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    ABSTRACT: Women with breast cancer (BC) and anti thyroid peroxidase (TPO) autoantibodies (TPOAb) have a better prognosis than women lacking TPOAb. Sera from women with TPOAb displayed immunoreactivity to BC tissue by immunofluorescence that was not apparent in women without TPOAb. We hypothesize a BC/thyroid shared antigen that provides a target for humoral or cell-mediated immune activity; candidates include the sodium/iodide symporter (NIS, expressed in thyroid and BC), cross-reacting epitopes in TPO and lactoperoxidase (LPO) or TPO itself. Since the association is with TPOAb, we investigated TPO expression in BC, breast peri-tumoral tissue (PT), other tissues (tumoral and not) and thyroid as positive control. Transcripts for known and novel TPO isoforms were detected in BC (n=8) and PT (n=8) but at approximately 10(4) fold lower than in thyroid while in non-BC tumours (n=5) they were at the limit of detection. TPO was expressed also in adipose tissue (n=17), 10(3) fold lower than in thyroid. Full length TPO (Mr 105-110 kDa) was detected in western blots in the majority of examined tissues; pre-absorption of the TPO antibody with recombinant TPO (but not LPO) reduced the signal, indicating specificity. The same occurred with some lower molecular weight bands, which could correspond to smaller TPO transcript isoforms, present in all samples. In conclusion TPO is weakly expressed in BC and other tissues; this could partly explain the high frequency and protective role of TPOAb in BC patients. Further studies will investigate tissue specificity, function and immunogenicity of the novel TPO variants (some BC-specific) identified. © 2013 Wiley Periodicals, Inc.
    Full-text · Article · Apr 2014 · International Journal of Cancer
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    ABSTRACT: The ability of neutrophils to rapidly change shape underlies their physiological functions of phagocytosis and spreading. A major problem in establishing the mechanism is that conventional microinjection of substances and indicators interferes with this dynamic cell behavior. Here we show that electroinjection, a “no-touch” point-and-shoot means of introducing material into the cell, is sufficiently gentle to allow neutrophils to be injected whilst undergoing chemokinesis and spreading without disturbing cell shape change behavior. Using this approach, a fluorogenic calpain-1 selective peptide substrate was introduced into the cytosol of individual neutrophils undergoing shape changes. These data showed that (i) physiologically elevated cytosolic Ca2 +concentrations were sufficient to trigger calpain-1 activation, blockade of Ca2 + influx preventing calpain activation and(ii) calpain-1 activity was elevated in spreading neutrophil.. These findings provide the first direct demonstration of a physiological role for Ca2 + elevation in calpain-1 activation and rapid cell spreading. Electroinjection of cells undergoing dynamic shape changes thus opens new avenues of investigation for defining the molecular mechanism underlying dynamic cell shape changes.
    No preview · Article · Jan 2014 · Biochimica et Biophysica Acta (BBA) - Molecular Cell Research
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    ABSTRACT: This review discusses the role that the sperm-specific phospholipase C zeta (PLCζ) is proposed to play during the fertilization of mammalian eggs. At fertilization, the sperm initiates development by causing a series of oscillations in cytosolic concentrations of calcium [Ca2] within the egg. PLCζ mimics the sperm at fertilization, causing the same pattern of Ca2+ release as seen at fertilization. Introducing PLCζ into mouse eggs also mimics a number of other features of the way in which the fertilizing sperm triggers Ca2+ oscillations. We discuss the localization of PLCζ within the egg and present a hypothesis about the localization of PLCζ within the sperm before the initiation of fertilization.
    No preview · Article · Jan 2014 · Biochemical and Biophysical Research Communications
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