Stillborn or liveborn? Comparing umbilical cord immunohistochemical expression of vitality markers (tryptase, α1-antichymotrypsin and CD68) by quantitative analysis and confocal laser scanning microscopy
Department of Forensic Pathology, University of Foggia, Ospedale Colonnello D'Avanzo, Foggia, Italy.Pathology - Research and Practice (Impact Factor: 1.4). 02/2009; 205(8):534-41. DOI: 10.1016/j.prp.2009.01.011
The distinction between stillborn and liveborn infants and the demonstration of a separate existence of fetuses are central issues in the daily practice of perinatologists and pathologists. The current knowledge about the chronology of responses of the tissue following the occurrence of a vital reaction, as well as the existence of numerous studies that aimed at identifying markers of vitality of cutaneous lesions, induced us to investigate the umbilical cord for the presence or absence of vitality indexes. We investigated 45 samples of umbilical cords obtained during post-mortem examinations of stillborns, as well as samples of umbilical cords taken from newborns after normal labor. On these samples, we performed a complete immunohistochemical study. Our results showed that some of the parameters investigated, such as tryptase for the mast cell, CD68, and alpha-1-antichymotrypsin, showed a statistically significant (p<0.0001) different expression in the two groups under study (stillborn and liveborn). Owing to the strong different expression of these markers in the samples of the umbilical cords from liveborns, compared to those from stillborns, one might regard them as reliable parameters, to which the pathologist may resort whenever he is dealing with the distinction between stillborns and liveborns.
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ABSTRACT: The proinflammatory cytokines interleukin-1beta (IL-1β), interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-α) hold important functions in the early and late courses of inflammation, trauma and wound healing. In the present study, human skin wounds due to sharp force (n=105) were collected during surgery and autopsy. The wound age mainly varied from several minutes to 5h, some specimens aged up to 6 weeks. Control specimens from uninjured skin were available in each case. After preparation of cryostat sections, immunohistochemistry was performed according to the APAAP technique, using monoclonal and polyclonal antibodies. The results were evaluated semiquantitatively. All markers were weakly expressed in normal human skin constitutively. However, the staining pattern changed significantly in vital wounds concerning epidermal layers, subepidermal cells, vessels and sweat glands. IL-1β and IL-6 showed enhanced expression after 15 and 20min at the earliest (increase of epidermal reactivity). After 30–60 and 60–90min, respectively, marked expression was observed with these markers. Similar alterations were detectable with TNF-α after 15 and 60–90min. The reactivity of all three markers persisted over several hours, then decreased to basal levels again and sometimes reappeared after days and in granulation tissue. Leukocytes reacting with IL-1β and IL-6 appeared after approximately 2h. Conclusion: proinflammatory cytokines can serve as a useful tool for the estimation of vitality and wound age, in particular in the early post-traumatic interval prior to leukocyte reaction. Autolysis did not play a role in the samples investigated (postmortem interval up to 8 days). Problems could sometimes rise from constitutive expression. Therefore, it is recommended to examine control samples from the same individual and to compare the reactivity with wound specimens.Forensic Science International 12/2002; 130(2):90-96. DOI:10.1016/S0379-0738(02)00342-0 · 2.14 Impact Factor
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ABSTRACT: The distribution of the proteinase inhibitors alpha-1-antichymotrypsin (alpha 1-act), alpha-2-macroglobulin (alpha-2-m) and lysozyme was analysed immunohistochemically in 27 intravitally acquired wounds, 3 postmortem skin lacerations and 9 specimens of undamaged skin. Intravitally acquired wounds demonstrated distinct positive reactions for all antibodies examined (alpha-1-act 66.6%; alpha-2-m 51.9%; lysozyme 25.9%). However the undamaged skin margins opposite the wound margins also gave positive reactions (alpha-1-act 51.8%; alpha-2-m 37.0%; lysozyme 25.9%). Nearly half of the control cases (specimens of undamaged skin) exhibited weak positive reactions for all 3 antibodies. These could be easily distinguished from the strong positive reactions observed in intravitally acquired wounds. False positive reactions were observed due to contamination resulting from contact with serum components, in cases of advanced autolysis of specimens, and as a result of fixation and drying artefacts. Even though immunohistochemical studies of alpha-1-act, alpha-2-m and lysozyme give some indications concerning wound vitality, they cannot be considered as proof because irrefutable differentiation of true positive and false positive reactions is not possible in all cases.Deutsche Zeitschrift für die Gesamte Gerichtliche Medizin 02/1994; 107(1):29-33. DOI:10.1007/BF01247271 · 2.71 Impact Factor
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ABSTRACT: Mast cells are multifunctional, tissue-dwelling cells capable of secreting a wide variety of mediators. They develop from bone marrow-derived progenitor cells, primed with stem cell factor (SCF), which mediates its actions by interacting with the SCF receptor or c-kit on the cell surface. Mast cells continue their maturation and differentiation in peripheral tissue, developing into two well described subsets of cells, MCT and MCTC cells, varying in content of tryptase and chymase as well as in immunobiology. Mast cells are activated by numerous stimuli, including antigen (acting via the high affinity IgE receptor, Fc?RI), superoxides, complement proteins, neuropeptides and lipoproteins resulting in activation and degranulation. Following activation, these cells express mediators such as histamine, leukotrienes and prostanoids, as well as proteases, and many cytokines and chemokines, pivotal to the genesis of an inflammatory response. Recent data suggests that mast cells may play an active role in such diverse diseases as atherosclerosis, malignancy, asthma, pulmonary fibrosis and arthritis. Mast cells directly interact with bacteria and appear to play a vital role in host defense against pathogens. Drugs, such as glucocorticoids, cyclosporine and cromolyn have been demonstrated to have inhibitory effects on mast cell degranulation or mediator release.Frontiers in Bioscience 10/2001; 6:D1109-27. DOI:10.2741/krishnas · 3.52 Impact Factor
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