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1.
Nat Commun ; 9(1): 1948, 2018 05 16.
Article in English | MEDLINE | ID: mdl-29769532

ABSTRACT

In the course of both innate and adaptive immunity, cytidine deaminases within the activation induced cytidine deaminase (AID)/apolipoprotein B editing complex (APOBEC) family modulate immune responses by mutating specific nucleic acid sequences of hosts and pathogens. The evolutionary emergence of these mediators, however, seems to coincide precisely with the emergence of adaptive immunity in vertebrates. Here, we show a family of genes in species within two divergent invertebrate phyla-the echinoderm Strongylocentrotus purpuratus and the brachiopod Lingula anatina-that encode proteins with similarities in amino acid sequence and enzymatic activities to the vertebrate AID/APOBECs. The expression of these invertebrate factors is enriched in tissues undergoing constant, direct interactions with microbes and can be induced upon pathogen challenge. Our findings suggest that AID/APOBEC proteins, and their function in immunity, emerged far earlier than previously thought. Thus, cytidine deamination is probably an ancient innate immune mechanism that predates the protostome/deuterostome divergence.


Subject(s)
APOBEC-1 Deaminase/genetics , Cytidine Deaminase/genetics , Immunity, Innate/genetics , Invertebrates/genetics , APOBEC-1 Deaminase/metabolism , Adaptive Immunity/genetics , Amino Acid Sequence , Animals , Cytidine Deaminase/metabolism , Gene Expression Regulation, Enzymologic , Humans , Invertebrates/classification , Invertebrates/enzymology , Mutation , Sequence Homology, Amino Acid , Species Specificity , Strongylocentrotus purpuratus/enzymology , Strongylocentrotus purpuratus/genetics
2.
PLoS One ; 12(11): e0187987, 2017.
Article in English | MEDLINE | ID: mdl-29125863

ABSTRACT

Coelomocytes represent the immune cells of echinoderms, but detailed knowledge about their roles during immune responses is very limited. One major challenge for studying coelomocyte biology is the lack of reagents to identify and purify distinct populations defined by objective molecular markers rather than by morphology-based classifications that are subjective at times. Glycosylation patterns are known to differ significantly between cell types in vertebrates, and furthermore they can vary depending on the developmental stage and activation states within a given lineage. Thus fluorescently labeled lectins that recognize distinct glycan structures on cell surface proteins are routinely used to identify discrete cell populations in the vertebrate immune system. Here we now employed a panel of fifteen fluorescently-labeled lectins to determine differences in the glycosylation features on the surface of Strongylocentrotus purpuratus coelomocytes by fluorescence microscopy and flow cytometry. Eight of the lectins (succinylated wheat germ agglutinin, Len culinaris lectin, Pisum sativum agglutinin, Saphora japonica agglutinin, Solanum tuberosum lectin, Lycopersicon esculentum lectin, Datura stramonium lectin, Vicia villosa lectin) showed distinct binding patterns to fixed and live cells of three major coelomocyte classes: phagocytic cells, red spherule cells, and vibratile cells. Importantly, almost all lectins bound only to a subgroup of cells within each cell type. Lastly, we established fluorescently-labeled lectin-based fluorescence activated cell sorting as a strategy to purify distinct S. purpuratus coelomocyte (sub-)populations based on molecular markers. We anticipate that this will become a routine approach in future studies focused on dissecting the roles of different coelomocytes in echinoderm immunity.


Subject(s)
Lectins/physiology , Strongylocentrotus purpuratus/cytology , Animals , Microscopy, Fluorescence
3.
Orthopedics ; 30(11): 959-64, 2007 11.
Article in English | MEDLINE | ID: mdl-18019991

ABSTRACT

This prospective, randomized trial compared treatment with Knowles pins and plates in 62 elderly patients (>50 years) with midclavicular fractures. The clinical outcomes were evaluated at 30 months postoperatively. The mean shoulder score of the Knowles pinning was 85 points and the plating was 84 points (P=.7). Knowles pinning requires significantly shorter operative time (P<.001), smaller wound size (P<.001), shorter hospital stay P=.03), less meperidine use (P=.02), lower complication rate (P=0.04), and less symptomatic hardware (P=.015). If surgery of mid-third clavicular fractures is indicated, fixation with a Knowles pin has more advantages than plate fixation in elderly patients.


Subject(s)
Bone Nails , Bone Plates , Clavicle/surgery , Fracture Fixation, Internal/methods , Fractures, Bone/surgery , Acetaminophen/therapeutic use , Aged , Aged, 80 and over , Analgesics, Non-Narcotic/therapeutic use , Analgesics, Opioid/therapeutic use , Clavicle/injuries , Drug Utilization/statistics & numerical data , Female , Fracture Fixation, Internal/adverse effects , Fracture Healing , Humans , Length of Stay/statistics & numerical data , Male , Meperidine/therapeutic use , Middle Aged , Pain, Postoperative/drug therapy , Prospective Studies , Time Factors , Treatment Outcome
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