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Background Preliminary data suggested that fat embolism could explain the importance of visceral obesity as a critical determinant of coronavirus disease-2019 (COVID-19). Methods We performed a comprehensive histomorphologic analysis of autoptic visceral adipose tissue (VAT), lungs and livers of 19 subjects with COVID-19 (COVID-19+), and 23 people without COVID-19 (controls). Human adipocytes (hMADS) infected with SARS-CoV-2 were also studied. Results Although there were no between-group differences in body-mass-index and adipocytes size, a higher prevalence of CD68+ macrophages among COVID-19+ VAT was detected (p = 0.005) and accompanied by crown-like structures presence, signs of adipocytes stress and death. Consistently, human adipocytes were successfully infected by SARS-CoV-2 in vitro and displayed lower cell viability. Being VAT inflammation associated with lipids spill-over from dead adipocytes, we studied lipids distribution by ORO. Lipids were observed within lungs and livers interstitial spaces, macrophages, endothelial cells, and vessels lumen, features suggestive of fat embolism syndrome, more prevalent among COVID-19+ (p < 0.001). Notably, signs of fat embolism were more prevalent among people with obesity (p = 0.03) independently of COVID-19 diagnosis, suggesting that such condition may be an obesity complication exacerbated by SARS-CoV-2 infection. Importantly, all infected subjects’ lungs presented lipids-rich (ORO+) hyaline membranes, formations associated with COVID-19-related pneumonia, present only in one control patient with non-COVID-19-related pneumonia. Importantly, transition aspects between embolic fat and hyaline membranes were also observed. Conclusions This study confirms the lung fat embolism in COVID-19+ patients and describes for the first time novel COVID-19-related features possibly underlying the unfavorable prognosis in people with COVID-19 and obesity.

Mycobacterium chimaera is a non-tuberculous mycobacterium, member of the Mycobacterium avium complex (MAC), which has become a global public health concern due to infection following cardiac surgery performed with contaminated heater-cooler units. M. chimaera infection is characterized by a long latency, non-specific signs and symptoms and high mortality rates. Thus, the diagnosis is still challenging both for forensic pathologists and for clinicians. Clinical manifestations of M. chimaera infection include endocarditis, hepatitis, nephritis, encephalitis and chorioretinitis. A constant histopathologic finding is the presence of non-caseating granulomas, with multinucleated giant cells and histiocytes. Hereby, we present two cases of fatal disseminated M. chimaera infection following aortic valve surgery reporting clinical history and post-mortem findings. Further, we provide a brief overview of the literature with a special focus on histopathological characteristics of M. chimaera infection. The aim of this article is to provide a complete synopsis of histopathological characteristics useful for forensic pathologists.

According to the EU Early Warning System (EWS), synthetic cathinones (SCs) are the second largest new psychoactive substances (NPS) class, with 162 synthetic cathinones monitored by the EU EWS. They have a similar structure to cathinone, principally found in Catha Edulis; they have a phenethylamine related structure but also exhibit amphetamine-like stimulant effects. Illegal laboratories regularly develop new substances and place them on the market. For this reason, during the last decade this class of substances has presented a great challenge for public health and forensic toxicologists. Acting on different systems and with various mechanisms of action, the spectrum of side effects caused by the intake of these drugs of abuse is very broad. To date, most studies have focused on the substances’ cardiac effects, and very few on their associated neurotoxicity. Specifically, synthetic cathinones appear to be involved in different neurological events, including increased alertness, mild agitation, severe psychosis, hyperthermia and death. A systematic literature search in PubMed and Scopus databases according to PRISMA guidelines was performed. A total of 515 studies published from 2005 to 2022 (350 articles from PubMed and 165 from Scopus) were initially screened for eligibility. The papers excluded, according to the criteria described in the Method Section (n = 401) and after full text analyses (n = 82), were 483 in total. The remaining 76 were included in the present review, as they met fully the inclusion criteria. The present work provides a comprehensive review on neurotoxic mechanisms of synthetic cathinones highlighting intoxication cases and fatalities in humans, as well as the toxic effects on animals (in particular rats, mice and zebrafish larvae). The reviewed studies showed brain-related adverse effects, including encephalopathy, coma and convulsions, and sympathomimetic and hallucinogenic toxidromes, together with the risk of developing excited/agitated delirium syndrome and serotonin syndrome.

To define the Y-chromosome genetic structure in Apennine populations, 17 Y-chromosome short tandem repeats (Y-STRs) and 37 Y-single nucleotide polymorphisms (Y-SNPs) were typed in 162 subjects living in the upland area of the Marches (Central Italy). A total number of 155 haplotypes (haplotype diversity was 0.9994) and 14 SNP haplogroups were observed. Testing high-resolution Y-chromosome data sets, e.g. using Yfiler and SNPs, increases the discriminatory capacity in individual identification for forensic purposes. It is also useful in autochthonous population and micro-population studies to highlight the most informative loci for evolutionary aims.

This work describes an efficient and rapid test for typing 37 single nucleotide polymorphisms (SNPs) of the non-recombining region of Y chromosome (NRY) from a minimal amount of DNA using six PCR multiplexes. Markers were drawn following a hierarchical strategy based on the phylogenetic tree of Y chromosome proposed by the Y Chromosome Consortium [The Y Chromosome Consortium, A nomenclature system for the tree of human Y-chromosomal binary haplogroups, Genome Res. 12 (2002) 339-348]. Two multiplexes--arbitrarily named MY1 and MY2--were developed to explore the basal branches of the tree encompassing all the major clades A-R: MY1 for markers M35, M89, M172, M170, M9, M173, M45 and MY2 for markers M52, M216, M174, M181, M201, M91, M96, M214. Four multiplexes able of typing the more superficial branches typical of most frequent European haplogroups E3b, J2, R1 and I, were also developed and named MY-E3b (M78, M107, M224, M165, M148, M81), MY-J2 (M158, M68, M47, M102, M137, M67), MY-R1 (M17, M269, M18, P25, SRY10831.2) and MY-I (M72, M223, M26, M21, M161). SNP genotyping was carried out by hot-start PCR amplification with primers yielding fragments between 63 and 210 nucleotides, followed by minisequencing reaction based on dideoxy single-base extension and capillary electrophoresis of extension products. The sequential application of these multiplexes is a robust and effective resource for typing the most frequent European Y-SNP haplogroups, and appears to be suitable for forensic purposes and evolutionary studies.

This study reports the selection of 22 autosomal SNPs and the setting of PCR and minisequencing multiplexes suitable for forensic purposes.

In order to define the Y-chromosome genetic structure in Apennine populations, 17 Y-chromosome microsatellites and 37 Y-single nucleotide polymorphisms were typed in 81 subjects living in Fabriano and Urbino, two small towns in the upland area of the Marches (central Italy), speaking different dialects and submitted to a limited genetic flow.

DNA recovered at a crime scene often results as damaged; this represents enormous difficulty for the correct typing because of fragmentation or the lack of DNA region of interest. In this work a set of biological samples was prepared and stored under different conditions; STRs and SNPs typing was performed at regular interval of time in order to study the effects of natural DNA degradation. Allelic/locus drop-out phenomenon for the higher molecular weight loci or no results were obtained for microsatellite analysis after 1 week. SNPs typing gave positive results depending on storage conditions and type of substrate; a nucleotide alteration (C to T) was observed for M269 locus in a sample after 3 months.

The aim of this study is to set-up multiplex PCR of NRY single nucleotide polymorphisms (SNPs) suitable for forensic purposes. A first multiplex has been developed with SNP loci defining the European haplogroups (M35, M89, M172, M170, M9, M173, M45). PCR was performed with primers designed to produce amplicons in a range between 96 and 136 bp starting from 1 ng of DNA template. PCR product was minisequenced with tailed primers of different length and run in an automated five-colour capillary electrophoresis sequencer.

Congenital long-QT syndrome (LQTS) is a hereditary cardiac disease characterized by a high risk of life-threatening arrhythmias. Until recently, LQTS was exclusively a cardiac channelopathy. It was observed that the LQT3 associated, SCN5A-encoded cardiac sodium channel localizes in caveolae and it was hypothesized that mutations in caveolin-3 may represent a novel pathogenetic mechanism for LQTS (LQT9). Caveolae are characterized by the presence of caveolins, scaffolding proteins that interact with cholesterol and provide the structural framework for macromolecular signaling complexes. Using polymerase chain reaction and direct DNA sequencing, mutational analysis on CAV3 gene was performed on DNA extracted from 50 patients with LQTS diagnosis, but with no mutations on the entire coding regions of the major LQTS associated genes – KCNQ1, KCNH2, SCN5A, KCNE1, KCNE2, and KCNJ2 – and targeted analysis of ANK2 and RyR2. Mutational analysis of CAV3 in 50 unrelated LQTS subjects identified 9 mutations; two of them were found in the coding region. They are missense mutations and are located in a very conserved region. Mutations in the intronic sequences were analyzed by SpliceAid (www.introni.it), a web resource to predict the effect of the DNA mutations at the level of the target sequences of the RNA-binding proteins that determine the pattern of mRNA splicing. Some of them may alter the correct mRNA splicing. Further studies are needed to characterize the impact of these mutations in vivo.

At the beginning of 2006 more than 301,000 immigrants resident in Italy resulted to come from Tunisia and Morocco, 66% of which are male subjects; in addition, it is estimated that some other thousand are clandestine. Our data show that there is an increasing involvement of Tunisian and Moroccan individuals in paternity testing and in individual identification cases. For these reasons, the aim of this work was to enrich forensic Y-chromosome databases with Northern Africa data to better know markers frequency and their distribution across these populations (in YHRD there are 246 Tunisian samples and 0 Moroccans, access date to www.yhrd.org: August 2007). 103 Tunisian and Moroccan healthy male donors were typed by 17 microsatellites extended haplotype and 41 Y-SNPs. A high-resolution level database was created, including both haplotype and haplogroup for each sample. This study confirmed that precious informations might come both from Y-SNPs haplogroup distribution besides Y-STRs data.

All forensic laboratories routinely use commercial kits and softwares for automated typing; in rare cases genotyping misinterpretations or mislabellings occur. This study refers to the investigation on a D2S1338 off-ladder allele mislabelling observed in DNA profile of murdered woman. The Identifiler ® revealed heterozygosity in the range of D16S539, with a presumptive microvariant allele "14.2", based on assigned size, while PowerPlex ® 16 resulted in a homozygosity of allele "11". Singleplex amplification of D16S539 locus confirmed homozigosity. D2S1338 locus, the closest to D16S1338 in Identifiler ® , genotyped as homozigote "19", was singleplex amplified. The off-ladder peak was gel-isolated, sequenced and designed as a rare "11" allele variant [(TGCC)6(TTCC)5]. Genotype was finally designed as D16S539 "11,11" and D2S1338 "11,19". To avoid genotyping misinterpretations or mislabelling, ambiguous genotypes should be established by two commercial kits at least. Furthermore, off ladder alleles as well as allele microvariants should be assigned by direct sequencing. This issue should be considered in Criminal DNA database requirements, that is still under debate in Italy.

Background Sudden cardiac death (SCD) is the clinical outcome of a lethal arrhythmia that can develop on the background of unrecognized channelopathies or cardiomyopathies. Several susceptibility genes have been identified for the congenital forms of these cardiac diseases, including caveolin-3 (Cav-3) gene. In the heart Cav-3 is the main component of caveolae, plasma membrane domains that regulate multiple cellular processes highly relevant for cardiac excitability, such as trafficking, calcium homeostasis, signal transduction and cellular response to injury. Here we characterized a new putative Cav-3 variant, Cav-3 V82I, found in a patient with SCD. Results In heterologous systems Cav-3 V82I was expressed at significantly higher level than Cav-3 WT and accumulated within the cells. Cells expressing Cav-3 V82I exhibited a decreased activation of extracellular-signal-regulated kinases (ERKs) and were more vulnerable to sub-lethal osmotic stress. Conclusion Considering that abnormal loss of myocytes can play a mechanistic role in lethal cardiac diseases, we suggest that the detrimental effect of Cav-3 V82I variant on cell viability may participate in determining the susceptibility to cardiac death.