TY - JOUR
T1 - A possible biomarker for methadone related deaths
AU - Farè, F; Roda, G; Gambaro, V; Karch, Sb
AU - Procaccianti, Paolo
AU - Argo, Antonina
AU - Pitruzzella, Alessandro
AU - Uzzo, Maria Laura
AU - Spatola, Giovanni Francesco
AU - Sortino, Cettina
AU - Zerbo, Stefania
AU - Lanzarone, Antonietta
PY - 2017
Y1 - 2017
N2 - Methadone (MTH) concentrations in those dying of MTH toxicity totally overlap concentrations where
the presence of MTH is only an incidental finding, making it very difficult to make distinctions in actual
cases. A biomarker, be it anatomical or biochemical for MTH toxicity is badly needed, particularly if that
markers were known to disrupt effective ventilation. Because the brainstem houses the regulatory
centers for cardiorespiratory-control enters, it would seem to be the most likely anatomical site to seek
abnormalities in cardiorespiratory control.
Objective: To locate and describe the cells of nucleus of the solitary tract (TS)(NTS) in human brainstem
and determine if neuronal cell death, either necrotic or apoptotic, within the TS of humans is more
common in deaths due directly to MTH toxicity than with in the solitary tract itself.
Design, setting, participants: This was a single cohort study of MTH related decedents autopsied at a large
university hospital. Each decedent had a recent history of non medical/illicit MTH use and had been
pronounced dead in the field, prior to ever reaching the hospital. Complete autopsy and complete
toxicology testing were performed on the formalin fixed brains of each individual. Multiple blocks were
prepared of the area of interest, namely the tissue lying immediately between the inferior and the super
colliculi. This volume, by definition, would have included the area of the Rostral Ventrolateral Medulla
(RVLM), the location of the TS. Immunohistochemistry studies utilizing caspase-9 reaction (a protease
enzyme involved in the process of preprogrammed death) were performed in order to estimate the
degree and proportion of neuronal apoptosis, and also access the degree of classical necrosis within the
NTS.
Main outcomes and measures: The primary outcome measure was the presence or absence of neuronal
apoptosis and/or necrosis within the NTS.
Results: Cells displaying evidence of early apoptosis and advanced apoptosis, consisting primarily of
nuclear fragmentation, admixed with other neurons displaying the features of classic necrosis were
found. Evidence of classic necrosis was identifiable in most of the controls, though minor degrees of
apoptosis were identifiable with Caspase staining and quantitative image analysis of immunohistochemical
stains.
Conclusions: and Relevance: Our study shows that neurons, primarily along the TS, but occasionally in
other cell nuclei (even controls) are vulnerable, both to direct MTH toxicity (via apoptosis) and indirectly
(via hypoxia leading to classical cell necrosis). When MTH is found to be present in significant concentrations,
but apoptotic lesions are absent, it would be reasonable to assume that MTH was not primarily
the cause of cardiorespiratory arrest.
AB - Methadone (MTH) concentrations in those dying of MTH toxicity totally overlap concentrations where
the presence of MTH is only an incidental finding, making it very difficult to make distinctions in actual
cases. A biomarker, be it anatomical or biochemical for MTH toxicity is badly needed, particularly if that
markers were known to disrupt effective ventilation. Because the brainstem houses the regulatory
centers for cardiorespiratory-control enters, it would seem to be the most likely anatomical site to seek
abnormalities in cardiorespiratory control.
Objective: To locate and describe the cells of nucleus of the solitary tract (TS)(NTS) in human brainstem
and determine if neuronal cell death, either necrotic or apoptotic, within the TS of humans is more
common in deaths due directly to MTH toxicity than with in the solitary tract itself.
Design, setting, participants: This was a single cohort study of MTH related decedents autopsied at a large
university hospital. Each decedent had a recent history of non medical/illicit MTH use and had been
pronounced dead in the field, prior to ever reaching the hospital. Complete autopsy and complete
toxicology testing were performed on the formalin fixed brains of each individual. Multiple blocks were
prepared of the area of interest, namely the tissue lying immediately between the inferior and the super
colliculi. This volume, by definition, would have included the area of the Rostral Ventrolateral Medulla
(RVLM), the location of the TS. Immunohistochemistry studies utilizing caspase-9 reaction (a protease
enzyme involved in the process of preprogrammed death) were performed in order to estimate the
degree and proportion of neuronal apoptosis, and also access the degree of classical necrosis within the
NTS.
Main outcomes and measures: The primary outcome measure was the presence or absence of neuronal
apoptosis and/or necrosis within the NTS.
Results: Cells displaying evidence of early apoptosis and advanced apoptosis, consisting primarily of
nuclear fragmentation, admixed with other neurons displaying the features of classic necrosis were
found. Evidence of classic necrosis was identifiable in most of the controls, though minor degrees of
apoptosis were identifiable with Caspase staining and quantitative image analysis of immunohistochemical
stains.
Conclusions: and Relevance: Our study shows that neurons, primarily along the TS, but occasionally in
other cell nuclei (even controls) are vulnerable, both to direct MTH toxicity (via apoptosis) and indirectly
(via hypoxia leading to classical cell necrosis). When MTH is found to be present in significant concentrations,
but apoptotic lesions are absent, it would be reasonable to assume that MTH was not primarily
the cause of cardiorespiratory arrest.
UR - http://hdl.handle.net/10447/227698
M3 - Article
VL - 49
SP - 8
EP - 14
JO - Journal of Forensic and Legal Medicine
JF - Journal of Forensic and Legal Medicine
SN - 1752-928X
ER -