Forensic species identification: practical guide for animal and plant DNA analysis

: The importance of non-human DNA in the forensic field has increased greatly in recent years, together with the type of applications. The molecular species identification of animal and botanical material may be crucial both for wildlife trafficking and crime scene investigation. However, especially for forensic botany, several challenges slow down the implementation of the discipline in the routine.Although the importance of molecular analysis of animal origin samples is widely recognized and the same value is acknowledged to the botanical counterpart, the latter does not find the same degree of application.The availability of molecular methods, especially useful in cases where the material is fragmented, scarce or spoiled preventing the morphological identification, is not well known. This work is intended to reaffirm the relevance of non-human forensic genetics (NHFG), highlighting differences, benefits and pitfalls of the current most common molecular analysis workflow for animal and botanical samples, giving a practical guide. A flowchart describing the analysis paths, divided in three major working areas (inspection and sampling, molecular analysis, data processing and interpretation), is provided. More real casework examples of the utility of non-human evidence in forensic investigations should be shared by the scientific community, especially for plants. Moreover, concrete efforts to encourage initiatives in order to promote quality and standardization in the NHFG field are also needed

Prolonged higher dose methylprednisolone vs. conventional dexamethasone in COVID-19 pneumonia: a randomised controlled trial (MEDEAS)

Dysregulated systemic inflammation is the primary driver of mortality in severe COVID-19 pneumonia. Current guidelines favor a 7-10-day course of any glucocorticoid equivalent to dexamethasone 6 mg·day-1. A comparative RCT with a higher dose and a longer duration of intervention was lacking

Persistence and learning effects in design innovation : evidence from panel data

This paper explores persistence and learning effects in the aesthetic and symbolic dimensions of design innovation. By combining insights from innovation economics and design studies, we discuss design innovation as the result of firm-specific cumulative learning. We then conceptualise design and product innovation as complementary processes whose interplay may lead to learning effects across different dimensions of knowledge creation. We provide quantitative evidence for these insights applying dynamic probit and bivariate probit models to a longitudinal dataset of manufacturing firms based in Spain for the period 2007-2016. Our findings confirm the presence of persistence effects in design innovation, offering novel evidence in support of the view whereby design is an iterative process shaped by the knowledge generated through firms’ previous engagement with design. In addition, the results contribute to our understanding of the role of design beyond its functional dimension, pointing to mutually reinforcing effects between aesthetic and symbolic design and product innovation

Forensic botany: species identification of botanical trace evidence using a multigene barcoding approach

Forensic botany can provide significant supporting evidence during criminal investigations. However, it is still an underutilized field of investigation with its most common application limited to identifying specific as well as suspected illegal plants. The ubiquitous presence of plant species can be useful in forensics, but the absence of an accurate identification system remains the major obstacle to the present inability to routinely and correctly identify trace botanical evidence. Many plant materials cannot be identified and differentiated to the species level by traditional morphological characteristics when botanical specimens are degraded and lack physical features. By taking advantage of a universal barcode system, DNA sequencing, and other biomolecular techniques used routinely in forensic investigations, two chloroplast DNA regions were evaluated for their use as "barcoding" markers for plant identification in the field of forensics. We therefore investigated the forensic use of two non-coding plastid regions, psbA-trnH and trnL-trnF, to create a multimarker system for species identification that could be useful throughout the plant kingdom. The sequences from 63 plants belonging to our local flora were submitted and registered on the GenBank database. Sequence comparison to set up the level of identification (species, genus, or family) through Blast algorithms allowed us to assess the suitability of this method. The results confirmed the effectiveness of our botanic universal multimarker assay in forensic investigations

Methods for characterization/manipulation of human corneal stem cells and their applications in regenerative medicine.

Cell therapy is an emerging therapeutic strategy aimed at replacing or repairing severely damaged tissues with cultured cells. Speci fi cally, ocular burns cause depletion of limbal stem cells, which leads to corneal opaci fi cation and visual loss. Corneal stem cells are segregated in the basal layer of the limbus, which is the transitional zone of the epithelium located between the cornea and the bulbar conjunctiva. Autologous cultured limbal epithelial cells can restore damaged corneas. We sought to establish a culture system that allows preservation of limbal stem cells and preparation of manageable epithelial sheets. We outline some quality criteria, which assure the clinical performance of keratinocyte culture: evaluation of the number of holoclones within a cultured epithelial graft, proportion of aborting colonies, and percentage of cells expressing high levels of D Np63 a

From helpers to vulnerable people: Consequences of the pandemic for older volunteers in an Italian study.

The Covid-19 pandemic had a devastating impact on the older population in many respects. One of the main consequences has been the strengthening of the dynamics of ageism. The research presented in this paper aims to investigate the consequences of the pandemic and the lockdown on the population of self-sufficient older citizens in a circumscribed area of Northern Italy, one of the territories that was affected the most by the pandemic. The research focuses on the conditions of people over 65 years who were carrying out voluntary work in various local organizations. Secondary analysis was conducted on data from a research carried out in a District of Northern Italy. The results of the study showed numerous consequences for this population group, suddenly driven to operate a profound shift in their routine. The main consequence was a transformation in their role: the volunteers were people whose helped other people and suddenly they found themselves labelled as fragile recipients of help. The paper examines how these conditions reinforce a stereotyped image of the elders and poses some possible strategies to counter it, with particular focus to the functions of social workers

PRELIMINARY RESULTS ON THE COMPARISON BETWEEN EMPIRICAL AND PHYSICALLY-BASED RAINFALL THRESHOLDS FOR SHALLOW LANDSLIDES OCCURRENCE

Rainfall induced shallow landslides are hazardous phenomena causing significant damages all over the world. Rainfall thresholds are the most used tool to predict the occurrence of such instabilities over large areas. Most of these thresholds are empirical, basing on past rainfall events triggered landslides. These present several limitations, due to the amount and the uncertainty of available data and neglecting antecedent soil hydrological conditions. Physically-based thresholds could take into account also for soil hydrological responses towards rainfalls. Thus, this work aimed to compare both these typologies of thresholds, reconstructed for a 250 km2 area of northern Italian Apennines prone to shallow landsliding. Preliminary results of this research show the significant differences between thresholds reconstructed by means of different approaches. Empirical threshold estimates a lower triggering cumulated amount of rainfall than the values for physically-based thresholds, considering the same duration of rainfall event. Antecedent pore water pressure conditions have significant effects on reconstructed threshold, reducing the amount of rainfall leading to instabilities according to conditions approaching soil complete saturation. These preliminary results will be improved in order to create threshold useful for early warning strategies

Species identification through DNA "barcodes

Conventional methods for forensic species identification are mainly based on immunological procedures, which have limited applications for old and degraded specimens. The mitochondrial cytochrome b gene sequence has emerged in forensics among molecular methods. Recent investigations in the taxonomic field have suggested that a DNA-based identification system may aid the resolution of animal diversity and classification using sequence analysis and phylogenetic links. Selected gene sequences can be viewed as a genetic "barcode," which is enclosed in every cell, and barcoding is a standardized approach for characterizing species using short DNA sequences as a diagnostic biomarker for organisms. The aim of this study was to evaluate the potential of barcode mitochondrial genes, such as the cytochrome c oxidase sub 1 (COI) and the 16S rRNA gene, as a forensic tool. We developed a new approach for species testing and identification with a singleplex PCR amplification that will be useful not only in criminal casework but also in biosecurity, food authentication, investigation against poaching or illegal trade of endangered species, and wildlife enforcement. Seven fragments ranging from 157 to 541 bp (base pairs) in humans were selected from COI and 16S rRNA genes by different redesigned sets of primers suitable for forensic purposes. The specificity of each primer pair was evaluated with a single PCR reaction on different substrates, and the diversity values were calculated by statistical tests to select a set of markers that could be useful in different caseworks. A case example of forensic species identification is also presented

Slow and fast evolving markers typing in Modena males (North Italy).

One hundred thirty male individuals, strictly selected for their geographical origin and for typical regional surnames were submitted to the analysis. 17 STRs (short tandem repeats) loci and 19 SNPs binary markers (single nucleotide polymorphisms) of male-specific region of the Y chromosome (MSY) were typed to well characterize the selected population of Modena province. The availability of joint distribution of MSY haplotypes and haplogroup frequencies is becoming an important tool for both human evolutionary studies and forensic investigation, but large databases of complete Y-lineages are needed for a better understanding of the power of the combined use of Y-specific polymorphisms. A total of 129 haplotypes and 9 haplogroups were found and R1b haplogroup with a frequency of 67.7% was the most frequent, as expected because of the geographical location of the sample (Northwestern Italy). The Modena Y-lineages (STRs and SNPs independently) were also compared with published data of other neighbouring populations' samples

Two caseworks for one gene: successful species identification from compromised bone materials with the 12S rRNA

The availability of a reliable molecular assay in species recognition in forensic cases is of paramount importance when visual inspection or morphological methods are not exhaustive, especially from challenging samples. Here, two different caseworks involving bone samples founded during medico-legal outdoor investigations are presented. In order to exclude the human nature of the specimens and to determine the exact species they belong to, we proceeded with the molecular approach trying to generate sequences from the classical mtDNA markers cyt b and COI. However, they both gave critical results. For this reason, a short amplicon of ~ 150 bp of the 12S rRNA gene was used as an alternative.This short fragment was sufficient to identify the biological origin of the bone specimens with a high degree of certainty leading to the exclusion of their human nature. This work highlights the utility of the 12S rRNA and underlines the importance of deepen the choice of alternative shorter markers with respect to the classical ones, in order to achieve species identification even from challenging and degraded material in forensic criminal and wildlife caseworks