Do Circulating Redox Biomarkers Have Diagnostic Significance in Alcohol-Intoxicated People?

The toxic properties of ethanol are inextricably linked to oxidative stress. Despite many reports on the effects of alcohol dependence on blood redox homeostasis, there are no data on the oxidative stress profile in alcohol-poisoned cases. There are also no data on the diagnostic usefulness of redox biomarkers determined post-mortem in various biological fluids. This work investigates the utility of enzymatic and non-enzymatic antioxidant barrier, redox status, and oxidative/nitrosative stress biomarkers in different biological fluids (such as blood, urine, vitreous humor, and cerebrospinal fluid) in the post-mortem study of patients with acute alcohol intoxication. The study group included those who died due to acute ethanol intoxication (n = 22). The research showed a significant increase in glutathione peroxidase activity, total antioxidant status, ferric reducing antioxidant power, and tryptophan concentration only in the study group's urine compared to the control. In other circulating fluids, both antioxidant enzyme activities and glycoxidation product concentrations were not significantly different in individuals who died of alcohol overdose compared with those who died suddenly. We also did not observe a connection between oxidation-reduction balance and the amount of alcohol consumed before death. These unexpected observations may be caused by irreversible post-mortem changes occurring at the cellular level due to autolysis and putrefaction. In summary, the use of circulating body fluids to assess redox homeostasis is limited in the post-mortem analysis. Our results indicate the increased stability of urine collected post mortem compared to other circulating bioliquids. Further studies are needed to assess the intensity of oxidative and carbonyl stress in ethanol-damaged organs and the effects of post-mortem processes on cellular redox balance.

In Vitro Animal Model for Estimating the Time since Death with Attention to Early Postmortem Stage.

Estimating the postmortem interval (PMI) has remained the subject of investigations in forensic medicine for many years. Every kind of death results in changes in metabolites in body tissues and fluids due to lack of oxygen, altered circulation, enzymatic reactions, cellular degradation, and cessation of anabolic production of metabolites. Metabolic changes may provide markers determining the time since death, which is challenging in current analytical and observation-based methods. The study includes metabolomics analysis of blood with the use of an animal model to determine the biochemical changes following death. LC-MS is used to fingerprint postmortem porcine blood. Metabolites, significantly changing in blood after death, are selected and identified using univariate statistics. Fifty-one significant metabolites are found to help estimate the time since death in the early postmortem stage. Hypoxanthine, lactic acid, histidine, and lysophosphatidic acids are found as the most promising markers in estimating an early postmortem stage. Selected lysophosphatidylcholines are also found as significantly increased in blood with postmortal time, but their practical utility as PMI indicators can be limited due to a relatively low increasing rate. The findings demonstrate the great potential of LC-MS-based metabolomics in determining the PMI due to sudden death and provide an experimental basis for applying this attitude in investigating various mechanisms of death. As we assume, our study is also one of the first in which the porcine animal model is used to establish PMI metabolomics biomarkers.

Experimental and Theoretical Approaches to Describing Interactions in Natural Cell Membranes Occurring as a Result of Fatal Alcohol Poisoning.

We propose herein a theoretical model describing the effect of fatal ethanol poisoning on the equilibria between cell membranes and the surrounding ions. Using this model, we determined the parameters characterizing the interaction between the electrolyte solution's ions and the functional groups on the blood cells' surface. Via the application of mathematical equations, we calculated the total surface concentrations of the acidic and basic groups, cA and cB, and their association constants with solution ions, KAH and KBOH. Using the determined parameters and mathematical equations' values, we calculated the theoretical surface charge density values. We verified the proposed model by comparing these values with experimental data, which were selected based on measurements of the electrophoretic mobility of erythrocyte and thrombocyte membranes. Compatibility of the experimental and theoretical surface charge density values was observed in the range of pH 2-8, while deviations were observed at higher pH values.

Applications of Metabolomics in Forensic Toxicology and Forensic Medicine.

Forensic toxicology and forensic medicine are unique among all other medical fields because of their essential legal impact, especially in civil and criminal cases. New high-throughput technologies, borrowed from chemistry and physics, have proven that metabolomics, the youngest of the "omics sciences", could be one of the most powerful tools for monitoring changes in forensic disciplines. Metabolomics is a particular method that allows for the measurement of metabolic changes in a multicellular system using two different approaches: targeted and untargeted. Targeted studies are focused on a known number of defined metabolites. Untargeted metabolomics aims to capture all metabolites present in a sample. Different statistical approaches (e.g., uni- or multivariate statistics, machine learning) can be applied to extract useful and important information in both cases. This review aims to describe the role of metabolomics in forensic toxicology and in forensic medicine.

The Activity of N-acetyl-ß-hexosaminidase in the Blood, Urine, Cerebrospinal Fluid and Vitreous Humor Died People Due to Alcohol Intoxication.

BACKGROUND: The article aimed to assess the activity of the hexosaminidase (HEX) and its HEX A and HEX B isoenzymes in persons who suddenly died due to ethanol poisoning and explain the cause of their death. METHODS: The research involved two groups of the deceased group A-22 people (20 males, 2 females; the average age 46 years) who died due to alcohol intoxication (with the blood alcohol content of 4‱ and above in all biological materials at the time of death-blood, urine, cerebrospinal fluid, and vitreous humor), and group B-30 people (22 males, 8 females; the average age 54 years), who died suddenly due to other reasons than alcohol. RESULTS: The highest activity of the HEX was found in the serum of A and B groups. A significantly lower activity of HEX, HEX A, and HEX B was observed in the urine of group A in comparison to the sober decedents. CONCLUSION: The lower activity of HEX and its isoenzymes in the dead's urine due to ethanol poisoning may suggest its usefulness as a potential marker of harmful alcohol drinking. Damage done to the kidneys by ethanol poisoning may be one of the possible mechanisms leading to death. Kidneys may be damaged intravitally via the inflammatory agent. Thus, it is necessary to conduct further research to evaluate the diagnostic usefulness of exoglycosidases while determining the death mechanisms of people who lost their lives due to ethanol poisoning.

Vitreous humour as an alternative material for the determination of alcohol concentration in human corpses.

AIM OF THE STUDY: The aim of the study was to assess the suitability of vitreous humour (VH) for determining alcoholaemia in deceased bodies. The study presents a correlation between the content of ethyl alcohol in vitreous humour and the concentration of ethyl alcohol in venous blood (femoral blood - FB). MATERIAL AND METHODS: The study material consisted of blood and vitreous humour samples taken from 62 individuals who died in the period 2012-2016 and whose post-mortem examinations were carried out at the Department of Forensic Medicine, Medical University of Bialystok. The content of alcohol was determined using the method of gas chromatography (GC). The findings of the study were analyzed statistically with MF Excel and Statistica 12.5, and then presented in a descriptive form, including figures and charts. RESULTS: Statistical analysis did not reveal significant differences between the concentration of ethanol in VH and FB. Spearman's correlation between the concentration of ethanol in the materials under study was r = 0.96, p < 0.01. CONCLUSIONS: The study demonstrated no statistically significant differences between the concentration of alcohol in VH and in FB. Consequently, vitreous humour can be used as a reliable alternative material in cases where there is no possibility of collecting venous blood for toxicological tests.

Analysis of suicide cases in post-mortem examination files of the Department of Forensic Medicine at the Medical University of Bialystok (2003 to 2015).

Aim of the study: The study presents the analysis of suicides based on the material consisting of the results of medico-legal autopsies and case files of the Department of Forensic Medicine of the Medical University of Bialystok. Material and methods: The material consisted of 6752 protocols of medico-legal autopsies carried out in the years 2003-2015 and case files, which enabled to classify cases as suicidal. The analysis included the dynamics of suicides over the years, basic demographic characteristics of victims, the ways, the time and place of suicide, the state of sobriety, the frequency of injuries in different regions of the body. The data were analyzed statistically using tests such as: Shapiro-Wilk test, Mann-Whitney U test, nonparametric Kruskal-Wallis test by programme Statistica 10.0 by StatSoft. Results: The research showed that suicides represented 11.09% of all post-mortem examinations. A statistically significant association was found between the age of the person committing suicide and the place chosen for suicide (p = 0.038). Post hoc analysis demonstrated statistically significant differences (p = 0.046) between suicide committed outdoors and in the supervised area. There was a statistically significant association between the sex and the preferable place of committing suicide (p = 0.0001). A high percentage of suicides were committed in the place of residence. There was a statistically significant difference in blood alcohol levels in women and in men at the time of suicide (p = 0.0029). Conclusions: The study revealed the increase of the percentage of suicides in relation to previous years. Hanging is still the most popular method of suicide.

Genetic variation of 15 autosomal STRs in a population sample of Bedouins residing in the area of the Fourth Nile Cataract, Sudan.

ABSTRACT: The purpose of the paper was to report allelic frequencies of 15 autosomal STR markers (AmpFlSTR NGM PCR Amplification Kit) for Bedouin inhabitants in the area of the Fourth Nile Cataract in Sudan, and compute commonly used population and forensic biostatistical parameters. Buccal swabs were collected from 117 unrelated individuals. DNA was extracted using DNA QIAamp® DNA Mini Kit, and quantitated with Quantifiler Human Quantification Kit in a 7500 Real-Time PCR System. Amplification of 15 AmpFlSTR NGM PCR Kit loci was performed in PCR System 9700. Electrophoresis and typing were performed in 3130 Genetic Analyzer. Arlequin v3.5 software and PowerStats v1.2 spreadsheet were used for statistical calculations. The STR frequency distributions showed no deviations from HWE. The combined values of Matching Probability and Power of Exclusion are 1.77 × 10-18 and 0.9999996, respectively. The average observed heterozygosity over 15 loci is 0.8069. Five different allelic microvariants were found. A significant linkage disequilibrium was observed in five pairs of loci. A 15 STR population database has been established for Sudanese Bedouins. The systems studied have been shown to be useful tool for personal identification in this population.

An Unusual Medicolegal Case of 32-Year-Old Mother and Her 29-Week Fetus With Hemolysis, Elevated Liver Enzymes, and Low Platelet Count Syndrome: Case Report and Short Literature Review.

Forensic medicine experts are well aware of blunt abdominal trauma, which often lead to death because of serious internal organ injury with severe blood loss. Sometimes careful examination of such cases may reveal some other underlying conditions, such as an existing disease that had contributed to the death or life-threatening complications. Our article presents a rare case of spontaneous hepatic rupture and intraperitoneal hemorrhage in a 32-year-old primigravida and her 29-week fetus, which was a result of hemolysis, elevated liver enzymes, and low platelet count syndrome. The said syndrome is a life-threatening obstetric complication usually considered to be a variant or complication of pre-eclampsia, which is characterized by hemolysis with microangiopathy, elevated liver enzymes, and low platelet count.

Typeability of DNA in Touch Traces Deposited on Paper and Optical Data Discs.

BACKGROUND: Nucleated epithelial cells that are transferred by casual touching and handling of objects are the primary source of biological evidence that is found in high-volume crimes. Cellular material associated with touch traces usually contains low levels of DNA template making it challenging to acquire an informative profile. OBJECTIVES: The main purpose of this study was to examine the efficacy of DNA typing in fingerprints deposited on optical data discs and the office paper. MATERIAL AND METHODS: Latent fingerprints were made by 60 subjects of both sexes (30 males and 30 females). A highly effective DNA extraction method with QIAamp DNA Mini Kit (Qiagen) and an increased sensitivity PCR by AmpFlSTR® NGM™ Amplification Kit (Applied Biosystems) carried out at standard 30 cycles and at increased 34 cycles were used. RESULTS: The mean value of total DNA recovery was 0.4 ng from CDs/DVDs and 0.3 ng from the office paper. Amplification of Low Template DNA (LT-DNA) resulted in improved analytical success by increasing the number of PCR cycles from standard 30 to 34. On the other hand, the increased PCR cycles resulted in allele drop-ins showing additional peaks, the majority of which were outside the stutter positions. CONCLUSIONS: Rigorous procedures and interpretation guidelines are required during LT-DNA for producing reliable and reproducible DNA profiles for forensic purposes.

Changes in Surface Charge Density of Blood Cells in Fatal Accidental Hypothermia.

The objective of this research was to evaluate postmortem changes concerning electric charge of human erythrocytes and thrombocytes in fatal accidental hypothermia. The surface charge density values were determined on the basis of the electrophoretic mobility measurements of the cells conducted at various pH values of electrolyte solution. The surface charge of erythrocyte membranes after fatal accidental hypothermia increased compared to the control group within whole range of experimental pH values. Moreover, a slight shift of the isoelectric point of erythrocyte membranes towards high pH values was observed. The surface charge of thrombocyte membranes in fatal accidental hypothermia decreased at low pH compared to the control group. However, at pH range 4-9, the values increased compared to the control group. The isoelectric point of thrombocyte membranes after fatal accidental hypothermia was slightly shifted towards low pH values compared to the control group. The observed changes are probably connected with the partial destruction and functional changes of the blood cell structure.

Analysis of the prevalence of medicines and psychoactive substances among drivers in the material of the Department of Forensic Medicine at the Medical University of Bialystok.

OBJECTIVES: To evaluate the prevalence of medicines and psychoactive substances in the blood of drivers. MATERIAL AND METHODS: The data, recorded in 2010-2011, was comprised of 274 blood samples of sober drivers (269 men and 5 women). 126 drivers within the group died immediately on the spot (mean age 29.7 years, range 19-53 years), 31 drivers survived the accident (mean age 32.4 years, range 24-69 years), and 117 drivers were stopped for traffic control (mean age 26.4 years, range 17-49 years). ELISA immuno-enzymatic blood tests detecting medicines and psychoactive substances were performed by the Neogen company. The presence of the tested substances was confirmed by gas chromatography and liquid chromatography with a mass detector (GC/MS and LC/MS). RESULTS: From the total number of 274 individuals, the presence of psychoactive substances was found in 132 cases, which accounted for 48.2% of all subjects. The drivers found to be under the influence of medicines and psychoactive substances were most often stopped for a roadside survey, among whom the percentage of positive results was 92.3% (108 cases out of 117 subjects). Among the total number of positive findings the psychoactive substances used, alone or in combination with other agents, were tetrahydrocannabinols (42.4%), amphetamines (26.5%), opiates (25.7%) and benzodiazepines (15.1%), and their concentration in the blood did not exceed toxic levels. CONCLUSIONS: In the group of psychoactive substances most often used, either alone or in combination with other agents, there were tetrahydrocannabinols, whose high percentage of positive results in drivers stopped for a roadside control may be explained by commissioning studies on early screening. Concentrations of the substances in most cases did not exceed toxic levels. Roadside examinations, on the basis of their alcohol-like effect, were performed primarily on male drivers.

The effect of fatal carbon monoxide poisoning on the surface charge of blood cells.

The objective of this investigation was to evaluate postmortem changes of electric charge of human erythrocytes and thrombocytes after fatal carbon monoxide (CO) poisoning. The surface charge density values were determined on the basis of the electrophoretic mobility measurements of the cells carried out at various pH values of electrolyte solution. The surface charge of erythrocyte membranes after fatal CO poisoning as well as after sudden unexpected death increased compared to the control group in the whole range of experimental pH values. Also, a slight shift of the isoelectric point of erythrocyte membranes to high pH values was observed. The surface charge of thrombocyte membranes after fatal CO poisoning decreased at low pH compared to the control group. However, at high pH, the values increased compared to the control group. The isoelectric point of thrombocyte membranes after fatal CO poisoning was considerably shifted toward low pH values compared to the control group. The observed changes are probably connected with the destruction of blood cell structure.

Population genetics of 30 INDELs in populations of Poland and Taiwan.

The Investigator DIPplex(®) kit (Qiagen) contain components for the simultaneous amplification and analysis of 30 biallelic autosomal INDELs and amelogenin. The objective of this study was to estimate the diversity of the 30 markers in Polish (N P = 122) and Taiwanese (N T = 126) population samples and to evaluate their usefulness in forensic genetics. All amplicon lengths were shorter than 160 base pairs. The DIPplex genotype distributions showed no significant deviation from Hardy-Weinberg rule expectations (Bonferroni corrected) except for DLH39 in the Taiwanese population. Among the Poles and the Taiwanese the mean observed heterozygosity values are 0.4385 and 0.4079, and the combined matching probability values are 7.98 × 10(-14) and 1.22 × 10(-11), respectively. The investigated marker set has been confirmed as a potential extension to standard short tandem repeat-based kits or a separate informative system for individual identification and kinship analysis. Eight INDELs have been selected as possible ancestry informative single-nucleotide polymorphisms for further analyses.

Fatal drowning as a result of an airplane crash--case report.

An ULM (Ultra Light Motorized) plane Aeroprakt 22-L with two men on board dived unexpectedly into the Dworackie lake near the town of Olecko in Warmia and Mazury district, Northeastern Poland. The pilot and the passenger had multiorgan blunt injuries, including subdural and subarachnoid hemorrhage, fractured ribs, fractured thigh (in one of the deceased) and pulmonary contusions as a result of the impact with the lake. The multiorgan injuries and unconsciousness resulting from head trauma prevented the casualties from saving themselves. The underlying cause of the death was the crash into the lake, however drowning was assessed as the proximate cause of death. The accident was analyzed together with an account prepared by State Commission on Aircraft Accident Investigation and with the Human Factor Analysis and Classification System - HFACS (the standardized method of analysis of flight accidents, the tool originally developed and tested within the U.S. military).

Changes in surface-charge density of blood cells after sudden unexpected death.

The objective of the investigation was evaluation of postmortem changes of electric charge of human erythrocyte and thrombocyte membranes after sudden unexpected death. The surface charge density values were determined on the basis of the electrophoretic mobility measurements of the cells carried out at various pHs of electrolyte solution. The interactions between both erythrocyte and thrombocyte membranes and electrolyte ions were studied. Values of parameters characterizing the membrane--that is, the total surface concentrations of both acidic and basic groups and their association constants with solution ions--were calculated on the basis of a four-equilibria mathematical model. The model was validated by comparison of these values to experimental data. We established that examined electric properties of the cell membranes are affected by sudden unexpected death. Postmortem processes occurring in the cell membranes can lead to disorders of existing equilibria, which in turn result in changes in values of all the above-mentioned parameters.

[Morphological, microscopic changes in the thyroid gland in the cases of hanging in vertical position with open and closed noose]. / Morfologiczne, mikroskopowe zmiany w tarczycy w przypadkach powieszen w pozycji stojacej w zamknietej i otwartej petli.

The aim of the morphological study was to search for traumatic changes in the thyroid in the cases of hanging in a vertical position with an open and closed noose. These changes would constitute an additional proof of intravital hanging and they would be useful in postmortem diagnosis of the hangings in which other commonly acknowledged features of intravitality are not discovered. In 33% of all cases, petechiae were observed in the thyroid gland. They appeared in 20% of the hangings with the closed noose, and in 13% of the hangings with the use of the open noose. The study showed that there was no difference between the open or closed, hard noose in terms of changes in the thyroid gland. There was also no correlation between the body position and the injury of the thyroid gland. In all the cases, hyperemia of the thyroid was observed and segmental expansion of blood vessels. The histopathological analysis showed that in one third of all the cases with negative macroscopic examination, it was discovered that microscopic changes indicated hanging alive.

[Microscopic diagnosis of pituitary gland damages resulting from cranio-cerebral trauma with the base of the skull fracture]. / Mikroskopowa diagnostyka uszkodzen przysadki u zmarlych w nastepstwie urazu czaszkowo-mózgowego ze zlamaniem podstawy czaszki.

The aim of this research was a macro- and microscopic investigation of pituitary glands in people who died after head injuries with the base of the skull fractures in the sella turcica. No macroscopic changes in the pituitary glands were discovered during an autopsy examinations. Histopathology demonstrated traumatic changes of the pituitary glands in all the cases, which were correlated with sella trurcica fractures. The evolution of traumatic alterations correlated with survival time after the injury. In the pituitary glands of people who died directly after head injuries, hyperemia and focal hemorrhages were discovered. Apart from hyperemia, focal necrosis was found in the pituitary glands among those who survived from 48 to 72 hours after head injury. The examined pituitary glands of those people who lived from 2 to 3 weeks after head injury showed considerable necrotic changes and numerous calcificationss.

[The analysis of neonatal deaths based on autopsy protocols of the Department of Forensic Medicine in Bialystok in the years 1955-2009]. / Analiza zgonów noworodków w oparciu o protokoly sekcyjne Zakladu Medycyny Sadowej w Bialymstoku w latach 1955-2009.

The authors performed an analysis of the causes of deaths of newborns with special emphasis placed on cases when neonaticide was suspected. In the discussed period, 17838 medico-legal autopsies were performed, 124 of them (0.695 per cent of all postmortem examinations) involved children who meet the WHO criteria for newborn - children under 28th day of life. The newborns were divided into two groups according to their exact age at the time of death - newborns who died within a short time after birth and newborns who died after the perinatal period. Their sex, body length and body weight were also taken into consideration. A total of 108 autopsies (87.1 per cent of all autopsied newborns) involved cases of suspected criminal infanticide within the first score of hours postnatally. In the majority of these cases, the cause of death was suffocation, rarely mechanical trauma. The case of death of older newborns was broadly understood trauma.