A case report of complex suicide in physician: attempt drugs poisoning and adhesive tape asphyxia.

Complex suicides are rare occurrences that can be categorized into planned (or primary) cases and unplanned (or secondary) cases. Additionally, individuals often select suicide methods based on their availability and accessibility. The body of a 58-year-old man was discovered deceased inside his medical office. He was found seated on the waiting room sofa, with his airways obstructed by several layers of adhesive tape wrapped around his head. An intravenous needle was observed in his left arm, and on the table in front of him, an empty 50 ml syringe, two empty vials of 10 ml potassium chloride, and an empty 10 mg vial of Valium (diazepam) were found. A roll of adhesive tape, similar to the one around his head, was also present. The autopsy, conducted 36 h after the body's discovery, revealed therapeutic concentrations of diazepam and its metabolite nordiazepam in the blood samples, while potassium chloride was not detected. Integrating forensic findings obtained from autopsy, histology, and other postmortem investigation, including toxicological analysis, can aid in defining suicidal behavior and preventing misinterpretation, particularly in differentiating diagnosis between homicide and suicide. It is crucial to consider circumstantial data and professional knowledge in such cases.

Morphological and molecular characterization of puparia of Piophilidae species of forensic relevance.

Piophilidae are a small family of Diptera with a worldwide distribution and which are historically associated with human activities. In addition to their economic importance, piophilid larvae can also be of medical and legal relevance. Within a medicolegal context, piophilids are frequently associated with cadavers in advanced stages of decomposition, thus being potentially useful forensic indicators and they have been reported also from archaeo-funerary contexts. An accurate species identification is therefore an essential prerequisite to ensure the reliable analysis of insect material in medical, forensic and archaeological investigations. Identification of the adult piophilid flies is possible because of the availability of identification keys, in contrast immature insects, especially puparia, have been poorly investigated and described. In this paper, puparia of 11 species of forensic interest (Piophila casei, Piophila megastigmata, Parapiophila atrifrons, Parapiophila flavipes, Parapiophila vulgaris, Protopiophila litigata, Liopiophila varipes, Prochyliza nigrimana, Prochyliza xanthosoma and Stearibia nigriceps in subtribe Piophilina and Centrophlebomyia furcata in subtribe Thyreophorina) are described and a molecular analysis, based on the COI sequencing, is presented to show the potential of the molecular approach in their identification.

Forensic investigative issues in a fireworks production factory explosion.

Since their discovery in ancient China, fireworks rapidly spread throughout the world, where they have always been used to celebrate either popular or private events. Their use is nonetheless related to several risks, especially within production factories, since several injuries or even death can occur following an accidental ignition. In cases of major disasters related to fireworks explosions, stating the accidental or intentional nature of the event might prove challenging, thus raising the need of a multidisciplinary approach. In this regard, we here discuss the case of an accidental explosion that occurred in a fireworks production factory, accountable for five deaths and two hospitalisations.

Mentha pulegium L.: A Plant Underestimated for Its Toxicity to Be Recovered from the Perspective of the Circular Economy.

The aim of the study was to investigate the micromorphology of Mentha pulegium leaves and flowers harvested in three different Sicilian (Italy) areas with peculiar pedo-climatic conditions, and to characterize the phytochemical profile, the phytotoxic activity, and the eco-compatibility of their essential oils (EOs) for potential use as safe bioherbicides. Light microscopy (LM) and scanning electron microscopy (SEM) highlighted that M. pulegium indumentum consists of non-glandular and glandular trichomes of different types. Peltate trichomes of plants from the different sites showed few significant differences in dimension and abundance, but they were characterized by a surprisingly high number of secretory cells both in leaves and flowers. Phytochemical analyses showed that oxygenated monoterpenes were the most abundant class in all the EOs investigated (92.2-97.7%), but two different chemotypes, pulegone/isomenthone and piperitone/isomenthone, were found. The complex of morphological and phytochemical data indicates that soil salinity strongly affects the expression of the toxic metabolite pulegone, rather than the EO yield. Phytotoxicity tests showed a moderate activity of EOs against the selected species as confirmed by α-amylase assay. Moreover, the low toxicity on brine shrimp provided a rationale for the possible use of investigated EOs as eco-friendly herbicides.

Potential treatment of leachate by Hermetia illucens (Diptera, Stratyomyidae) larvae: Performance under different feeding conditions.

In this study, the ability of H. illucens larvae (black soldier fly (BSF)) to metabolise different semisolid biowastes (e.g. kitchen waste, animal manure) has been applied to the treatment of landfill leachate. A testing programme has been developed by mixing leachate with three different solid supports: wheat bran, a biodegradable nutrient substrate, brewers' spent grain, a biodegradable nutrient residue from the brewery industry and sawdust, a low biodegradable residue from the wood industry. Larvae growth rate was monitored in terms of weight variation, mortality and time to reach the prepupal stage. Prepupal biomass composition was analysed in terms of crude protein, lipids and fatty acids. Substrates were monitored at the beginning and the end of tests for total solids (TS), total organic carbon (TOC), total Kjeldahl nitrogen (TKN), ammonia and (whenever significant) the 7-day Respirometric Index (RI7). The best performance was observed with wheat bran and brewers' spent grain, achieving an average larval weight ranging from 155.1 to 226.1 mg (w/w) with prepupation of more than 80% over 21 days. The initial TS, TOC and nitrogen content in feeding substrates had been metabolised (gasified and accumulated in prepupal biomass) by approximately 55%, 60% and 48%, respectively. Dry mass characterisation displayed a significant content of fats and proteins. The analysis demonstrated the suitability of BSF prepupal biomass for the production of biodiesel; however, the potential use of proteins as an animal feed needs further studies for assessing the presence of contaminants.

The iliac crest in forensic age estimation: evaluation of three methods in pelvis X-rays.

Age assessment in children and young adults is a relevant forensic issue. It is requested not only in evaluating criminal responsibility in youths without proper identification documents, often in relation to age thresholds, but also for release of a residency permit, and asylum seekers of minors. The analysis of the Risser sign and the iliac crest ossification process has been proposed as a useful tool for forensic age estimation. We evaluated the applicability of three different age estimation methods on a sample of 497 pelvic radiographs of Italian individuals between 10 and 25 years of age. Each method showed high reliability for both reproducibility and repeatability. The staging technique inspired by Kreitner and Kellinghaus methods (KK-MS) is easier than the Risser method in applicability, as it is not affected by the variations of ossification. We observed that all subjects who attained stage 3c of KK-MS and 5 of Risser Fr were >14 years, suggesting the benefits of these methods for that age threshold. The applicability of the area measurement method, inspired by Cameriere's approach, ranged between 12 and 20 years, but the statistical analysis showed only a moderate correlation with age. In order to evaluate the possible use of this approach it is therefore necessary to clarify and exclude external factors influencing the parameter. In conclusion, the iliac crest ossification is of interest in age estimation for forensic purposes. The evaluation of the pelvis X-ray, in addition to the other common dental and skeletal methods, could become a useful supplementary tool in age estimation for the 14 year threshold in order to fulfill forensic-level requirements.

Unexpected features of Drosophila circadian behavioural rhythms under natural conditions.

Circadian clocks have evolved to synchronize physiology, metabolism and behaviour to the 24-h geophysical cycles of the Earth. Drosophila melanogaster's rhythmic locomotor behaviour provides the main phenotype for the identification of higher eukaryotic clock genes. Under laboratory light-dark cycles, flies show enhanced activity before lights on and off signals, and these anticipatory responses have defined the neuronal sites of the corresponding morning (M) and evening (E) oscillators. However, the natural environment provides much richer cycling environmental stimuli than the laboratory, so we sought to examine fly locomotor rhythms in the wild. Here we show that several key laboratory-based assumptions about circadian behaviour are not supported by natural observations. These include the anticipation of light transitions, the midday 'siesta', the fly's crepuscular activity, its nocturnal behaviour under moonlight, and the dominance of light stimuli over temperature. We also observe a third major locomotor component in addition to M and E, which we term 'A' (afternoon). Furthermore, we show that these natural rhythm phenotypes can be observed in the laboratory by using realistic temperature and light cycle simulations. Our results suggest that a comprehensive re-examination of circadian behaviour and its molecular readouts under simulated natural conditions will provide a more authentic interpretation of the adaptive significance of this important rhythmic phenotype. Such studies should also help to clarify the underlying molecular and neuroanatomical substrates of the clock under natural protocols.

Drosophila circadian rhythms in seminatural environments: Summer afternoon component is not an artifact and requires TrpA1 channels.

Under standard laboratory conditions of rectangular light/dark cycles and constant warm temperature, Drosophila melanogaster show bursts of morning (M) and evening (E) locomotor activity and a "siesta" in the middle of the day. These M and E components have been critical for developing the neuronal dual oscillator model in which clock gene expression in key cells generates the circadian phenotype. However, under natural European summer conditions of cycling temperature and light intensity, an additional prominent afternoon (A) component that replaces the siesta is observed. This component has been described as an "artifact" of the TriKinetics locomotor monitoring system that is used by many circadian laboratories world wide. Using video recordings, we show that the A component is not an artifact, neither in the glass tubes used in TriKinetics monitors nor in open-field arenas. By studying various mutants in the visual and peripheral and internal thermo-sensitive pathways, we reveal that the M component is predominantly dependent on visual input, whereas the A component requires the internal thermo-sensitive channel transient receptor potential A1 (TrpA1). Knockdown of TrpA1 in different neuronal groups reveals that the reported expression of TrpA1 in clock neurons is unlikely to be involved in generating the summer locomotor profile, suggesting that other TrpA1 neurons are responsible for the A component. Studies of circadian rhythms under seminatural conditions therefore provide additional insights into the molecular basis of circadian entrainment that would otherwise be lost under the usual standard laboratory protocols.

A combined protocol for identification of maggots of forensic interest.

In Forensic Entomology the estimation of the age of insects is used for the estimation of the minimum post-mortem interval. As insect development is temperature dependent and species specific, a correct species identification is therefore fundamental. In the majority of cases the molecular identification is based on a destructive approach. In this paper a working protocol for molecular identification of fly larvae without affecting the anatomical characters used for morphological identification is presented. The suggested technique allows the preservation of the larval exoskeleton and of the unused soft tissues in the same vial allowing a repetition of both the morphological and molecular identification and reducing the risk of loss of the evidence. This method also allows the possibility of measuring the size of the specimens before their morphological and biomolecular characterization. In order to demonstrate that this technique can be applied on maggots of a large spectrum of dimensions it has been tested and validated using larvae of different size from ~1.7-1.3cm [Calliphora vomitoria and Lucilia sericata (Diptera: Calliphoridae)] to ~10-6.5mm [Musca domestica (Diptera: Muscidae) and Megaselia scalaris (Diptera: Phoridae)]. The importance of a unique identifier and of a complete database with all the specimen information (origin, sample size, identification, etc.) is also discussed.

Splitting hairs: differentiating between entomological activity, taphonomy, and sharp force trauma on hair.

PURPOSE: The analysis of hair can provide useful information for the correct evaluation of forensic cases, but studies of trauma on hair are extremely rare. Hair may present lesions caused by traumatic events or by animals: in fact, signs of sharp force weapons on hair may provide important information for the reconstruction of the manner of death, and, for example, may suggest fetishist practice. This study stemmed from a judicial case where it was fundamental to distinguish between sharp force lesions and insect activity on hair. METHODS: In order to highlight differences between sharp force lesions and insect feeding activity, different experiments were performed with high power microscopy: hair samples were subjected to several lesions by blunt and sharp force trauma; then samples were used as pabulum for two taxa of insects: the common clothes moth (Tineola bisselliella Lepidoptera, Tineidae) and the carpet beetle (Anthrenus sp., Coleoptera, Dermestidae). Hairs were examined from a macroscopic and microscopic point of view by stereomicroscopy and scanning electron microscopy (SEM): the morphological characteristics of the lesions obtained from the different experimental samples were compared. RESULTS: Results show that sharp force trauma produces lesions with regular edges, whereas insects leave concave lesions caused by their "gnawing" activity. These two types of lesions are easily distinguishable from breaking and tearing using SEM. CONCLUSIONS: This study demonstrates that insect activity leaves very specific indications on hair and sheds some light on different hair lesions that may be found in forensic cases.

New Insights into Geometric Morphometry Applied to Fish Scales for Species Identification.

The possibility of quick and cheap recognition of a fish species from a single dermal scale would be interesting in a wide range of contexts. The methods of geometric morphometry appear to be quite promising, although wide studies comparing different approaches are lacking. We aimed to apply two methods of geometric morphometry, landmark-based and outline-based, on a dataset of scales from five different teleost species: Danio rerio, Dicentrarchus labrax, Mullus surmuletus, Sardina pilchardus, and Sparus aurata. For the landmark-based method the R library "geomorph" was used. Some issues about landmark selection and positioning were addressed and, for the first time on fish scales, an approach with both landmarks and semilandmarks was set up. For the outline-based method the R library "Momocs" was used. Despite the relatively low number of scales analyzed (from 11 to 81 for each species), both methods achieved quite good clustering of all the species. In particular, the landmark-based method used here gave generally higher R2 values in testing species clustering than the outline-based method, but it failed to distinguish between a few couples of species; on the other hand, the outline-based method seemed to catch the differences among all the couples except one. Larger datasets have the potential to achieve better results with outline-based geometric morphometry. This latter method, being free from the problem of recognizing and positioning landmarks, is also the most suitable for being automatized in future applications.

Decomposition and entomological colonization of charred bodies - a pilot study.

AIM: To use forensic entomological approach to estimate the post mortem interval (PMI) in burnt remains. METHODS: Two experiments were performed in a field in the outskirts of Milan, in winter and summer 2007. Four 60-kg pigs were used: two for each experiment. One pig carcass was burnt until it reached the level 2-3 of the Glassman-Crow scale and the not-burnt carcass was used as a control. In order to describe the decomposition process and to collect the data useful for minimum PMI estimation, macroscopic, histological, and entomological analyses were performed. RESULTS: In the winter part of the experiment, the first insect activity on the burnt carcass began in the third week (Calliphora vomitoria) and at the beginning of the fourth week an increase in the number of species was observed. In the summer part, adult flies and first instar maggots (Phormia regina) appeared a few minutes/hours after the carcass exposure. Both in winter and summer, flies belonging to the first colonization wave (Calliphoridae) appeared on burnt and control pigs at the same time, whereas other species (Diptera and Coleoptera) appeared earlier on burnt pigs. CONCLUSION: In forensic practice, burnt bodies are among the most neglected fields of entomological research, since they are supposed to be an inadequate substratum for insect colonization. Entomological approach for PMI estimation proved to be useful, although further studies on larger samples are needed.

A New, Non-Invasive Methodology for the Molecular Identification of Adult Sarcophagidae from Collections.

Correct species identification is the cornerstone of all scientific studies that involve insects. Alongside traditional morphological identification techniques, molecular identification based on the characterization and analysis of specific mitochondrial or nuclear gene regions is becoming commonplace. Despite the good results that can be achieved, DNA extraction usually involves invasive techniques that lead to the partial or total destruction of specimens. In this work, a non-invasive DNA extraction technique is described. The technique was tested on the abdomens of dry-preserved Sarcophagidae (Diptera) specimens collected between 1889 and 2015. This allowed for the correct identification of species without impairing diagnostic morphological structures useful for further studies.

[Diagnostic atypique d'une myiase]. / Atypical diagnosis of myiasis.

Muscoid larvae were observed on self-medicated dressing material loaded with purulent material taken from a 91-year-old hospital patient. These larvae were identified as Lucilia sericata. However, no larvae were found in the patient's tissues. The observation of larvae on dressings should not automatically lead to a diagnosis of cutaneous myiasis.

On-Site Inspection Form in Veterinary Cases: The Parma Veterinary Form.

The on-site inspection of the scene of an animal cadaver is crucial for a correct interpretation of the autopsy results, to determine the manner, method, and cause of death. This information plays a crucial role in the control of public health including the prevention of zoonoses. It is also fundamental for the recognition and the contrast of crimes against animals and to animal abuse phenomena, considered an alert sign of an anti-social or violent behavior of humans. Today the best veterinary procedure requires an accurate collection of the evidence at the scene that can be then handed to experts belonging to other forensic disciplines for further evaluation and data interpretation. In this paper authors suggest a form aiming to facilitate either the on-site and the autopsy activities, as a guarantee of the quality of the forensic process starting from the discovery scene up to the reconstruction of the case. Essential is training of non-medical personnel who often represent the first responder to be present on the scene. The form is inspired by the interdisciplinary form developed by the European Council of Legal Medicine and represents an initial tool to stimulate a multidisciplinary activity in close synergy with other forensic experts.

Multidisciplinary Forensic Approach in "Complex" Bodies: Systematic Review and Procedural Proposal.

The recovery of severely altered cadavers (i.e., extensively decomposed, mummified, charred or dismembered) can be a challenge for forensic pathologists due to the difficulties in identification, PMI estimation and manner and cause of death determination. In such cases, integrating routine approaches (autopsy, histology, toxicology) to more specific forensic branches can be fundamental to improving the investigative process. In this paper a systematic review using PubMed, Scopus and Web of Science databases has been performed. The aim was to evaluate the forensic approaches implemented in the management of severely altered bodies due to decomposition, mummification, skeletonization, charring or dismemberment (to which we refer to as "complex"), and the role of each approach in the solution of a case. Then, the literature revision results were used to propose a schematic flowchart summarizing the post mortem activities that can be performed in forensic practice, adaptable in relation to each case.

A review of morphological characters for the identification of three common European species of Sarcophaga s. str. (Diptera: Sarcophagidae), with an emphasis on female terminalia.

The subgenus Sarcophaga Meigen, 1824 (s. str.) currently comprises over 30 species distributed in the West Palearctic Region, the identification of which is normally based on characters of the male terminalia. Females of the three closely-related species Sarcophaga (Sarcophaga) carnaria (Linnaeus, 1758), S. (S.) subvicina Rohdendorf, 1937 and S. (S.) variegata (Scopoli, 1763), which are especially widespread and abundant in NW Europe, are considered morphologically indistinguishable by most authors. However, a few authors have proposed keys to separate females of these three species based on external and internal characters of the terminalia. Following a preliminary molecular identification using DNA barcode sequences (COI, cytochrome c oxidase subunit I), we herein revise the morphological characters used to differentiate female S. carnaria, S. subvicina and S. variegata in existing identification keys as well as search for additional diagnostic characters. Our results suggest that only one previously-proposed female character, namely the length to width ratio of abdominal sternite 7, can be used to separate S. subvicina from the other two species (Mann-Whitney U test: p < 0.0001), at least in a majority of cases. Other characters, such as the degree of sclerotisation and setation of tergite 8, show a high degree of overlap that does not allow to reliably separate females of these three species. Nevertheless, we propose a combination of characters that should allow the separation of female S. carnaria from female S. variegata in most cases. An additional analysis of males of the same species showed that the distribution of pruinosity and setation on syntergosternite 7+8, a character mentioned in a previously-published key, is also not reliable for identification.

Back to the Middle Ages: Entomological and Botanical Elements Reveal New Aspects of the Burial of Saint Davino of Armenia.

The natural mummy of Saint Davino Armeno (11th century) is preserved in the church of Saint Michele in Foro in the city of Lucca (Tuscany, Central Italy). The body of Davino is one of the oldest Italian mummies of a Saint, and his paleopathological study was performed in 2018. In the present research, we investigated the arthropod fragments and botanical remains collected from the body, coffin, and fabrics of Saint Davino. Entomological analyses outlined the presence of 192 arthropod fragments. Among these, Diptera, Muscidae (Hydrotaea capensis and Muscina sp.), and Phoridae (Conicera sp.) puparia were the most abundant. Regarding Coleoptera, Ptinidae (Anobium punctatum) were the most frequent, followed by Cleridae (Necrobia sp.), Trogidae (Trox scaber), Curculionidae (Sitophilus granarius), and Histeridae (Gnathoncus). Cocoons of Tineidae and Pyralidae moths were found, along with a propodeum joined to the petiole and a mesopleuron of an Ichneumoninae parasitoid. Numerous metamera of Julida and three scorpion fragments were also found. Botanical samples indicated the presence of a quite broad botanical community, including gramineous species, olives, evergreen oaks, and grapevine. Overall, entomological data allow us to argue that Saint Davino was first buried into the soil, probably in a wooden coffin, thus supporting the historical-hagiographic tradition according to which he was buried sub divo in the cemetery of Saint Michele. The preservation of the body as a natural mummy may have been facilitated by burial in a coffin that prevented direct contact of the corpse with the earth. Botanical remains offer confirmation of a late medieval urban environment rich in horticultural areas and trees, giving us a landscape that is very different from the current Tuscan city.

Puparia Cleaning Techniques for Forensic and Archaeo-Funerary Studies.

Diptera puparia may represent both in forensic and archaeo-funerary contexts the majority of the entomological evidence useful to reconstruct the peri and post-mortem events. Puparia identification is quite difficult due to the lack of identification keys and descriptions. In addition, external substances accumulated during the puparia permanence in the environment make the visualization of the few diagnostic characters difficult, resulting in a wrong identification. Six different techniques based on physical and chemical treatments have been tested for the removal of external substances from puparia to make identification at species level feasible. Furthermore, the effects of these methods on successful molecular analyses have also been tested as molecular identification is becoming an important tool to complement morphological identifications. The results of this study indicate that cleaning via warm water/soap, the sonication and treatment with a sodium hydroxide solution are the best methods to achieve a good quality of the samples.

Comparison of Accumulated Degree-Days and Entomological Approaches in Post Mortem Interval Estimation.

Establishing the post mortem interval (PMI) is a key component of every medicolegal death investigation. Several methods based on different approaches have been suggested to perform this estimation. Among them, two methods based their evaluation on the effect of the temperature and time on the considered parameters: total body score (TBS)/accumulated degree-days (ADDs) and insect development. In this work, the two methods were compared using the results of minPMI and PMI estimates of 30 forensic cases occurring in northern Italy. Species in the family Calliphoridae (Lucilia sericata, Calliphora vomitoria and Chrysomya albiceps) were considered in the analyses. The results highlighted the limits of the TBS/ADD method and the importance of the entomological approach, keeping in mind that the minPMI is evaluated. Due to the fact that the majority of the cases occurred in indoor conditions, further research must also be conducted on the different taxa to verify the possibility of increasing the accuracy of the minPIM estimation based on the entomological approach.