Combining Oxidative Stress Markers and Expression of Surfactant Protein A in Lungs in the Diagnosis of Seawater Drowning.

BACKGROUND AND OBJECTIVES: The diagnosis of seawater drowning (SWD) remains one of the most complex and contentious. It is one of the leading causes of unintentional death around the world. In most cases, the forensic pathologist must reach an accurate diagnosis from the autopsy findings and a series of complementary tests such as histopathological, biological, and chemical studies. Despite the lung being the most affected organ in death by submersion, there are few studies on this type of death's impact on this organ. The aim was to investigate human lung cadavers of forensic cases due to different causes of death, the concentration of the oxidative stress markers malondialdehyde (MDA) and γ-glutamyl-l-cysteinyl glycine (GSH), and the relationship with the expression of surfactant protein A (SP-A) to try to discriminate SWD from other types of causes of death. MATERIALS AND METHODS: A total of 93 forensic autopsy cases were analyzed. Deaths were classified into three major groups based on the scene, cause of death, and autopsy findings (external foam, frothy fluid in airways, overlapping medial edges of the lungs): (a) drowning in seawater (n = 35), (b) other asphyxia (n = 33), such as hangings (n = 23), suffocations (n = 6), and strangulation (n = 4), and (c) other causes (n = 25), such as multiple suffocations. Oxidative stress markers (MDA and GSH) and the immunohistochemical expression of SP-A were determined in both lungs. RESULTS: MDA levels were statistically higher in both lungs in cases of SWD than in other causes of death (p = 0.023). Similarly, significantly higher levels of GSH were observed in SWD compared to the rest of the deaths (p = 0.002), which was more significant in the right lung. Higher immunohistochemical expression of SP-A was obtained in the cases of SWD than in the other causes of death, with higher levels in both lungs. The correlation analysis between the levels of oxidative stress (MDA and GSH) in the lung tissue and the expression level of SP-A showed positive and significant results in SWD, both in the alveolar membrane and the alveolar space. CONCLUSIONS: Determining the levels of MDA and GSH in lung tissue and the expression level of SP-A can be of great importance in diagnosing SWD and the circumstances of death. A better understanding of the physiology of submersion is essential for its possible repercussions in adopting measures in the approach to patients who have survived a submersion process. It is also necessary for forensic pathology to correctly interpret the events that lead to submersion.

Trace elements in forensic human lung: A new approach to the diagnosis of seawater drowning. A preliminary study.

The diagnosing of drowning remains one of the most challenging activities for the forensic pathologist. There is little information on the impact on the lung as a target organ in death by drowning. We aimed to investigate the concentration of trace elements in the lungs of people who had suffered different types of death to evaluate the discriminating ability of trace elements to identify seawater drowning (SWD). A total of 11 trace elements were analyzed by Inductively Coupled Plasma-Mass Spectrometry in 74 forensic cases. Sampler scanning electron microscopy and Energy Dispersive X-ray spectroscopy (EDX) were used to identify ultrastructural lung alterations. A Principal Component Analysis (PCA) of trace elements was carried out. The trace elements in SWD lungs were detected in the following order of concentration: Br˃Zn˃Sr˃Cr˃Cu˃As˃Pb˃Se˃Mn˃Ni˃Cd. Our results showed significantly higher concentrations of Br and Sr (P = 0.010 and P = 0.000) and significantly lower concentrations of Zn, Pb, Cu, Cd, and Se in SWD compared with other causes of death. After adjusting by confounder factors, Sr and Br remained as predictive independent factors for diagnosis of drowning (p = 0.042, in both cases). These results were confirmed by PCA, which revealed a wide separation between SWD and the rest of the causes of death. Our SWD cohort was characterized by high concentrations of the trace elements Br and Sr and low concentrations of Zn, Pb, Cu, Cd, and Se in lung tissue, while PCA showed its discriminatory capacity to identify death by seawater drowning. These findings, together with those obtained using other techniques, can be of great importance in the diagnosis of SWD.

Proteomics in Deaths by Drowning: Diagnostic Efficacy of Apolipoprotein A1 and α-1Antitrypsin, Pilot Study.

Drowning is one of the leading causes of death worldwide. The pathophysiology of drowning is complex and, sometimes, interpretation of the circumstances of death in the autopsy becomes the main source of information in its diagnosis. New advances in medical research, such as proteomics, especially in forensic pathology, are still in the development. We proposed to investigate the application of Mass Spectrometry-based technologies, to identify differentially expressed proteins that may act as potential biomarkers in the postmortem diagnosis of drowning. We performed a pilot proteomic experiment with the inclusion of two drowned and two control forensic cases. After applying restrictive parameters, we identified apolipoprotein A1 (ApoA1) and α-1 antitrypsin as differentially expressed between the two diagnostic groups. A validation experiment, with the determination of both proteins in 25 forensic cases (16 drowned and 9 controls) was performed, and we corroborated ApoA1 higher values in the drowning group, whereas α-1 antitrypsin showed lower levels. After adjusting by confounder factors, both remained as predictive independent factors for diagnosis of drowning (p = 0.010 and p = 0.022, respectively). We constructed ROC curves for biomarkers' levels attending at the origin of death and established an ApoA1 cut-off point of 100 mg/dL. Correct classification based on the diagnosis criteria was reached for 73.9% of the cases in a discriminant analysis. We propose apolipoprotein A1 (with our cutoff value for correct classification) and α-1 antitrypsin as valuable biomarkers of drowning. Our study, based on forensic cases, reveals our proteomic approach as a new complementary tool in the forensic diagnosis of drowning and, perhaps, in clinical future implications in drowned patients. However, this is a pilot approach, and future studies are necessary to consolidate our promising preliminary data.

Spatial distribution analysis of strontium in human teeth by laser-induced breakdown spectroscopy: application to diagnosis of seawater drowning.

The diagnosis of drowning can be extremely difficult, especially when the typical morphological signs of drowning are not present, or when the body is in an advanced stage of putrefaction. The main aim of this work is to demonstrate the applicability of laser-induced breakdown spectroscopy (LIBS) to the diagnosis of seawater drowning. Ten teeth samples were selected from eight medico-legal autopsies. A Nd:YAG laser operating at its fundamental wavelength (1,064 nm) was used to generate microplasmas at the sample surface. Strontium (Sr) concentration in tooth samples has been found to be a key factor for the diagnosis of seawater drowning. Spectral differences between the dentin and the enamel were observed. Greater Sr abundance was located in the dentin, with relative standard deviations in the range of 30 to 35%. In addition, chemical images were generated to study the spatial distribution of Sr along the piece. In all cases, Sr content was higher when the cause of the individual death was drowning. A blind experiment was performed to exclude the possibility that the increase of Sr is due to passive diffusion in the blood. The detection of Sr as well as the determination of its distribution by LIBS in dentin seems to be a promising complementary tool for the diagnosis of death by seawater drowning.