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Healthcare workers experienced high degree of stress during COVID-19. Purpose of the present article is to compare mental health (depressive and Post-Traumatic-Stress-Disorders—PTSD—symptoms) and epigenetics aspects (degree of methylation of stress-related genes) in front-line healthcare professionals versus healthcare working in non-COVID-19 wards. Sixty-eight healthcare workers were included in the study: 39 were working in COVID-19 wards (cases) and 29 in non-COVID wards (controls). From all participants, demographic and clinical information were collected by an ad-hoc questionnaire. Depressive and PTSD symptoms were evaluated by the Patient Health Questionnaire-9 (PHQ-9) and the Impact of Event Scale—Revised (IES-R), respectively. Methylation analyses of 9 promoter/regulatory regions of genes known to be implicated in depression/PTSD (ADCYAP1, BDNF, CRHR1, DRD2, IGF2, LSD1/KDM1A, NR3C1, OXTR, SLC6A4) were performed on DNA from blood samples by the MassARRAY EpiTYPER platform, with MassCleave settings. Controls showed more frequent lifetime history of anxiety/depression with respect to cases (χ2 = 5.72, p = 0.03). On the contrary, cases versus controls presented higher PHQ-9 (t = 2.13, p = 0.04), PHQ-9 sleep item (t = 2.26, p = 0.03), IES-R total (t = 2.17, p = 0.03), IES-R intrusion (t = 2.46, p = 0.02), IES-R avoidance (t = 1.99, p = 0.05) mean total scores. Methylation levels at CRHR1, DRD2 and LSD1 genes was significantly higher in cases with respect to controls (p p = 0.03 and p = 0.03, respectively). Frontline health professionals experienced more negative effects on mental health during COVID-19 pandemic than non-frontline healthcare workers. Methylation levels were increased in genes regulating HPA axis (CRHR1) and dopamine neurotransmission (DRD2 and LSD1), thus supporting the involvement of these biological processes in depression/PTSD and indicating that methylation of these genes can be modulated by stress conditions, such as working as healthcare front-line during COVID-19 pandemic.

Urinary bladder hamartoma is a rare benign proliferation with only 14 cases reported in the literature at present. Urinary bladder hamartoma is composed of a disorderly admixture of normal urinary bladder components, essentially represented by glands lined by transitional epithelium and a variable percentage of fibrous stroma, smooth muscle bundles, and adipose tissue. Urinary bladder hamartomas do not exhibit cytological or architectural abnormalities and show no necrosis or increase in mitotic activity. Clinical manifestations are usually represented by lower urinary tract symptoms, more or less frequently paired with gross hematuria. Several pediatric cases of urinary bladder hamartoma have been reported, sometimes with syndromic associations. Transurethral resection has been curative in all cases reported, with no evidence of recurrence. Here we report an additional rare urinary bladder hamartoma, clinically mimicking urothelial carcinoma, providing a review of the literature regarding this unusual entity.

Background and ObjectivesCoenzyme Q10(CoQ10)–deficient cerebellar ataxia can be due to pathogenic variants in genes encoding for CoQ10biosynthetic proteins or associated with defects in protein unrelated to its biosynthesis. Diagnosis is crucial because patients may respond favorably to CoQ10supplementation. The aim of this study was to identify through whole-exome sequencing (WES) the pathogenic variants, and assess CoQ10levels, in fibroblasts from patients with undiagnosed cerebellar ataxia referred to investigate CoQ10deficiency.MethodsWES was performed on genomic DNA extracted from 16 patients. Sequencing data were filtered using a virtual panel of genes associated with CoQ10deficiency and/or cerebellar ataxia. CoQ10levels were measured by high-performance liquid chromatography in 14 patient-derived fibroblasts.ResultsA definite genetic etiology was identified in 8 samples of 16 (diagnostic yield = 50%). The identified genetic causes were pathogenic variants of the genesCOQ8A(ADCK3) (n = 3 samples),ATP1A3(n = 2),PLA2G6(n = 1),SPG7(n = 1), andMFSD8(n = 1). Five novel mutations were found (COQ8An = 3,PLA2G6n = 1, andMFSD8n = 1). CoQ10levels were significantly decreased in 3/14 fibroblast samples (21.4%), 1 carrying compound heterozygousCOQ8Apathogenic variants, 1 harboring a homozygous pathogenicSPG7variant, and 1 with an unknown molecular defect.DiscussionThis work confirms the importance ofCOQ8Agene mutations as a frequent genetic cause of cerebellar ataxia and CoQ10deficiency and suggestsSPG7mutations as a novel cause of secondary CoQ10deficiency.

Purpose To report the unique case of a pair of phenotypically discordant monozygotic twins, with one of them affected by unilateral Coats disease. Case report Both patients underwent a complete ophthalmologic evaluation and were genetically tested with whole-exome sequencing (WES). Any known or unknown potential genetic determinant of Coats disease wasn't found. Conclusion It may suggest a non-genetic etiology for this disorder. This represents, to the best of our knowledge, the first case of genetic analysis of monozygotic twins, one of whom is affected by Coats disease. Further studies are warranted, including performing genetic analysis directly on retinal biopsy tissue.

Human and animal hairs have been used in forensic investigations for over a century. Hair is stable under adverse natural conditions; hence, it is often recovered at the crime scene, and it is necessary to determine whether the hair is of human or animal origin. Morphological and genetic characteristics are useful to differentiate human hair from animal hair. In the present study, we analyzed the distinguishing characteristics of hair of various species. In addition, we explore species identification by cytochrome c oxidase I mitochondrial gene analysis. We confirm that both the microscopic and molecular analyses of hairs are useful in forensic investigations.

The identification of human remains using DNA analysis can be extremely challenging and its success is certainly influenced by the time elapsed since death. In that context, intact teeth have been shown to be highly successful in DNA analysis. However, restored teeth are usually available and, surprisingly, these specimens have been poorly studied. In fact, there are no reports regarding forensic DNA analysis of those types of samples in real cases. Therefore, the aim of this study was to perform DNA typing on healthy and restored teeth from exhumed human remains, which had been buried for 46 years. A powder-free DNA extraction protocol specifically designed for teeth was followed and human DNA quantitation and degradation assessment was performed using an in-house qPCR assay. Samples were amplified with commercial human identification kits for autosomal and Y chromosome markers. The obtained DNA profiles were compared to those of a previously processed femur sample as well as a buccal swab from a putative son. One healthy and one restored tooth yielded complete, concordant and compatible DNA profiles with previously typed samples from the femur and the putative son. Biostatistical calculations supported the paternity relationship with a likelihood ratio greater than 11 million. The present study highlights the use of restored teeth in a real exhumation case and the powder-free approach specifically designed for the extraction of DNA from teeth is discussed.

The typing of short tandem repeats (STRs) is a common method used in forensic laboratories based on its good performance. However, because of the relatively large size of amplicons (150–500 bp), most of them are unsuitable for degraded DNA and low copy number samples. So, there is a need to find new genetic makers with smaller amplicon size. Recently, some forensic researchers focused their attention to alternative and supplementary genetic markers in the human genome: insertion-deletion polymorphisms (InDels). They are diallelic, of smaller size, widely distributed throughout the genome and with a lower mutation rate compared to STRs. Due to their characteristics, InDels can aid forensic analysis even when a very small amount of DNA is available in a sample. In this study we tested the ability of a set of 38 InDel markers on genotyping different forensic samples containing low DNA concentration, already typed using the Identifiler kit but obtaining poor results. Statistical analysis was performed using the STRs results with appropriated softwares. After, InDel typing results were added to the STRs statistic calculations. The purpose of this work was to verify the actual usefulness of InDels analysis in order to improve the weight of the statistical analysis of challenging samples.

For over 10 years, quantitative PCR (qPCR) for DNA quantitation has been reported in forensics. However, assays have not been described for small qPCR platforms. Thus, technological advancement is not always implemented in small forensic genetics laboratories. A duplex qPCR assay is reported, using a StepOne instrument and targeting a short and a long human DNA region. This study was performed according to international validation guidelines, including sensitivity, repeatability, reproducibility, precision, accuracy, contamination assessment, known and case-type samples, and degradation studies. Characterization of the genetic markers, species specificity, and population studies had already been conducted. Moreover, case-type samples were quantified, amplified using commercial kits and the number of alleles detected was recorded. Sensitivity was shown to be 10 pg/µL. Standard curve replicates demonstrated the assay is accurate, precise, as well as fairly repeatable and reproducible. The NGM Detect kit was shown to yield higher peaks than Identifiler Plus and NGM Select for degraded samples. Moreover, quality sensors were always present and proved useful. The quantification values of the large target showed a correlation with the number of alleles detected in the STR profiles for known and casework samples. The degradation index was shown to be informative, with a value of 10 or higher indicating dropout. It is suggested that after quantitation, samples with low or degraded DNA be amplified using newer amplification kits containing quality sensors to confirm that the low-quality profile was not affected by inhibition.

DNA profiling can identify an individual from a sample of biological material but it does not reveal what body fluid or tissue source the DNA profile originated from. In many cases it is important to know from what body fluid or tissue the DNA profile originated in order to provide crucial information necessary to the investigation, especially in cases where the victims are not able to give information about the dynamics of the event. For this purpose messenger RNA (mRNA) analysis has been shown to be a suitable method for the identification of body fluids, resulting in a trend to overcome the conventional approaches. Here we present the first report about case regarding a three-year-old child supposedly victim of a sexual assault with digital penetration. Thanks to the use of the combined DNA profiling and RNA analysis it was possible to demonstrate the sexual assault suffered by the victim.

The Applied Biosystems™ NGM Detect™ PCR Amplification Kit is a new Human Identification solution with increased sensitivity in comparison to NGM SElect Kit. The purpose of this study is to evaluate the performance of the NGM Detect™ kit compared to AmpFlSTR® NGM Select™ and AmpFlSTR® Identifiler® Plus kits, using a variety of casework samples. The study is being carried out on samples obtained from forensic cases such as sexual violence, exhumations, paternity, homicides and suspicious deaths. The sample typologies refer to: buccal swabs, nail swabs, swabs deriving from sexual violence, human biological tissues and liquids, clothes and objects of common use. The preliminary results show that the combined use of NGM Detect and NGM Select kits provide maximum information in a dual amplification strategy, as the size of several markers are complementary. Furthermore, the NGM Detect and NGM Select kits appear more sensitive than the Identifiler Plus kit. In conclusion, together with the NGM Select™ kit, the NGM Detect™ kit offers the possibility of recovering a lot of genetic information from degraded samples, delivering the best likelihood ratio value of identity compared to other ESS-focused solutions in the market today.

The discovery of human corpses in urban domestic settings does not constitute an unusual case in criminal casework. These scenarios can be very challenging to investigate since the uninformative evidences encountered also demand a multidisciplinary effort among several specialties in the forensic sciences field. The occurrence of this incident is usually accompanied by social isolation, which is an emblematic aspect of urban modern society. The elderly population is especially susceptible to being socially isolated, which is associated with higher mortality. We present a case report of an elderly woman who had been living with her husband's dead body, contributing to the scarce literature on the "Living with the Dead" phenomenon. The use of a multidisciplinary approach and the challenges that social isolation presents to forensic sciences and the contemporary society are discussed.

Atherosclerotic calcifications, as calcified atheromatous elements, are markers of cardiovascular disease. However, the literature gives little information regarding their morphological aspect, making their identification very rare in skeletonized cases. In this paper, we document the morphological, histological, and SEM aspects of atherosclerotic plaques collected from unclaimed cemeterial skeletal remains from an identified osteological collection and extracted from well-preserved cadavers autopsied at the medico-legal institute of Milan. Each of the three analyses provided similar results: atherosclerotic calcifications are convex-concave plaques with a stratified structure, a pale-yellow coloration in autopsy cases and yellow to brown when recovered in dry bone. Histologically, undecalcified and decalcified sections showed a stratified aspect formed by superimposed layers. Lastly, the SEM analysis showed a precise view of the stratified structure of the plaques in transverse section. As markers of disease, atherosclerotic calcifications can provide important antemortem information on the deceased that may be compared to antemortem data.

Objective The aim of this paper is to provide information on the morphology and composition of gallstones based on clinical samples in order to assist paleopathologists and bioarchaeologists in recognizing their presence in archaeological contexts. Materials and Methods 270 gallstones were extracted and macerated from autopsies conducted at the Istituto di Medicina Legale in Milan (Italy) in order to simulate a dry bone recovered from archaeological contexts. Morphological, histological, and elemental variation was documented. Results Gallstones vary in size, shape, color and texture. The cross-sectional surface correlates with chemical composition and is a valuable tool for classification into subcategories of stones. Histological analysis can confirm the classification. Elemental analysis yielded a higher frequency of carbon, calcium and phosphorus. Conclusions Although identification of gallstones in archaeological contexts can be challenging, familiarity with morphological, histological, and elemental variation can assist researchers in the field and laboratory. Significance Identifying gallstones in archaeological populations will assist researchers in estimating their frequency in the past and the environmental, cultural, and biological conditions leading to their presence. Limitations Small sample size derived from a modern and limited autopsy population may minimize the types and degree of variation present in the past. Effects of climate, soil, and taphonomy were not evaluated. Suggestions for Further Research Examination of larger samples derived from diverse populations may reveal greater variation or more diagnostic aspects of stones.