Immunodeficiency as a side effect of anabolic androgenic steroid abuse: a case of necrotizing myofasciitis.

Even if there are well-known consequences of anabolic androgenic steroid (AAS) abuse, their full pathway of action is still being investigated. In this context, the presented case report aims to discuss and provide evidence of unusual adverse effects linked to immunodeficiency in an AAS abuser. In fact, this kind of chronic complication, even if not usually considered, may lead sudden death. In this case a 31-year-old aesthetic weightlifter, who presented to the emergency department due to an accidental fall that resulted in left thigh trauma. This subsequently developed into left thigh necrotizing myofasciitis in the following few days. Although surgery and hyperbaric therapy were carried out, the man died. An autopsy with complete biological sampling for toxicological studies was performed. This case highlights the close relationship between AAS abuse and immunodeficiency and highlights it's importance for further studies. However, it should be considered that of all the dangerous effects produced by AAS use, necrotizing fasciitis is not such an unusual consequence.

Decomposing Human Blood: Canine Detection Odor Signature and Volatile Organic Compounds.

The admissibility of human "odor mortis" discrimination in courts depends on the lack of comprehension of volatile organic compounds (VOCs) during the human decay process and of the lack in standardized procedures in training cadaver dogs. Blood was collected from four young people who died from traffic accidents and analyzed using HS-SPME/GC-MS at different decompositional stages. Two dogs, professionally trained, were tested to exactly locate blood samples, for each time point of the experiment. We found a long list of VOCs which varied from fresh to decomposed blood samples, showing differences in specific compounds. Dog performance showed a positive predictive value between 98.96% and 100% for DOG A, and between 99.47% and 100% for DOG B. Our findings demonstrated that decomposing human blood is a good source of VOCs and a good target for canine training.

Inhibition of aldose-reductase-2 by a benzofuroxane derivative bf-5m increases the expression of kcne1, kcnq1 in high glucose cultured H9c2 cardiac cells and sudden cardiac death.

Long QT syndrome (LQTS) is characterized by prolonged QT interval, leading to sudden cardiac death. Hyperglycemia is an important risk factor for LQTS, inhibiting the cardiac rapid component delayed rectifier K+ current (Iks), responsible for QT interval. We previously showed that the new ALR2 inhibitor BF-5m supplies cardioprotection from QT prolongation induced by high glucose concentration in the medium, reducing QT interval prolongation and preserving morphology. Here we investigated the effects of BF-5m on cell cytotoxicity and viability in H9c2 cells, and on cellular potassium ion channels expression. H9c2 cells were grown in medium with high glucose and high glucose plus the BF-5m by assessing the cytotoxic effects and the cell survival rate. In addition, KCNE1 and KCNQ1 expression in plasma and mitochondrial membranes were monitored. Also, the expression levels of miR-1 proved to suppress KCNQ1 and KCNE1, were analyzed. BF-5m treatment reduced the cytotoxic effects of high glucose on H9c2 cells by increasing cell survival rate and improving H9c2 morphology. Plasmatic KCNE1 and KCNQ1 expression levels were restored by BF-5m in H9c2 exposed to high glucose, down-regulating miR-1. These results suggest that BF-5m exerts cardioprotection from high glucose in rat heart ventricle H9c2 cells exposed to high glucose.

Neurotoxicity by synthetic androgen steroids: oxidative stress, apoptosis, and neuropathology: A review.

Anabolic-androgenic steroids (AAS) are synthetic substances derived from testosterone that are largely employed due to their trophic effect on muscle tissue of athletes at all levels. Since a great number of organs and systems are a target of AAS, their adverse effects are primarily on the following systems: reproductive, hepatic, musculoskeletal, endocrine, renal, immunological, infectious, cardiovascular, cerebrovascular, and hematological. Neuropsychiatric and behavioral effects as a result of AAS abuse are well known and described in the literature. Mounting evidence exists suggesting that in addition to psychiatric and behavioral effects, non-medical use of AAS carries neurodegenerative potential. Although, the nature of this association remains largely unexplored, recent animal studies have shown the recurrence of this AAS effect, ranging from neurotrophin unbalance to increased neuronal susceptibility to apoptotic stimuli. Experimental and animal studies strongly suggest that apoptotic mechanisms are at least in part involved in AAS-induced neurotoxicity. Furthermore, a great body of evidence is emerging suggesting that increased susceptibility to cellular oxidative stress could play a pivotal role in the pathogenesis of many neurodegenerative disorders and cognitive impairment. As in other drug-evoked encephalopathies, the key mechanisms involved in AAS - induced neuropathology could represent a target for future neuroprotective strategies. Progress in the understanding of these mechanisms will provide important insights into the complex pathophysiology of AAS-induced neurodegeneration, and will pave the way for forthcoming studies. Supplementary to abandoning the drug abuse that represents the first step in reducing the possibility of irreversible brain damage in AAS abusers, neuroprotective strategies have to be developed and implemented in future.

Confocal laser scanning microscope, Raman microscopy and Western blotting to evaluate inflammatory response after myocardial infarction.

Cardiac muscle necrosis is associated with inflammatory cascade that clears the infarct from dead cells and matrix debris, and then replaces the damaged tissue with scar, through three overlapping phases: the inflammatory phase, the proliferative phase and the maturation phase. Western blotting, laser confocal microscopy, Raman microscopy are valuable tools for studying the inflammatory response following myocardial infarction both humoral and cellular phase, allowing the identification and semiquantitative analysis of proteins produced during the inflammatory cascade activation and the topographical distribution and expression of proteins and cells involved in myocardial inflammation. Confocal laser scanning microscopy (CLSM) is a relatively new technique for microscopic imaging, that allows greater resolution, optical sectioning of the sample and three-dimensional reconstruction of the same sample. Western blotting used to detect the presence of a specific protein with antibody-antigen interaction in the midst of a complex protein mixture extracted from cells, produced semi-quantitative data quite easy to interpret. Confocal Raman microscopy combines the three-dimensional optical resolution of confocal microscopy and the sensitivity to molecular vibrations, which characterizes Raman spectroscopy. The combined use of western blotting and confocal microscope allows detecting the presence of proteins in the sample and trying to observe the exact location within the tissue, or the topographical distribution of the same. Once demonstrated the presence of proteins (cytokines, chemokines, etc.) is important to know the topographical distribution, obtaining in this way additional information regarding the extension of the inflammatory process in function of the time stayed from the time of myocardial infarction. These methods may be useful to study and define the expression of a wide range of inflammatory mediators at several different timepoints providing a more detailed analysis of the time course of the infarct.

Cadaver dogs: unscientific myth or reliable biological devices?

Dogs are commonly used to detect explosives, narcotics, and other illegal materials. In the forensic setting, cadaver dogs are trained to detect and locate concealed human remains or fluids due to the high sensitivity and selectivity of the canine olfactory system and the relative ease with which dogs can be trained and handled. The need for international and scientifically validated standards has long been outlined by the literature. It is important, therefore, to establish the reliability of the handler/dog team. Our study aimed to detect the real effectiveness of dogs trained to locate human cadaveric blood in very low concentrations, through an optimized and rigorously controlled design which would rule out any possible sources of bias. The study was designed to determine the dogs' olfactory sensitivity to human cadaveric blood and how this capacity might change as the dilution of blood increases from pure blood to very low concentrations. The further step was to examine the dogs' ability to discriminate among target (human cadaveric blood) and non-target (confounding substances) odors (discriminative capability). Our results revealed that well trained dogs were able to detect human cadaveric blood samples even when very low concentrations of blood were stored in the tubes, showing high levels of olfactory sensitivity and to discriminate the target odor even when the non-target odor was orders of magnitude higher in concentrations. Although our results are based only on two dogs, the procedure we used may provide a comprehensive answer to the need for a scientifically unassailable tool for quantifying and objectifying the performance of well-trained specific search dogs in detecting human cadaveric blood traces.

Cytokines, chaperones and neuroinflammatory responses in heroin-related death: what can we learn from different patterns of cellular expression?

Heroin (3,6-diacetylmorphine) has various effects on the central nervous system with several neuropathological alterations including hypoxic-ischemic brain damage from respiratory depressing effects and neuroinflammatory response. Both of these mechanisms induce the release of cytokines, chemokines and other inflammatory mediators by the activation of many cell types such as leucocytes and endothelial and glial cells, especially microglia, the predominant immunocompetent cell type within the central nervous system. The aim of this study is to clarify the correlation between intravenous heroin administration in heroin related death and the neuroinflammatory response. We selected 45 cases among autopsies executed for heroin-related death (358 total cases); immunohistochemical studies and Western blotting analyses were used to investigate the expression of brain markers such as tumor necrosis factor-α, oxygen-regulated protein 150, (interleukins) IL-1ß, IL-6, IL-8, IL-10, IL-15, cyclooxygenase-2, heat shock protein 70, and CD68 (MAC387). Findings demonstrated that morphine induces inflammatory response and cytokine release. In particular, oxygen-regulated protein 150, cyclooxygenase-2, heat shock protein 70, IL-6 and IL-15 cytokines were over-expressed with different patterns of cellular expression.

A novel macabre ritual of the Italian Mafia ('Ndrangheta): covering hands with gloves and burying the corpse with burnt lime after execution.

'Ndrangheta is one of the famous "4 Mafias," which typically insists in Southern Italy. A particular case of 'Ndrangheta-related homicide double execution by means of multiple gunshots is presented. To the best of our knowledge, this is the first report of `Ndrangheta's homicidal modality (covering hands with gloves and burying) as a macabre ritual. In homicides committed by criminal organization, the method used as well as the positioning of the body follow a macabre ritual laden with significance and intending to be an admonition to other persons.

A multidisciplinary approach to the investigation of a collapsed building.

Investigating the collapse of a building poses multiple and complex forensic challenges. Large numbers of specialized personnel and equipment are required, as are the combined technical skills of many different kinds of forensic investigators. Forensic pathology teams are integral to these efforts. This report describes the investigation that occurred after a building collapsed in southern Italian location. Several families were still living in small, and abandoned building built in the early 20th century. The buildings were located over cellars 3 meters underground, known locally as "the caves." Eight people were found dead under the debris of one of the collapsed houses and 6 were brought out alive. A team of forensic pathologists and engineers was appointed to investigate the causes of death and of the collapse, respectively. A complete autopsy was performed in every case, along with radiologic assessment and toxicological analysis. Autopsy findings were coded using the Abbreviated Injury Scale and the New Injury Severity Score. Systems for victim identification, arrangements for human remains, management of dead bodies, evaluation of the different patterns of injuries and, finally, detailed identification of the cause of death all played an important role.

Myocardial expression of TNF-alpha, IL-1beta, IL-6, IL-8, IL-10 and MCP-1 after a single MDMA dose administered in a rat model.

Indirect effects of 3,4-methylenedioxy-N-methylamphetamine (MDMA) and metabolites on the cardiac cells are well-known, the mechanism(s) underlying direct MDMA-induced cardiotoxicity remaining to be clarified. To better understand the immuno-inflammatory phenomena accompanying the cardiac alterations during MDMA administration, we conducted a study in an in vivo animal model to evaluate the cellular morphological alterations related to the biological response between MDMA administration and inflammatory cytokines (tumor necrosis factor-alpha, IL-1beta, IL-6, 8, 10, and monocyte chemotactic protein-1). A total of 25 male rats were used. The effects were evaluated at 6, 16 and 24 hours after a single dose MDMA administered (20 mg/kg i.p.). We found high levels of the cardioinhibitory cytokines in rat heart after 3 and 6 hs from MDMA administration. Strongest reaction was observed at 24 hs for TNF-alpha, IL-1beta, IL-6, 8, 10 and for MCP-1. Furthermore, we still determined the presence of MDMA and MDA in the plasma of rats treated with MDMA intra-peritoneal single injection; it was present as early at 6 hs and still present 24 hs after treatment. Western blot analysis in cardiac samples demonstrated the IL-1beta and IL-6 reactions in rats died spontaneously at fourth hour. The rise of the selective cardioinhibitory cytokines may be interpreted as the adaptive response of jeopardized myocardium to the cardiac dysfunction resulting from MDMA injection.

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.

Reconstruction of the weapon in a case of homicidal decapitation.

Decapitation as homicidal mode of death is relatively rare. In most cases of decapitation, the differentiation between the modes of death might be difficult to some extent, particularly in cases where essential investigative elements, like the decedent's head and the weapon, are unavailable. Our report concerns a case of homicide by decapitation without any further mutilation of the victim, where only the combination of autopsy results, histologic findings, and engineering technical reconstruction allowed us to identify with certainty the mode of death as vital decapitation. The technical reconstruction of the alleged weapon allowed the identification and the discovery of the hand saw used by the murderer to decapitate the victim.

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.

Cardiac oxidative stress determination and myocardial morphology after a single ecstasy (MDMA) administration in a rat model.

Experimental and clinical data indicate that 3,4-methylenedioxy-N-methylamphetamine (MDMA) abuse can produce significant cardiovascular toxicity. A mechanism may be a direct toxic effect of redox active metabolites of MDMA. To evaluate the effect of a single MDMA dose on cellular antioxidant defence system and to investigate the morphology in male albino rats, total glutathione (GSH), oxidised glutathione (GSSG), ascorbic acid (AA), glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD) and malondialdehyde (MDAL) were studied. The effects were evaluated at 3, 6, 16 and 24 h after MDMA administration. Antioxidant enzymes activity was significantly reduced: GPx (-24%) and SOD (-50%) after 3 h and GR (-19%) after 6 h from treatment. AA levels decrease (-37%) after 3 h and (-30%) after 6 h; MDAL level increased (+119%) after 3 h; GSH levels decreased after 3 (31.3%) and 6 h (37.9%) from MDMA treatment. GSSG content was not affected by ecstasy administration. Myocardial contraction band necrosis (CBN) was already visible in rats killed at 6 h. After 16 h, macrophagic monocytes around the necrotic myocardial cells were observed, and within 24 h, this infiltrate became more widespread with an early removal of the necrotic material. Calcium deposits were observed within ventricular cardiomyocytes with intact nuclei and sarcomeres. Single administration of MDMA can significantly alter the cellular antioxidant defence system and produce oxidative stress which may result in lipid peroxidation and disruption of Ca(2 +) homeostasis. The depression in Ca(2+) regulatory mechanism by reactive oxygen species ultimately results in intracellular Ca(2 +) overload, CBN and cell death.

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.

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 beta1-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-alpha), 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-alpha, 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.

Morphology, TNFalpha expression and apoptosis in the hearts of patients who died of abdominal compartment syndrome: an immunohistochemical study.

Most of the existing reports on abdominal compartment syndrome (ACS) deal with pathophysiology, focusing on its clinical picture and course, and knowledge of the histopathological features of ACS is very poor. Since the heart, kidneys and lungs are the organs primarily targeted for injury in multi-organ failure, we investigated the expression of TNFalpha and apoptosis in tissue specimens of the heart. This retrospective study was conducted at the Department of Forensic Pathology, University of Foggia. A retrospective review of records extending over a period of 4 years, from 1 April 2001 to 31 March 2005, was carried out and all cases subjected to medico-legal autopsy during this period, whose detailed history and case records were available, were the subjects of our study. Over a 4-year period, on 848 cases subjected to medico-legal autopsy, three cases qualified for inclusion into the study. The immunohistochemical study revealed an intensive positive result for TNFalpha in heart specimens. The TUNEL assay was positive in heart specimens too. The presented study can contribute to elucidate the pathophysiology of fatal ACS and also define efficient markers for future therapeutic approaches suggesting anti-TNFalpha strategies may be useful in the management of ACS.