Lethal hemopericardium caused by infection of mitral annular calcification: An autopsy case report.

Mitral annular calcification (MAC) is a chronic and degenerative condition involving calcification of the mitral annulus. MAC is a risk factor for coronary artery disease, cardiovascular events, stroke, and cardiovascular death. However, patients with MAC are often asymptomatic. Herein, we present the first case of cardiac tamponade due to infection of MAC in forensic pathology. An 80-year-old woman was found in cardiopulmonary arrest shortly after experiencing fatigue. She was transferred to a hospital, and despite chest compression and ventilation, she was pronounced dead due to no response. Postmortem computed tomography, autopsy, and histological examination showed MAC, abscess formation involving Gram-positive cocci on the MAC, and fistulation of the abscess into the intracardial pericardial cavities, resulting in a massive lethal hemopericardium.

Quantitative estimation of TNF-α and IL-3 by using ELISA from human lung tissue in fatal asphyxial deaths.

Asphyxia-related deaths have always been a challenging task in the specialty of forensic pathology. Apart from helpful macroscopical signs (e.g., strangulation marks, cyanosis, petechial haemorrhage, and lung edema), recent literature indicates that prolonged asphyxia is sufficient to induce an increase in mast cells (MC). Inflammatory cells migrate from the bone marrow to the lungs, aiding in the diagnosis of fatal asphyxial death. The present study analyzed human lung tissue samples from 90 medico-legal autopsy cases, including 45 asphyxial deaths and 45 controls (non-asphyxial deaths). The cases ranged from 2 to 68 years, with a mean age of 33.23 years. In 90 cases, 74 cases were of males, and 16 were of females. Human lung tissue samples were analyzed by using the sandwich ELISA method. The results indicated a statistically significant increase in TNF-α and IL-3 concentration in fatal asphyxial deaths, including those caused by hanging, drowning, and smothering. Mean ± SD in asphyxial and non-asphyxial cases for the TNF-α and IL-3 concentration statistically analysed. In asphyxial cases, the average IL-3 concentration (Conc.) was 1558.50 ± 350.53 pg/ml, and the average TNF-α concentration (Conc.) was 499.75 ± 479.41 pg/ml. In contrast, in non-asphyxial cases, the average IL-3 concentration (Conc.) was found to be 849.73 ± 484.99 pg/ml, and the average TNF-α concentration (Conc.) was 208.08 ± 81.23 pg/ml. The mean change in IL-3 and TNF-α (Conc.) values are found to significant (<0.01) in asphyxial cases as compared to non-asphyxial cases. The ROC (Receiver operating characteristic curve) analysis revealed that TNF-α (AUC = 0.89) and IL-3 (AUC = 0.87) concentration (conc.) were stronger predictors of asphyxial deaths with an optimal cut-off value of 455.20 pg/ml for TNF-alpha and 1700.62 pg/ml for IL-3 respectively. Our findings imply that mast cells (MC) are critical in fatal hypoxia-related mortality and that TNF-α and IL-3 can be reliable markers for detecting mast cells in asphyxial deaths. It could be very beneficial to forensic pathologists tasked with differentiating fatal asphyxial fatalities from other causes of death.

A method for bone marrow extraction of diatoms for forensic science applications.

Diatoms show potential as trace evidence indicators, particularly as evidence of drowning. Often, the diatom test to diagnose drowning is done on soft tissue or bone marrow from a recently deceased individual. This method presented here combines elements from previous forensic literature and methods of diatom isolation in phycology to extract diatoms from bone marrow of skeletal remains for forensic use. This diatom extraction method is time-efficient, minimizes contamination risk, and produces samples of intact diatoms. This method is designed to complete sample preparation within 24 h, sampling the bone for diatoms internally and externally. This method was developed using porcine long bones submerged in water with live diatoms for up to 3 months. Three marrow samples were extracted from each bone so the method was developed using 102 marrow samples. Additionally, 132 surficial bone and environmental samples were collected and prepared during method development. To briefly summarize the method, the bone joints were cut off with an angle grinder in a biosafety hood to expose the marrow, which was removed from the hip, knee, and shaft as separate samples. The marrow was digested with nitric acid at 400°C in glass beakers before being centrifuged with DI water, plated onto microscope slides, and observed with a compound microscope. Observation found good preservation of unbroken diatom cell walls throughout the process. This method can be used to prepare diatoms as forensic trace evidence.

Potential of photoautotrophic microbial organisms in deciphering forensic issues.

In forensic pathology, solving the crime mystery of death due to drowning still remains a challenging issue. The amalgamation of autopsy findings and comparative study of diatoms recovered from the victim's body organs and suspected drowning site help to decipher the cause of death due to drowning or post-mortem immersion. Since the correct interpretation of the cause of death is an important criterion to provide justice to the victim, therefore, the main objective of our study is to throw light on the application of photoautotrophic micro-algal organisms, known as Diatoms, in solving seven cases of victims whose bodies were recovered from various water bodies of Himachal Pradesh, India. The diatom test was conducted by using reverse aqua regia solution (15 ml HNO3: 5 ml HCl) on the bone marrow extracted from the organs and water samples respectively. The informative outcomes of the experimental analysis demonstrated that the diatom test acts as a beneficial adjunct to solve drowning-related crimes where the exact cause of death remains hidden even after performing an autopsy of the victims. The protocol followed by the authors can be used conveniently to recover diatoms from bone marrow as well as from water samples. Our results showed that the maximum cases were of death due to accidental drowning but for one case of suicidal drowning in extremely cold water.

Postmortem computed tomography differentiation between intraperitoneal decomposition gas and pneumoperitoneum.

PURPOSE: The aim of this study is to investigate the ability of postmortem computed tomography (PMCT) to distinguish intraperitoneal decomposition gas from pneumoperitoneum due to intestinal perforation. METHODS: This retrospective study investigated the factors affecting intraperitoneal gas in two groups of 14 decedents as detected by postmortem CT performed in the Department of Legal Medicine of Hamburg University. The first group died with a cause of death associated with intestinal perforation, and the second group with other different natural causes of death. These factors include postmortem interval, gas volume, gas distribution, radiology alteration index (RAI), and pneumoperitoneum-associated pathology. RESULTS: The findings of this study showed the appearance of specific gas distribution patterns and a significant increase in gas volumes in the cases of intestinal perforation. Moreover, postmortem interval and the pneumoperitoneum-associated pathology could help distinguish postmortem-generated gas from pneumoperitoneum. CONCLUSION: Based on the findings of this study, we propose that these findings can improve the proper detection of intestinal perforation cases in the future.

Evaluation of gingival tissue samples for predicting the time of death using histological and biochemical tests.

Thanatochemistry also known as chemistry of death and is used to determine post mortem interval (PMI). It is arguably one of the critical steps in forensic investigation. Recent addition of analyzing biochemical changes along with the traditional methods have gained importance, as they help us to record very early changes in the tissue specimens. In this view, our study aimed to correlate both histological changes and enzymatic changes in gingival tissue samples at intervals of immediate, 1 h, 5 h, 24 h and 48 h after death. Histologic changes noted were loss of epithelial architecture, chromatin clumping, nuclear vacuolation, karryopyknosis, eosinophilia and wide intercellular junctions. Two enzymes which differentiate between the autolytic phase (acid phosphatase) and putrefactive phase (ammonia) of decomposition were evaluated using UV spectrometer. Results in our study demonstrated there were variations as in gradual increase in ammonia levels (1.13±0.24-26.6±2.09) and gradual decrease in acid phosphatase levels (5.61±0.67-1.25±0.53) at different time intervals till 48 h. The cellular changes in gingival tissue could also be related to time. The result of our study helps us to identify potential of enzymatic changes which when correlated with histological reports helps us to predict the time of death accurately. Replicating this experiment in various known taphonomic conditions and other enzymes could highlight the usefulness of gingival tissue samples in determining time of death.

An algorithm for automatically generating gas, bone and foreign body visualizations from postmortem computed tomography data.

Post mortem computed tomography (PMCT) can aid in localizing foreign bodies, bone fractures, and gas accumulations. The visualization of these findings play an important role in the communication of radiological findings. In this article, we present an algorithm for automated visualization of gas distributions on PMCT image data of the thorax and abdomen. The algorithm uses a combination of region growing segmentation and layering of different visualization methods to automatically generate overview images that depict radiopaque foreign bodies, bones and gas distributions in one image. The presented method was tested on 955 PMCT scans of the thorax and abdomen. The algorithm managed to generate useful images for all cases, visualizing foreign bodies as well as gas distribution. The most interesting cases are presented in this article. While this type of visualization cannot replace a real radiological analysis of the image data, it can provide a quick overview for briefings and image reports.

Analysis of non-derivatized 2-(4-R-2,5-dimethoxyphenyl)-N-[(2-hydroxyphenyl)methyl]ethanamine using short column gas chromatography - mass spectrometry.

The 25R-NBOH family is a group of thermally labile compounds that are relevant for forensic sciences and traditionally analyzed by GC-MS after derivatization - a step that is time consuming in a routine work. In this paper, the use of short analytical columns (4 and 10 m) showed to decrease compound degradation in the GC oven during chromatographic separation and to allow the analysis of non-derivatized 25R-NBOH compounds by GC-MS. A shorter column demanded a higher gas flow rate, and both factors decreased residence time of the analytes in the column and their degradation. The inlet temperature (250° C or 280°C) did not impact the response of 25R-NBOH. A 25R-NBOH fragmentation pathway by electron ionization was also presented for the first time. The GC-MS method with a 4 m column was successfully applied to other compounds of forensic interest, and it can be tested in the analysis of biological samples in toxicological investigations.

Open source software tool for the automated detection and characterization of epithelial cells from trace biological samples.

This paper presents a strategy for an unsupervised workflow for identifying epithelial cells in microscopic images and characterizing their morphological and/or optical properties. The proposed method can be used on cells that have been stained with fluorescent dyes and imaged using conventional optical microscopes. The workflow was tested on cell populations that were imaged directly on touch/contact surfaces and stained with nucleic acid dyes to visualize genetic content. Our results show that this approach could be a useful strategy for characterizing differences in staining efficiency and/or morphological properties of individual cells or aggregate populations within a biological sample. Further, they can potentially reduce the laborious nature of microscopic analysis and increase throughput and reproducibility of similar studies.

The use of torus palatinus in the identification of unknown skeletal remains.

Torus palatinus (TP) is a benign, bony outgrowth located on the hard palate. Variation in this trait occurs, ranging from absent to a protuberance several millimeters in length. If a TP becomes too large, it can interfere with daily activities and thus warrant medical attention. This case utilised the unique morphology of a TP after surgical invention to identify the unknown skeletal remains of a woman missing for six years. Even though there is still much unknown regarding the aetiology, occurrence and prevalence of TP, this study demonstrates the usefulness of anatomical variants in the identification of unknown skeletal remains.

Post-mortem cerebrospinal fluid diagnostics: cytology and immunocytochemistry method suitable for routine use to interpret pathological processes in the central nervous system.

Due to its protected anatomical location, cerebrospinal fluid (CSF) is a very stable fluid which undergoes comparatively little change in the early post-mortem phase. While many immunohistochemical markers already established for clinical diagnostic issues in tissue samples obtained by biopsy could meanwhile be translated also to post-mortem tissue, no systematic immunocytochemical investigations have generally been conducted on post-mortem body fluids and for CSF specifically, have not been established at all. CSF as the fluid directly surrounding the brain should also be examined to allow a more detailed characterization of processes in the central nervous system. Comparing traumatized tissue and CSF can complete forensic assessment and complement neuropathological evaluation.

Discordance of Gross and Histologic Findings in Estimating the Range of Fire of Gunshot Wounds.

Forensic pathologist use soot and/or stippling surrounding entrance gunshot wounds in categorizing range of fire. If absent, some pathologists suggest utilizing histology when the range is diagnostically critical. This study investigates the concordance of macroscopic and microscopic findings in estimating range of fire by evaluating gunshot entrance and exit wounds made through human tissue analogs at defined distances using two handgun calibers. Examination of over 150 entrance wounds verified the ease of visually detecting soot from a muzzle distance of contact to 1 foot (30.5 cm), and its absence at 9 feet (274.3 cm). Distinctly apparent was bullet wipe surrounding the entrance wounds regardless of muzzle distance. Although variations existed, dark material was histologically identified in many skin, soft tissue, and bone sections at all ranges with both calibers. These nonparallel results decrease the dependability of histology for range of fire estimation and reinforce using gross observation.

Histological dating of subdural hematoma in infants.

BACKGROUND: After infant deaths due to non-accidental head injury (NAHI) with subdural hematoma (SDH), the magistrates ask experts to date the traumatic event. To do so, the expert only has tools based on adult series of NAHI. We aimed to develop an SDH dating system applicable to infants aged under 3 years. METHODS AND RESULTS: We studied a retrospective multicenter collection of 235 infants who died between the ages of 0 and 36 months, diagnosed with SDH by forensic pathological examination and with known posttraumatic interval (PTI). Two pathologists assessed blindly and independently 12 histomorphological criteria relating to the clot and 14 relating to the dura mater in 73 victims (31 girls, 42 boys) whose median age was 3.8 months. Histopathological changes were significantly correlated with PTI for the appearance of red blood cells (RBCs) and the presence or absence of siderophages, and regarding the dura mater, the quantity of lymphocytes, macrophages, and siderophages; presence or absence of hematoidin deposits; collagen and fibroblast formation; neomembrane thickness; and presence or absence of neovascularization. Dating systems for SDH in adults are not applicable to infants. Notably, neomembrane of organized connective tissue is formed earlier in infants than in adults. CONCLUSION: Our dating system improves the precision and reliability of forensic pathological expert examination of NAHI, particularly for age estimation of SDH in infants. However, the expert can only define a time interval. Histopathology is indispensable to detect repetitive trauma.

[The forensic medical estimation of the biological age of the corpse based on the morphological changes in the skin]. / Sudebno-meditsinskaia otsenka biologicheskogo vozrasta trupa po morfologicheskim izmeneniiam kozhi.

The objective of the present work was the immunohistochemical evaluation of the age-related dynamics of apoptosis-associated proteins contained in the human skin epidermis. We studied the samples of skin tissues obtained during the forensic medical expertises of 100 corpses of the subjects who died in the consequence of acute blood loss in the absence of injuries or pathological changes in the skin (56 men and 44 women at the age varying from 5 to 85 years). The results of this study give evidence that the histoimmunological investigations of the skin tissues can be carried out with the use of the mmunohistochemical Ki67, bcl-2, and p53 markers for the more accurate determination of the biological age of the deceased subjects. The quantitative characteristics thus obtained serve as reliable indicators of the chronic changes in the skin and can be used as the components of the combined diagnostic procedures.

Lethal cardiac amyloidosis: Modification of the Congo Red technique on a forensic case.

Congo Red staining is usually used in diagnosing amyloidosis, a pathology characterized by the storage of abnormal proteins in several human organs. When assessed on samples fixated in formalin and embended in paraffin, this staining can undergo several artefacts, causing diagnostic and interpretative difficulties due to its weak stainability and a consequent reduced visibility of the amyloid. These complications, in time, requested several variations of this staining technique, especially in clinical practice, while in the forensic field no protocols has ever been adapted to cadaveric samples, a material that is already characteristically burdened by a peculiar stainability. In our work, studying a sudden death caused by cardiac amyloidosis and diagnosed only with post-mortem exams, we present a modified Congo Red staining used with the purpose to demonstrate amyloid in cadaveric material after the unsuccessfully use of all standard protocols.

[The principal and auxiliary immunohistochemical markers of intravital mechanical strangulation asphyxia]. / Osnovnye i dopolnitel'nye immunogistokhimicheskie markery prizhiznennosti stranguliatsionnoi mekhanicheskoi asfiksii.

The objective of the present study was the evaluation of the auxiliary methods for the diagnostics of the intravital formation of the constriction marks; the secondary objective was to determine the pace at which the death and asphyxia occur. MATERIAL AND METHODS: The materials on which the study was based included 17 cases of mechanical strangulation asphyxia involving 13 men and 4 women at the age from 8 to 28 years. All cases of hanging were associated with different blood alcohol levels. Their characteristic feature was the formation of the obliquely ascending constriction marks. The group of comparison was comprised of three cases of death by drowning and one case of manual strangulation. The control group consisted of 10 patients who died from the acute form of coronary heart disease and 5 cases of death from traumatic shock. All the corpses were examined with the use of the traditional methods within the first 24 hours after death. The special laboratory studies were performed by means of the standard histological and immunohistochemical methods with the use of the polyclonal antibodies against total cytokeratin, fibrinogen, immunoglobulin-lambda, fibronectin, and CD-117 antigen. The histological preparations were stained by the method of Spielmeyer and with toluidine blue. RESULTS: The results of the study give evidence of the possibility of diagnostics of mechanical strangulation asphyxia making use of the reaction with anti-fibrinogen antibodies in the stromal tissue of the constriction mark even in the absence of other intravital signs of death. Such diagnostics is also possible with the use of the CD-117 antigen in the pulmonary tissue. The expression of this antigen is characteristic of the cases of alveolar hypoxia. The possibility of application of other markers for the differential diagnostics of mechanical strangulation asphyxia from other causes of death is discussed.

[The forensic medical characteristic of sudden death associated with metabolic syndrome]. / Sudebno-meditsinskaia diagnostika vnezapnoi smerti pri metabolicheskom sindrome.

The detection of grade II-III central obesity on a corpse in conjunction with the identification of two additional criteria (such as arterial hypertension and impaired glucose tolerance) provides, if combined with the autopsy data (including the visual reduction of muscular mass, the increased total amount of the adipose tissue, gynecomastia in men together with hypertrophied abdominal adipose tissue accumulation, decreased face and body pilosis), a basis for diagnostics of metabolic syndrome (MS). The objective parameters for this purpose are waist circumference measurements, corpse weight and height, the degree of visceral obesity, narrowing of the renal arteries as a result of their compression by the surrounding adipose tissue, and accumulation of epicardial fat confirmed by the results of the biochemical analysis. The signs of plasmorrhagia combined with fibrinoid degeneration of the vascular walls in the microcirculatory bed make it possible to suspect, with a high degree of probability, the development of hypertensive crisis that may result in a sudden death of the patients presenting with metabolic syndrome.

A valid method to determine the site of drowning.

The diatom test is considered a useful aid in determining the site of death from drowning. Nevertheless, there is disagreement within the scientific community concerning its reliability, and its findings have been challenged and sometimes overturned in courts of law. Using a model based on animal experimentation, we have developed a diatom test to discriminate between the locations of drowning sites from different aquatic systems. We carried out a complementary combination of quantitative and qualitative analyses together with a statistical analysis based on the Kullback-Leibler distance of the samples. A restrictive selection of exclusive diatom species from each reservoir was also made. This approach allowed us to validate the usefulness of the diatom test in determining the location of the site of drowning.

Histological criteria for age determination of fatal venous thromboembolism.

In clinical and forensic practice, fatal thromboembolism is a major problem, particularly in a patient with no pre-existing risk factors. Out of a recent case, we discuss here the criteria for age determination of venous thrombi in a 46-year-old female with concomitant deep vein thrombosis in the common femoral left vein and in the right heart and fatal pulmonary embolism. At autopsy, histopathology and immunohistochemistry evidenced the different composition of thrombi in different sites and permitted to define the different timing. We discuss and review the histopathological criteria for age estimation of venous thrombi starting from the case and in relation to the acquired and inherited thrombophilic risk factors. The appropriateness of clinical management is also discussed.

[Alcohol-associated mortality in Russia (based on the materials for the period from 2011 till 2016)]. / Alkogol'-assotsiirovannaia smertnost' v Rossii (po materialam 2011-2016 gg.).

The objective of the present study was to evaluate the frequency of alcoholic drunkenness documented during forensic medical expertises (investigations) of the corpses carried out in this country throughout the period from 2011 till 2016. The investigations were conducted with the use of medical statistics methods by calculating the fractional difference, dynamics, and rates of detection of the cases of alcoholic intoxication depending on the cause of death. The study has demonstrated the high frequency of the cases of alcoholic drunkenness revealed during forensic medical expertises (investigations) of the corpses that amounted to 30.5% [15, 16]. The total number of the corpses examined in 2016 was 8.6% higher than in 2011. The frequency of the documented cases of alcoholic drunkenness during the same period decreased by 19.7%. The frequency of the documented cases of alcoholic drunkenness in the cases of violent death was 2.8 times that in the cases of death from various diseases (52.8 and 19.0% respectively). The enhanced frequency of alcoholic drunkenness in relation to the number of the conducted forensic medical expertises was documented in the cases of death by drowning and from hypothermia whereas the lowest frequency of alcoholic intoxication was recorded for the corpses of the people who had died from malignant tumours and diseases of the nervous system. Various regions of Russia differed in terms of the frequency of alcoholic drunkenness recorded among the recently deceased people.