Estimation of Human Chronological Age from Buccal Swab Samples through a DNA Methylation Analysis Approach of a Five-Locus Multiple Regression Model.

Recent advancements in forensic genetics have facilitated the extraction of additional characteristics from unidentified samples. This study delves into the predictive potential of a five-gene (ELOVL2, FHL2, KLF14, C1orf132, and TRIM59) methylation rate analysis for human age estimation using buccal swabs collected from 60 Italian volunteers. The methylation levels of specific CpG sites in the five genes were analyzed through bisulfite conversion, single-base extension, and capillary electrophoresis. A multivariate linear regression model was crafted on the training set, then the test set was employed to validate the predictive model. The multivariate predictive model revealed a mean absolute deviation of 3.49 years in the test set of our sample. While limitations include a modest sample size, the study provides valuable insights into the potential of buccal swab-based age prediction, aiding in criminal investigations where accurate age determination is crucial. Our results also highlight that it is necessary to investigate the effectiveness of predictive models specific to biological tissues and individual populations, since models already proven effective for other populations or different tissues did not show the same effectiveness in our study.

Defining medical liability when artificial intelligence is applied on diagnostic algorithms: a systematic review.

Artificial intelligence (AI) in medicine is an increasingly studied and widespread phenomenon, applied in multiple clinical settings. Alongside its many potential advantages, such as easing clinicians' workload and improving diagnostic accuracy, the use of AI raises ethical and legal concerns, to which there is still no unanimous response. A systematic literature review on medical professional liability related to the use of AI-based diagnostic algorithms was conducted using the public electronic database PubMed selecting studies published from 2020 to 2023. The systematic review was performed according to 2020 PRISMA guidelines. The literature review highlights how the issue of liability in case of AI-related error and patient's damage has received growing attention in recent years. The application of AI and diagnostic algorithm moreover raises questions about the risks of using unrepresentative populations during the development and about the completeness of information given to the patient. Concerns about the impact on the fiduciary relationship between physician and patient and on the subject of empathy have also been raised. The use of AI in medical field and the application of diagnostic algorithms introduced a revolution in the doctor-patient relationship resulting in multiple possible medico-legal consequences. The regulatory framework on medical liability when AI is applied is therefore inadequate and requires urgent intervention, as there is no single and specific regulation governing the liability of various parties involved in the AI supply chain, nor on end-users. Greater attention should be paid to inherent risk in AI and the consequent need for regulations regarding product safety as well as the maintenance of minimum safety standards through appropriate updates.

An mRNA Profiling Study of Vaginal Swabs from Pre- and Postmenopausal Women.

Body fluid identification by means of mRNA profiling provides valuable supplementary information in forensic investigations. In particular, the detection of vaginal mucosa mRNA markers is highly relevant in sexual assault cases. Although the vagina undergoes characteristic age-related physiological changes over a lifetime, few studies have evaluated the efficacy of vaginal mRNA markers in women of different ages. In this multicentric study, a 19-plex mRNA profiling assay including vaginal-specific markers (CYP2B7P1, MUC4, MYOZ1) was tested in a collection of 6-20-month-old vaginal swabs obtained from pre- (n = 84) and postmenopausal (n = 55) female volunteer donors. Overall, participating laboratories were able to correctly identify ~85% of samples as vaginal mucosa by mRNA profiling. The assay's success rate did not differ between the two age groups and was not affected by the time interval between swab collection and RNA analysis. MYOZ1 resulted a less sensitive vaginal marker compared to MUC4 and CYP2B7P1. A significant relative increase in the contribution to the total amplification signal was observed for MUC4, compared to CYP2B7P1 and MYOZ1, in postmenopausal women. Observation of other body fluids and tissues different from vaginal mucosa was also evaluated in connection to information on previous sexual activity and menstrual cycle phase at the time of sampling.

Lessons Learned from the COVID-19 Pandemic: A Survey-Based Study on a Sample of Italian Physicians' Opinions on Telemedicine.

Telemedicine was born out of the need to ensure clinical evaluation and personal care regardless of the physical presence of the healthcare professional nearby. Information technologies have been vital during the COVID-19 pandemic to ensure medical care and avoid the contagion between patients and clinicians. Accordingly, telecare services multiplied worldwide and gained paramount importance. The present work aims to collect field-based opinions about Telemedicine and ethics among Italian physicians. We developed a web-based questionnaire that was administered to Italian physicians from 1 May to 15 June 2022. The questionnaire was distributed as a link to Google Forms via social networks/instant messaging applications to groups of graduated and qualified physicians. A total of 180 physicians answered the questionnaire (with an age range from 25 to 68 years old). Physicians belonging to the medical area of expertise appear to more frequently use new technologies in comparison to other specialties. The vast majority believe that it is appropriate to use Telemedicine for monitoring and follow-up but not for evaluating a new patient. Concerns about changes in the physician-patient relationship, informed consent, digital barrier, and privacy and data protection also emerged. Finally, telehealth is thought to be a potential useful tool for the future by the majority of respondents but proper training for physicians is therefore needed.

Digital Biobanking and Big Data as a New Research Tool: A Position Paper.

Big data analytics in medicine is driving significant change, as it offers vital information for improving functions, developing cutting-edge solutions and overcoming inefficiencies. With the right archiving and analysis tools, all players in the healthcare system, from hospitals to patients and from medical personnel to the pharmaceutical industry, can yield numerous benefits. Therefore, to analyze and interpret these analytics effectively, so that they can be useful for the advancement of scientific knowledge, we require information sharing, specific skills, training, integration between all system players, unique infrastructures and security. All these characteristics will make it possible to establish and harmonize real big data biobanks, for which it will be appropriate to consider new forms of governance compared to those traditionally conceived for large-sample biobanks.

Forensic Age Estimation through a DNA Methylation-Based Age Prediction Model in the Italian Population: A Pilot Study.

DNA methylation is one of the epigenetic marks which has been studied intensively in recent years for age predicting purposes in the forensic area. In order to integrate age prediction into routine forensic workflow, the purpose of this study was to standardize and optimize a DNA methylation-based protocol tailored to the Italian context. A previously published protocol and age-predictive method was implemented for the analysis of 84 blood samples originating from Central Italy. The study here presented is based on the Single Base Extension method, considering five genes: ELOVL2, FHL2, KLF14, C1orf132, now identified as MIR29B2C, and TRIM59. The precise and specific steps consist of DNA extraction and quantification, bisulfite conversion, amplification of converted DNA, first purification, single base extension, second purification, capillary electrophoresis, and analysis of the results to train and test the tool. The prediction error obtained, expressed as mean absolute deviation, showed a value of 3.12 years in the training set and 3.01 years in the test set. Given that population-based differences in DNA methylation patterns have been previously reported in the literature, it would be useful to further improve the study implementing additional samples representative of the entire Italian population.

Touch DNA Sampling Methods: Efficacy Evaluation and Systematic Review.

Collection and interpretation of "touch DNA" from crime scenes represent crucial steps during criminal investigations, with clear consequences in courtrooms. Although the main aspects of this type of evidence have been extensively studied, some controversial issues remain. For instance, there is no conclusive evidence indicating which sampling method results in the highest rate of biological material recovery. Thus, this study aimed to describe the actual considerations on touch DNA and to compare three different sampling procedures, which were "single-swab", "double-swab", and "other methods" (i.e., cutting out, adhesive tape, FTA® paper scraping), based on the experimental results published in the recent literature. The data analysis performed shows the higher efficiency of the single-swab method in DNA recovery in a wide variety of experimental settings. On the contrary, the double-swab technique and other methods do not seem to improve recovery rates. Despite the apparent discrepancy with previous research, these results underline certain limitations inherent to the sampling procedures investigated. The application of this information to forensic investigations and laboratories could improve operative standard procedures and enhance this almost fundamental investigative tool's probative value.

Human microbiome and microbiota identification for preventing and controlling healthcare-associated infections: A systematic review.

Objective: This systematic review describes the role of the human microbiome and microbiota in healthcare-associated infections (HAIs). Studies on the microbiota of patients, healthcare environment (HE), medical equipment, or healthcare workers (HCW) and how it could be transmitted among the different subjects will be described in order to define alarming risk factors for HAIs spreading and to identify strategies for HAIs control or prevention. Methods: This review was performed in adherence to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. After retrieval in databases, identification, and screening of available records, 36 published studies were considered eligible and included in the review. Results: A multifaceted approach is required and the analyses of the many factors related to human microbiota, which can influence HAIs onset, could be of paramount importance in their prevention and control. In this review, we will focus mainly on the localization, transmission, and prevention of ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) bacteria and Clostridium difficile which are the most common pathogens causing HAIs. Conclusions: Healthcare workers' microbiota, patient's microbiota, environmental and medical equipment microbiota, ecosystem characteristics, ways of transmission, cleaning strategies, and the microbial resistome should be taken into account for future studies on more effective preventive and therapeutic strategies against HAIs.

The role of SPATA2 in TNF signaling, cancer, and spermatogenesis.

The activation of TNF receptors can lead to cell death with a mechanism of cell necrosis regulated genetically and distinct from apoptosis which is defined as necroptosis. Necroptosis has been one of the most studied emerging cell death/signaling pathways in recent years, especially in light of the role of this process in human disease. However, not all regulatory components of TNF signaling have been identified in relation to both physiological and pathological conditions. In 2008, Spata2 (Spermatogenesis-associated protein 2) was identified as one of the seven fundamental genes for the cellular signaling network that regulates necroptosis and apoptosis. This gene had been cloned by our group and named Spata2 as its expression was found to be elevated in the testis compared to other tissues, localized at the Sertoli cell level and FSH-dependent. More recently, it has been demonstrated that deletion of Spata2 gene causes increased inhibin α expression and attenuated fertility in male mice. However, more importantly, five recently published reports have highlighted that SPATA2 is crucial for recruiting CYLD to the TNFR1 signaling complex thus promoting its activation leading to TNF-induced cell death. Loss of SPATA2 increases transcriptional activation of NF-kB and limits TNF-induced necroptosis. Here we will discuss these important findings regarding SPATA2 and, in particular, focus attention on the evidence that suggests a role for this protein in the TNF signaling pathway.

Post-Mortem Interval and Microbiome Analysis through 16S rRNA Analysis: A Systematic Review.

The determination of the Post-Mortem Interval (PMI) is an issue that has always represented a challenge in the field of forensic science. Different innovative approaches, compared to the more traditional ones, have been tried over the years, without succeeding in being validated as successful methods for PMI estimation. In the last two decades, innovations in sequencing technologies have made it possible to generate large volumes of data, allowing all members of a bacterial community to be sequenced. The aim of this manuscript is to provide a review regarding new advances in PMI estimation through cadaveric microbiota identification using 16S rRNA sequencing, in order to correlate specific microbiome profiles obtained from different body sites to PMI. The systematic review was performed according to PRISMA guidelines. For this purpose, 800 studies were identified through database searching (Pubmed). Articles that dealt with PMI estimation in correlation with microbiome composition and contained data about species, body site of sampling, monitoring time and sequencing method were selected and ultimately a total of 25 studies were considered. The selected studies evaluated the contribution of the various body sites to determine PMI, based on microbiome sequencing, in human and animal models. The results of this systematic review highlighted that studies conducted on both animals and humans yielded results that were promising. In order to fully exploit the potential of the microbiome in the estimation of PMI, it would be desirable to identify standardized body sampling sites and specific sampling methods in order to align data obtained by different research groups.

Skin Microbial Changes during Space Flights: A Systematic Review.

Background. Sixty years after the launch of the first human into space, different studies on the physiological changes that humans undergo during dynamic flight phases and prolonged weightlessness have been undertaken. Understanding these changes is important for the creation of the preventative measures that are essential to ensuring astronaut health. Among these changes, those of the skin are frequent, despite being rarely treated during missions. The skin is a physical barrier that protects the body from pathogen invasion and environmental changes, and it harbors diverse microbial communities that form the skin microbiota. Methods. A systematic literature review of skin microbiome changes during space flight was conducted using public electronic databases (PubMed and Scopus) selecting studies published from 2015 to 2022. The systematic review was performed according to 2020 PRISMA guidelines. Results. A total of 17 studies were collected and, after screening for inclusion and exclusion criteria, eight studies were included in this review. According to the examined literature, some skin microbiota changes seems to be only temporary, in particular Gamma- and Betaproteobacteria abundance tends to decrease, while the occurrence of the Malassezia species and Firmicutes, including Staphylococcus and Streptococcus, tends to increase. At the same time, there seems to be an exchange of microorganisms between astronauts and between the confined environment and a single astronaut, with alterations in the proportion of microorganisms maintained during the flight, in particular for species such as Corynebacterium spp., Staphylococcus spp., Streptococcus spp. and Cloacibacterium spp. Given that skin contributes both to protecting the body from pathogen invasion and environmental changes and to maintaining human homeostasis, changes in the skin microbiota of astronauts might result in skin diseases. Discussion. The skin microbiota of astronauts seems to influence the microbial composition of the International Space Station, but further studies should be performed to better understand skin microbiota dynamics and to prevent the development of dermatologic conditions during space flight.

Family and Educational Strategies for Cyberbullying Prevention: A Systematic Review.

Cyberbullying can be described as a form of bullying carried out by an individual or a group through digital media with the intention to harm others. It has been recognized as a public health issue recently; however, of the vast literature published in recent years on the phenomenon, only a small part concerns strategies adopted to prevent and combat cyberbullying, and the effectiveness of these strategies appears to be scarce. We conducted a systematic review of the literature published in the last five years about different interventions studied to prevent and contrast cyberbullying. Our results show how most of the strategies currently developed focus on the educational aspect, involving schools and families. Other authors describe technology-based practices to set programs to reduce and prevent cyberbullying through the usage of digital instruments, the same used by minors themselves. Finally, remaining tactics use a more comprehensive approach, mixing tools already in use in the aforementioned strategies. Cyberbullying requires wide-ranging methods to combat it, involving the contribution of mental health professionals, educators, and digital experts cooperating synergically. Prevention and contrast instruments should be defined, implemented, tested, and combined in order to deal with cyberbullying.

An Update on Planned Oocyte Cryopreservation (POC) in Italy: Medical, Epidemiological and Legal Consideration.

Starting with a brief socioeconomic analysis of the phenomenon of female fertility, this narrative review aims to provide an analysis of the use and possibilities of medically assisted reproductive technology in combating fertility issues, adopting socioeconomic, legal and medical perspectives in Italy. The authors mainly employ data from the annual reports of the National Registry of Medically Assisted Reproduction (PMA Registry) and the Italian Statistical Institute (ISTAT) to understand the evolution of oocyte use in medically assisted reproductive technology in Italy from 2015 to 2018 and in particular to dissect the possibilities of oocyte cryopreservation as a measure to counteract age-related infertility, specifically through Planned Oocyte Cryopreservation (POC), also known as "social freezing". It seems that the best course of action in the context of medically assisted reproduction would be the use of young and healthy cryopreserved oocytes (autologous or donated), preferably before the age of 40, while encouraging donation of oocytes whenever possible. Italy's dependence on foreign biobanks for donated oocytes calls for the institution of a national biobank and further specific regulation of gamete donation. For this reason, it would be useful to encourage the acceptance of Planned Oocyte Cryopreservation to allow greater availability of healthy, younger oocytes.

Leonardo Botallo (1530-1587) and his pioneering contributions to traumatology, cardiology and deontology.

Leonardo Botallo (1530-c. 1587) is widely known for the eponymous "foramen Botalli" and "ductus Botalli". The first, most commonly named "foramen ovale", allows blood in the fetal heart to enter the left atrium from the right atrium. The second, named "ductus arteriosus", consists of a blood vessel in the developing fetus connecting the trunk of the pulmonary artery to the proximal descending aorta. However, Botallo was a multifaceted figure who studied many aspects of human anatomy and physiology, also making important contributions to clinical and surgical practices. Moreover, as we will see in the last section of this paper, Botallo wrote a book on medical deontology having significant features in relationship to the history of medical ethics. Botallo's multidisciplinary approach is a typical characteristic of Renaissance physicians and scientists, who contributed to making this period a fundamental prelude to the scientific revolution of the 17th century.

External visible characteristics prediction through SNPs analysis in the forensic setting: a review.

Numerous major advances have been made in forensic genetics over the past decade. One recent field of research has been focused on the analysis of External Visible Characteristics (EVC) such as eye colour, hair colour (including hair greying), hair morphology, skin colour, freckles, facial morphology, high myopia, obesity, and adult height, with important repercussions in the forensic field. Its use could be especially useful in investigative cases where there are no potential suspects and no match between the evidence DNA sample under investigation and any genetic profiles entered into criminal databases. The present review represents the current state of knowledge of SNPs (Single Nucleotide Polymorphisms) regarding visible characteristics, including the latest research progress in identifying new genetic markers, their most promising applications in the forensic field and the implications for police investigations. The applicability of these techniques to concrete cases has stoked a heated debate in the literature on the ethical implications of using these predictive tools for visible traits.

[New and old parenting: reflections about assisted reproductive technologies in view of recent Italian law.] / Nuove e vecchie genitorialità: riflessioni sulla procreazione medicalmente assistita alla luce della recente giurisprudenza italiana.

The constant advancement of reproduction technologies has broadened the panorama of possible techniques and has led to a profound change in the concept of the family that has so far been widespread. Assisted reproductive techniques (ART) currently allow to conceive a human being in the absence of physical connection between man and woman. In light of the possibility of conception using heterologous techniques, the concept of parent/child relationship may also be changed. This contribution is intended to highlight how the concept of family is changing and may change in the future in the light of the new ART and the principles recently affirmed in Italian jurisprudence. In the light of the above considerations, it appears of primary importance to definitively clarify the position and rights of the involved parties, underlining the need for an adequate and timely intervention aimed at resolving, among the various issues, the conflict between the donor's interest in protect his/her anonymity and their child's interest in knowing his/her their origins.

Microbiome Forensic Biobanking: A Step toward Microbial Profiling for Forensic Human Identification.

In recent years many studies have highlighted the great potential of microbial analysis in human identification for forensic purposes, with important differences in microbial community composition and function across different people and locations, showing a certain degree of uncertainty. Therefore, further studies are necessary to enable forensic scientists to evaluate the risk of microbial transfer and recovery from various items and to further critically evaluate the suitability of current human DNA recovery protocols for human microbial profiling for identification purposes. While the establishment and development of microbiome research biobanks for clinical applications is already very structured, the development of studies on the applicability of microbiome biobanks for forensic purposes is still in its infancy. The creation of large population microbiome biobanks, specifically dedicated to forensic human identification, could be worthwhile. This could also be useful to increase the practical applications of forensic microbiology for identification purposes, given that this type of evidence is currently absent from most real casework investigations and judicial proceedings in courts.

"Touch microbiome" as a potential tool for forensic investigation: A pilot study.

Human skin hosts a variety of microbes that can be transferred to surfaces ("touch microbiome"). These microorganisms can be considered as forensic markers similarly to "touch DNA". With this pilot study, we wanted to evaluate the transferability and persistence of the "touch microbiome" on a surface after the deposition of a fingerprint and its exposure for 30 days at room temperature. Eleven volunteers were enrolled in the study. Skin microbiome samples were collected by swabbing the palm of their hands; additionally, donors were asked to touch a glass microscope slide to deposit their fingerprints, that were then swabbed. Both human and microbial DNA was isolated and quantified. Amelogenin locus and 16 human STRs were amplified, whereas the V4 region of 16 S rRNA gene was sequenced using Illumina MiSeq platform. STR profiles were successfully typed for 5 out of 22 "touch DNA" samples, while a microbiome profile was obtained for 20 out of 22 "touch microbiome" samples. Six skin core microbiome taxa were identified, as well as unique donor characterizing taxa. These unique taxa may have relevance for personal identification studies and may be useful to provide forensic intelligence information also when "touch DNA" fails. Additional future studies including greater datasets, additional time points and a greater number of surfaces may clarify the applicability of "touch microbiome" studies to real forensic contexts.

The Usage of Mobile Apps to Fight Violence against Women: A Survey on a Sample of Female Students Belonging to an Italian University.

The prevalence of violence against women continues to grow and this plague has had a huge impact from a clinical, social and judicial point of view. For this reason, alongside the efforts made at the legislative level to prevent the phenomenon and to improve assistance to victims in recent years, efforts to contain and better manage this phenomenon have also grown in the extra-legislative sphere: for example, through the application of new technological solutions and safety planning. In recent years, there has been an increase in the marketing of mobile phone apps dedicated to the prevention of violence against women, with different functions and different objectives. The purpose of this study is to investigate the knowledge and propensity to download this type of app in a group of 1782 Italian female university students. This research was performed using an online questionnaire administered to female students attending four different courses (law, medicine, healthcare professionals and political sciences) at one Italian university. Chi-square or Fisher's exact test was used to analyze associations between responses to questionnaire and the type and the year of course. The results show that 62.6% of our sample are unaware of the existence of these apps and that 79.5% of the sample would be willing to download one in the future. With regard to whom to turn to after a violent incident, the majority of those interviewed (43.9%) would turn to the police and not to health facilities. According to our findings, law female students (52.7%) think, more than any other category, that the most effective way to improve public safety and reduce the number of victims lies in legislative solutions. Our results suggest that, although this type of technology may be promising, it is necessary to improve the knowledge and dissemination of these apps in order to make them a useful tool for prevention, education and assistance in cases of violence against women.

Rehabilitation healthcare professionals' perceptions of professional responsibility: a focus group study in Italy.

BACKGROUND AND AIM OF THE WORK: Law no. 24/17 has updated the Italian legislation regarding healthcare professional responsibility. In practice, all eight groups of healthcare professionals have well defined areas of activity and responsibility. The aim of this research is to investigate how rehabilitation healthcare professionals perceive responsibility in the professional life. METHODS: a qualitative pilot study was conducted using a focus group consisting of rehabilitation professionals enrolled in the Master's Degree program in Rehabilitation Sciences at the University of Padova. RESULTS: the analysis (performed at micro, meso and macro-level) highlighted that respondents perceived professional responsibility as a dilemma, because it is identified as a continuous balancing between external and internal pressures regarding professional competence, professional autonomy, teamwork, and social matters. CONCLUSIONS: rehabilitation professionals seem to experience a great deal of dilemmas because they felt that their responsibility was a constant challenge of weighing pros and cons. Rehabilitation professionals need a genuine autonomy recognition at every level of their practice to better cope with the professional responsibility on a daily basis. Responsibility has to be continuously and effectively implemented in all its aspects throughout the care pathway as to allow rehabilitation professionals to provide quality care to all patients. This analysis was useful for highlighting the emerging issues that need to be addressed in everyday practice.