1,720,976 research outputs found
Analisi delle Vescicole Extracellulari come potenziali biomarkers nelle patologie tumorali
No full textExtracellular Vesicles (EVs) are non-replicating particles released by cells, crucial for cellular communication, and implicated in cancer progression. Their content mirrors their parent cells, making them potential candidates for cancer biomarker discovery. While most studies relied on two-dimensional (2D) cell cultures, these do not accurately represent in vivo tumor microenvironments. In contrast, three-dimensional (3D) cultures provide a more realistic model, though standardized EV isolation protocols remain undeveloped.
This study aimed to develop a 3D culture model using the human ovarian cancer cell line CABA I to isolate EVs. Using the hanging drop method, 3D spheroids were optimized for size and morphology, evaluated for EV release, and showed multilayered structures and vasculogenic mimicry phenomenon. The study was then extended to 2 breast cancer cell lines with different aggressiveness grades (MDA-MB-231 and MCF-7), adapting methods to optimize 3D culture conditions and EV isolation.
Since the main purpose of the present study was to identify potential biomarkers associated with EVs, concurrently, studies were initiated on 3 blood samples from healthy donors, aiming to identify the best biological fluid between serum and plasma for EV isolation: both biofluids contained EVs but each one carries pros and cons. In the end, plasma was chosen for further study.
Preliminary in vitro evaluations compared 2D and 3D breast cancer cell lines-derived EVs using Fourier Transform Infrared Spectroscopy (FT-IR) and ELISA for glycan profiling.
Notably, FT-IR analysis revealed differences in the molecular composition of EVs from MDA-MB-231 2D and 3D models, reflecting, hypothetically, different biological activities, whereas MCF-7 showed no significant differences.
Glycan profiling indicated variations in 2D and 3D breast cancer-derived EVs, with differences in galactose, fucose, and N-acetylglucosamine levels. Parallel analyses on the ES-2 ovarian cancer cell line supported these findings, highlighting the dynamic EV glycan composition.
FT-IR and glycan profiling were also applied to plasma-EVs from breast cancer patients and healthy donors. FT-IR requires further refinement for plasma-derived EV analysis, while the glycan profile evaluation revealed few differences between patients and healthy donors in the level expression of N-acetyl galactosamine and fucose, respectively, which were higher and more stable in healthy donors compared to patients. However, further investigations with larger cohorts are needed to achieve statistical significance.
In summary, the results of this thesis showed that the molecular composition of EVs varies in the different cell culture models, emphasizing the importance of selecting an appropriate in vitro model to accurately reflect in vivo conditions. FT-IR spectroscopy and glycan profiling are promising techniques for identifying EV-related biomarkers, but further research is required to refine FT-IR protocols, particularly for plasma-derived EVs. Nevertheless, glycan analysis offers promising initial results for potential clinical applications
Extracellular Vesicles as Matrix Messengers
No full textThe extracellular matrix (ECM) is a dynamic, acellular framework that maintains homeostasis and supports the preservation of tissues’ structural integrity. From early embryonic stages forward, the ECM organizes itself through a variety of changes, including degradation, synthesis, and rearrangement of matrix elements. Although the mechanisms that underlie these changes are strictly controlled, the ECM can undergo structural and functional alterations that result in diseases or even promote the growth of tumors. Several elements mediate communication between the cells that make up the tissue and the matrix, with extracellular vesicles (EVs) playing a significant role. EVs are spherical particles surrounded by a phospholipid bilayer, secreted by all living organisms and present in all organs, tissues, and bio
logical fluids, both in physiological and in pathological conditions. In this review, we will present a comprehensive understanding of the matrix’s structure and the roles in which it is engaged, as well as the implications of EVs. Several aspects of EVs–ECM interaction under healthy and pathological settings will be covered, with an emphasis on EVs’ function in matrix degradation
Unleash the potential of Fourier-Transform Infrared (FT-IR) spectroscopy to study extracellular vesicles cancer
No full textBreast cancer remains a primary health concern among women worldwide, and insight into its initiation, progression,
and metastasis are of the utmost importance. The role of Extracellular Vesicles (EVs), cell-derived membrane- surrounded vesicles, in cell-to-cell communication has long been recognized as a crucial component in these processes. A wide range of studies based on EVs are carried out on in vitro models, based on bi-dimensional (2D) cultures; these studies led to a greater understanding not only of the different mechanisms by which EVs contribute to tumor progression but also of their potential clinical use in diagnostics as biomarkers. Thus, investigating the molecular composition of EVs is of considerable importance, since it reflects the molecular composition of the parental cells [1]. Among all the techniques already set up, Fourier Transform Infrared spectroscopy (FTIR) has recently been introduced to the study of EVs. This innovative approach, commonly used in the material sciences field, enables fast label-free profiling, obtaining a distinctive molecular ‘fingerprint’ of EVs from minimal sample quantities, allowing the discovery
of potential differences in molecular composition [2]. For this reason, for our study, EVs were isolated from two breast cancer cell lines, with different aggressiveness levels, cultured in 2D; EVs were isolated by ultracentrifugation technique and, once resuspended in a saline solution, they were analyzed for their FTIR spectra, to evaluate whether there could be differences that could reflect the different tumor grades. The results showed that there are some differences in several spectral regions of the two EVs samples, suggesting that the FTIR could allow the recognition of EVs, based on cell origin. However, the 2D models aren’t able to truthfully mimic the true complexity of the in vivo tumor microenvironment and the interactions mediated by EVs in it. Thus, more recent studies concerning EVs were focused on in vitro three-dimensional (3D) cell cultures, using tumor spheroids models [3]. To evaluate if there could be a difference in the molecular fingerprint of EVs derived from cells cultured in 2D or 3D, we isolated the EVs from breast tumor cells cultured by these two models and FTIR was performed on them. The results showed that the EVs isolated
from 3D culture have different content in some molecular components compared to the EVs isolated from the respective cell lines cultured on 2D, suggesting that the composition of EVs is profoundly influenced by the cell culture methods and leading us to reflect on a careful evaluation of the most suitable cellular models in the in vitro studies. In conclusion, our preliminary data suggest that the FTIR offers new directions for EVs molecular fingerprints, making this tool potentially useful for biomarker discovery in liquid biopsy based on EVs
Development of a new fetal growth curve from a large sample of Italian population
No full textBACKGROUND: 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
Maternal smoking and socio-demographic characteristics in correlation with low birth weight: A Turin (piedmont) study
No full textAbstract
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)
Metastatic Dissemination: Role of Tumor-Derived Extracellular Vesicles and Their Use as Clinical Biomarkers
Full text linkCancer is a major cause of mortality in humans; often, rather than the primary tumor, it is the presence of metastases that are the cause of death. Extracellular vesicles (EVs) are small structures released by both normal and cancer cells; regarding the latter, they have been demonstrated to modulate almost all cancer-related processes, such as invasion, angiogenesis induction, drug resistance, and immune evasion. In the last years, it has become clear how EVs are widely involved in metastatic dissemination as well as in pre-metastatic niche (PMN) formation. Indeed, in order to achieve a successful metastatic process, i.e., penetration by cancer cells into distant tissues, the shaping of a favorable environment into those distant tissue, i.e., PMN formation, is mandatory. This process consists of an alteration that takes place in a distant organ and paves the way for the engraftment and growth of circulating tumor cells derived from the tumor primary site. This review focuses on the role of EVs in pre-metastatic niche formation and metastatic dissemination, also reporting the last studies suggesting the EVs role as biomarkers of metastatic diseases, possibly in a liquid biopsy approach
The Trend of Long Pentraxin 3 and Other Inflammatory Serum Markers in the 30 Days After Total Hip Arthroplasty
Full text linkOne of the most dangerous and difficult side effects to treat after total hip arthroplasty (THA) is periprosthetic or superficial site infection. Blood and synovial fluid biomarkers have recently come into focus in addition to conventional systemic indicators of inflammation in order to assess their potential utility in the diagnosis of infections. Long pentraxin 3 (PTX3) appears to be a sensitive biomarker of acute-phase inflammation. The purpose of this study is to determine plasma PTX3 in patients undergoing THA and compare its trend with other common serum markers, such as CRP, D-dimer, procalcitonin, and ESR up to 30 days post-operatively. Patients with hip arthritis or avascular necrosis of the femoral head were consecutively enrolled in a single-center study. Each patient underwent blood testing for ESR, CRP, procalcitonin, D-dimer, and PTX3 levels before surgery and at 1, 3, 5, 15, and 30 days after THA. PTX3 was measured using the ELISA method. Other markers’ values and trends were compared with PTX3’s. A total of 50 patients met our inclusion criteria. When different trends were evaluated, PTX3 was found to have a trajectory and sensitivity comparable to other inflammatory markers. Notably, PTX3 changed more quickly than the other markers, with a sharp increase immediately post-operatively, followed by normalization at the 5-, 15-, and 30-day follow-ups, corresponding to the resolution of the inflammatory condition. However, 30 days post surgery, no patients exhibited signs or symptoms of early prosthetic infection. PTX3 is confirmed as a reliable and promising serum biomarker for tracking the level of inflammation in patients undergoing total hip replacements. Blood PTX3 values rise even more rapidly than CRP and procalcitonin and then quickly return to normal values when the inflammatory process resolves. One of the primary barriers to PTX3’s inclusion in routine studies on early periprosthetic infections is the waiting period for PTX3 sample analysis
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
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