Deep Convolutional Neural Networks for Histological Image Analysis in Gastric Carcinoma Whole Slide Images

Introduction/ Background In this paper, histopathological whole slide images of gastric carcinoma are analyzed using deep learning methods. A convolutional neural network architecture is proposed for two classification applications in H&E stained tissue images, namely, cancer classification based on immunohistochemistry (IHC) into classes Her2/neu+ tumor, Her2/neu- tumor and non-tumor, and necrosis detection based on existence of necrosis into classes necrotic and non-necrotic. The studies in [1] and [2] explored computer-aided classification using graphbased methods and necrosis detection by textural approach respectively, which are extended using deep convolutional neural networks. Performance is quantitatively compared with established handcrafted image features, namely Haralick GLCM, Gabor filter-banks, LBP histograms, Gray histograms, RGB histograms and HSV histograms followed by classification by random forests, another well-known machine learning algorithm. Aims Convolutional neural networks (CNN) have recently gained tremendous attention in general image analysis [3-5]. There has also been an emergence of deep learning in digital histopathology for diverse classification and detection problems [6-8]. The prime motivation behind this work is that no previous study has explored deep learning for the specified goals in gastric cancer WSI. Automated cancer classification can assist pathologists in computer-aided diagnosis in H&E stained WSI without the requirement of IHC staining, thereby reducing preparation and inspection times, and decreasing inter- and intra-observer variability. Necrosis detection can play an important role in prognosis, as larger necrotic areas indicate a smaller chance of survival and vice-versa. Moreover, most deep learning studies have used smaller image sizes mainly due to memory restrictions of GPU, however, we consider larger regions in order to preserve context i.e. neighborhood information and tissue architecture at higher magnification. Further, this method is independent of nuclei segmentation, hence its performance is not limited by segmentation performance as in [1] (evaluation details in [9]). Methods Firstly, standard data augmentation techniques are applied on the available gastric cancer WSI dataset and thousands of images of size 512x512 are generated. Different CNN architectures are empirically studied to observe the behavior of variation in model characteristics (network depth, layer properties, training parameters, etc.) by training them from scratch on a representative subset of whole data for cancer classification. One of these is the Imagenet model [4], however it doesn’t perform desirably on the representative dataset. The self-designed CNN architecture with best classification rates is selected. Later, the proposed CNN is also applied for necrosis detection. Performance is compared with state of the art methods using handcrafted features and random forests. For evaluation, randomized three-fold stratified shuffle split and leave-one-patient-out cross validations are used. Results Conclusion: A self-designed CNN architecture is proposed for image analysis (cancer classification based on IHC and necrosis detection) in H&E stained WSI of gastric cancer. Quantitative evaluation shows that deep learning methods mostly compare favorably to state of the art methods, especially for necrosis detection. In future the aim is to expand the current WSI dataset and to improve the CNN architecture for optimal performance

Prevention of Birch Pollen-Related Food Allergy by Mucosal Treatment with Multi-Allergen-Chimers in Mice

Among birch pollen allergic patients up to 70% develop allergic reactions to Bet v 1-homologue food allergens such as Api g 1 (celery) or Dau c 1 (carrot), termed as birch pollen-related food allergy. In most cases, specific immunotherapy with birch pollen extracts does not reduce allergic symptoms to the homologue food allergens. We therefore genetically engineered a multi-allergen chimer and tested if mucosal treatment with this construct could represent a novel approach for prevention of birch pollen-related food allergy.BALB/c mice were poly-sensitized with a mixture of Bet v 1, Api g 1 and Dau c 1 followed by a sublingual challenge with carrot, celery and birch pollen extracts. For prevention of allergy sensitization an allergen chimer composed of immunodominant T cell epitopes of Api g 1 and Dau c 1 linked to the whole Bet v 1 allergen, was intranasally applied prior to sensitization.Intranasal pretreatment with the allergen chimer led to significantly decreased antigen-specific IgE-dependent β-hexosaminidase release, but enhanced allergen-specific IgG2a and IgA antibodies. Accordingly, IL-4 levels in spleen cell cultures and IL-5 levels in restimulated spleen and cervical lymph node cell cultures were markedly reduced, while IFN-γ levels were increased. Immunomodulation was associated with increased IL-10, TGF-β and Foxp3 mRNA levels in NALT and Foxp3 in oral mucosal tissues. Treatment with anti-TGF-β, anti-IL10R or anti-CD25 antibodies abrogated the suppression of allergic responses induced by the chimer.Our results indicate that mucosal application of the allergen chimer led to decreased Th2 immune responses against Bet v 1 and its homologue food allergens Api g 1 and Dau c 1 by regulatory and Th1-biased immune responses. These data suggest that mucosal treatment with a multi-allergen vaccine could be a promising treatment strategy to prevent birch pollen-related food allergy

eDNA metabarcoding biodiversity of freshwater fish in the Alpine area

Environmental DNA (eDNA) based methods are proving to be a promising tool for freshwater fish biodiversity assessment in Europe within the Water Framework Directive (WFD, 2000/60/EC) especially for large rivers and lakes where current fish monitoring techniques have known shortcomings. Many freshwater fish are experiencing critical population declines with risk of local or global extinction because of intense anthropogenic pressure and this can have serious consequences on freshwater ecosystem functioning and diversity. Within the EU project Eco-AlpsWater, advanced high throughput sequencing (HTS) techniques are used to improve the traditional WFD monitoring approaches by using environmental DNA (eDNA) collected in Alpine waterbodies. An eDNA metabarcoding approach specifically designed to measure freshwater fish biodiversity in Alpine lakes and rivers has been extensively evaluated by using mock samples within an intercalibration test. This eDNA method was validated and used to study fish biodiversity of eight lakes and six rivers of the Alpine region including four EC countries (Austria, France, Italy, Slovenia) and Switzerland. More in detail, this metabarcoding approach, based on HTS sequencing of a section of the 12S rRNA gene, was used to assess freshwater fish biodiversity and their distribution in the different habitats. These data represent the first attempt to provide a comprehensive description of freshwater fish diversity in different ecosystems of the Alpine area confirming the applicability of eDNA metabarcoding analyses for the biomonitoring of fish inhabiting Alpine and perialpine lakes and rivers

Block-Centric Visualization Of Histological Whole Slide Images With Application To Revealing Growth-Patterns Of Early Colorectal Adenomas And Aberrant Crypt Foci

Introduction/ Background Comfortable navigation through diagnostic images is a prospective challenge for the acceptance of virtual microscopy applications in routine pathology [1],[2]. Tracing different regions of interest through multiple sections on one or several slides is a typical task in diagnostic slide examination. This laborious and time-consuming co-localization is currently executed by pathologists. Retaining the relative positions of tissue structures while alternating between multiple slides is still not feasible in a satisfactory manner in conventional nor virtual microscopy. Aims To address this issue we present a more comfortable and intuitive method to read slides using computer-assisted navigation. Furthermore, we demonstrate the strengths of our method by applying it to large series of serial colorectal tissue sections, creating new kinds of visualizations of different adenomatous mucosal architectures in human tissue, while looking for human correlates of lesions recently described in mice [3]. Methods Histological images contain multiple distortions from different sources in the laboratory and digitalization process. An interconnection model was created to describe distortions by several layers, providing a normalized tissue representation. Layers were associated with specific distortions with each layer serving as a new level of abstraction. The first layers enabled a coarse alignment of tissue sections. Further alignment is achieved by piecewise, multi-resolution, SIFT-based [4] correspondence extraction and refinement. Inside the convex hull of all fiducial points local affine transformations were applied whereas a global affine transformation was used on the outside. Animated stacks were generated for regions of interest using local rigid transformations to preserve exact morphological coherences. For subsequent creation of 3D models, the relevant histological objects within these images were annotated by pathologists, partly using computer assisted segmentation based on active contours [5]. These annotations were used subsequently to create simplified 3D models by applying VTK [6].  Results The presented methods provide an efficient means to retrieve correspondences and additional spatial information from serial sections of histological slides. They also show good applicability for specimen from different origin. Alignment methods can be applied to generate block-centric visualizations such as parallel and transparent viewing of multiple stains. Moreover, the generated stack videos and 3D models demonstrate the very good accuracy of section alignment even in large series. The visualizations enable pathologists and researchers to grasp the 3D structural relationships in the tissue at a glance, providing an excellent tool to communicate more complex histomorphological findings. Interestingly, we see two kinds of tubular adenomas, which could imply multiple ways to tubular adenoma formation in FAP-patients, possibly akin to the recent observations in mice [3]

A forest typology for monitoring sustainable forest management: The case of European Forest Types

Sustainable forest management (SFM) is presently widely accepted as the overriding objective for forest policy and practice. Regional processes are in progress all over the world to develop and implement criteria and indicators of SFM. In continental Europe, a set of 35 Pan-European indicators has been endorsed under the Ministerial Conference on the Protection of Forests in Europe (MCPFE) to measure progress towards SFM in the 44 countries of the region. The formulation of seven indicators (forest area, growing stock, age structure/diameter distribution, deadwood, tree species composition, damaging agents, naturalness) requires national data to be reported by forest types. Within the vast European forest area the values taken by these indicators show a considerable range of variation, due to variable natural conditions and anthropogenic influences. Given this variability, it is very difficult to grasp the meaning of these indicators when taken out of their ecological background. The paper discusses the concepts behind, and the requirements of, a classification more soundly ecologically framed and suitable for MCPFE reporting than the three (un-informative) classes adopted so far: broadleaved forest, coniferous forest, mixed broadleaved and coniferous forest. We propose a European Forest Types scheme structured into a reasonably higher number of classes, that would improve the specificity of the indicators reported under the MCPFE process and its understanding.L'articolo è disponibile sul sito dell'editore www.tandf.co.uk/journals

Sequence Capture and Next Generation Resequencing of the MHC Region Highlights Potential Transplantation Determinants in HLA Identical Haematopoietic Stem Cell Transplantation

How cells coordinate the immune system activities is important for potentially life-saving organ or stem cell transplantations. Polymorphic immunoregulatory genes, many of them located in the human major histocompatibility complex, impact the process and assure the proper execution of tolerance-versus-activity mechanisms. In haematopoietic stem cell transplantation, on the basis of fully human leukocyte antigen (HLA)-matched donor–recipient pairs, adverse effects like graft versus leukaemia and graft versus host are observed and difficult to handle. So far, high-resolution HLA typing was performed with Sanger sequencing, but for methodological reasons information on additional immunocompetent major histocompatibility complex loci has not been revealed. Now, we have used microarray sequence capture and targeted enrichment combined with next generation pyrosequencing for 3.5 million base pair human major histocompatibility complex resequencing in a clinical transplant setting and describe 3025 variant single nucleotide polymorphisms, insertions and deletions among recipient and donor in a single sequencing experiment. Taken together, the presented data show that sequence capture and massively parallel pyrosequencing can be used as a new tool for risk assessment in the setting of allogeneic stem cell transplantation

Condensed Matter Theory of Dipolar Quantum Gases

Recent experimental breakthroughs in trapping, cooling and controlling ultracold gases of polar molecules, magnetic and Rydberg atoms have paved the way toward the investigation of highly tunable quantum systems, where anisotropic, long-range dipolar interactions play a prominent role at the many-body level. In this article we review recent theoretical studies concerning the physics of such systems. Starting from a general discussion on interaction design techniques and microscopic Hamiltonians, we provide a summary of recent work focused on many-body properties of dipolar systems, including: weakly interacting Bose gases, weakly interacting Fermi gases, multilayer systems, strongly interacting dipolar gases and dipolar gases in 1D and quasi-1D geometries. Within each of these topics, purely dipolar effects and connections with experimental realizations are emphasized.Comment: Review article; submitted 09/06/2011. 158 pages, 52 figures. This document is the unedited author's version of a Submitted Work that was subsequently accepted for publication in Chemical Reviews, copyright American Chemical Society after peer review. To access the final edited and published work, a link will be provided soo

Induction and processing of the radiation-induced gamma-H2AX signal and Its link to the underlying pattern of DSB: A combined experimental and modelling study

We present here an analysis of DSB induction and processing after irradiation with X-rays in an extended dose range based on the use of the γH2AX assay. The study was performed by quantitative flow cytometry measurements, since the use of foci counting would result in reasonable accuracy only in a limited dose range of a few Gy. The experimental data are complemented by a theoretical analysis based on the GLOBLE model. In fact, original aim of the study was to test GLOBLE predictions against new experimental data, in order to contribute to the validation of the model. Specifically, the γH2AX signal kinetics has been investigated up to 24 h after exposure to increasing photon doses between 2 and 500 Gy. The prolonged persistence of the signal at high doses strongly suggests dose dependence in DSB processing after low LET irradiation. Importantly, in the framework of our modelling analysis, this is related to a gradually increased fraction of DSB clustering at the micrometre scale. The parallel study of γH2AX dose response curves shows the onset of a pronounced saturation in two cell lines at a dose of about 20 Gy. This dose is much lower than expected according to model predictions based on the values usually adopted for the DSB induction yield (≈ 30 DSB/Gy) and for the γH2AX foci extension of approximately 2 Mbp around the DSB. We show and discuss how theoretical predictions and experimental findings can be in principle reconciled by combining an increased DSB induction yield with the assumption of a larger genomic extension for the single phosphorylated regions. As an alternative approach, we also considered in our model the possibility of a 3D spreading-mechanism of the H2AX phosphorylation around the induced DSB, and applied it to the analysis of both the aspects considered. Our results are found to be supportive for the basic assumptions on which GLOBLE is built. Apart from giving new insights into the H2AX phosphorylation process, experiments performed at high doses are of relevance in the context of radiation therapy, where hypo-fractionated schemes become increasingly popular

Ecological commonalities among pelagic fishes: comparison of freshwater ciscoes and marine herring and sprat

Systematic comparisons of the ecology between functionally similar fish species from freshwater and marine aquatic systems are surprisingly rare. Here, we discuss commonalities and differences in evolutionary history, population genetics, reproduction and life history, ecological interactions, behavioural ecology and physiological ecology of temperate and Arctic freshwater coregonids (vendace and ciscoes, Coregonus spp.) and marine clupeids (herring, Clupea harengus, and sprat, Sprattus sprattus). We further elucidate potential effects of climate warming on these groups of fish based on the ecological features of coregonids and clupeids documented in the previous parts of the review. These freshwater and marine fishes share a surprisingly high number of similarities. Both groups are relatively short-lived, pelagic planktivorous fishes. The genetic differentiation of local populations is weak and seems to be in part correlated to an astonishing variability of spawning times. The discrete thermal window of each species influences habitat use, diel vertical migrations and supposedly also life history variations. Complex life cycles and preference for cool or cold water make all species vulnerable to the effects of global warming. It is suggested that future research on the functional interdependence between spawning time, life history characteristics, thermal windows and genetic differentiation may profit from a systematic comparison of the patterns found in either coregonids or clupeids