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      Characterization of longitudinal canal tissue in the acorn barnacle Amphibalanus amphitrite

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          Abstract

          The morphology and composition of tissue located within parietal shell canals of the barnacle Amphibalanus amphitrite are described. Longitudinal canal tissue nearly spans the length of side shell plates, terminating near the leading edge of the specimen basis in proximity to female reproductive tissue located throughout the peripheral sub-mantle region, i. e. mantle parenchyma. Microscopic examination of stained longitudinal canal sections reveal the presence of cell nuclei as well as an abundance of micron-sized spheroids staining positive for basic residues and lipids. Spheroids with the same staining profile are present extensively in ovarioles, particularly within oocytes which are readily identifiable at various developmental stages. Mass spectrometry analysis of longitudinal canal tissue compared to tissue collected from the mantle parenchyma reveals a nearly 50% overlap of the protein profile with the greatest number of sequence matches to vitellogenin, a glycolipoprotein playing a key role in vitellogenesis–yolk formation in developing oocytes. The morphological similarity and proximity to female reproductive tissue, combined with mass spectrometry of the two tissues, provides compelling evidence that one of several possible functions of longitudinal canal tissue is supporting the female reproductive system of A. amphitrite, thus expanding the understanding of the growth and development of this sessile marine organism.

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          The prophenoloxidase-activating system in invertebrates.

          A major innate defense system in invertebrates is the melanization of pathogens and damaged tissues. This important process is controlled by the enzyme phenoloxidase (PO) that in turn is regulated in a highly elaborate manner for avoiding unnecessary production of highly toxic and reactive compounds. Recent progress, especially in arthropods, in the elucidation of mechanisms controlling the activation of zymogenic proPO into active PO by a cascade of serine proteinases and other factors is reviewed. The proPO-activating system (proPO system) is triggered by the presence of minute amounts of compounds of microbial origins, such as beta-1,3-glucans, lipopolysaccharides, and peptidoglycans, which ensures that the system will become active in the presence of potential pathogens. The presence of specific proteinase inhibitors prevents superfluous activation. Concomitant with proPO activation, many other immune reactions will be produced, such as the generation of factors with anti-microbial, cytotoxic, opsonic, or encapsulation-promoting activities.
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            C-type lectins in immunity and homeostasis

            The C-type lectins are a superfamily of proteins that recognize a broad repertoire of ligands and that regulate a diverse range of physiological functions. Most research attention has focused on the ability of C-type lectins to function in innate and adaptive antimicrobial immune responses, but these proteins are increasingly being recognized to have a major role in autoimmune diseases and to contribute to many other aspects of multicellular existence. Defects in these molecules lead to developmental and physiological abnormalities, as well as altered susceptibility to infectious and non-infectious diseases. In this Review, we present an overview of the roles of C-type lectins in immunity and homeostasis, with an emphasis on the most exciting recent discoveries.
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              Rapid mass spectrometric conversion of tissue biopsy samples into permanent quantitative digital proteome maps

              Clinical specimens are each inherently unique, limited and non-renewable. As such, small samples such as tissue biopsies are often completely consumed after a limited number of analyses. Here we present a method that enables fast and reproducible conversion of a small amount of tissue (approximating the quantity obtained by a biopsy) into a single, permanent digital file representing the mass spectrometry-measurable proteome of the sample. The method combines pressure cycling technology (PCT) and SWATH mass spectrometry (MS), and the resulting proteome maps can be analyzed, re-analyzed, compared and mined in silico to detect and quantify specific proteins across multiple samples. We used this method to process and convert 18 biopsy samples from 9 renal cell carcinoma patients into SWATH-MS fragment ion maps. From these proteome maps we detected and quantified more than 2,000 proteins with a high degree of reproducibility across all samples. The identified proteins clearly separated tumorous kidney tissues from healthy tissue, and differentiated distinct histomorphological kidney cancer subtypes.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – original draftRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: ConceptualizationRole: Formal analysisRole: Writing – review & editing
                Role: Data curationRole: InvestigationRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: Investigation
                Role: Data curationRole: Formal analysisRole: Investigation
                Role: ConceptualizationRole: Data curationRole: InvestigationRole: Writing – review & editing
                Role: Investigation
                Role: InvestigationRole: Writing – review & editing
                Role: SupervisionRole: Writing – review & editing
                Role: ConceptualizationRole: Project administrationRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                10 December 2018
                2018
                : 13
                : 12
                : e0208352
                Affiliations
                [1 ] National Research Council Research Associateship Program, Washington, D.C., United States of America
                [2 ] Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, Washington, D.C., United States of America
                [3 ] Naval Research Enterprise Internship Program, Washington, D.C., United States of America
                [4 ] Duke University Marine Laboratory, Beaufort, N.C., United States of America
                [5 ] Chemistry Division, Naval Research Laboratory, Washington, D.C., United States of America
                University of Split, Faculty of science, CROATIA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                [¤a]

                Current address: Molecular Genetics and Microbiology, Duke University, Durham, N.C., United States of America

                [¤b]

                Current address: University of Virginia, Charlottesville, VA, United States of America

                [¤c]

                Current address: Lantham & Watkins LLP, Washington, D.C., United States of America

                Author information
                http://orcid.org/0000-0002-4132-5341
                http://orcid.org/0000-0002-9894-6640
                http://orcid.org/0000-0002-6825-4133
                http://orcid.org/0000-0003-4307-108X
                Article
                PONE-D-18-23653
                10.1371/journal.pone.0208352
                6287898
                30532169
                76bb51b3-117a-47b7-b374-e5388cd6b7f6

                This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

                History
                : 10 August 2018
                : 15 November 2018
                Page count
                Figures: 8, Tables: 0, Pages: 19
                Funding
                Work under C.S. and K.W. was funded through an internal basic research program at NRL. C.W. and J.N.S. were both supported through National Research Council Post-Doctoral Associateships. Work at Duke University Marine Laboratory under D.R. was performed under the Office of Naval Research Grant N00014-16-1-3112. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Invertebrates
                Arthropoda
                Crustaceans
                Barnacles
                Biology and Life Sciences
                Cell Biology
                Cellular Types
                Animal Cells
                Germ Cells
                OVA
                Oocytes
                Research and analysis methods
                Specimen preparation and treatment
                Staining
                Nuclear staining
                DAPI staining
                Biology and Life Sciences
                Anatomy
                Reproductive System
                Medicine and Health Sciences
                Anatomy
                Reproductive System
                Biology and Life Sciences
                Biochemistry
                Lipids
                Research and Analysis Methods
                Specimen Preparation and Treatment
                Staining
                Group-Specific Staining
                Hematoxylin Staining
                Biology and Life Sciences
                Anatomy
                Histology
                Medicine and Health Sciences
                Anatomy
                Histology
                Biology and Life Sciences
                Molecular Biology
                Macromolecular Structure Analysis
                Lipid Analysis
                Custom metadata
                The mass spectrometry proteomics data, "Amphibalanus amphitrite Longitudinal Canal Tissue," are publically available at the ProteomeXchange via the PRIDE database with the dataset identifier PXD010743, Website: http://www.ebi.ac.uk/pride.

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