About Polish forensic medicine based on archival material described in scientific literature. / O polskiej medycynie sadowej na kanwie materialu archiwalnego opisanego w literaturze naukowej.

The need for medical assessment of murder victims and the establishment of rules for this assessment became the driving force behind the creation of forensic medicine, while the non-medical area of crime assessment, including crime scene examination, became the driving force behind the creation and development of criminalistics. Polish forensic medicine has been developing for over 200 years, and the Kraków Chair of Forensic Medicine is the oldest unit of this type. It is therefore not surprising that the archive which collects protocols of expert and scientific research is quite extensive. This material is a valuable foundation for scientific work including not only numerous publications in national and international journals, but also books. The author of the work selected several books on medical and forensic topics that appeared in recent decades on the Polish publishing market, indicating the value and importance of archival material in the literature of this area of science.

Lower dose of plant substance more effective in repelling Rhyzopertha dominica F . (Coleoptera, Bostrichidae) and Sitophilus granarius L. (Coleoptera, Dryophthoridae).

INTRODUCTION AND OBJECTIVE: The study compares the effect of two concentrations (0.1% and 1%) of plant material on repelling two dangerous stored pests - Rhyzopertha dominica F. and Sitophilus granarius L. Both species were treated with the essential oils of Foeniculum vulgare Mill., Carum carvi L., Mentha pulegium L., and Cananga odorata (Lam.) Hook.f. & Thomson, as well as the chemicals contained in these oils: anethole, L-carvone, pulegone and thymoquinone. MATERIAL AND METHODS: Laboratory tests were carried out at 29±1 °C with 60±5% relative humidity (RH). Repellence of insects were noted after 1, 2, 3, 4, 5, 24 and 48 h. RESULTS: A lower dose of essential oils and some of the tested compounds caused a stronger repellence in the specified species of beetles. In the case of R. dominica, the concentration of 0.1% pulegone and L-carvone had the greatest repellent effect. Differences in the repellence of R. dominica were found between the concentrations of 0.1%-1% among all the tested substances in each time interval. This difference ranged from 5.0%-52.92%. All substances used in the tests had a strong repellent effect on S. granarius (30%-100%). There were differences in the repellency of S. granarius between 0.1%-1% concentration in most of the tested substances in each time interval, but they were not as large as for R. dominica.. CONCLUSIONS: The most interesting results were obtained concerning the relationship: the lower tested substance concentration caused a much stronger repellent effect of R. dominica and S. granarius..

Review of ticks attacking domestic dogs and cats, and their epidemiological role in the transmission of tick-borne pathogens in Poland.

INTRODUCTION AND OBJECTIVE: Progressive climate change plays a major role in the expansion of tick populations, as well as the transmission of tick-borne disease pathogens (TBPs) to humans and animals throughout the world. Zoonoses are a growing environmental problem of great importance for public health. In Poland, domestic dogs and cats are mainly infested by Ixodes ricinus, Ixodes crenulatus, Ixodes hexagonus, Ixodes rugicollis and Dermacentor reticulatus ticks from the Ixodidae family. Other tick species, such as I. apronophorus and Haemaphysalis concinna, recorded sporadically on domestic dogs or cats, may expand their range in the future and successfully infest pets. Individual cases of infestations in Poland with foreign tick species, such as Rhipicephalus sanguineus, are known and may soon be recorded more frequently. OBJECTIVE: The aim of this review is to summarize the occurrence and identification of tick species and TBDs that are present or are likely to spread in Poland, which, given the medical and veterinary importance of ticks, may help in the development of public health strategies. REVIEW METHODS: A comprehensive literature review of publications and own research and data analysis was performed, obtained from reports and scientific descriptions on the epidemiology of tick-borne diseases. BRIEF DESCRIPTION OF THE STATE OF KNOWLEDGE: The ecology of ticks and hosts in urban and suburban environments is critical for quantifying the parameters necessary for an initial risk assessment, and identifying public health strategies to control and prevent TBDs. In the future, these species may expand their range of occurrence and their host, becoming common members of the Polish tick fauna. SUMMARY: Anaplasma, Babesia, Borrelia and Rickettsia spp. are the major TBPs in Poland, and their prevalence is usually higher in dogs than cats.

The Identification of Potential Immunogenic Antigens in Particular Active Developmental Stages of the Rice Weevil (Sitophilus oryzae).

BACKGROUND: The rice weevil (Sitophilus oryzae) originates from subtropical and tropical areas of Asia and Africa, but it also appears on other continents, mostly as a result of trade in rice. It may occur in grain fields as well as in storage facilities, and cause allergenic reactions. The aim of this study was to identify the potential antigens in all developmental stages of S. oryzae, which may cause an allergic response in humans. METHODS: Sera of 30 patients were tested for the presence of IgE antibodies to antigens from three life stages of the rice weevil. To identify protein fractions containing potential allergens, proteins collected from larvae, pupae, and adults separated by sex of S. oryzae were fractionated by SDS-PAGE. Then, they were probed with anti-human, anti-IgE monoclonal antibodies, fractionated by SDS-PAGE and detected by Western blotting. RESULTS: In total, 26 protein fractions of males and 22 fractions of other life stages of S. oryzae (larvae, pupae, and females) positively reacted with the examined sera. CONCLUSIONS: The conducted study showed that S. oryzae may be a source of many antigens which may cause the potential allergic reactions in humans.

Post-aggression suicide under the influence of new psychoactive substances AMB-FUBINACA and U-47700.

The objective of the present study was the development and validation of the method for determining AMB-FUBINACA and its metabolite - AMB-FUBINACA O-desmethyl acid - in blood samples, followed by verification of the method in toxicological judicial and forensic medicine practice employing the example of post-aggression suicide. Most likely in consequence of development of adverse effects resulting in psychotic symptoms, a male being under the influence of the synthetic cannabinoid AMB-FUBINACA and the new synthetic opioid U-47700, mortally wounded his female partner and subsequently committed suicide. Identification and determination of the afore-mentioned xenobiotics in blood samples collected from the male and female victims were performed employing high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). The analytes were isolated from blood samples using the solid phase extraction (SPE) method. The blood samples collected from the male and female demonstrated respectively 110 and 196 ng/mL of AMB-FUBINACA O-desmethyl acid metabolite, 1935 and 357 ng/mL of U-47700, 250 and 200 ng/mL of N-desmethyl-U-47700, as well as 410 and 200 ng/mL of N,N-didesmethyl-U-47700. The concentration values of new psychoactive substances (NPS's) in blood samples originating from the male and female were within the ranges encountered in cases of poisoning, including these resulting in death. Nevertheless, the evident signs of exsanguination proof that the woman was alive when she sustained lethal injuries. The presented cases illustrate the difficult to be anticipated effect exerted on the users by NPS's.

[The importance of the quality of toxicological expertise for the need of the judiciary in the historical and contemporary aspect]. / Znaczenie ekspertyzy toksykologicznej dla wymiaru sprawiedliwosci w aspekcie historycznym i wspólczesnym.

The history of humanity is closely related to the use of various poisons, different in each epoch. They served different purposes for centuries. In addition to being a remedy for various ailments and diseases, they also helped to bear the hard life of a person thanks to the possibility of causing mental elation, making it more bearable. They were also used to kill other people, most often for very low motives. The number of poisons existing in particular stages of civilization has been systematically increasing, reaching the number of 100,000 - 500,000 toxicologically active compounds in modern times, with the generally estimated number of known chemical compounds at the level of about 240 million. The research work of thinkers and people of progress is a counterbalance to the evil deeds of poisoners in antiquity and the Middle Ages. These works appeared in the late Middle Ages and are continued in various forms until the present day. As a result of these works, modern toxicological forensic expertise has been developed. However, before it appeared in its modern shape, it had to go through a very difficult development path, which lasted continuously for several centuries. Modern toxicological expertise, based on a highly specialized instrumental methodology, operating with high methodological standards, is the achievement of many generations. These standards have now become a requirement for expert works of our times, and failure to comply with them is treated as a malpractice. This work is a review of the types of poisons and reports in terms of the development and application of toxicological forensic expertise for the purposes of the judiciary.

Repellent Effect of the Caraway Carum carvi L. on the Rice Weevil Sitophilus oryzae L. (Coleoptera, Dryophthoridae).

The aim of the study was to check the effect of Carum carvi L. essential oil and L-carvone on the emigration, repellence and mortality of Sitophilus oryzae L. In the experiments with caraway essential oil, concentrations of 0.1%, 0.5% and 1% were used, and concentrations of 0.05, 0.1, 0.5 and 1% were used with L-carvone. We assessed whether, in what concentrations and after what exposure time the substances acted on S. oryzae as repellents and/or insecticides. The laboratory tests were carried out at 29 ± 1 °C with 60 ± 5% relative humidity (RH). The deterrence, mortality and abundance of insects were noted after 1, 2, 3, 4, 5, 24 and 48 h. For caraway essential oil and L-carvone, the highest repellency was not shown in the highest concentrations that were used in the tests but in the lower concentrations of 0.5% and 0.1%, respectively. In all used concentrations, caraway essential oil showed repellent effects on S. oryzae. The highest repellency (60-98%) was caused by 0.5% caraway essential oil after 1, 2, 3, 4 and 5 h of research and by 0.1% L-carvone (16-100%). The highest mortality of S. oryzae was caused by 0.5% caraway essential oil and 1% L-carvone. L-carvone at a concentration of 0.05% did not cause mortality in S. oryzae. In conclusion, the greatest repellent effects on S. oryzae were caused by lower doses of caraway essential oil and L-carvone. These compounds do not show the normal relationship described in the previous literature on warehouse pests, in which the repellency increased with increasing concentrations of the tested plant material.

Molecular detection of tick-borne pathogens in ticks collected from pets in selected mountainous areas of Tatra County (Tatra Mountains, Poland).

The mountainous and foothill areas, in which the city of Zakopane, the capital of Tatra County, is located are characterized by continuous weather changes, lower air temperature, persistent snow cover, and poorer vegetation than in the lowlands. Ixodes ricinus and Ixodes hexagonus are vectors of tick-borne diseases and play an important role in the persistence of tick-borne diseases. The aim of the study was to determine the risk of exposure of domestic cats and dogs to the attacks of Ixodid ticks, to tick-borne infections with Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Babesia microti and Toxoplasma gondii in the city of Zakopane and the surrounding area. In 2017-2018 ticks were collected from a total of 10 domestic cats and 88 domestic dogs. Selected pathogens of tick-borne diseases were detected by PCR and nested PCR. The study material contained 119 I. ricinus and 36 I. hexagonus. The molecular examinations showed the presence of A. phagocytophilum in 3.8%, B. microti in 24.5% and T. gondii in 4.5% of the all ticks. In addition, in the study area, there is a high potential risk of tick-borne infection by B. microti, and a low potential risk of exposure to A. phagocytophilum and T. gondii infection.

Aspects related to analysis and medico-legal assessment in the context of the presence of synthetic cannabinoids in biological material based on the example of AB-CHMINACA.

AIM OF THE STUDY: The aim of this study was to develop and validate a method for the determination of the synthetic cannabinoid AB-CHMINACA in blood, followed by its verification in forensic toxicological practice. MATERIAL AND METHODS: The case of a 41-year-old man admitted to hospital because of scheduled cardiac surgery was discussed. The man died after 12 hours of hospitalization. Based on collected evidence, AB-CHMINACA poisoning was suspected. The identification and determination of AB-CHMINACA in the man's blood was performed by high-performance liquid chromatography coupled with tandem mass spectrometry using electrospray ionization (HPLC-ESI-MS-MS), after prior solid phase extraction. RESULTS: The concentration of AB-CHMINACA determined in the man's blood sample was 0.5 ng/ml. CONCLUSIONS: In the interpretation of the case, it was concluded that AB-CHMINACA had no direct effect on the patient's death, the cause of which was ascertained as chronic heart failure secondary to aortic valve disease, decompensated by pneumonia. However, an indirect impact of side effects resulting from the use of synthetic cannabinoids cannot be ruled out. They might have exacerbated the man's disease process leading to sudden cardiac arrest caused by asystole.

A new challenge in forensic toxicology exemplified by a case of murder under the influence of a synthetic cannabinoid - AM-2201.

Among new psychoactive substances (NPS) available on the narcotic market, a significant number consists of synthetic cannabinoids commonly known as smokable herbal "spice" and "K2", and which are legally treated as a legal alternative to marijuana. The dearth of information on the pharmacology of these intoxicants as they are introduced into the market has created the urgent need among healthcare providers for case studies on the substances belonging to this group, both in terms of the consequences of using such intoxicants, and in methods of detection. The subject of the present report is a multi-parameter analysis of a criminal case of an 18-year-old male who was charged with murder of his female relative and attempted murder of two other victims by stabbing. The defendant pleaded guilty, but he claimed that he had been acting without volition, because he was under the influence of the synthetic cannabinoid AM-2201, which had been purchased from a dealer as a 10g package labelled "Mr Green - No bad trip". Analytical methods including gas chromatography - electron ionization - quadrupole ion trap mass spectrometry (GC-EI-QIT/MS) and liquid chromatography, electrospray ionization, tandem mass spectrometry (LC-ESI-MS-MS) were developed to determine the presence of AM-2201 in the Mr Green - No Bad Trip, and in the blood of the perpetrator, respectively. Toxicological findings are discussed in the context of psychoactive and adverse physical effects resulting from the presence of AM-2201 in the human body; the observations were also analyzed in conjunction with data from the literature.

Population pharmacokinetic modelling of valproic acid and its selected metabolites in acute VPA poisoning.

BACKGROUND: Valproic acid (VPA) is a first-line antiepileptic drug. It is used in the treatment of many different types of partial and generalized epileptic seizures. Though the clinical pharmacokinetics of VPA has been well defined, information about pharmacokinetics after overdoses is rare. The aim of this study was to try to build a population pharmacokinetic model that would describe the time course of VPA and its selected metabolites when the drug is ingested in an overdose situation. METHODS: Blood samples were collected during admission to the hospital and several times during treatment for poisoning (10 men and 10 women). The concentration of VPA and its metabolites were determined by liquid chromatography coupled with mass spectrometry. For population pharmacokinetic evaluation of VPA and its metabolites, the two-compartment-model was applied. RESULTS: The estimated doses of VPA taken ranged from 6 to 65g, while the time after ingestion ranged from 1 to 30h. Results showed that the ß-oxidation process exhibited Michaelis-Menten kinetics becoming saturated during acute intoxication. The same could not be said for the desaturation route. VPA therapy increased the Vmax for ß-oxidation by 59% while decontamination appeared to be of moderate efficacy lowering the F value by 34% on the average. CONCLUSIONS: None of the models perfectly described the experimental data. Important factors like the variable degree of protein binding by VPA could not be included in the models. The small number of subjects used in the study made the analysis of more covariates impossible.

New psychoactive substances in substantive evidence in expert practice of the Department of Forensic Medicine, UJCM in the years 2010-2015.

AIM OF THE STUDY:: Aim of the study was to analyse of 2075 evidences containing new psychoactive substances (NPS). MATERIAL AND METHODS: The prepared samples were identified employing an analytical procedure where the analytes were investigated by gas chromatography-electron impact mass spectrometry (GC-EI-MS) using a created library of mass spectra. RESULTS: The analysis revealed the following substances in the investigated products: piperazine derivatives (including BZP, MPMP, TFMPP), cathinone derivatives (including: pentedrone, 3-MMC, butylone, 4-MEC), pyrovalerone derivatives (MDPV, naphyrone, α-PVP, α-PVT), synthetic cannabinoids (such as AM-2201, UR-144, XLR-11, JWH073, JWH081, PB-22, AB-CHMINACA). Research conducted in 2010-2015 made it possible to track changes in the composition of investigated preparations. CONCLUSIONS: The following relationships has been shown: number of components decreased and in the end of 2015 dominated preparations with single component; introduction of amendments Act on Preventing Drug Addictionn affect the elimination from the market of one compounds and replacing them by their derivatives; since 2011 on the market of 'legal highs' we did not observe occurrence of compounds of piperazine group.

New challenges in toxicology of new psychoactive substances exemplified by fatal cases after UR-144 and UR-144 with pentedrone administration determined by LC-ESI-MS-MS in blood samples.

The topic of this paper relates to the study of cases involving the use of new psychoactive substances (NPS) from the classes of synthetic cannabinoids and cathinones, analyzed from multiple viewpoints including clinical and medico-legal perspectives. The paper investigates three fatal cases in which UR-144 and UR-144 with pentedrone identified in the bodies of victims during post-mortem examinations were responsible for the tragic consequences and proved to be the indirect cause of death. The victims were men aged 16, 22 and 40 years who used drugs, for example they smoked marijuana or its substitutes in the form of synthetic cannabinoids. In addition, all of them had behavioural problems. On account of emotional imbalance attributable probably to the presence of UR-144 (in one case) and a mixture of UR-144 and pentedrone (in the other two cases), two men committed suicide by jumping from a height and hanging, and one man had fatal accidental poisoning with pentedrone which was used to enhance the effect of previously used UR-144. The presence of UR-144 and pentedrone in the post-mortem material was analyzed by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS). The results of toxicological tests were analyzed with a focus on possible tragic side effects caused by the presence of UR-144 and UR-144 with pentedrone in the body.

Medicolegal aspects of PMA-related deaths.

Unlike amphetamine, amphetamine-like substances accessible on the drug market are less expensive and more easily available; they also produce hallucinogenic effects expected by the users. Such properties render them more attractive as compared to amphetamine. On the other hand, the knowledge of the toxicity of these compounds is very limited, what in consequence generates problems that create ever-expanding research areas, including analytical, clinical and medicolegal issues, thus leading to development of systemic databases. An example here is paramethoxyamphetamine (PMA), which appeared on the drug market in recent years as a result of creative inventiveness of producers of psychoactive substances, who aimed at PMA replacing the popular ecstasy (MDMA) as a less expensive and more available product. It is more potent than MDMA, but has a slower onset of action, which encourages users to take more. The problem is illustrated in the present paper by three fatal cases involving PMA, which were comprehensively investigated taking into consideration case histories, pathological and toxicological findings obtained with the use of LC-MS-MS method. In blood samples taken from all the three victims, very high concentrations of PMA were found (in the range of 10-27mg/L) and thus the cause of deaths was determined as overdoses of PMA with the underlying mechanism of acute cardiorespiratory failure.

New psychoactive substance α-PVP in a traffic accident case.

The problems of new psychoactive substances (NPSs), especially related to drivers, constitute an open research area. In this case report, we present a traffic accident case, in which two passengers of five individuals died instantly, while the other three persons survived the accident with minor injuries only. From the blood samples of the driver and the passengers, α-pyrrolidinovalerophenone (α-PVP), an NPS belonging to the category of cathinone derivatives, was disclosed. Therefore, we established a detailed procedure for analysis of α-PVP in blood samples by liquid chromatography-tandem mass spectrometry. After careful validation tests of this method, α-PVP concentration in blood samples from the surviving driver and passengers, and from the two deceased, were measured. The concentrations varied from 20 to 650 ng/mL. Access to detailed information originating from the court files and from explanations provided by the driver and eye witnesses revealed extremely valuable illustrative details addressing the symptoms and pharmacological effects of α-PVP on the human organism, thus contributing to enriching the body of knowledge of α-PVP abuse.

The threats for human health induced by food pests of Plodia interpunctella as reservoirs of infectious microbiota / Zagrozenia zdrowia czlowieka indukowane przez szkodniki zywnosci Plodia interpunctella jako rezerwuary infekcyjnych microbiota

INTRODUCTION: World-wide distributed pests of Plodia interpunctella occur with increasing frequency also in Poland, in areas where food is prepared and stored, in dwellings, buildings of public use, hospitals. Larvae damage various products causing economic losses. There were no data about microbiota transmission by pests. The aim of our systematic studies firstly conducted in Poland was to explain a role of pests as reservoirs of microbiota and assess health risk induced by them in human environments. MATERIAL AND METHODS: 300 adults and 200 larvae, collected in households and health facilities by traps and directly from products, were examined by light microscopy, in vitro cultivations, molecular techniques; the susceptibility /resistance of microbiota to chemicals was also assessed. RESULTS: Gram+ bacteriae of genera Enterococcus, Micrococcus, Bacillus, Gram-: Klebsiella, Escherichia, mold fungi: Aspergillus, Penicillium and yeast-like fungi were identified, including strains potentially pathogenic for humans. CONCLUSIONS: In the European Union countries, the food circulation is audited by the law; chemicals are applied to eliminate P.interpunctella pests causing economic losses. Our successive studies showed that pyralids may generate health problems as food pests and as reservoirs of microbiota. Sources of the pathogenic, drug-resistant strains revealed by us, not identified earlier, may be particularly dangerous for elder persons, with weakened immune system, persons from groups of high risk of infections. The increased awareness of the problem is necessary for more efficacy of preventive measures. A monitoring of consequences of the health risk induced by the pests may supply data useful for adequate practical approach.

[Evaluation of chemical compounds acute poisoning risk among Kraków inhabitants in 1966, 1983 and 2015].

The total number of out-patients and hospitalized poisoned patients above 14 years old in Krakow in 1966 was 520 and 2132 in 1983. The incidence of poisoning per 10 000 of Kraków population in 1966 was 25.0 and increased to 36.2 in 1983. The mortality in 1966 was 0.75%. It was low also in 1983 ­ 1.01%, whatever reached 7.8% if forensic data were included (the fatalities on the spot). There were 847 poisoned patient hospitalized in Clinic of Toxicology UJ CM in 2015 and mortality in this group was 1.06%. Forensic analysis shown decreasing number of deaths on the spot (154 in 1983 and 89 in 2015). The percentage of poisoned with ethanol was declining (78.9% in 1983 and 57.3% in 2015). The frequency of patients treated in intensive care unit were increased from 6% in 1966 to 21.7% in 2015. The percentage of addicted acute intoxicated patients was lower (8%) in 1983 and achieved 21.7% in 2015. Poisoning structure, particularly pharmaceuticals, significantly influenced medical procedures performed in intensive care unit, especially administration of antidotes (5% in 1966 and 48.9% in 2015). Social evolution and changes of culture habits increased percentage of addiction but the number of deaths caused by ethanol on the spot was smaller.

Opium alcaloids in toxicological medico-legal practice of Department of Forensic Medicine, Jagiellonian University Medical College / Alkaloidy opium w toksykologicznej praktyce medyczno-sadowej Zakladu Medycyny Sadowej UJ CM.

Most likely, opium was the first narcotic substance discovered at the dawn of mankind. Contemporary drug abuse predominantly poses a social and clinical problem and encompasses among other aspects emergency procedures in cases of intoxication and treatment of addictions. On the other hand, this is also a problem of the judicial system, which implements the rule of apt punishment in criminal cases (rapes, robberies, drivers, production and trade in narcotic substances) and of the necessity of monitoring drug-associates deaths. In all drug-associated cases, investigative capabilities have increased with the introduction of extremely sensitive and specific analytical methods (GC-MS, LC/MS, HPLC/DAD) allowing for detection and identification of multi-component mixtures of xenobiotics found at low concentration levels in complex biological matrices. The history of the Krakow Department of Forensic Medicine dates back to the year 1877, since archival materials have been kept since that time. Isolated deaths resulting from morphine poisoning, mostly involving individuals employed in the health care sector, constituted the subject of medico-legal expert opinions starting at the beginning of the 20th century, but only the eighties did bring the need for multidirectional toxicological examinations of opiates and their metabolites in diversified biological and non-biological materials. The present report, in addition to the historical background of opiate addiction, discusses selected problems derived from published by Department reports on opiates, including cases of fatal intoxication, hair analysis of drug addicts in its various aspects, interactions in cases of poisoning and others.