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.

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.

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.

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.

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.

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.

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.

Post-mortem detection of six human herpesviruses (HSV-1, HSV-2, VZV, EBV, CMV, HHV-6) in trigeminal and facial nerve ganglia by PCR.

BACKGROUND: Among over 100 types of Herpesviridae viruses, eight can infect humans: herpes simplex viruses (HSV-1, HSV-2), varicella zoster virus (VZV), cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human herpesviruses 6, 7, and 8 (HHV-6, HHV-7, HHV-8). After initial infection, the viruses remain latent for the lifetime of the host. The aim of this study was to determine the distribution of six different herpesviruses: HSV-1, HSV-2, VZV, EBV, CMV, and HHV-6 in trigeminal and facial nerve ganglia among a random group of Polish population. METHODS: The studied group consisted of 47 individuals (40 male, seven female); mean age of 47.4 ± 16.5 years) who died of independent causes (suicide, traffic accident, and poisoning, among others). Bilateral trigeminal and facial nerve ganglia of each cadaver were collected during the autopsy. Herpesviruses were detected using multiplex polymerase chain reaction technique. RESULTS: Herpesviruses were found in trigeminal and/or facial ganglia in 30/47 (63.8%) of cadavers. HHV-6 was the most prevalent of the herpesviruses and was found in nearly half of cadavers (n = 22; 46.8%), followed by HSV-1 (n = 7; 14.9%), VZV (n = 4; 8.5%), EBV (n = 4; 8.5%), HSV-2 (n = 2; 4.3%), and CMV (n = 1; 2.1%). Facial nerve ganglia (n = 23; 48.9%) were more often infected than trigeminal ganglia (n = 13; 27.7%). DISCUSSION: The results of this study have revealed a common presence of the herpesviruses in trigeminal and facial nerve ganglia among a random group of Polish population. Furthermore, the data also demonstrate simultaneous infection of the ganglia with different herpesviruses. This study has contributed to the knowledge of prevalence and localization of herpesviruses in different structures of the nervous system.

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 paternity exclusions in the material collected by the Department of Forensic Medicine, Medical University of Bialystok in 2008-2017.

AIM OF THE STUDY: Analysis of frequency and structure of paternity exclusions in the material collected by the Department of Forensic Medicine, Medical University of Bialystok in 2008-2017. MATERIAL AND METHODS: The paper is based on paternity test reports involving alleged father-child-mother trios. In a total of reviewed 958 cases, 187 exclusions were identified. The analysis was carried out on the basis of the results of DNA tests. DNA extraction was performed using QIAamp DNA Mini Kit (Qiagen) and DNA quantitation using Quantifiler Human DNA Quantification Kit and 7500 Real-Time PCR System (Applied Biosystems). AmpFLSTR Identifiler PCR Amplification Kit and a PCR System 9700 thermal cycler (Applied Biosystems) were used for DNA amplification. RESULTS: Over the analyzed period, the number of paternity tests was nearly halved, whereas the percentage of exclusions in individual years varied significantly (33.9-13.3%), with the average of 26.3%. The highest efficiency of exclusions was observed for D18S51 (0.7166) and FGA (0.7059), and the least effective system was TPOX (0.3048). CONCLUSIONS: The applied set of markers has been demonstrated to be an efficient tool in genetic paternity tests in the context of the recommended rules of exclusion.

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.

The rs2228145 polymorphism in the interleukin-6 receptor and its association with long-term prognosis after myocardial infarction in a pilot study.

INTRODUCTION: Interleukin-6 (IL-6) is a cytokine with a complex function that is described as both pro- and anti-inflammatory. One factor that influences its function is the rs2228145 A/C single nucleotide polymorphism (SNP) of the IL-6 receptor (IL6R) gene. C allele carriers have a decreased inflammatory response and decreased prevalence of ischemic heart disease. The aim of the study was to investigate the association of the rs2228145 SNP of the IL6R gene with long-term total mortality in patients with ST-elevation myocardial infarction (STEMI) treated invasively. MATERIAL AND METHODS: We analyzed the data of consecutive patients with ST elevation myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PCI). Genotyping was performed with the TaqMan method. The analyzed end-point was total long-term mortality (median: 2875 days). RESULTS: The registry comprised 553 patients (mean age: 62.4 ±11.9 years; 25.6% females, n = 142; TIMI 3 obtained in 91.7% of patients, n = 507). No significant differences in baseline characteristics were found between the genotypes. During long-term follow-up 171 (30.9%) patients died. There was non-significantly higher mortality in the rs2228145 AA homozygotes compared to C allele carriers (OR = 1.34, 95% CI: 0.93-1.93, p = 0.1). CONCLUSIONS: The rs2228145 polymorphism of IL6R was not significantly associated with long-term mortality after STEMI. However, AA homozygotes (high-risk genotype for ischemic heart disease) showed a trend towards adverse outcome compared to C allele carriers. The observed trend is promising, but it requires independent replication studies.

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.

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.

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.