Stojan Petković

Postmortem Production of Ethanol in Different Tissues under Controlled Experimental Conditions

The aim of this study was to follow the postmortem ethanol production phenomenon under controlled experimental conditions (temperature, time interval) in different tissues. Specimens of blood, liver, skeletal muscle and kidney were taken from 30 corpses and no chemical preservatives were used in the specimens collected. Ethanol concentrations were detected by gas chromatography. All specimens stored at -20 degrees C and 4 degrees C did not show any change in ethanol concentration in an eight-day time interval. At 20 degrees C and 30 degrees C, all tissues, except blood, showed statistically significant ethanol production over the time interval tested. However, blood sample kept at 30 degrees C, showed statistically significant increase in ethanol production on the 2nd and 4th day comparing to the controls. Thus, we can state that postmortem ethanol production occurs in different tissues, and is increased at higher temperatures and, in general, it is in accordance with the course of time.

The influence of essential oil of aniseed (Pimpinella anisum, L.) on drug effects on the central nervous system

Anise (Pimpinella anisum L.; Apiaceae) and its essential oil have been widely used in folk medicine, pharmacy and food industry. Since there are some data about the impact of anise on functions of central nervous system (CNS), the issue of possible interactions with drugs acting in CNS should be considered. This survey aimed to examine the influence of aniseed essential oil (EO) intake on the effects of drugs that act in CNS. The chemical profile of essential oil determined by GC-MS revealed as the main components: trans-anethole (88.49%), γ-himachalene (3.13%), cis-isoeugenol (1.99%), and linalool (1.79%). The effects of codeine, diazepam, midazolam, pentobarbital, imipramine and fluoxetine were tested in mice after 5days of peroral pretreatment with human equivalent dose of aniseed EO (0.3mg/kg). The intake of EO led to significant increase of analgesic effect of codeine. The motor impairment caused by midazolam was enhanced in the group treated by EO. The application of diazepam decreased the number and percentage of entries in open arm in elevated maze plus test in the group pretreated with EO indicating augmented effect of drug on motor activity. EO pretreatment caused significant shortage of pentobarbital induced sleeping time when compared to control. The decrease in antidepressant effect of imipramine and fluoxetine was diminished by the pretreatment with aniseed EO. Based on the results of this study we conclude that concomitant intake of aniseed EO preparations and drugs that act on CNS should be avoided due to potential herb-drug interactions, which also need further clinical confirmation.

Forensic aspects of water intoxication: Four case reports and review of relevant literature

Water intoxication (WI) is a rare condition that originates from over-consumption of water, with a potentially fatal outcome. Increased water intake (polydipsia) is followed by urination of high amount of diluted urine (polyuria) which are the main initial symptoms of WI. We present four case reports of WI. Two of them are unusual pediatric clinical cases using medical documentation and police case files, one of which is related to child abuse, and the other to a psychiatric disorder. The other two cases are fatal adult cases submitted to autopsy from a psychiatric hospital. Also, we present a diagnostic algorithm for polydipsia and polyuria before death. WI is usually seen in patients with psychiatric disorders, victims of child abuse or torture, drug abusers or it can be iatrogenically induced.

Multiple stabbing in sex-related homicides

It is possible that sexually driven homicides are the consequence of sexual deviation, but more often than not the main cause of such homicides stems from a non-paraphilic persons emotional component. It is known that homicides which involve multiple stabbing are very often the result of the assailants highly expressed affect. This study tries to establish whether such homicides are essentially related to the sexual motives of the murderer. This paper is conducted through a retrospective autopsy study that includes 766 cases of homicides. These were reviewed and analysed according to the motive of the homicide, as well as by method, age, and gender of the victim, and the relationship between the victim and the assailant. The motives of homicides are classified as non-sexual and sexual, including homicides related to rape, jealousy, amorous affairs outside of an established relationship, deviant sexual behaviour of psychiatric patients, paraphilia, and disturbed emotional relationships between the victim and the assailant. Cases include both hetero- and homosexual relationships. Multiple stabbing occurs significantly more frequently in sex-related homicides (≥ 3 stab wounds) than in other homicide types. When the criteria for multiple stabbing are changed (≥ 4, 5, or 25 stab wounds), the percentage of sex-related homicides rises in every group in relation to the increased number of wounds. Sensitivity and specificity are represented by ROC curves for 3, 5 and 7 stab wounds. The correlation coefficients between multiple stabbing and sex-related homicides regarding gender are all near 0.9. For female victims, all homicides committed by 25 and more stab wounds were found to be sex-related. Statistically, jealousy was the most frequent motive for sex-related multiple stabbing homicides. Regardless of age, homicides involving multiple stabbings should be considered sex-related, especially when the victim is female.

Acute and Chronic Pretreatment with Essential Oil of Peppermint (Mentha × piperita L., Lamiaceae) Influences Drug Effects

The appearance of common and self‐initiative usage of various herbal preparations in everyday practice and life imposes the question of possible interactions with drugs. This survey examined the influence of acute and chronic peppermint oil (PO – Mentha × piperita L., Lamiaceae; prepared as emulsion for oral use) on pentobarbitone‐induced sleeping time, analgesic effect of codeine and impairment of motor coordination caused by midazolam in mice. The chemical profile of essential oil was determined by GC‐MS. Applied doses of PO were 0.1 and 0.2 mL/kg. Chronic PO intake (in both doses) led to significant decrease of analgesic effect of codeine, while acute intake of PO did not change this effect. Acute PO pretreatment in higher dose caused significant prolongation of pentobarbitone‐induced sleeping time, while it was significantly shortened by chronic PO pretreatment at the same dose. Midazolam effect was enhanced and prolonged significantly by chronic PO intake at higher dose, while acute intake of PO did not change this effect. Gut motility was increased only by acute intake of higher PO dose. Regarding the fact that PO produces changes in tested drug effects, the interaction between drugs and phytopreparations containing PO should be additionally followed/confirmed in humans. Copyright

Complex Suicide: An Unusual Case with Six Methods Applied

Abstract:  Complex suicides (CSs) are committed by using more than one method. They account for 1.5–5% of all suicides. We present a case of CSs of a 44‐year‐old man, found dead in the vicinity of his car, in a deserted frozen field. Police investigation excluded homicide, and no medical data confirmed mental illnesses. Autopsy revealed wrist cuts, neck cuts, acid burns in the GI tract, multiple stab wounds to the head by a screwdriver, and several uncertain signs of hypothermia. Toxicology analysis (gas chromatography–mass spectrometry) confirmed ingestion of insecticide. We concluded that stab wounds to the head were the cause of death, while external hemorrhage and hypothermia were contributing factors. This is the first case of CSs reviewed in the literature where six suicide methods were applied. This particular case is interesting because the victim used a screwdriver as a tool for inflicting stab wounds to the head, which is a rare suicidal method.

Ethanol Concentrations in Antemortem Blood Samples Under Controlled Conditions

AIM The change of antemortem blood alcohol concentration (BAC) in inadequately processed samples was examined. METHODS The study was performed on nine healthy, sober volunteers after overnight fasting. Blood samples were divided into groups and stored, with or without NaF, for varying time periods (12, 24 and 48 h) and at different temperatures (4 degrees C and 20 degrees C). BAC analysis was performed by the gas-chromatography method. RESULT All groups showed very low values of BAC. CONCLUSION Surprisingly, this study showed no ethanol production in improperly processed antemortem blood samples in healthy and sober individuals who were subjected to overnight fasting.

Methadone-Related Deaths – Epidemiological, Pathohistological, and Toxicological Traits in 10-year Retrospective Study in Vojvodina, Serbia†

The number of methadone‐related deaths (MRDs) during a 10‐year period (2002–2011) in the region of Vojvodina, Serbia, was increased. The cases were evaluated according to epidemiological parameters, pathohistological findings, and toxicological screening. The majority of victims were men, aged from 20 to 38. Pathohistologically, the signs of acute focal myocardial damage were present in the heart of victims with drug abuse history shorter than 2 years, while both signs of recent and chronic focal myocardial damage were developed among victims with longer drug abuse history (2–5 years). In postmortem blood samples of 54.84% of victims, methadone was detected in combination with diazepam, both in therapeutic range. Alcohol was absent in most cases. Other detected drugs were antipsychotics and antidepressants in therapeutic concentrations. These findings raise the attention to the concomitant use of methadone and benzodiazepines with the need for further studies to clarify the mechanism of death in such cases.

Genetic characterization of 27 Y-STR loci with the Yfiler ® Plus kit in the population of Serbia

Recently, a new 6-dye multiplex, AmpFlSTR Yfiler Plus PCR Amplification Kit (Yfiler Plus, Applied Biosystems, USA) has been developed, which can co-amplify 27 Y-STR loci. 17 Y-STRs widely used loci included in the AmpFlSTR Yfiler Amplification Kit (Yfiler, Applied Biosystems, USA) are included in this kit, besides 10 new additional Y-STR loci (DYS481, DYS460, DYS533, DYS449, DYS576, DYS627, DYS518, DYS570, and DYF387S1a/b), with the last seven being rapidly mutating Y-STRs (RM Y-STRs) [1]. These RM YSTRs are very useful for the discrimination between closely related and unrelated males [1–4]. In this study, we used the Yfiler Plus kit to type 203 unrelated individuals from 3 geographically different areas of Serbia; Northern Serbia, Central Serbia and Southern Serbia (Fig. S1). DNA was obtained from bloodstains or buccal swabs by using the Chelex 100 extraction method [5] from healthy unrelated individuals; 68 from Northern Serbia, 68 from Central Serbia and 67 from Southern Serbia, born and residing in the selected geographical areas, following informed consent. PCR amplification was performed using a GeneAmp PCR System 9700 Gold Plate (Applied Biosystems, USA) using Yfiler Plus kit, according to manufacturer’s recommendations with modifications in the reaction volume (12.5 ml). The electrophoresis was carried out on an Applied Biosystems 3500 Genetic Analyzer (Applied Biosystems, USA) and the fragment analysis was performed with GeneMapper IDX v.1.4 software (Applied Biosystems, USA). Allele designations were made using the allelic ladders provided with the Yfiler Plus kit, and following the recommendations of the DNA Commission of the ISFG on Y-STR analysis [6,7]. Following the guidelines of Forensic Science International Genetics, data were submitted to the YHRD (Y-chromosomal haplotype reference database; https://yhrd.org [8]) and received the accession number YA004149. The contributor successfully passed the YHRD quality control test. The Y-STR haplotypes observed in the three Serbian geographic areas is reported in Table S1. A total of 200 different haplotypes was observed in the sample of 203 Serbian individuals. A total of 197 unique haplotypes (singletons) were obtained from 203 individuals using the Yfiler Plus kit (Table S2). All haplotypes in Northern, Central and Southern Serbia were unique, but three haplotypes shared between different Serbian regions were found (Table S1). A null allele was observed in one individual at DYS391. In all the samples, four different intermediate alleles were found in two loci (DYS390 and DYS458) whereas locus DYS458 showed the highest number (three different intermediate alleles with

Fatal Intoxication Because of Trihexyphenidyl

Abstract:  Trihexyphenidyl (THP) is an anticholinergic agent with forensic toxicological interest. We present a case of a 59‐year‐old woman with a history of paranoid disorder, who was found dead in the house where she lived alone. The autopsy findings revealed no marked pathological changes. Toxicological analysis based on gas chromatography–mass spectrometry (GC–MS) analysis revealed THP and its major metabolite (hydroxy—THP) in blood and urine, with THP concentrations of 0.053 and 0.560 mg/L, respectively. The blood and urine ethanol concentrations were low 0.096 and 0.100 g/L, respectively. Based on these results, we determined the cause of death to be THP poisoning. It is suggested that rare case of death associated with THP overdosage should be taken in conjunction with central nervous system depressants (benzodiazepines, ethanol) and/or with other pathological disorders. Thus, our case could not be supportive for this allegation.