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New psychoactive substances (NPS) are present on the Italian illicit markets, but data from the analysis of biological samples to evaluate their real consumption are rare. For this reason, an epidemiological study was carried out by means of a ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS-MS) method for the determination of 115 NPS on the keratin matrix. A total of 847 hair samples were collected in 2020 and 2021 and analyzed. The sample donors were in the age range of 18–40 years, from both genders, and were tested either for driving relicensing or for drug withdrawal monitoring. The UPLC–MS-MS system consisted of a Waters ACQUITY UPLC® I-Class, coupled with a Waters XEVO TQ-XS triple quadrupole mass spectrometer. The method was developed and fully validated according to international guidelines. Limits of detection were set as the minimum criterion to identify positive samples. Overall, 56 samples resulted positive for ketamine, 35 for norketamine, 6 for fentanyl, 3 for norfentanyl, 3 for 4-ANPP, 3 for MDMB-4en-PINACA, 2 for N , N -DMT, 2 for 5-chloro AB-PINACA, 1 for α-PHP and 1 for methcathinone. NPS were detected in a small part of samples (8.4%), which seems in contrast with their apparent wide diffusion in Italy, yet it is congruent with similar investigations based on hair analysis. Future studies will be performed to expand the investigated population, especially in terms of age and origin. [ABSTRACT FROM AUTHOR]

A retrospective study was carried out on postmortem examination data of 131 sharp force-related casualties recorded by the Brescia Institute of Forensic Medicine between 1982 and 2012. The objective was to identify relevant parameters that may be used to distinguish the manner of deaths between homicide, suicide, and accident. The following variables were considered: manner of death; demographic data; scene; type of sharp object; location and numbers of wounds; the presence of hesitation marks/defense wounds; toxicological findings; psychiatric history. There were 92 homicides, 28 suicides, and 11 accidents. Most victims were male, with an average age of 43. Injuries in isolation were present in 9.8% of homicides, in 35.7% of suicides, and in 54.5% of accidents. Most injuries involved the left anterior chest in homicides and the forearms in suicides. This study underlines the importance of a meticulous postmortem examination of injuries on the body and their relationship with other results from the death scene investigation. [ABSTRACT FROM AUTHOR]

Abstract: One hundred twenty-five unrelated individuals (69 females and 56 males) from Sassari (Northern Sardinia) and Orgosolo (Central Sardinia) were typed for 15 STRs loci. The 56 males were typed for 12 Y chromosome STRs loci too. Frequency distribution is described. [Copyright &y& Elsevier]

Abstract: Twelve Y-chromosome STRs—DYS19, DYS389-I, DYS389-II, DYS390, DYS391, DYS392, DYS393, DYS385, DYS437, DYS438, DYS439—were typed in a population sample (n =104) of unrelated males from Brescia (northern Italy). A total of 91 haplotypes were identified by the 12 Y-STR loci. The haplotype diversity (98.68%), discrimination capacity (87.50%) and gene diversity were calculated. [Copyright &y& Elsevier]

Abstract: The morphology evaluation of the sternal end of the fourth rib through the İşcan’s “phases method” is considered one of the most reliable method for age-at-death estimation of corpses of unknown identity. This study aims to develop a component model for age estimation alternative to İşcan’s method. A numerical score, ranging from 3 to 9, was assigned based on the morphology of surface (S), antero/posterior walls (W) and articular margins (M) of the sternal end of the fourth rib in 71 white female corpse. For each component a regression formula was determined. The best correlation between end rib morphology and age-at-death was obtained through the formula of multiple regression with two independent variables (articular surface and antero/posterior walls): Age (years)=6.37(S)+5.33(W)−17.15±3.3. [Copyright &y& Elsevier]

Abstract: One hundred and thirty unrelated individuals (80 females and 50 males) from Brescia (northern Italy) were typed for DXS8378, DXS7132, HPRTB and DXS7423 loci. Allele, genotype and haplotype frequencies were calculated. A comparison between our population data and others from Caucasians and Asians populations was performed. [Copyright &y& Elsevier]

Bones are among the structures most likely to be recovered after death. However, the low quantity of preserved DNA and complex processing from sample to DNA profile make forensic DNA analysis of bones a challenging task. Raman spectroscopy and gas chromatography-mass spectrometry (GC/MS), have the potential to be useful as screening tools for DNA analysis and in decomposition studies. The objective of this research was to assess the usefulness of such molecular investigations. Femur samples collected from 50 decomposing human bodies were subjected to Raman spectroscopy and GC/MS. Assessment of nuclear DNA quantity and short tandem repeat (STR) genotyping efficiency were also performed. Raman parameters (crystallinity, carbonate-to-phosphate ratio, mineral-to-matrix ratio) and detected lipids were recorded. Background fluorescence proved problematic for Raman analysis of forensic bones. Regardless, it was not associated with less preserved DNA or less detected STR alleles. Fatty acids, hydrocarbons, and five types of fatty acid methyl esters (FAMEs) were detected. The main phosphate peak position in Raman spectra was significantly correlated with preserved DNA (p = 0.03713), while significantly more STR alleles were detected in bones containing methyl hexadecenoate (p = 0.04236). Detection of FAMEs in the bone matrix suggests a reaction between methanol produced by bacteria and free fatty acids, which are not associated with the level of preservation of endogenous DNA. The techniques assessed have shown to be useful in molecular taphonomy studies and forensic genetics. • Raman spectroscopy and gas chromatography-mass spectrometry, are useful in decomposition studies. • the main phosphate peak position in Raman spectra was significantly correlated with preserved DNA. • significantly more STR alleles were detected in bones containing methyl hexadecenoate. • bacterial methanol and free fatty acids form esters during decomposition. • Raman analysis of forensic bones can be hampered by background fluorescence. [ABSTRACT FROM AUTHOR]

• "Complex suicides" means two or more methods of death are applied by the victim. • Two suicides methods may be planned in advance to prevent failure (planned). • A second suicide method may occur when the first one is not effective or too painful (unplanned). • In complicated suicides the chosen method fails and it's followed by a secondary and unforeseen trauma. • Multiple lesions on the victim's body are not in themselves indicative of the intervention of third parties in death. The role of the forensic pathologist is central to both identifying the cause and determining the manner of death. Distinguishing a suicide from a homicide or accidental event is essential to define whether third parties are involved in death. Suicides are most frequently performed using a single method; therefore, they can be defined as simple. The term "complex suicide" refers to a form of suicide in which two or more methods are applied by the victim, simultaneously or in chronological succession, to achieve the death. The different methods may have been planned in advance to prevent failure of the first method or may occur because the first method was not effective or was too painful, so the victim quickly seeks another way to complete the suicide. "Complicated suicides", on the other hand, are characterised by an unintentional secondary trauma following the suicidal act. This study analyses 35 complex suicides and 4 complicated suicides investigated at the Institute of Forensic Medicine in Brescia (Italy) during the period 1983–2022. Some data about the cases are discussed. This study aims to demonstrate how multiple lesions on the victim's body are not in themselves indicative of the intervention of third parties in their production, but complex and complicated suicides must always be considered. [ABSTRACT FROM AUTHOR]

Human decomposition in sea water poses several challenges to forensic practitioners tasked with the analysis of drowned bodies. Postmortem changes in the marine environment have not been extensively investigated and the current literature is mainly based on bodies retrieved from shallow waters or on limited samples. On 18 April 2015, a fishing boat carrying allegedly 1,000 migrants sank in the Mediterranean Sea. In a fifteen-month time span, humanitarian missions were carried out to recover the bodies from the sea. The present study investigates postmortem changes on the drowned victims in a non-sequestered environment in the Mediterranean Sea. A retrospective study was performed by two observers on the autopsy photographic records in the series of bodies recovered from the open sea. For 184 bodies, the postmortem changes were evaluated according to facial, body, limb, and total aquatic decomposition score (FADS, BADS, LADS, TADS, respectively). Furthermore, a modification to the current scoring system that divides upper and lower limbs was suggested. The interobserver agreement was assessed using Krippendorff's alpha coefficient. Possible relations between the decomposition scores and PMSI were investigated with Pearson correlation analysis. According to the sequence of the recovery missions, increasing FADS, BADS, LADS, and TADS were observed. The modified scoring system showed a strong agreement between observers, allowing a more accurate description of the actual extent of decomposition. The scores showed a significant relation with the PMSI (p < 0.01). Prolonged submersion in the open marine environment was confirmed to show increasing decomposition rates, from moderate decay to full disarticulation. This study provides a descriptive unicum of the postmortem changes in the open sea, which may contribute to strengthen the discipline and aid description of bodies recovered in similar circumstances, especially if a body needs to be associated to a disaster or period with respect to another, thus facilitating families or authorities in the search for specific victims. [ABSTRACT FROM AUTHOR]

Aim To investigate allele distribution and genetic parameters of a population-based sample from Maghreb region. Methods Allele frequencies for 5 new autosomal short tandem repeat (STR) markers (D10S1248, D22S1045, D2S441, D1S1656, and D12S391) and several forensic parameters were determined for 95 unrelated individuals. Results The combined power of discrimination and power of exclusion for the 5 loci were high (0.9999991 and 0.9954757, respectively). Allele frequencies were compared with previously published population data. Significant differences were found between Maghreb population and all other populations at the locus D2S441. Also, significant differences were found between the Maghreb and the African American population at the D22S1045, D1S1656, and D12S391 loci, between Maghreb and Caucasian population at the D1S1656 locus, and between Maghreb and Hispanic population at the D22S1045 locus. Conclusions Typing of the 5 new STR loci may provide a useful addition to the previously established sets of autosomal STRs. [ABSTRACT FROM AUTHOR]

In forensic genetics, the analysis of postmortem bones is one of the most challenging due to the low quantity of degraded endogenous DNA. The most widely used approach for sample preparation, in those cases, is pulverizing the bone. However, processing pulverized bone is extremely delicate, requiring strict laboratory conditions and operating procedures. In fact, several recent publications have focused on non-powder approaches. The objectives of this study were, thus, to validate a non-powder protocol for DNA extraction from forensic bones and an alternative pretreatment, large fragment demineralization (LFD). Thirty human femurs and tibiae received by the Legal Medicine Institute of Brescia, Italy, were included in the study. Bone powder and one transversal section of the diaphysis were sampled from each bone. DNA extraction from the powder was carried out using PrepFiler BTA (BTA), while the transversal section was submitted to the alternative demineralizing pretreatment (LFD) followed by DNA extraction using the QIAamp DNA Investigator. DNA extracts were assessed for human DNA quantity and degradation by means of a validated in-house qPCR assay and amplified with commercial kits. Inhibition assessment was carried out through Quality Sensor analysis using 24plex QS Kit. The differences in quantity, quality of human DNA, and number of alleles detected between both methods were comparable and not statistically significant. We propose the use of the LFD protocol as a complementary approach capable of confirming the genotypes or detect alleles not observed using BTA, without the need for pulverization. [ABSTRACT FROM AUTHOR]

Complete decapitation as a result of a suicidal hanging is rarely observed in forensic practice. The authors report a case of a 38-year-old man hanging from a bridge with a 4.2 m nylon rope. The man, weighing 70 kg, was wearing a 6.5 kg rucksack. The headless body was found lying in the river below the bridge and his head was found the next day in the river 6 m away from the bridge. Autopsy findings recorded a regular and concentric skin abrasion pattern and cervical spine transection between C2 and C3. Radiologic study depicted C2-C3 neck transection and the fracture of the left lamina and the spinous process of the axis. The force applied to the neck and the kinetic energy were calculated. [ABSTRACT FROM AUTHOR]

Aim To report on the successful analysis of amplicons obtained with PowerPlex® Fusion 6C System, a highly robust 27-plex genotyping kit developed for human identification laboratories, on the Applied Biosystems® 3500 Genetic Analyzer. Method We performed characterization and evaluation studies following the Scientific Working Group on DNA Analysis Methods (SWGDAM) validation guidelines, examining several critical areas of kit performance. We report the results of sensitivity, robustness, heterozygous peak height ratio, precision, concordance, caseworks, and mixture interpretations. We tested sensitivity, using serial dilutions of control DNA. Results The minimum amount of input DNA resulting in a full profile was 125 pg. Inhibition, inducted by urea, showsed a progressively fragmentation of DNA and a full profile was obtained until 1M of inhibitor factor. To test the profile quality, casework samples were extracted with different extraction methods: Chelex®100, QIAmp DNA Micro Kit and Phenol-Chloroform extraction. The results demonstrated that extraction chemistries do not have affect on amplification performance. Concordance check was performed by typing some casework samples and comparing the typing results with those obtained with other available kits. Thus, concordance was expected and supported by the data. Conclusion Reliable DNA typing results can be obtained using this new kit, demonstrating its effectiveness and utility in forensic analysis. [ABSTRACT FROM AUTHOR]

• The medico-legal assessment of physical violence is crucial. • The medico-legal limits are important and well-known by practitioners. • The Istanbul Protocol's invitation to a multidisciplinary approach is desirable. • The correct assessment of injuries guarantees a proper and consistent help for refugees. The Istanbul Protocol section on medicolegal evaluation delineates the guidelines for the correct management of the physical examination and the methods of assigning the degrees of consistency. Considering that most cases exhibit highly heterogeneous lesions, the examiner is forced to rely on his own experience, and their evaluation may be very subjective. The purpose of this work is to understand how subjective such an evaluation may be and whether the "experience" factor, interpreted as years dedicated to this profession and the number of cases evaluated, is statistically significant. To this end, a survey containing eleven cases of pre-evaluated asylum seekers was sent to thirty Italian clinical forensic practitioners. The participants were invited to assign a degree of consistency to each case according to the Istanbul Protocol guidelines, besides answering a few questions regarding their professional record. The doctors were divided into groups based on the number of cases evaluated and the experience collected expressed in years, and then interobserver analysis was performed. Results showed that the Fleiss' Kappa coefficient acquired significant values when attention was turned to the sub-samples composed of more experienced participants. Therefore, the introduction of appropriately trained health professionals – "experts in migrations and torture" – could lower the risks of misinterpretation and make the assessment as reproducible as possible. [ABSTRACT FROM AUTHOR]

• This study evaluates the assessment protocol of mesothelioma with SEM-EDS. • The quantity of asbestos fibres was evaluated by SEM. • The SEM-EDS allows to identify the mineralogical species of each fibre. • The microscopic analysis should be introduced in the evaluation protocol. This study evaluates the assessment protocol that allows the correlation between the development of mesothelioma to a specific exposure, with particular focus on investigations with Scanning Electron Microscope with Energy Dispersion Spectroscopy. This retrospective study includes 80 subjects who died from mesothelioma in the period 2001–2019. A judicial autopsy was performed for each case to confirm cause of death and correlate the disease with specific asbestos exposure. In 28 cases investigations were carried out to determine the pulmonary load of the asbestos fibres and corpuscles in the lung tissue through microscopic investigations, in order to confirm the suspicion of occupational exposure. Our data agree with the scientific literature reported, but it is interesting to underline how the present study uses a different systematic approach than others, which are mainly based on epidemiological and environmental studies without considering the lung content of fibres and corpuscles. It would be desirable that the use of the microscopic analysis was introduced in the evaluation protocol: it should always be carried out if the suspicion of asbestos-related disease is raised and not only as a possible integration to the less expensive anamnestic evaluation, even more so if the work or personal history should be suggestive of exposure to asbestos fibres. [ABSTRACT FROM AUTHOR]

Violence against women is a violation of human rights, crossing all cultures, classes, levels of education, earnings, ethnic and age groups. We conducted a retrospective study to review forensic records of sexual assault examinations carried out in different Italian health facilities and to correlate these findings with the results of the forensic DNA analyses. The goal was to determine which factors could have affected the obtained results, to identify the fundamental aspects to search for while examining a sexual assault victim in order to gather useful evidence to identify the offender and reconstruct the dynamics of the fact. We analysed 102 cases that occurred between 2006 and 2017, coming from ten participating laboratories. Despite a relatively limited number of cases, this study shows that the ability to ascertain the presence of male biological material in the samples collected is not a problem for forensic laboratories and seems to be influenced by other factors, such as how much time elapsed between the event and the sampling, the availability of the aggressor's biological material on the victim and the identification of biological fluids/stains. Therefore, the need for health structures to adopt specific protocols has been highlighted. It is necessary for health structures to define specific pathways and adopt homogeneous procedures or operational protocols, and it is essential to provide adequate training for health personnel. The results of the study could be useful in drafting and revising protocols/guidelines implemented in Italian hospital. Issues related to the limited number of analyses requested by Italian Authorities are also discussed. [ABSTRACT FROM AUTHOR]

Inferring the ethnic origin of individuals by means of short tandem repeat (STR) profiles has received considerable attention recently. Gene frequency variation among human populations has been extensively documented, and it has been suggested that the differences in allele proportions between ethnic groups could form the basis of an inferential system. We report the use of DNA profiles from 13 STR loci for inferring the ethnic origin of samples of unknown provenance using five populations of immigrants. This preliminary work shows that a population assignment test can be already used in real casework studies. [Copyright &y& Elsevier]