Evaluation of eye and serum findings in different waters in rabbits by drowning and submersion modeling.

Background/aim: This study investigated serum, vitreous, and anterior chamber fluid electrolyte changes, corneal thickness (CT), corneal volume (CV), anterior chamber volume (ACV), and anterior chamber depth (ACD) as an auxiliary diagnostic method in the identification of drowning in fresh or salt water. Materials and methods: The study used 35 healthy, adult, male, white New Zealand rabbits, seperated into five groups (control, saltwater drowning (SWD), saltwater immersion (SWI), freshwater drowning (FWD), freshwater immersion (FWI)). CT, CV, ACV, and ACD measurements were made with Pentacam topography at 0, 2, and 4 h in all groups. Magnesium (mg), sodium (Na), and chlorine (Cl) were measured in the blood at 0 and 2 h, and in blood, vitreous fluid, and humor aqueous at 4 h. Results: It was determined that CT, CV, ACV, and ACD are not of great value in drowning diagnosis and are affected by the fresh or salt water rather than drowning. Vitreous Na, Cl, and Mg levels are ineffective in determining drowning after one h. Anterior chamber fluid may provide valuable information in the differentiation freshwater - saltwater drownings at the 4th h in corpses retrieved from water. Conclusion: Anterior chamber fluid Na and Cl levels, especially in corpses removed from salt water, can be an easily used test that can help diagnose drowning.

Hemolytic staining of the endocardium of the left heart chambers: a new sign for autopsy diagnosis of freshwater drowning.

Despite the availability of modern imaging and molecular tools, traditional autopsy, and laboratory findings remain the gold standard for the diagnosis of drowning. This article presents two cases of freshwater drowning in which hemolytic staining of the endocardium of the left heart chambers was observed at autopsy. One case was a suicidal drowning of an 84-year-old man, and the other case was an accidental drowning of an 86-year-old woman. In both cases, there was marked hemolytic staining of the endocardium of the left atrium and ventricle. The endocardium of the right heart chambers was clear and transparent in appearance. Hemolytic intimal staining of the aortic root was observed in one case. Gettler's test was positive in both cases. Hemolytic discoloration of the endocardium of the left heart chambers after freshwater drowning is analogous to hemolytic staining of the aortic root. Both staining patterns result from the hypo-osmolar hemolysis that occurs in the left heart chambers and systemic circulation after hypotonic fluid passes across the alveolocapillary membrane. Hemolytic discoloration of the endocardium of the left heart chambers at autopsy may support a diagnosis of freshwater drowning.

A simple DNA coprecipitation method for the detection of diatoms in heart blood.

We developed a method for detecting and enumerating diatoms in the heart blood of drowning victims and evaluate its utility for diagnosing death by drowning. For purification of diatoms from blood, the DNA binding ability of the diatom frustule in the presence of a chaotropic agent was utilized. The procedure is basically the same as the commonly used method for DNA purification from blood using Proteinase K treatment and denaturation by a chaotropic agent. DNA adsorbed to the diatom (DNA/diatom complex) is recovered by ethanol precipitation, and the DNA is subsequently digested using DNase. Purified diatoms could be clearly observed under a microscope. Diatoms spiked in the blood of non-drowned cadavers (n=15) were well recovered, and were detected in heart blood from all drowning victims (n=22). The mean number of diatoms found in 5 ml of blood from drowning victims was 7.8±5.8 (mean±SD), and the number of diatoms detected in the blood of the left ventricle (6.1±5.8) was approximately two times higher than that of the right ventricle (3.0±2.7, p<0.05). These results suggest that this simple and safe method can become an effective tool for diagnosing the cause of death as drowning.

Virtual autopsy using multislice computed tomography in forensic medical diagnosis of drowning.

PURPOSE: This study evaluated the usefulness of multidetector computed tomography (MDCT) in the postmortem diagnosis of death by drowning in fresh water by measuring the difference of blood density within the cardiac chambers. MATERIALS AND METHODS: Twenty-two corpses including six cases of fresh-water drowning (group A) and 16 deaths by other causes (group B), among which were also different forms of mechanical asphyxia other than drowning, underwent MDCT and conventional autopsy. Blood density within the right and left heart chambers, the aorta and the pulmonary trunk was measured and values compared between groups and within each group between heart chambers. RESULTS: Blood density in all cardiac chambers was lower in group A than in group B. The difference was statistically significant within the left atrium and ventricle and was significantly lower in the left than in the right heart chambers in group A only. CONCLUSIONS: MDCT, together with conventional autopsy, may contribute to the diagnosis of drowning, by measuring blood density in the heart chambers.

Postmortem serum tenascin-C (TN-C) concentrations in forensic autopsy cases: A pilot study.

In forensic investigations, autopsy findings offer major clues for the diagnosis of the cause of death. Thus, various clinical biochemical markers are now being tested to complement conventional investigation in the field of forensic medicine. In this study, we focused on tenascin-C (TN-C), a glycoprotein present in the extracellular matrix and expressed in pathological states. We reviewed autopsy cases for a 4-year period (2006-2009) using autopsy records, and analyzed the blood serum concentrations of TN-C and C-reactive protein (CRP) in these cases (N=101). The TN-C levels were relatively higher in the postmortem serum samples than in the samples from healthy individuals, and in cases of head injury, both TN-C and CRP levels were high in the postmortem serum sample. Moreover, high TN-C levels were observed particularly in cases with a long survival period. These findings indicate that postmortem serum TN-C levels may represent a useful tool for identifying the cause of specific fatal traumas.

Postmortem serum levels of amylase and gamma glutamyl transferase (GGT) as markers of systemic tissue damage in forensic autopsy.

Serum amylase (AMY) and gamma glutamyl transferase (GGT) are routine clinical markers for investigating pancreatic and hepatobiliary disorders, respectively, but are also increased in systemic deterioration following critical trauma and diseases. The present study investigated the postmortem levels in bilateral cardiac blood of medicolegal autopsy cases without decomposition (n=163), excluding those with pancreatic or hepatic injury, or preexisting pathologies, as well as prolonged death cases, to evaluate the changes due to systemic deterioration in the death process after fatal insults with special regard to intoxication, hyperthermia (heatstroke) and hypothermia (cold exposure). Serum AMY and GGT levels were virtually independent of postmortem interval. Serum AMY level was mostly higher than the clinical reference range, predominantly including salivary fractions, but was usually below 1000U/L except for fatal intoxication, which showed significant increases of total AMY as well as salivary and pancreatic fractions in bilateral cardiac blood. Serum levels of salivary and pancreatic AMY fractions showed tendencies to be related to pancreatic subcapsular and interstitial bleeding, respectively, which were relatively frequent and evident in mechanical asphyxiation, intoxication and hyperthermia (heatstroke). Serum GGT was often elevated (mostly below 300U/L) in cases other than hypothermia (cold exposure). These findings suggest postmortem serum AMY and GGT as indicators of the severity of systemic organ damage in death processes, especially in intoxication; however, elevated serum AMY and GGT levels over 1000 and 300U/L might indicate significant pancreatic and hepatobiliary pathologies, respectively, except for an elevated serum AMY level in intoxication.

Serum biochemical markers in drowning: diagnostic efficacy of Strontium and other trace elements.

The aim of our study was to analyse several serum biochemical markers in order to evaluate the discriminant capacity of trace elements individually and jointly in drowning (seawater drowning and freshwater drowning) and their correlation with age, sex, postmortem interval, time in water and concentrations of the trace elements in the drowning medium. Sixty-seven cases of drowning (53 seawater drownings (SWD); 14 freshwater drownings (FWD) and 73 control cases (other asphyxias, n=44, and other causes of death, n=29) were selected according to the scene, cause and circumstances of death, together with autopsy findings. Serum strontium (Sr), magnesium (Mg), sodium (Na), chloride (Cl), calcium (Ca), iron (Fe), urea, creatinine (Cr) and cardiac Troponine T (cTn-T) were measured in the left ventricle (Lv), right ventricle (Rv) and peripheral blood. Lv-Rv differences for each marker and Sr, Mg, Na, Cl, Ca and Fe concentrations in the drowning medium were determinated. Mean concentrations of Sr, Cl and Mg in both ventricles and peripheral serum and Lv-Rv differences and Ca Lv and Na Rv were significantly higher in cases of drowning than for other causes of death. In SWD, Sr, Mg, Ca, Na and Cl were significantly higher in Lv than in Rv as a result of aspirating water. In contrast, haemodilution is evident from the significantly higher levels of Fe and urea in Rv than in Lv in cases of SWD, and from the higher Mg and Cr levels in Rv in FWD. In the case of SWD, serum levels of Sr are confirmed as the best parameter for diagnosis, although other trace elements may also be useful, such as the serum concentrations of Mg and Cl. In the case of FWD, the joint determination of Sr and other biochemical markers, especially Fe, may increase correct diagnosis.

[Altered water fractions in the blood and its components after drowning].

Previous studies have shown that signs of drowning in fresh water are of low diagnostic value. Regardless of the mechanism of drowning death, a certain amount of water enters the blood and somehow changes its composition. The objective of this study was to determine water fractions in the blood of drowned subjects. It included 30 cases of drowning deaths. The control group comprised 30 cases of hanging and 7 bodies found in water after the death on land. The water content in the blood was measured in accordance with the universally accepted diagnostic procedure from the difference of freezing temperatures of free and bound water. It was shown that arterial blood and its constituent components as well as whole venous blood and concentrated red cells mass of drowned subjects contained significantly more water than the blood of those who died on land. The rise was in the first place attributable to the selective increase of the free water fraction. Changes of water content in the blood were virtually identical in the drowned subjects who died from asphyxia and from aspiration of water. The data obtained in this study may be used as additional diagnostic signs of drowning death.

von Willebrand factor in cadaveric urine for forensic investigation.

Preliminary experimental study of urinary von Willebrand factor (VWF) concentration was undertaken to evaluate the utility of this parameter in forensic investigations. ELISA was used to measure VWF concentration. Correlations of urinary VWF with cause of death and postmortem interval (PMI) were ascertained. As PMI advanced, urinary VWF increased but plasma VWF did not. Cause of death was not significantly correlated with VWF. This study indicated the possibility that urinalysis would be helpful to estimate PMI.

Influence of cause of death and blood ethanol concentration on the coagulability of cadaveric blood.

It is well-known that cadaveric blood tends to retain their fluidity in acute deaths. However, our experience indicated that blood clots are occasionally seen in such cases, especially with antemortem consumption of ethanol. We reviewed autopsy reports from 2001 to 2006 in our division retrospectively. We made evaluations of blood coagulability based on the descriptions in the autopsy reports, and classified them into three categories; "none", "some amount of clot", and "large amount of clot". Out of 782 cases except for putrefactive or charred bodies, 202 cases showed coagulation of heart bloods. Among them, large amount of clots were observed in 140 cases, in which the cause of death included so-called slow deaths such as hypothermia, infections and in-hospital deaths. Additionally, some reputed acute deaths exhibited large amount of clots; fire death (18 cases), drowning (17 cases), other types of asphyxia (5 cases) and cardiac deaths (9 cases). We adopted 0.1 mg/ml of blood ethanol concentration (BEC) as a cut off value, and a correlation of blood coagulability and BEC was tested with chi-square test. With exclusion of slow deaths, the number of cases with blood clot was significantly higher in cases with positive BEC (180 cases) than those with negative BEC (367 cases) (p=0.000085). However, after raising cut off value of BEC (>1.0 mg/ml), the p value was elevated (p=0.00066). Based on this study, we considered that postmortem blood coagulability was influenced not only by simple pharmacodynamic actions of ethanol but also by other factors including coexisting pathological conditions.

Postmortem serum erythropoietin levels in establishing the cause of death and survival time at medicolegal autopsy.

Circulating erythropoietin (EPO) is mainly produced in the kidneys, depending on blood oxygen level. The present study investigated the postmortem serum EPO levels with regard to the cause of death and survival time. Serial medicolegal autopsy cases of postmortem time within 48 h (n = 536) were examined. Serum EPO levels were within the clinical reference range in most cases. Uremic patients with medical administration of an EPO agent (n = 11) showed a markedly high level (140-4,850 mU/ml; median, 1,798 mU/ml). Otherwise, an elevation in serum EPO level (>30 mU/ml) was mainly seen in protracted deaths due to blunt injury and fire fatality, depending on the survival time (r = 0.69, p < 0.0001, and r = 0.45, p < 0.0001, respectively), and in subacute deaths from gastrointestinal bleeding and infectious diseases. However, mildly to moderately elevated serum EPO levels were sporadically found in acute deaths due to mechanical asphyxiation, fire fatality, and acute ischemic heart disease, and in fatal hypothermia cases, especially for elderly subjects. Protracted deaths due to mechanical asphyxiation and ischemic heart disease did not show any survival time-dependent increase in serum EPO level (p > 0.05). EPO was immunohistochemically detected in the tubular epithelia and interstitial cells, showing no evident difference among the causes of death, independent of survival time or serum level. These findings suggest that serum EPO can be used as a marker for investigating anemia and/or hypoxia as a consequence of fatal insult in subacute or prolonged deaths, or a predisposition to traumatic deaths or fatal heart attacks in acute deaths.

Strontium levels in different causes of death: diagnostic efficacy in drowning.

Trace element determination can be applied in forensic medicine to diagnose the cause of death. Drowning is the second leading cause of death from unintentional injury. Despite the many diagnostic methods used, the post-mortem diagnosis of drowning continues to be one of the most difficult in forensic pathology. Strontium is a highly sensitive marker of water aspiration in a liquid medium rich in this metal. The aims of this study were to confirm the diagnostic value of strontium in cases of drowning compared with other causes of death, to analyse factors that could affect its concentration and to ascertain the sensitivity and specificity of strontium in right and left ventricles and peripheral serum for the post-mortem diagnosis of drowning. We studied 120 cadavers selected from medico-legal autopsies with different causes of death. Strontium (Sr) levels were measured in the serum (left and right ventricles and peripheral vein) of all cadavers and, in the case of drowning, in the water medium itself, by using Zeeman AAS. Our results confirm the usefulness of blood Sr levels for diagnosing seawater and freshwater drowning, although great care should be exercised in the latter case.

Evaluation of freshwater submersion in small animals: 28 cases (1996-2006).

OBJECTIVE: To determine clinical characteristics, treatments, and outcome in dogs and cats evaluated after submersion in freshwater. DESIGN: Retrospective case series. ANIMALS: 25 dogs and 3 cats. PROCEDURES: Medical records were reviewed for signalment; causes, location, and month of submersion; physical examination findings at admission; results of blood gas analysis; treatments administered; duration of hospitalization; and outcome, including evidence of organ failure or compromise. RESULTS: All submersions involved bodies of freshwater. Fourteen animals were submerged in man-made water sources, 13 were submerged in natural water sources, and the body of water was not recorded in 1 case. Twenty (71%) submersions occurred from May through September. Cause was identified in 16 animals and included extraordinary circumstances (n = 6), falling into water (5), breaking through ice (3), and intentional submersion (2). Twelve animals were found submerged in water with unclear surrounding circumstances. Treatment included administration of supplemental oxygen, antimicrobials, furosemide, corticosteroids, and aminophylline and assisted ventilation. Respiratory dysfunction was detected in 21 animals. Neurologic dysfunction was detected in 12 animals, hepatocellular compromise was detected in 6 animals, and cardiovascular dysfunction was detected in 4 animals. Three dogs had hematologic dysfunction, and 2 dogs had acute renal dysfunction. Eighteen (64%) animals survived to hospital discharge, but all of the cats died. In 9 of 10 nonsurvivors, respiratory tract failure was the cause of death or reason for euthanasia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that submersion is an uncommon reason for veterinary evaluation but is associated with a good prognosis in dogs in the absence of respiratory tract failure.

Postmortem serum nitrogen compounds and C-reactive protein levels with special regard to investigation of fatal hyperthermia.

The present study analyzed serum levels of urea nitrogen (UN), creatinine (Cr), and C-reactive protein (CRP), which are very stable during the early postmortem period, for investigation of the cause of death with special regard to hyperthermia (heat stroke) in serial medico-legal autopsy cases (n = 429), excluding fatal injury, intoxication, and fire fatality. In this series, mechanical asphyxiation, drowning, and sudden cardiac death cases (n = 56, n = 43, and n = 212, respectively) usually showed low levels within postmortem reference ranges for these serum markers, although UN and CRP levels were mildly elevated in cases of sudden cardiac death and cerebrovascular stroke. There were concomitant significant elevations in serum levels of UN (>50 mg/dL), Cr (>2 mg/dL), and CRP (>2 mg/dL) for chronic renal failure, gastrointestinal bleeding, pneumonia, and hypothermia (cold exposure). UN and CRP were especially high for chronic renal failure and pneumonia, respectively. However, hyperthermia cases showed an isolated elevation in the serum Cr level, suggesting an influence of systemic skeletal muscle damage. These serum markers may be practically useful for postmortem investigation of death due to hyperthermia (heat stroke), for which specific pathological and toxicological evidence may not be available.

Factors affecting strontium absorption in drownings.

This study examines the effects of age, gender, a cold water medium versus warm water medium, and salinity on strontium levels determined in left ventricular blood in drownings. Significant differences in the amount of strontium absorbed into the bloodstream (p<0.001) were detected between individuals who drowned in fresh water versus those drowning in seawater, and between those drowning in cold water versus warm water (p=0.030). However, no significant differences were noted in the strontium concentrations of left ventricular blood according to gender or age.

Comparison between lung weight and blood strontium in bodies found in seawater.

This paper examines the use of lung weight increase as an indicator of seawater drowning compared to the amount of Sr absorbed by the blood. The study population was limited to male victims older than 20 years. Significant differences between cases of drowning and "non-drowning" were detected in terms of the lung-heart weight ratio (L/H) (p<0.001) or lung-body weight ratio (L/B) (p=0.005). However, using lung weight (L), L/H or L/B to distinguish between seawater drownings and saltwater non-drownings some overlap was produced. The factor rendering least overlap was L/B, which also appeared to be non-dependent on the victim's age. Our findings suggest that a value of L/B higher than 19.5 g/kg could be a useful indicator of death by drowning, but that when a lower value is found, additional drowning diagnoses would be needed to establish the manner of death.

Drowning, haemodilution, haemolysis and staining of the intima of the aortic root-- preliminary observations.

In order to demonstrate that hyponatraemia due to haemodilution occurs within the left ventricle following freshwater drowning, and to determine whether lysed blood resulting from left ventricular haemodilution may cause staining of the aortic intima, the following studies were undertaken. Measurements of left ventricular sodium levels were performed in 74 consecutive coronial cases where death was attributed to drowning, consisting of 44 and 30 deceased who were believed to be victims of freshwater and saltwater drowning, respectively. Left ventricular sodium levels differed significantly between the two groups (p<0.001), with a range of 93-147 mmol/L in freshwater drowning (mean=117+/-14.2 mmol/L) and 123-183 in saltwater drowning (mean=153+/-14.4 mmol/L). In addition, the mean sodium level of 117 mmol/L in freshwater drowning was significantly lower than the standard range of 137-145 mmol/L. In a second study, portions of aorta and pulmonary trunk from a euthanised pig were soaked in lysed blood resulting in marked haemolytic staining of the intima of both vessels after 20 min. Water and a mixture of blood and water were then injected into the left ventricles in two further pig carcasses, respectively, resulting in haemolytic staining of the intima of the aortic roots, with no staining of the pulmonary trunks. These studies have confirmed that significant hyponatraemia secondary to haemodilution may occur within the left ventricle in freshwater drowning, and that haemolysed blood is capable of causing staining of the aortic root in an animal model. These results provide further data to support haemolytic staining of the aortic root intima as a possible manifestation of freshwater drowning.

Postmortem serum protein S100B levels with regard to the cause of death involving brain damage in medicolegal autopsy cases.

The protein S100 is an acidic calcium-binding protein, and the subunit S100B is the most abundantly found in the brain. The aim of the present study was a comprehensive analysis of serum S100B levels in medicolegal autopsy cases (within 48 h postmortem, n = 283) including victims with head and non-head injuries and also non-injury fatalities with regard to the cause of death involving brain damage. The serum level was usually higher in the subclavian vein than in the right heart and external iliac vein, and the lowest in the left heart blood, showing no significant postmortem influence. The serum level in the right heart and subclavian vein was markedly higher for acute deaths from head injury and asphyxiation due to neck compression (strangulation and hanging), and moderately and mildly elevated for other blunt and sharp instrument injury cases, respectively. For head injury, the serum levels were lower for subacute deaths than for acute deaths. These observations suggest that the elevation of serum S100B may mainly be caused by leakage following massive brain damage due to injury and cerebral hypoxia/ischemia, and in part by systemic hypoxic/traumatic tissue damage, depending on the survival time.

Study of the diagnostic value of iron in fresh water drowning.

The aim of our study was to test the diagnostic value of iron (Fe) in fresh water drowning by investigating the postmortem levels of hemodilution in drowning cases compared to control cases. Twenty-six typical fresh water drowning cases were selected from 128 immersion cases autopsied in our Department of Forensic Pathology between 1998 and 2004. The exclusion criteria were a long postmortem interval and other causes of death than drowning. For all selected cases, the diagnosis of drowning was based on the presence of autopsy findings and positive diatom test. A control population of 12 cases was also selected. For each drowning and control case, iron blood levels were measured in the left ventricle (LV) and right ventricle (RV) of the heart. The mean difference of iron concentration (RVFe-LVFe) between the drowning group and the control group was statistically compared. Furthermore, iron measurements were performed in 19 drowning cases showing advanced putrefaction. The mean difference of iron concentration was significantly higher in the drowning cases compared with controls (P<0.001). All drowning cases showed hemodilution. No overlap was found in the RVFe-LVFe levels between the two groups. Resuscitation attempts seemed to have no effect on the results. In cases of drowning showing advanced putrefaction, the iron test was not reliable because biochemical iron measurement was often prevented by no sufficient blood in the heart or postmortem clots. In conclusion, according to our results, iron seems to be a good biochemical marker in fresh water drowning with a short postmortem interval.