracking solutions based on different physical principles, which mainly define their performance parameters. However, special requirements have to be considered for rehabilitation purposes. This paper studies and compares the accuracy and jitter of three tracking solutions (optical, electromagnetic, and skeleton tracking) in a practical scenario and analyzes the subjective perceptions of 19 healthy subjects, 22 stroke survivors, and 14 physical therapists. The optical tracking system provided the best accuracy (1.074 ± 0.417 cm) while the electromagnetic device provided the most inaccurate results (11.027 ± 2.364 cm). However, this tracking solution provided the best jitter values (0.324 ± 0.093 cm), in contrast to the skeleton tracking, which had the worst results (1.522 ± 0.858 cm). Healthy individuals and professionals preferred the skeleton tracking solution rather than the optical and electromagnetic solution (in that order). Individuals with stroke chose the optical solution over the other options. Our results show that subjective perceptions and preferences are far from being constant among different populations, thus suggesting that these considerations, together with the performance parameters, should be also taken into account when designing a rehabilitation system.

The goal of the software proposed in this paper is to assist ophthalmologists in diagnosis and disease prevention, helping them to determine cardiovascular risk or other diseases where the vessels can be altered, as well as to monitor the pathology progression and response to different treatments. The performance of the tool has been evaluated by means of a double-blind study where its sensitivity, specificity, and reproducibility to discriminate between health fundus (without cardiovascular risk) and hypertensive patients has been calculated in contrast to an expert ophthalmologist opinion obtained through a visual inspection of the fundus image. An improvement of almost 20% has been achieved comparing the system results with the clinical visual classification.

The VESSEL12 (VESsel SEgmentation in the Lung) challenge objectively compares the performance of different algorithms to identify vessels in thoracic computed tomography (CT) scans. Vessel segmentation is fundamental in computer aided processing of data generated by 3D imaging modalities. As manual vessel segmentation is prohibitively time consuming, any real world application requires some form of automation. Several approaches exist for automated vessel segmentation, but judging their relative merits is difficult due to a lack of standardized evaluation. We present an annotated reference dataset containing 20 CT scans and propose nine categories to perform a comprehensive evaluation of vessel segmentation algorithms from both academia and industry. Twenty algorithms participated in the VESSEL12 challenge, held at International Symposium on Biomedical Imaging (ISBI) 2012. All results have been published at the VESSEL12 website http://vessel12.grand-challenge.org. The challenge remains ongoing and open to new participants. Our three contributions are: (1) an annotated reference dataset available online for evaluation of new algorithms; (2) a quantitative scoring system for objective comparison of algorithms; and (3) performance analysis of the strengths and weaknesses of the various vessel segmentation methods in the presence of various lung diseases., (Copyright © 2014 Elsevier B.V. All rights reserved.)

There are few fully automated methods for liver segmentation in magnetic resonance images (MRI) despite the benefits of this type of acquisition in comparison to other radiology techniques such as computed tomography (CT). Motivated by medical requirements, liver segmentation in MRI has been carried out. For this purpose, we present a new method for liver segmentation based on the watershed transform and stochastic partitions. The classical watershed over-segmentation is reduced using a marker-controlled algorithm. To improve accuracy of selected contours, the gradient of the original image is successfully enhanced by applying a new variant of stochastic watershed. Moreover, a final classifier is performed in order to obtain the final liver mask. Optimal parameters of the method are tuned using a training dataset and then they are applied to the rest of studies (17 datasets). The obtained results (a Jaccard coefficient of 0.91 ± 0.02) in comparison to other methods demonstrate that the new variant of stochastic watershed is a robust tool for automatic segmentation of the liver in MRI., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Background: Tick Subolesin and its ortholog in insects and vertebrates, Akirin, have been suggested to play a role in the immune response through regulation of nuclear factor-kappa B (NF-kB)-dependent and independent gene expression via interaction with intermediate proteins that interact with NF-kB and other regulatory proteins, bind DNA or remodel chromatin to regulate gene expression. The objective of this study was to characterize the structure and regulation of subolesin in Ixodes scapularis. I. scapularis is a vector of emerging pathogens such as Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti that cause in humans Lyme disease, anaplasmosis and babesiosis, respectively. The genome of I. scapularis was recently sequenced, and this tick serves as a model organism for the study of vector-host-pathogen interactions. However, basic biological questions such as gene organization and regulation are largely unknown in ticks and other arthropod vectors., Principal Findings: The results presented here provide evidence that subolesin/akirin are evolutionarily conserved at several levels (primary sequence, gene organization and function), thus supporting their crucial biological function in metazoans. These results showed that NF-kB (Relish) is involved in the regulation of subolesin expression in ticks, suggesting that as in other organisms, different NF-kB integral subunits and/or unknown interacting proteins regulate the specificity of the NF-kB-mediated gene expression. These results suggested a regulatory network involving cross-regulation between NF-kB (Relish) and Subolesin and Subolesin auto-regulation with possible implications in tick immune response to bacterial infection., Significance: These results advance our understanding of gene organization and regulation in I. scapularis and have important implications for arthropod vectors genetics and immunology highlighting the possible role of NF-kB and Subolesin/Akirin in vector-pathogen interactions and for designing new strategies for the control of vector infestations and pathogen transmission.

The algorithm proposed in this paper allows to automatically segment the optic disc from a fundus image. The goal is to facilitate the early detection of certain pathologies and to fully automate the process so as to avoid specialist intervention. The method proposed for the extraction of the optic disc contour is mainly based on mathematical morphology along with principal component analysis (PCA). It makes use of different operations such as generalized distance function (GDF), a variant of the watershed transformation, the stochastic watershed, and geodesic transformations. The input of the segmentation method is obtained through PCA. The purpose of using PCA is to achieve the grey-scale image that better represents the original RGB image. The implemented algorithm has been validated on five public databases obtaining promising results. The average values obtained (a Jaccard's and Dice's coefficients of 0.8200 and 0.8932, respectively, an accuracy of 0.9947, and a true positive and false positive fractions of 0.9275 and 0.0036) demonstrate that this method is a robust tool for the automatic segmentation of the optic disc. Moreover, it is fairly reliable since it works properly on databases with a large degree of variability and improves the results of other state-of-the-art methods.

Purpose: This work presents the protocol carried out in the development and validation of an augmented reality system which was installed in an operating theatre to help surgeons with trocar placement during laparoscopic surgery. The purpose of this validation is to demonstrate the improvements that this system can provide to the field of medicine, particularly surgery., Method: Two experiments that were noninvasive for both the patient and the surgeon were designed. In one of these experiments the augmented reality system was used, the other one was the control experiment, and the system was not used. The type of operation selected for all cases was a cholecystectomy due to the low degree of complexity and complications before, during, and after the surgery. The technique used in the placement of trocars was the French technique, but the results can be extrapolated to any other technique and operation., Results and Conclusion: Four clinicians and ninety-six measurements obtained of twenty-four patients (randomly assigned in each experiment) were involved in these experiments. The final results show an improvement in accuracy and variability of 33% and 63%, respectively, in comparison to traditional methods, demonstrating that the use of an augmented reality system offers advantages for trocar placement in laparoscopic surgery.

The success of oral surgery is subject to accurate advanced planning. In order to properly plan for dental surgery or a suitable implant placement, it is necessary an accurate segmentation of the jaw tissues: the teeth, the cortical bone, the trabecular core and over all, the inferior alveolar nerve. This manuscript presents a new automatic method that is based on fuzzy connectedness object extraction and mathematical morphology processing. The method uses computed tomography data to extract different views of the jaw: a pseudo-orthopantomographic view to estimate the path of the nerve and cross-sectional views to segment the jaw tissues. The method has been tested in a groundtruth set consisting of more than 9000 cross-sections from 20 different patients and has been evaluated using four similarity indicators (the Jaccard index, Dice's coefficient, point-to-point and point-to-curve distances), achieving promising results in all of them (0.726±0.031, 0.840±0.019, 0.144±0.023 mm and 0.163±0.025 mm, respectively). The method has proven to be significantly automated and accurate, with errors around 5% (of the diameter of the nerve), and is easily integrable in current dental planning systems., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

The impact of PET on radiation therapy is held back by poor methods of defining functional volumes of interest. Many new software tools are being proposed for contouring target volumes but the different approaches are not adequately compared and their accuracy is poorly evaluated due to the illdefinition of ground truth. This paper compares the largest cohort to date of established, emerging and proposed PET contouring methods, in terms of accuracy and variability. We emphasise spatial accuracy and present a new metric that addresses the lack of unique ground truth. 30 methods are used at 13 different institutions to contour functional VOIs in clinical PET/CT and a custom-built PET phantom representing typical problems in image guided radiotherapy. Contouring methods are grouped according to algorithmic type, level of interactivity and how they exploit structural information in hybrid images. Experiments reveal benefits of high levels of user interaction, as well as simultaneous visualisation of CT images and PET gradients to guide interactive procedures. Method-wise evaluation identifies the danger of over-automation and the value of prior knowledge built into an algorithm.

Breast Magnetic Resonance Image skin has similar intensity levels than dense tissue, and may produce segmentation errors if not managed correctly. In this work a novel skin segmentation method is presented and validated by experts, aimed to obtain as many pixels belonging to the real skin as possible. Segmented skin will be used to build a breast biomechanical model to register X-Ray Images with Magnetic Resonance Images in the future, using a virtually deformed Magnetic Resonance Image.

Two experiments, each consisting of 2 trials, were conducted to determine the effect of salmon protein hydrolysate (SPH) and spray-dried plasma protein (SDPP) fed during the first week postweaning and their subsequent effect on the growth performance of weanling pigs. Pigs were fed in a 3-phase feeding program with durations of 7 d for phase 1 in both Exp. 1 and 2; 14 or 15 d for phase 2 in Exp. 1 and 2, respectively; and 7 or 8 d for phase 3 in Exp. 1 and 2, respectively. Dietary treatments were fed only during phase 1, whereas the same diet was fed to all pigs in phases 2 and 3. Pigs were blocked by initial BW and sex, and littermates were balanced across treatments. Data from the 2 trials within each experiment were combined and analyzed together; no treatment × trial interactions (P > 0.10) were observed. In Exp. 1, a total of 324 weanling pigs (10 replications of 5 or 6 pigs per pen) with an average initial BW of 6.4 ± 1.3 kg were assigned to 1) a control diet with no SPH or SDPP, 2) 1.5% SPH, 3) 3.0% SPH, 4) 1.5% SDPP, 5) 3.0% SDPP, or 6) 1.5% SPH + 1.5% SDPP. Experiment 2 was similar to Exp. 1, but red blood cells were removed from all diets to reduce diet complexity. In Exp. 2, weanling pigs (n = 320, 14 replications of 5 or 6 pigs per pen) with an average initial BW of 5.4 ± 1.2 kg were assigned to 1) a control diet with no SPH or SDPP, 2) 1.5% SPH, 3) 1.5% SDPP, or 4) 1.5% SPH + 1.5% SDPP. Three batches of SPH were used, and each batch was analyzed for AA composition. In Exp. 1, the inclusion of SDPP or SPH during phase 1 did not affect (P > 0.10) ADG, ADFI, or G:F compared with those of pigs fed the control diet. No carryover effects on growth performance were observed in any of the subsequent phases. Overall, G:F was greater (P = 0.08) in pigs fed the 1.5% diets compared with those fed the 3.0% diets. In Exp. 2, no differences (P > 0.10) were observed in ADG, ADFI, or G:F among pigs fed the SPH or SDPP diets compared with those of pigs fed the control diet. Pigs fed the combined diet had greater (P < 0.10) overall ADFI compared with that of pigs fed the control diet, but ADFI was similar to that of pigs fed the SPH and SDPP diets. These results indicate that inclusion of up to 3% SDPP or SPH in diets fed during the first week postweaning did not affect the growth performance of weanling pigs, and no subsequent carryover effects were observed. Salmon protein hydrolysate did not affect the growth performance of weanling pigs and may be considered an alternative protein source in diets for weanling pigs.

Most dental implant planning systems use a 3D representation of the CT scan of the patient under study as it provides a more intuitive view of the human jaw. The presence of metallic objects in human jaws, such as amalgam or gold fillings, provokes several artifacts like streaking and beam hardening which makes the reconstruction process difficult. In order to reduce these artifacts, several methods have been proposed using the raw data, directly obtained from the tomographs, in different ways. However, in DICOM-based applications this information is not available, and thus the need of a new method that handles this task in the DICOM domain. The presented method performs a morphological filtering in the polar domain yielding output images less affected by artifacts (even in cases of multiple metallic objects) without causing significant smoothing of the anatomic structures, which allows a great improvement in the 3D reconstruction. The algorithm has been automated and compared to other image denoising methods with successful results., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Experiments were conducted to assess the effect of dried whey (DW; 70% lactose) or milk chocolate product (MCP; 20% lactose and 60% sugars) on wk-1 feed intake and growth performance of pigs. Diets contained 1.60, 1.40, 1.40, and 1.20% total Lys for phase 1 (d 0 to 7), 2 (d 7 to 14), 3 (d 14 to 21), and 4 (d 21 to 35), respectively. Pigs were blocked by initial BW; sex and littermates were balanced across treatment; treatments were replicated with a minimum of 5 pens of 4 pigs each. Pigs were weaned at 24, 19, and 24 d of age with an initial BW of 6.5, 6.0, or 6.3 kg for Exp. 1 to 3, respectively. In Exp. 1 and 2, the treatments were 1) negative control (NC), no lactose added, 2) positive control (PC) with DW, 3) 25% replacement of the level of DW of the PC diet with MCP (25MCP), and 4) 50% replacement of the level of DW of the PC diet with MCP (50MCP). The level of DW or combinations of DW and MCP were 20, 10, and 5% for phases 1, 2, and 3 respectively. A common diet with no lactose was fed during phase 4. In Exp. 3, the treatments were 1) NC, 2) PC, and 3) 100% replacement of the level of DW of the PC diet with MCP (100MCP). In the combined data of Exp. 1 and 2, daily collected feed intake during wk 1 was increased (P < 0.10) from d 3 to 7 for the PC pigs; on d 2, 3, 4, and 7 for the 25MCP pigs; and from d 2 to 7 for the 50MCP pigs compared with the NC pigs. There was no difference (P > 0.10) on any day of wk 1 among pigs fed the PC and MCP diets. During phase 1, ADG and ADFI were increased (P < 0.10) for the PC, 25MCP, and 50MCP pigs compared with the NC pigs, but G:F was not affected (P > 0.10). During phases 2, 3, 4, and overall, there were no differences (P > 0.10) in growth performance among pigs fed the PC and MCP diets. In Exp. 3, collected daily feed intake during wk 1 was increased (P < 0.10) from d 2 to 5 for PC pigs and on d 1 and 2 for the 100MCP pigs compared with the NC pigs. However, there was no difference (P > 0.10) on any day of wk 1 between the PC and 100MCP pigs. Growth performance was not affected (P > 0.10) during any phase of the experiment. Combined data from Exp. 1, 2, and 3 using the NC and PC diets indicated that dietary DW increased (P < 0.10) final BW and increased ADG and ADFI during phases 1, 2, 3, and overall. These results indicate that partial or total replacement of DW with MCP had no effect on wk-1 feed intake or growth performance of weanling pigs and that MCP could be considered as a formulation alternative to DW.

Three experiments were conducted to compare dried whey permeate (DWP; 80% lactose) and a carbohydrate product (CHO; 40% lactose, 30% sucrose, and 10% glucose) for nursery pigs. Pigs were fed in a 3-phase feeding program, and diets contained 1.6, 1.4, and 1.2% total Lys for phases 1 (d 0 to 7), 2 (d 7 to 21), and 3 (21 to 28). Dietary treatments included 1) control (no lactose), 2) low level of DWP, 3) high level of DWP, 4) low level of CHO, and 5) high level of CHO. In Exp. 1 (4 reps of 4 pigs per pen; initial BW = 7 kg and 23 d of age), the low and high levels used for each source in each phase were phase 1 (12.5 and 25%), phase 2 (10 and 20%), and phase 3 (6 and 12%). In Exp. 2 (6 reps of 5 pigs per pen; initial BW = 8 kg and 26 d of age) and 3 (4 reps of 4 pigs per pen; initial BW = 6 kg and 21 d of age), the inclusion levels were phase 1 (6 and 12%), phase 2 (3 and 6%), and phase 3 (common diet with no lactose). In Exp. 1, pigs fed diets with DWP or CHO had increased ADG (P = 0.02 and P = 0.01) and ADFI (P = 0.01) compared with pigs fed the control diet during phase 1. Gain:feed was reduced (P = 0.08) for pigs fed diets with CHO. During phases 2, 3, and overall, ADG, ADFI, and G:F were not affected (P > 0.10) by diet. In Exp. 2, pigs fed diets with CHO had increased ADG (P = 0.08 and P = 0.07) and ADFI (P = 0.04 and P = 0.01) compared with pigs fed the control diet during phases 1 and 2. Pigs fed diets with CHO had increased ADFI (P = 0.08 and P = 0.07) in phases 1 and 2 and increased ADG (P = 0.02) in phase 2 compared with pigs fed diets with DWP. Overall, pigs fed diets with DWP and CHO had increased ADFI (P = 0.06 and P = 0.01) compared with pigs fed the control diet, but ADG was increased (P = 0.07) for pigs fed diets with CHO. In Exp. 3, ADG, ADFI, and G:F were not affected (P > 0.10) by DWP or CHO during phase 1. Daily BW gain was increased (P = 0.02 and P = 0.07) for pigs fed diets with DWP or CHO during phase 2 compared with pigs fed the control diet. Overall, ADG was increased (P = 0.05) for pigs fed diets with DWP, but ADFI and G:F were not affected. Results from the combined data of Exp. 2 and 3, indicated that overall ADG (P = 0.05 and P = 0.04) and ADFI (P = 0.04) were increased in pigs fed diets with DWP or CHO compared with pigs fed the control diet. These data suggest that DWP or CHO improve growth performance of weanling pigs.

Transcranial Doppler (TCD) has been widely used to monitor cerebral blood flow velocity (BFV) during the performance of cognitive tasks compared with repose periods. Although one of its main advantages is its high temporal resolution, only some of the previous functional TCD studies have focused on the analysis of the temporal evolution of the BFV signal and none of them has performed a spectral analysis of the signal. In this study, maximum BFV data in both posterior cerebral arteries was monitored during a visual perception task (10 cycles of alternating darkness and illumination) for 23 subjects. A peak was located in the low-frequency band of the spectrum of the maximum BFV of each subject both during visual stimulation and repose periods. The frequency of this peak was in the range between 0.037 and 0.098Hz, depending on the subject, the vessel and the experimental condition. The component of the signal at this frequency, which is associated with the slow variations caused by the visual stimuli, was estimated. That way, the variations in BFV caused by the experimental stimuli were isolated from the variations caused by other factors. This low-frequency estimation signal was used to obtain parameters about the temporal evolution and the magnitude variations of the BFV in a reliable way, thus, characterizing the neurovascular coupling of the participants., (Copyright 2010 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

The lone star tick, Amblyomma americanum, vectors pathogens of emerging diseases of humans and animals in the United States. Currently, measures are not available for effective control of A. americanum infestations. Development of vaccines directed against tick proteins may reduce tick infestations and the transmission of tick-borne pathogens. However, the limiting step in tick vaccine development has been the identification of tick protective antigens. Herein, we report the application of RNA interference (RNAi) for screening an A. americanum cDNA library for discovery of tick protective antigens that reduce tick survival and weights after feeding. Four cDNA clones, encoding for putative threonyl-tRNA synthetase (2C9), 60S ribosomal proteins L13a (2D10) and L13e (2B7), and interphase cytoplasm foci protein 45 (2G7), were selected for vaccine studies in cattle, along with subolesin, a tick protective protein identified previously. In vaccinated cattle, an overall efficacy (E)>30% was obtained when considering the vaccine effect on both nymphs and adults, but only 2D10, 2G7 and subolesin affected both tick stages. The highest efficacy of control for adult ticks (E>55%) was obtained in cattle vaccinated with recombinant 2G7 or subolesin. These collective results demonstrated the feasibility of developing vaccines for the control of lone star tick infestations. The use of RNAi for identification of tick protective antigens proved to be a rapid and cost-effective tool for discovery of candidate vaccine antigens, and this approach could likely be applied to other parasites of veterinary and medical importance., (Copyright (c) 2009 Elsevier Ltd. All rights reserved.)

The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes several tick-transmitted pathogens that impact veterinary and human health. Tick-borne pathogens cycle between tick vectors and vertebrate hosts and their interaction is mediated by molecular mechanisms at the tick-pathogen interface. These mechanisms have evolved characteristics that involve traits from both the tick vector and the pathogen to insure their mutual survival. Herein, we review the information obtained from functional genomics and genetic studies to characterize the tick-Anaplasma interface and evolution of A. marginale and A. phagocytophilum. Anaplasma and tick genes and proteins involved in tick-pathogen interactions were characterized. The results of these studies demonstrated that common and Anaplasma species-specific molecular mechanism occur by which pathogen and tick cell gene expression mediates or limits Anaplasma developmental cycle and trafficking through ticks. These results have advanced our understanding of the biology of tick-Anaplasma interactions and have opened new avenues for the development of improved methods for the control of tick infestations and the transmission of tick-borne pathogens.

Transcranial Doppler is a tool to measure blood flow velocity (BFV) in the main arteries of the brain that has been used in previous studies to analyze brain activity during virtual reality (VR) experiences. Increments in BFV were found during the exposure to virtual environments in comparison with baseline periods. However, due to the complexity of VR experiences, there are several factors that can be having an influence in these variations, so it is necessary to separately analyze those different aspects. In this work, we summarize our results related to visual perception. A method based on spectral analysis was used to analyze the magnitude and temporal evolution of the maximum BFV signal. Results showed that, in the presence of visual stimuli, BFV quickly rises to a maximum that is achieved after a few seconds. The contribution of the visual stimuli factor to the observed BFV variations during a VR experience can be estimated from the results of the developed work.

Host genetic diversity plays an important role in buffering populations against pathogens. We characterized the allelic diversity at the second exon of the b (DRB-2) chain of the major histocompatibility complex class II (MHC-II) locus in a population of Iberian red deer (Cervus elaphus hispanicus) and its impact on parasitism by macroparasites, on a microparasite causing tuberculosis, and on relevant life history traits (spleen size and body condition). No DRB-2 haplotype conferred general resistance or susceptibility against all parasites. However, specific significant correlations were found between some DRB-2 haplotypes and specific parasites. We also detected associations between DRB-2 haplotypes and body condition and spleen size after controlling for body size, sex and age. Our results evidenced a functional significance of MHC-II genes in the defence of Iberian red deer against parasites. These results also support a role of MHC-II as a fitness-enhancing genetic element which can be mediated by parasite effects on life traits with a genetic basis. We conclude that MHC immunogenetic studies may assess management decisions in Iberian red deer because (i) loss of genetic diversity may lead to increased disease occurrence, and (ii) MHC genes are ecologically relevant since they underlie host infection rates and life history traits.

Background: The tick-borne pathogen Anaplasma marginale, which is endemic worldwide, is the type species of the genus Anaplasma (Rickettsiales: Anaplasmataceae). Rhipicephalus (Boophilus) microplus is the most important tick vector of A. marginale in tropical and subtropical regions of the world. Despite extensive characterization of the genetic diversity in A. marginale geographic strains using major surface protein sequences, little is known about the biogeography and evolution of A. marginale and other Anaplasma species. For A. marginale, MSP1a was shown to be involved in vector-pathogen and host-pathogen interactions and to have evolved under positive selection pressure. The MSP1a of A. marginale strains differs in molecular weight because of a variable number of tandem 23-31 amino acid repeats and has proven to be a stable marker of strain identity. While phylogenetic studies of MSP1a repeat sequences have shown evidence of A. marginale-tick co-evolution, these studies have not provided phylogeographic information on a global scale because of the high level of MSP1a genetic diversity among geographic strains., Results: In this study we showed that the phylogeography of A. marginale MSP1a sequences is associated with world ecological regions (ecoregions) resulting in different evolutionary pressures and thence MSP1a sequences. The results demonstrated that the MSP1a first (R1) and last (RL) repeats and microsatellite sequences were associated with world ecoregion clusters with specific and different environmental envelopes. The evolution of R1 repeat sequences was found to be under positive selection. It is hypothesized that the driving environmental factors regulating tick populations could act on the selection of different A. marginale MSP1a sequence lineages, associated to each ecoregion., Conclusion: The results reported herein provided the first evidence that the evolution of A. marginale was linked to ecological traits affecting tick vector performance. These results suggested that some A. marginale strains have evolved under conditions that support pathogen biological transmission by R. microplus, under different ecological traits which affect performance of R. microplus populations. The evolution of other A. marginale strains may be linked to transmission by other tick species or to mechanical transmission in regions where R. microplus is currently eradicated. The information derived from this study is fundamental toward understanding the evolution of other vector-borne pathogens.

Background: The cattle pathogen, Anaplasma marginale, undergoes a developmental cycle in ticks that begins in gut cells. Transmission to cattle occurs from salivary glands during a second tick feeding. At each site of development two forms of A. marginale (reticulated and dense) occur within a parasitophorous vacuole in the host cell cytoplasm. However, the role of tick genes in pathogen development is unknown. Four genes, found in previous studies to be differentially expressed in Dermacentor variabilis ticks in response to infection with A. marginale, were silenced by RNA interference (RNAi) to determine the effect of silencing on the A. marginale developmental cycle. These four genes encoded for putative glutathione S-transferase (GST), salivary selenoprotein M (SelM), H+ transporting lysosomal vacuolar proton pump (vATPase) and subolesin., Results: The impact of gene knockdown on A. marginale tick infections, both after acquiring infection and after a second transmission feeding, was determined and studied by light microscopy. Silencing of these genes had a different impact on A. marginale development in different tick tissues by affecting infection levels, the densities of colonies containing reticulated or dense forms and tissue morphology. Salivary gland infections were not seen in any of the gene-silenced ticks, raising the question of whether these ticks were able to transmit the pathogen., Conclusion: The results of this RNAi and light microscopic analyses of tick tissues infected with A. marginale after the silencing of genes functionally important for pathogen development suggest a role for these molecules during pathogen life cycle in ticks.

The control of arthropod vectors of pathogens that affect human and animal health is important for the eradication of vector-borne diseases. The ortholog of the tick-protective antigen, subolesin, was identified in Aedes albopictus and found to have conserved epitopes in ticks and mosquitoes. RNA interference with the tick and mosquito double-stranded RNA in three tick species resulted in significant gene knockdown and decreased tick weight and/or survival. Feeding Anopheles atroparvus, Aedes caspius, and Culex pipiens female mosquitoes on an A. albopictus subolesin hyperimmune serum resulted in 11 +/- 5% to 29 +/- 6% survival inhibition when compared to controls fed on preimmune serum. Feeding sand flies, Phlebotomus perniciosus, on antimosquito subolesin ortholog protein antibodies inhibited female survival and the number of larvae and adults obtained after hatching by 28 +/- 22% and 16 +/- 3%, respectively, when compared to controls. Vaccination with tick and mosquito subolesin ortholog proteins significantly reduced Ixodes scapularis tick infestation and weight in a similar way. However, vaccination with the recombinant mosquito subolesin ortholog antigen did not protect against Amblyomma americanum and Rhipicephalus sanguineus tick infestations. Collectively, these preliminary results provided the first evidence that development of vaccines may be possible for control of multiple arthropod vectors using subolesin orthologs but suggested that multiple antigens may be required to produce an effective vaccine.

Global gene expression profiles were analyzed in European wild boar naturally infected with Mycobacterium bovis. Spleen RNA was extracted from 23 M. bovis-infected and 17 uninfected animals and analyzed using a Pigoligoarray representing 20,400 genes. Differentially expressed sequences (N=161) were identified affecting cellular processes such as apoptosis, cell communication and signal transduction, cell growth and/or maintenance, cytoskeleton organization and biogenesis, DNA repair, immune response, metabolism and energy pathways, protein metabolism, regulation of cell proliferation, regulation of gene expression, regulation of nucleic acid metabolism, regulation of physiological processes, and transport. Real-time RT-PCR analysis of mRNA levels was used to corroborate microarray results of selected genes. Immune response genes were among the most represented differentially expressed sequences and were selected for further discussion. Beta-defensin 129, T-cell surface glycoprotein CD8 and B-cell receptor-associated protein 29 were overexpressed in infected animals. Lower expression levels of the immune response genes galectin-1, complement component C1qB and certain HLA class I and class II histocompatibility antigens and immunoglobulin chains were found in infected animals. This study identified new mechanisms by which naturally infected European wild boar respond to M. bovis infection and how the pathogen circumvents host immune responses to establish infection. Gene expression studies in naturally infected wildlife reservoirs of bovine tuberculosis are important for functional genomics and vaccine studies to aid in disease control in wildlife.

The tick protective antigen, subolesin, is a regulatory protein involved in the control of multiple cellular pathways. Subolesin is evolutionary conserved in invertebrates and vertebrates with sequence homology to akirins, a recently renamed group of proteins that were proposed to function as transcription factors in Drosophila and mice. The objective of this research was to provide evidence of the sequence and functional homology between tick subolesin and akirins. The phylogenetic analysis of subolesin and akirins showed that they are evolutionary conserved. The effect of subolesin and akirin2 knockdown was compared in adult ticks and mice, respectively. The results demonstrated that tick subolesin is an ortholog of insect and vertebrate akirins and suggested that these proteins function in the regulation of NF-kappaB-dependent and independent expression of signal transduction and innate immune response genes. These results suggest that these proteins have an important role in host-pathogen interactions.