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STUDY QUESTION Can chromosomal abnormalities beyond copy-number aneuploidies (i.e. ploidy level and microdeletions (MDs)) be detected using a preimplantation genetic testing (PGT) platform? SUMMARY ANSWER The proposed integrated approach accurately assesses ploidy level and the most common pathogenic microdeletions causative of genomic disorders, expanding the clinical utility of PGT. WHAT IS KNOWN ALREADY Standard methodologies employed in preimplantation genetic testing for aneuploidy (PGT-A) identify chromosomal aneuploidies but cannot determine ploidy level nor the presence of recurrent pathogenic MDs responsible for genomic disorders. Transferring embryos carrying these abnormalities can result in miscarriage, molar pregnancy, and intellectual disabilities and developmental delay in offspring. The development of a testing strategy that integrates their assessment can resolve current limitations and add valuable information regarding the genetic constitution of embryos, which is not evaluated in PGT providing new level of clinical utility and valuable knowledge for further understanding of the genomic causes of implantation failure and early pregnancy loss. To the best of our knowledge, MDs have never been studied in preimplantation human embryos up to date. STUDY DESIGN, SIZE, DURATION This is a retrospective cohort analysis including blastocyst biopsies collected between February 2018 and November 2021 at multiple collaborating IVF clinics from prospective parents of European ancestry below the age of 45, using autologous gametes and undergoing ICSI for all oocytes. Ploidy level determination was validated using 164 embryonic samples of known ploidy status (147 diploids, 9 triploids, and 8 haploids). Detection of nine common MD syndromes (-4p=Wolf-Hirschhorn, -8q=Langer-Giedion, -1p=1p36 deletion, -22q=DiGeorge, -5p=Cri-du-Chat, -15q=Prader-Willi/Angelman, -11q=Jacobsen, -17p=Smith-Magenis) was developed and tested using 28 positive controls and 97 negative controls. Later, the methodology was blindly applied in the analysis of: (i) 100 two pronuclei (2PN)-derived blastocysts that were previously defined as uniformly euploid by standard PGT-A; (ii) 99 euploid embryos whose transfer resulted in pregnancy loss. PARTICIPANTS/MATERIALS, SETTING, METHODS The methodology is based on targeted next-generation sequencing of selected polymorphisms across the genome and enriched within critical regions of included MD syndromes. Sequencing data (i.e. allelic frequencies) were analyzed by a probabilistic model which estimated the likelihood of ploidy level and MD presence, accounting for both sequencing noise and population genetics patterns (i.e. linkage disequilibrium, LD, correlations) observed in 2504 whole-genome sequencing data from the 1000 Genome Project database. Analysis of phased parental haplotypes obtained by single-nucleotide polymorphism (SNP)-array genotyping was performed to confirm the presence of MD. MAIN RESULTS AND THE ROLE OF CHANCE In the analytical validation phase, this strategy showed extremely high accuracy both in ploidy classification (100%, CI: 98.1–100%) and in the identification of six out of eight MDs (99.2%, CI: 98.5–99.8%). To improve MD detection based on loss of heterozygosity (LOH), common haploblocks were analyzed based on haplotype frequency and LOH occurrence in a reference population, thus developing two further mathematical models. As a result, chr1p36 and chr4p16.3 regions were excluded from MD identification due to their poor reliability, whilst a clinical workflow which incorporated parental DNA information was developed to enhance the identification of MDs. During the clinical application phase, one case of triploidy was detected among 2PN-derived blastocysts (i) and one pathogenic MD (-22q11.21) was retrospectively identified among the biopsy specimens of transferred embryos that resulted in miscarriage (ii). For the latter case, family-based analysis revealed the same MD in different sibling embryos (n = 2/5) from non-carrier parents, suggesting the presence of germline mosaicism in the female partner. When embryos are selected for transfer based on their genetic constitution, this strategy can identify embryos with ploidy abnormalities and/or MDs beyond aneuploidies, with an estimated incidence of 1.5% (n = 3/202, 95% CI: 0.5–4.5%) among euploid embryos. LIMITATIONS, REASONS FOR CAUTION Epidemiological studies will be required to accurately assess the incidence of ploidy alterations and MDs in preimplantation embryos and particularly in euploid miscarriages. Despite the high accuracy of the assay developed, the use of parental DNA to support diagnostic calling can further increase the precision of the assay. WIDER IMPLICATIONS OF THE FINDINGS This novel assay significantly expands the clinical utility of PGT-A by integrating the most common pathogenic MDs (both de novo and inherited ones) responsible for genomic disorders, which are usually evaluated at a later stage through invasive prenatal testing. From a basic research standpoint, this approach will help to elucidate fundamental biological and clinical questions related to the genetics of implantation failure and pregnancy loss of otherwise euploid embryos. STUDY FUNDING/COMPETING INTEREST(S) No external funding was used for this study. S.C., M.F., F.C., P.Z., I.P., L.G., C.P., M.P., D.B., J.J.-A., D.B.-J., J.M.-V., and C.R. are employees of Igenomix and C.S. is the head of the scientific board of Igenomix. A.C. and L.P. are employees of JUNO GENETICS. Igenomix and JUNO GENETICS are companies providing reproductive genetic services. TRIAL REGISTRATION NUMBER N/A.

Il volume curato da Mario Brogi, Elisabetta Novello e Remy Simonetti rappresenta il primo contributo di TerrEvolute, collana che ha l’obiettivo di proporre una riflessione sul rapporto tra società e ambiente, analizzando non solo le conseguenti trasformazioni del territorio, ma anche le ripercussioni economiche e sociali sulle comunità lì residenti. In particolare, l’attenzione di questo contributo si focalizza sui territori di bonifica che rappresentano scenari privilegiati per indagare la complessa e secolare relazione tra componente antropica e ambientale. In queste aree, infatti, è possibile rintracciare quelle pratiche umane che nel corso del tempo si sono sedimentate sul territorio ed hanno contribuito ad assegnare una nuova connotazione al paesaggio, in molti casi completamente stravolto nel suo aspetto originario dalle opere idrauliche e ingegneristiche dei primi decenni del Novecento. L’evoluzione dei luoghi di bonifica si può cogliere attraverso un ricco patrimonio materiale e immateriale che costituisce un inestimabile valore per le comunità locali e per questo motivo deve essere oggetto di conservazione, valorizzazione e diffusione., Semestrale di studi e ricerche di geografia, No. 1 (2022)

Study question Can chromosomal abnormalities beyond aneuploidies (i.e., ploidy and microdeletions, MD) be detected on a single trophectoderm (TE) embryo biopsy using a next-generation sequencing (NGS)-based workflow? Summary answer This NGS-based integrated approach allows accurate detection of ploidy status and the most common microdeletions from a single TE-biopsy,expanding PGT-A clinical validity and diagnostic capabilities. What is known already Standard methodologies employed in PGT-A do not determine embryo ploidy status due to the normalization process during copy-number-variation analysis. Transferring embryos with abnormal ploidy variations is expected to result in miscarriage or molar pregnancy. Common pathogenic MD are undetected as they fall below the PGT-A resolution limit ( Study design, size, duration Ploidy determination was validated using 244 embryo samples of known ploidy status (226 diploids, 10 triploids, 8 haploids). We analysed nine common MD syndromes (-4p=Wolf-Hirschhorn, -8q=Langer-Giedion, -1p=1p36 deletion, -22q=DiGeorge, -5p=Cri-du-Chat, -15q=Prader-Willi/Angelman, -11q=Jacobsen, -17p=Smith-Magenis) using 24 positive controls (amniocentesis DNA from MD cases or TE biopsies from autosomal monosomy mimicking MD) and 96 negative controls (healthy newborns). Overall, the dataset included 72 cases of individual chromosomal abnormalities and 576 negative cases across the eight MD regions. Participants/materials, setting, methods PGT-A products were reamplified and sequenced (IonTorrentS5-ThermoFisher) using a custom AmpliSeq panel targeting 384 regions with at least one Single Nucleotide Polymorphism (SNP) of high B-allelic frequency. A bioinformatic algorithm based on gaussian-mixture modelling of sequencing data was developed. This algorithm calculates the conditional probability of the observed B-allelic ratio for each SNP, depending on the copy number, then estimates the likelihood of ploidy and the presence of MD based on the sequencing outcomes. Main results and the role of chance Ploidy was correctly determined in 233/234 cases (Accuracy=99.4%), with only one diploid sample misclassified as triploid (PPV=94.1%, NPV=100%, Non-informative rate=9/243=3.1%). Microdeletions could be consistently detected with high reliability in 6 out of the 8 considered regions (-8q,-22q,-5p,-15q,-11q and -17p; PPV=98.5%, NPV=99.5%). Detection of microdeletions of 1p and 4p were less reliable due to the presence of recurrent haplotype blocks in the population at those genomic regions, as confirmed by the analysis of a dataset of 2504 whole genome sequencing from One Thousand Genome Project database (1kGP). The only MD false positive case showed extended loss of heterozygosity in the microdeletion region (-22q), which might be related to uniparental disomy or consanguinity and requires further testing in the family. This analytical framework was blindly applied to: (i) the analysis of 9 embryos from a family affected by DiGeorge syndrome (female partner was carrier of del22.q11.21(20754422-21440514), resulting in all embryos classified consistently with the conventional PGT-M results (using indirect linkage analysis); (ii) the analysis of samples from 99 transferred human euploid embryos resulting in pregnancy losses. No ploidy alteration was detected in miscarried euploid embryos, but 2 microdeletions (-8q, -22q) were found, with an estimated prevalence of 2/99 in the miscarriage population. Limitations, reasons for caution Larger cohort studies will be required to accurately assess the incidence of ploidy alterations and microdeletions in preimplantation embryos and particularly in euploid miscarriages. Despite the high accuracy of the assay developed, the use of parental DNA to support diagnostic calling can further increase the precision of the assay. Wider implications of the findings This study provides, for the first time, detection of common pathogenic microdeletions and ploidy status from a single TE biopsy, expanding PGT-A clinical validity. This new assay will also help elucidate fundamental biological and clinical questions related to the genetics of implantation failure and pregnancy loss of apparently euploid embryos. Trial registration number not applicable

Microplastics pose a threat to all the environmental compartments and specifically to aquatic ecosystems. Lakes are particularly exposed to it as they act as accumulator of pollution from their watersheds. Unfortunately, the fate of microplastics in lake ecosystems remains poorly understood because of the multiplicity of sources and transfer pathways. For the first time, the Plastilac project focuses on the contamination of high altitude lakes (from 1300 m to 2800 m above the sea level) by microplastics. Remote lakes constitute easier-to-investigate ecosystems because there are fewer potential sources of microplastics in their watershed, namely the atmospheric deposit, the supply from the watershed through the tributaries and the tourist attendance. Thus, both water column and sediment from 10 lakes located across the French Alps were sampled. The results showed that no lake was free from microplastics, proving the ubiquity of this pollution at a regional scale. The abundance of microplastics varied significantly from one lake to another and the concentrations measured in high altitude lakes (around 10 MP.m-3) were approximately 100 times lower than those reported in the literature for lowland lakes. The water column contamination was not correlated to the vicinity of potential sources (urban areas). On the contrary, higher sediment contaminations were observed in lakes located nearby urban areas. Our analyses also showed that the residence times of microplastics in the water column of these lakes were relatively short, of the order of a few days. In contrast, the residence times of microplastics in the sediments were much longer and lake bottoms retain traces of past contamination. This work constitutes a first for understanding the fate of microplastics in mountainous environments. It provides important information on their dynamics and, in particular, on the temporal dimension of this pollution.

The role of distributor system operators is experiencing a gradual but relevant change to include enhanced ancillary and energy dispatch services needed to manage the increased power provided by intermittent distributed generations in medium voltage networks. In this context, the paper proposes the insertion, in strategic points of the network, of specific power electronic systems, denoted as active nodes, which permit the remote controllability of the active and reactive power flow. Such capabilities, as a further benefit, enable the distributor system operators to provide ancillary network services without requiring any procurement with distributed generation systems owners. In particular, the paper highlights the benefits of active nodes, demonstrating their capabilities in reducing the inverse power flow issues from medium to high voltage lines focusing on a network cluster including renewable energy resources. As a further novelty, this study has accounted for a real cluster operated by the Italian distributor system operator Areti. A specific simulation model of the electrical lines has been implemented in DigSilent PowerFactory (DIgSILENT GmbH–Germany) software using real operating data obtained during a 1-year measurement campaign. A detailed cost-benefit analysis has been provided, accounting for different load flow scenarios. The results have demonstrated that the inclusion of active nodes can significantly reduce the drawbacks related to the reverse power flow.

Grazie all'abbattimento dei loro costi e allo sviluppo delle loro funzionalità, i Sistemi Informativi Geografici (GIS) vengono applicati in sempre nuovi ambiti e settori. Incentivando una didattica partecipata e laboratoriale, le geo-tecnologie comportano un rinnovamento anche dell'offerta formativa della geografia: il loro uso aumenta infatti la capacità dello studente di condurre una ricerca strutturata, analizzando il territorio e derivando informazioni elaborate; allo stesso modo, l'attività dell'insegnante aumenta il suo coinvolgimento, svolgendosi in un'atmosfera di cooperazione e partecipazione tra gli alunni. In questo senso, l'esperienza dell'Officina Didattica "GIS e geotecnologie per una moderna geografia", tenutasi nell'ambito del 59° Convegno Nazionale AIIG presso "La Sapienza" Università di Roma, fornisce una dimostrazione dell'interesse dei partecipanti e delle metodologie dei facilitatori intervenuti in questo ambito.

Lo scopo di questo lavoro è lo studio delle trasformazioni di un territorio che, nel corso di duecento anni, è passato da avere una connotazione fortemente naturale ad essere intensamente frammentato a causa della intensa antropizzazione. Fino ai primi decenni del ’900, canali, boschi e pascoli connotavano il paesaggio della Pianura Pontina; in pochi decenni, le bonifiche integrali degli anni ’30 e la consecutiva nascita di agglomerati urbani hanno stravolto completamente il territorio. Oggi alla crescita esponenziale del tessuto edilizio si è affiancata un’agricoltura di tipo intensivo e lo sviluppo di un apparato industriale lungo le principali arterie viarie. Per la ricostruzione dell’uso del suolo di una parte della Pianura Pontina sono state utilizzate tre fonti eterogenee e diacroniche (cartografia storica ottocentesca, foto aeree di metà ’900 e dati attuali ricavati da immagini satellitari) che, opportunamente elaborate attraverso i GIS, sono servite per produrre cartografie digitali con differenti livelli di dettaglio. In quest’ottica, i GIS si dimostrano imprescindibili strumenti per l’indagine geostorica perché permettono di effettuare analisi multitemporali di tipo qualitativo e quantitativo analizzando i cambiamenti avvenuti in un determinato territorio. The aim of this work is the study of the landscape changes suffered by the Pontine Region. The region was originally characterized by strong natural features and became heavily fragmented over the last two hundred years because of the extensive human activity. Until the first decades of the 1900s, the Pontine Region was characterized by wetlands, woods and pastures. In a short time, the reclamation during the Fascism period led to the growth of new urban areas and a radical and fast change of a landscape. Today, this region is characterized by intensive cultivation areas and the growth of industrial sector along the main roads. Heterogeneous and diachronic sources used to land use and cover reconstruction of a part of Pontine Region (historical cartography of 19th century, aerial photographs of the half of 20th century and current data from satellite imagery) are been elaborated trough GIS to create digital cartographies with different levels of detail. For this reason, GIS are essential tools for geo-hisrorical studies because allow to conduct qualitative and quantitative analyses and show change detection of territory.