Stefano D'Errico

Correlation between cardiac oxidative stress and myocardial pathology due to acute and chronic norepinephrine administration in rats

Background: To investigate the cardiotoxic role of reactive oxygen species (ROS) and of products derived from catecholamines auto‐oxidation, we studied: (1) the response of antioxidant cardiac cellular defence systems to oxidative stress induced by norepinephrine (NE) administration, (2) the effect of NE administration on cardiac β1‐adrenergic receptors by means of receptor binding assay, (3) the cellular morphological alterations related to the biologically cross‐talk between the NE administration and cytokines [tumor necrosis factor‐alpha (TNF‐α), monocyte chemotactic protein‐1 (MCP‐1), interleukins IL6, IL8, IL10]Methods and Results: A total of 195 male rats was used in the experiment. All animals underwent electrocardiogram (EKG) before being sacrificed. The results obtained show that NE administration influences the antioxidant cellular defence system significantly increasing glutathione peroxidase (GPx) activity, glutathione reductase (GR) and superoxide dismutase (SOD). The oxidized glutathione (GSH/GSSG) ratio significantly decreases and malondialdehyde (MDA) levels increase showing a state of lipoperoxidation of cardiac tissue. We describe a significant apoptotic process randomly sparse in the damaged myocardium and the effect of ROS on the NE‐mediated TNF‐α, MCP‐1, and IL6, IL8, IL10 production. Conclusions: Our results support the hypothesis that catecholamines may induce oxidative damage through reactive intermediates resulting from their auto‐oxidation, irrespective of their interaction with adrenergic receptors, thus representing an important factor in the pathogenesis of catecholamines‐induced cardiotoxicity. The rise of the cardioinhibitory cytokines may be interpreted as the adaptive response of jeopardized myocardium with respect to the cardiac dysfunction resulting from NE injection.

Cardiac pathology in death from electrocution

To better characterize the morphologic changes in electrocution, morphologic changes in the hearts of 21 subjects, who died instantaneously of electrocution, were compared to the hearts of decedents with different types of death. Sixteen myocardial samples per heart were processed for histological examination, and sections were prepared with a variety of specific stains. The frequency, location and extent of myocellular segmentation (stretching and/or rupture) of intercalated discs and associated changes of myocardial bundles and single myocells [myofibre break-up (MFB)] were recorded, quantitatively analysed and statistically evaluated. The frequency of MFB was maximal in cases of electrocution (90%). The findings show that MFB is an ante-mortem change and may be a distinct finding in electrocution.

Enzymatic–nonenzymatic cellular antioxidant defense systems response and immunohistochemical detection of MDMA, VMAT2, HSP70, and apoptosis as biomarkers for MDMA (Ecstasy) neurotoxicity

3,4‐Methylenedioxymethamphetamine (MDMA)‐induced neurotoxicity leads to the formation of quinone metabolities and hydroxyl radicals and then to the production of reactive oxygen species (ROS). We evaluated the effect of a single dose of MDMA (20 mg/kg, i.p.) on the enzymatic and nonenzymatic cellular antioxidant defense system in different areas of rat brain in the early hours (<6 hr) of the administration itself, and we identified the morphological expressions of neurotoxicity induced by MDMA on the vulnerable brain areas in the first 24 hr. The acute administration of MDMA produces a decrease of reduced and oxidized glutathione ratio, and antioxidant enzyme activities were significantly reduced after 3 hr and after 6 hr in frontal cortex. Ascorbic acid levels strongly increased in striatum, hippocampus, and frontal cortex after 3 and 6 hr. High levels of malonaldehyde with respect to control were measured in striatum after 3 and 6 hr and in hippocampus and frontal cortex after 6 hr. An immunohistochemical investigation on the frontal, thalamic, hypothalamic, and striatal areas was performed. A strong positive reaction to the antivesicular monoamine transporter 2 was observed in the frontal section, in the basal ganglia and thalamus. Cortical positivity, located in the most superficial layer was revealed only for heat shock protein 70 after 24 hr.

Anabolic Steroid - and Exercise - Induced Cardio-Depressant Cytokines and Myocardial β1 Receptor Expression in CD1 Mice

Few animal model studies have been conducted in order to evaluate the impact of androgenic anabolic steroids (AAS) supraphysiological doses on the cardiovascular system and myocardial injury. Twenty-five male CD1 mice (8-10 weeks old; 35g initial body weight) were randomized into three AAS treated groups and two control groups. The AAS mice received intramuscular Nandrolone Decanoate (DECA-DURABOLIN), vehicled in arachidis oil, for 42 days, twice per week, with different dosages, studying plasma lipid analysis, cardiac histopathological features, cardiac β (1) adrenergic receptor expression, and the effects of the myocardial expression of inflammatory mediators (IL-1β, TNF-α) on the induction of cardiomyocytes apoptosis (HSP 70, TUNEL), using proteomic and immunohistochemical analysis. The mice had free movements in their animal rooms (two groups) or exercised by running on a motor-driven treadmill the others three groups. Recurring high dose AAS administration and physical training in mice produce significant increase in body weight and for total cholesterol. A moderate increase of the heart weight, cardiac hypertrophy and wide colliquative myocytolysis, were observed in high dose AAS administration and physical training group. The expression of HSP70 and inflammatory cytokine IL-1β, increased in the three AAS-treated groups. TNF- α showed a more extensive expression in the AAS-high dose group. A significant apoptotic process randomly sparse in the myocardium was described. Our data support the hypothesis that the combined effects of vigorous training, anabolic steroid abuse and stimulation of the sympathetic nervous system, may predispose to myocardial injury.

Calcium oxalate crystals in acute ethylene glycol poisoning: A confocal laser scanning microscope study in a fatal case

Introduction. The severity of ethylene glycol toxicity is related to the metabolic acidosis resulting from the biotransformation of ethylene glycol into toxic metabolites. Glycolic acid causes severe acidosis and oxalate precipitates as calcium oxalate in the kidneys and other tissues. Case Report. An adult male was taken to the local hospital by the team rescue and was apparently unconscious; severe metabolic acidosis and renal failure led to death a few hours after the arrival. Confocal laser scanning microscopy demonstrated oxalate crystals deposition within the tubular epithelial cells and widespread necrosis of the tubular epithelium in the proximal tubules. Toxicological examinations revealed ethylene glycol; the blood level was 250 mg/L and in urine the concentration was 0.3%. Discussion. In cases of ethylene glycol poisoning, calcium oxalate may be excreted not only as dihydrate crystals, but also as monohydrate crystals. Direct toxicity, cortical edema, and inhibition of mitochondrial activity, as evidenced by decreased succinate dehydrogenase activity, are possible mechanisms of crystal damage. Since calcium oxalate monohydrate crystals are transported intracellularly by kidney cells, the renal toxicity of ethylene glycol may result from inhibition of mitochondrial respiratory function in proximal tubular cells by calcium oxalate monohydrate crystals. Conclusions. The histologic diagnosis of acute renal failure secondary to ethylene glycol poisoning depends on the recognitions of the changes of acute tubular damage in association with calcium oxalate crystals deposition within the tubular epithelial cells and the widespread necrosis of the tubular epithelium in the proximal tubules.

Cave Canem: Bite Mark Analysis in a Fatal Dog Pack Attack

Deaths resulting from animal attacks are rare, and according to The Humane Society of the United States, more than 300 individuals died of dog attacks in the United States from 1979 to 1996. The case of a fatal dog-pack attack on an 83-year-old woman is presented. Wide lacerations of the scalp, several tooth puncture wounds, and bruises reproducing bite marks were recorded on the whole body. Exsanguinations due to brachial artery laceration subsequent to multiple dog bites were indicated as the main cause of death. An integrated study in association with a veterinary doctor was performed on 27 dogs of different breed (24 Cane Corso, 1 Dalmatian, 2 German Shepherds) collecting dental formula and dental casts. Dental casts were superimposed on the victims wound samples collected at autopsy and analyzed for compatibility-the patterns taken from the jaws of 3 suspected dogs could be clearly adapted on the bite marks. At the end of investigation, the son of the victim indicated the 3 dogs of his own as the responsible ones and he was condemned for manslaughter. Bite marks analysis provided conclusive evidences in identifying the offending animals. The results may be important to give details about bite circumstances and predisposition of specific breeds of dogs to bite or inflict severe bites.

Renal heat shock proteins over-expression due to anabolic androgenic steroids abuse.

Chronic use of anabolic adrogenic steroids (AAS) has been known to cause serious adverse effects. While the effects of AASs on cardiovascular system are well known, toxicity on other organs has received less attention. A doserelated nephrotoxic effect has been proposed and a wide variety of morpho-functional damages have been observed, but the exact pathophysiological mechanism of action is still not well known. In the present minireview, we highlight the remaining issues through an analysis of the pertinent literature. As with HSPs toxic agents their overexpression could be considered a protective reaction against AAS abuse however, comprehensive studies concerning the whole range of Hsps/chaperones expressions in all organs after long term use of AAS are needed.

Blast overpressure after tire explosion: A fatal case

Fatal blast injuries are generally reported in literature as a consequence of the detonation of explosives in war settings. The pattern of lesion depends on the position of the victim in relation to the explosion, on whether the blast tracks through air or water, and whether it happens in the open air or within an enclosed space and the distance from the explosion. Tire explosion-related injuries are rarely reported in literature. This study presents a fatal case of blast overpressure due to the accidental explosion of a truck tire occurring in a tire repair shop. A multidisciplinary approach to the fatality involving forensic pathologists and engineers revealed that the accidental explosion, which caused a series of primary and tertiary blast wave injuries, was due to tire deterioration.

Stillborn or liveborn? Comparing umbilical cord immunohistochemical expression of vitality markers (tryptase, alpha(1)-antichymotrypsin and CD68) by quantitative analysis and confocal laser scanning microscopy.

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.