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.

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.

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.

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.

Battered child syndrome in the records of the Department of Forensic Medicine, Medical University of Bialystok.

Violence against children has been until now recognised as physical aggression escalating from corporal punishment, hard physical labour to homicide. Nowadays child abuse is considered as maltreatment occurring in four different aspects: physical abuse, psychological abuse, sexual abuse and neglect. Child abuse is difficult to disclose, as it is concealed both by the perpetrators and the victims. The child is afraid and at the same time, almost always ashamed of looking for help. The victim frequently feels guilty, considers the violence as a norm, but in many cases is not able to speek precisely and very often has problems in contacting the required person. The objective of this study is to analyse the phenomenon of child abuse on the basis of Department of Forensic Medicine records. The victims were characterized in terms of their age, sex, place of residence and the type and location of injuries. The kind of abuse and perpetrators were also determined by researchers.

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.

The influence of renal function on the association of rs854560 polymorphism of paraoxonase 1 gene with long-term prognosis in patients after myocardial infarction.

Paraoxonase 1 (PON1) is an enzyme responsible for the antioxidant properties of high density lipoprotein (HDL). The activity of PON1 is decreased in patients with coronary artery disease, myocardial infarction or chronic kidney disease. rs662 and rs854560 are single nucleotide polymorphisms (SNPs) associated with PON1 activity and 10-year cardiovascular mortality of patients with stable coronary artery disease. We investigated the association of rs662 and rs854560 SNPs of the PON1 gene with 5-year mortality in patients with ST-elevation myocardial infarction (STEMI) treated invasively. We analyzed the data of consecutive patients with STEMI treated with primary PCI. Genotyping was performed with the TaqMan method. The analyzed end-point was total 5-year mortality. Additional subgroup analysis was performed for survival of patients depending on their eGFR. The study group comprised 634 patients (mean age 62.3 ± 11.85 years; 25.2% of women, n = 160; PCI successful in 92.3%, n = 585). No clinically relevant differences in baseline characteristics were found between the genotypes. No association between either genotype and 5-year mortality was found: p = 0.4 for the rs662 SNP, p = 0.73 for the rs854560 one (log-rank test). However, in a subgroup of patients with eGFR below median value (78.6 ml/min/1.73m2) the rs854560 AA homozygotes had a significantly lower probability of survival (p = 0.047, log-rank test). The AA genotype of the rs854560 SNPs of the PON1 gene is associated with increased mortality in patients after myocardial infarction in the subpopulation of patients with lowered eGFR. This phenomenon may be explained by potentially lower PON1 activity in kidney disease.

The 9p21 polymorphism is linked with atrial fibrillation during acute phase of ST-segment elevation myocardial infarction.

The aim of the study was to find whether patients carrying polymorphic allele of the rs10757278 polymorphism from 9p21 locus have changed risk of arrhythmia (atrial fibrillation, AF; sustained ventricular tachycardia or ventricular fibrillation, sVT/VF) during acute phase of myocardial infarction. Retrospective analysis of data collected prospectively from two independent centers was performed. The clinical data were pooled from two independent cardiac registries: (1) the Warsaw ACS genetic registry (STEMI and NSTEMI/UA patients hospitalized in the years 2008-2011; only STEMI patients were analyzed); (2) the Bialystok STEMI genetic registry (STEMI patients hospitalized in years 2001-2005, who survived the first 48 h from hospital admission). Data regarding sVT/VF and AF within first 24 h were analyzed. The patients were genotyped with rs10757278 polymorphism. 1083 patients were included in the analysis; 62 (5.7 %) patients had sVT/VF during acute phase and 78 (7.2 %) patients had AF, 46 (4.2 %) patients had new-onset AF. Minor allele frequency in all patients with AF was significantly different from those without AF (0.40 vs 0.51, p = 0.0096). When only new-onset AF was analyzed, the trend was the same, with significant protective effect in recessive model [OR 0.41 (95 % CI 0.17-0.97), p = 0.025]. The effect was independent of age and GRACE score. No relationship was found between sVT/VF and rs10757278. Patients with STEMI, who survived until hospitalization with polymorphic allele of 9p21 rs10757278 SNP have less AF during acute phase of STEMI. SNP rs10757278 is not linked with sVT/VF in acute phase of STEMI.

Review of fatal gunshot cases in the files of the Department of Forensic Medicine in Bialystok, Poland, in the years 1964-2015.

Aim of the study was to assess gunshot fatalities on the basis of post-mortem examination reports collected at the Department of Forensic Medicine in the Medical University of Bialystok. Material and methods: The research covered 87 gunshot fatalities occurring in the Podlasie district (north-eastern Poland) over the period from 1964 to 2015. The analysis included demographic data such as the victims' gender, age, place of residence and such data as time of year, circumstances of the event and causes of death. The study also investigated blood alcohol level in the victims. The results were analysed statistically using the Mann-Whitney and Kruskal-Wallis tests. Results: The majority of victims were young males, most of whom committed suicide. The most common cause of death among the suspected suicides was gunshot wounds in the skull area. The second most frequent gunshot wound location among non-suicides was the chest. The majority of male victims were intoxicated at the time of death. Conclusions: In the area of the city of Bialystok and its surroundings, there are approximately two gunshot fatalities per year. In most cases, the victims were urban-dwelling young males (aged 21-30) committing suicide. The most common cause of death among the suspected suicides was gunshot wounds in the skull area and the chest. The majority of male victims were intoxicated at the time of death.

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.

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.

Diagnosis and Treatment of Lower Extremity Arterial Disease-A Survey among Family Medicine Trainees in Poland.

Guidelines point to the ankle-brachial index (ABI) as a non-invasive tool for the initial diagnosis of lower extremity artery disease (LEAD). Questions have been raised whether primary practices should perform ABI. An online questionnaire was distributed among family medicine trainees in two academic centers in Poland. The questionnaire aimed to establish their knowledge about LEAD management and their opinion on the usefulness of ABI measurement and other LEAD diagnostic methods in primary care. ABI measurement was found either very or moderately useful in LEAD diagnosis by 94.5% of the respondents. Among the three most important elements of LEAD management, lifestyle changes, secondary prevention of atherosclerosis and exercise treatment were chosen, respectively, by 98.6%, 83.6% and 72.6% of them. ABI was seen as a useful diagnostic method at the primary care by 74% of the participants; however, 82.2% of them do not have access to ABI measurement in their workplace. The residents have good knowledge of the diagnostic methods of LEAD and consider ABI measurement as useful in LEAD diagnosis. However, most of them do not have access to ABI measurements in their clinical practices. Future discussion and potential financial changes will be needed for the introduction of ABI measurements into Polish primary care.

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.

Y-STR data of the Yfiler Plus panel in population of north-eastern Poland. / Dane populacyjne dla markerów zestawu Yfiler Plus w populacji Polski pólnocno-wschodniej.

Background: The use of new high-resolution and forensic identification capabilities for population studies offered by new multiplex methods (such as Yfiler Plus) is crucial in forensic genetics cases. The development of haplotype frequency databases is essential to take full advantage of the new Y chromosome determination capabilities. Purpose: Development of the haplotype database of the Yfiler Plus kit for a population-based sample of 534 males from northeastern Poland and calculation of suitability parameters for forensic genetics studies. Materials and methods: The study was conducted on a population sample of 534 unrelated males from the area of northeastern Poland using the Yfiler Plus panel of 27 markers located on the Y chromosome. Results: Four haplotypes appeared twice. The Discrimination Capacity (DC) of the entire set was 0.9925. The highest Gene Diversity (GD) value was calculated for DYS518 (0.86) belonging to the fast-mutation markers, while the lowest GD was calculated for DYS392 (0.42). Conclusion: The results indicate the need for further research and observation of changes, both in different regions of Poland and across Europe.

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.

Population Genetic Data of 30 Insertion-Deletion Markers in the Polish Population.

(1) Background: Insertion-deletion (InDel) markers show the advantages of both short tandem repeats (STRs) and single nucleotide polymorphisms (SNPs) and are considered alternative markers in forensic genetics. (2) Methods: Allelic frequencies and corresponding forensic efficiency parameters of 30 autosomal polymorphic InDel loci included in the Investigator DIPplex kit (Qiagen) were obtained in a sample of 631 unrelated Polish individuals. Allelic frequency data were compared with those reported for selected populations (3) Results: All the loci conformed with Hardy-Weinberg equilibrium after applying a Bonferroni correction and no pair-wise significant linkage disequilibrium was detected. (4) Conclusions: DIPplex Kit differences were high among populations worldwide. The InDel markers are highly discriminating for human identification purposes in the Polish population.

[Validation and evaluation of a five miniSTRs kit in forensic genetics]. / Walidacja i ocena przydatnosci zestawu pieciu markerów miniSTR w genetyce sadowej.

The newly designed and optimized miniplex contains the following markers: D3S3053, D6S474, D9S2157, D20S482 and sex-determining marker - amelogenin. The target amplicon lengths for the developed multiplex are 71-135 bp. Amplification products were detected in a fluorescence based automated genetic analyzer. A minimal DNA sample required to obtain full genetic profiles was 250 pg. The usefulness of these miniSTRs in genotyping of severely degraded forensic samples, such as stains of blood and semen, saliva on cigarette butts and telogen hair has been confirmed in validation studies. The designed pentaplex offers a new potential screening tool in cases of old crime scenes, mass disasters, mass graves, etc., where DNA degradation, body fragmentation or large numbers of victims occur. The use of additional non-CODIS markers may increase typeability of severely degraded samples and ensure a higher potential for genetic discrimination.

[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.