Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Cancer Three-Dimensional Spheroids Mimic In Vivo Tumor Features, Displaying “Inner” Extracellular Vesicles and Vasculogenic Mimicry

The role of extracellular vesicles (EVs) as mediators of cell-to-cell communication in cancer progression is widely recognized. In vitro studies are routinely performed on 2D culture models, but recent studies suggest that 3D cultures could represent a more valid model. Human ovarian cancer cells CABA I were cultured by the hanging drop method to form tumor spheroids, that were moved to low adhesion supports to observe their morphology by Scanning Electron Microscopy (SEM) and to isolate the EVs. EVs release was verified by SEM and their identity confirmed by morphology (Transmission Electron Microscopy, TEM), size distribution (Nanoparticles Tracking Analysis), and markers (CD63, CD9, TSG-101, Calnexin). CABA I form spheroids with a clinically relevant size, above 400 μm; they release EVs on their external surface and also trap “inner” EVs. They also produce vasculogenic mimicry-like tubules, that bulge from the spheroid and are composed of a hollow lumen delimited by tumor cells. CABA I can be grown as multicellular spheroids to easily isolate EVs. The presence of features typical of in vivo tumors (inner entrapped EVs and vasculogenic mimicry) suggests their use as faithful experimental models to screen therapeutic drugs targeting these pro-tumorigenic processes

Maternal Smoking and Socio-Demographic Characteristics in Correlation with Low Birth Weight: A Turin (Piedmont) Study

Uvod. Med nosečnostjo kadi 15 % do 25 % žensk. Znanstveni dokazi kažejo, da izpostavljenost kajenju povzroča nižjo porodno težo. Cilj te študije je oceniti povezavo med kajenjem med nosečnostjo, socialnodemografskimi značilnostmi matere in nizko porodno težo. Metode. Podatki so bili pridobljeni iz 1572 vprašalnikov, ki so jih izpolnile vse ženske, ki so v obdobju od leta 2008 do leta 2010 rodile v učni bolnišnici za ginekologijo »S. Anna« v Torinu (Italija). Za oceno povezave med socialno-demografskimi značilnostmi in porodno težo je bila uporabljena multipla logistična analiza; sledil je postopni pristop z »vzvratnim izločanjem« in ocena ustreznosti modela s pomočjo Hosmer- Lemeshowega testa. Rezultati. Univariatna analiza je razkrila, da kajenje cigaret (17 %), nižja stopnja izobrazbe (13 %) in ženski spol dojenčka (13 %) predstavljajo dejavnik tveganja, saj povečujejo tveganje, da bo imel otrok nizko porodno težo. Analiza z logistično regresijo je pokazala, da sta gestacijska starost in kajenje matere statistično povezani spremenljivki. Zaključki. Rezultati potrjujejo, da se porodna teža povečuje sorazmerno z gestacijsko starostjo ter da kajenje matere in spol otroka (ženski) povečujeta tveganje za nižjo porodno težo ob rojstvu. Logistična regresija dokazuje, da povezava med kajenjem matere in nizko porodno težo pomeni povečano tveganje za celotno populacijo (RO = 2,85), tako za novorojenčke moškega (RO = 3,45) kot tudi ženskega spola (RO = 2,44

Maternal smoking and socio-demographic characteristics in correlation with low birth weight: A Turin (piedmont) study

Abstract Introduction. 15 to 25% of women smoke during pregnancy. Scientific evidence suggests that exposure to smoking causes decreased birth weight. The aim of this study was to assess the correlation between smoking during pregnancy, maternal sociodemographic characteristics, and low birth weight. Methods. Data were derived from 1572 questionnaires administered to each woman that gave birth at the Gynecology Teaching Hospital "S. Anna" in Turin (Italy) during the period from 2008 to 2010. Multiple logistic analysis was used to evaluate the association between socio-demographic characteristics and birth weight; the stepwise approach with a "backward elimination" procedure was followed, and the goodness of fit of the model was estimated using the Hosmer-Lemeshow test. Results. The univariate analysis revealed that smoking cigarettes (17%), having a lower educational level (13%), and female sex of the infant (13%) seem to be risk factors, as they increase the risk of having a low birth weight child. Logistic regression analysis showed that gestational age and maternal smoking are the statistically associated variables. Conclusions. The results confirmed that birth weight increases proportionally with the length of the gestational age and that maternal smoking and the child's sex (female) increase the risk of having a lower birth weight. Logistic regression demonstrated that the association between maternal smoking and low birth weight shows an increased risk for the whole population (OR=2.85), for male (OR=3.45) and for female newborns (OR=2.44).Abstract Introduction. 15 to 25% of women smoke during pregnancy. Scientific evidence suggests that exposure to smoking causes decreased birth weight. The aim of this study was to assess the correlation between smoking during pregnancy, maternal sociodemographic characteristics, and low birth weight. Methods. Data were derived from 1572 questionnaires administered to each woman that gave birth at the Gynecology Teaching Hospital "S. Anna" in Turin (Italy) during the period from 2008 to 2010. Multiple logistic analysis was used to evaluate the association between socio-demographic characteristics and birth weight; the stepwise approach with a "backward elimination" procedure was followed, and the goodness of fit of the model was estimated using the Hosmer-Lemeshow test. Results. The univariate analysis revealed that smoking cigarettes (17%), having a lower educational level (13%), and female sex of the infant (13%) seem to be risk factors, as they increase the risk of having a low birth weight child. Logistic regression analysis showed that gestational age and maternal smoking are the statistically associated variables. Conclusions. The results confirmed that birth weight increases proportionally with the length of the gestational age and that maternal smoking and the child's sex (female) increase the risk of having a lower birth weight. Logistic regression demonstrated that the association between maternal smoking and low birth weight shows an increased risk for the whole population (OR=2.85), for male (OR=3.45) and for female newborns (OR=2.44)

Studio multicentrico sul protocollo di revisione dell'uso dell'ospedale in ostetricia (pruo ostetrico).

INTRODUZIONE: Il concetto di appropriatezza rappresenta uno dei principi guida della qualità in ambito sanitario. L’esigenza di razionalizzare i costi dell’assistenza porta alla necessità di valutare l’appropriatezza delle ammissioni ospedaliere e della durata della degenza. Nei ricoveri ostetrici in particolare si è ritenuto opportuno sviluppare uno specifico strumento, realizzato nell’ospedale S. Anna di Torino, denominato PRUO Ostetrico, che è stato impiegato nel presente studio per valutare l’applicabilità in differenti contesti del territorio nazionale e documentare il fenomeno dell’inappropriatezza in tale ambito. MATERIALI E METODI: Lo studio, coss-sectional, è stato condotto in 8 strutture sanitarie di Roma, Torino, Palermo, Avezzano e Salerno. Ciascun centro ha valutato le cartelle cliniche dei reparti di ostetricia avvalendosi del PRUO Ostetrico, per l’anno 2009. Il protocollo suddiviso in giornata di ammissione e degenza, prevede 49 criteri di appropriatezza ed inappropriatezza. Le singole valutazioni sono state riportate su schede cartacee e inserite in un database. L’analisi statistica, condotta utilizzando il software SPSS 19.00, si è focalizzata preliminarmente sulla prevalenza dell’inappropriatezza dell’ammissione e delle singole giornate di degenza, sulla completezza dell’anamnesi e dell’esame obiettivo e valutando possibili associazioni riguardo a modalità e periodo di ricovero e tipo di struttura. RISULTATI: Sono state valutate 986 cartelle cliniche. L’età media delle donne ricoverate è 33 anni (SD=10.7); il 73% è di nazionalità italiana, il 7% rumena, il 3% marocchina e il 17% di altra cittadinanza. Il 90% dei ricoveri è di tipo urgente. Le percentuali di inappropriatezza sulla giornata di ammissione e la prime tre giornate di degenza sono rispettivamente: 4%, 4.7%, 15% e 24.5%. L’anamnesi è presente nell’86.5% delle cartelle, mentre l’esame obiettivo è completo nel 62%. Maggiore inappropriatezza si segnala nei policlinici rispetto ai presidi ospedalieri (7% vs 3.5%; p=0.025) CONCLUSIONI: Alla luce dell’esperienza maturata il PRUO Ostetrico sembra essere uno strumento affidabile che permette di apprezzare l’appropriatezza di tali ricoveri che per peculiarità di ordine clinico e sociale differiscono da quelli per adulti o pediatrici. L’inappropriatezza sembra essere direttamente proporzionale con il protrarsi della degenza, e ciò evidenzia la necessità di una strategia che comprenda discussioni dei risultati con i medici, incontri dedicati nei reparti, al fine di migliorare gli aspetti organizzativi dell’assistenza sanitaria

Development of a new fetal growth curve from a large sample of Italian population

BACKGROUND: Intrauterine growth curves are considered an essential instrument in prenatal medicine for an appropriate auxological classification of fetuses and they have a great importance in clinical practice. Nowadays, in Italy a national curve published in 1975, is the most used. It Is based on birth weights of 8458 newborns from physiological pregnancies. The aim of the present study was to develop a modern fetal growth curve based on accurately selection of 35 240 physiological singleton Italian pregnancies with sure gestational age confirmed by ultrasound. METHODS: This is a retrospective analysis of 35,240 pregnancies from "A. Gemelli" University Hospital in Rome and "S. Anna" University Hospital in Turin from January 2001 to December 2006. Non-resident pregnant women or coming from other countries, women with diabetes, hypertensive disorders of pregnancy, multiple pregnancies, fetuses with major malformations and/or chromosomal disorders and stillborn fetuses were excluded. RESULTS: An increasing trend of median neonatal weight, in comparison with the previous Italian National Curve drawn up in 1975, was found. CONCLUSIONS: Combining data from two centers, a new fetal growth curve, in which the 10th and the 90th percentiles are clinically reliable, was performed, in order to have a better tool to evaluate the Italian fetal population. A trend towards an increase of birth weight was observed if compared to previous growth curve drawn up more than 30 years ago

A multicenter study on the appropriateness of hospitalization in obstetric wards: application of Obstetric Appropriateness Evaluation Protocol (Obstetric AEP)

Abstract The cross-sectional study has been based on the implementation of the Obstetric Appropriateness Evaluation Protocol (OAEP) in seven hospitals to determine inappropriate hospital admissions and days of stay. The outcomes were: inappropriateness of admission and "percentage of inappropriateness" for one hospitalization. A total number of 2196 clinical records were reviewed. The mean percentage of inappropriateness for hospitalization was 22%. The percentage of inappropriateness for the first 10 d of hospitalization peaked in correspondence of the fourth (42%). The logistic regression model on inappropriated admission reported that emergency admission was a protective factor (OR = 0.4) and to be hospitalized in wards with ≥30 beds risk factor (OR = 5.12). The second linear model on "percentage of inappropriateness" showed that inappropriated admission and wards with ≥30 beds increased the percentage (p < 0.001); whereas the admission in Teaching Hospitals was inversely associated (p < 0.001). The present study suggests that the percentage of inappropriate admission depends especially on the inappropriate admission and the large number of beds in obstetric wards. This probably indicates that management of big hospitals, which is very complex, needs improving the processes of support and coordination of health professionals. The OAEP tool seems to be an useful instrument for the decision-makers to monitor and manage the obstetric wards