198 research outputs found
OMICS-DRIVEN CHARACTERIZATION OF PEDIATRIC CD30-POSITIVE LYMPHOMAS: FROM TUMOR BIOPSY TO TUMOR MICROENVIRONMENT
No full textIl linfoma di Hodgkin (HL) e il Linfoma anaplastico a grandi cellule (ALCL) sono entrambi linfomi CD30 positivi che rendono conto rispettivamente del ~10% e 2%-3% di tutti i casi di linfomi pediatrici. Gli attuali trattamenti di prima linea riescono a curare la maggior parte di questi pazienti ma tra il 10% e il 30% ricade o risulta essere resistente/refrattario alla terapia. La biopsia liquida è uno strumento che ha il potenziale di aiutare i clinici a controllare la malattia e a monitorare la risposta alla terapia. In questo contesto, gli approcci OMICI sono stati utilizzati per investigare il ruolo delle small extracellular vescicles (s-EVs) nella disseminazione e progressione tumorale.
Abbiamo condotto uno studio di trascrittomica in una serie di biopsie tumorali di pazienti affetti da ALCL. Abbiamo identificato 19 miRNA maggiormente espressi nei pazienti con una prognosi migliore. In questi pazienti la sovraespressione di miR-939 può contribuire nel regolare finemente il signaling oncogenico di JUNB modulando l’espressione di PDGFRB, suo target trascrizionale.
Un’analisi di small RNA sequencing è stata condotta nelle s-EVs plasmatiche e abbiamo identificato 7 miRNA caricati selettivamente in quelle dei pazienti ricaduti. MiR-146a-5p, miR-378a-3p e let-7g-5p hanno dimostrato avere valore prognostico negativo, sia in analisi univariata sia multivariata, rappresentando quindi un possibile pannello di miRNA che possono essere valutati per l’identificazione di pazienti con prognosi infausta. In base ai dati di letteratura, abbiamo concentrato la nostra attenzione su miR-146a-5p dato il suo ruolo nel modulare il differenziamento in senso M2 pro-tumorale dei macrofagi.
Abbiamo quindi confermato questo ruolo dimostrando la capacità di questo miRNA di stimolare la migrazione dei macrofagi verso le cellule tumorali e di potenziare l’aggressività tumorale. Un'altra importante componente del cargo esosomiale è costituita dalle proteine. Abbiamo caratterizzato il cargo delle s-EVs di pazienti con ALCL e i nostri risultati hanno indicato la presenza di proteine facenti parte del pathway di PI3K/AKT in particolare HSP90. Livelli anormali di un'altra proteina, l’osteopontina (OPN/SPP1), sono risultati essere più alti nei pazienti ricaduti, mentre la tenascina (TNC) è risultata essere molto espressa in generale nelle vescicole dei pazienti rispetto ai donatori sani. Nel complesso, dall’analisi proteomica del cargo delle s-EVs si evidenzia che TNC, OPN/SPP1 e HSP90 rappresentano potenziali biomarcatori per i pazienti pediatrici con ALCL.
Anche per HL abbiamo analizzato i miRNA caricati nelle s-EVs isolate dal plasma dei pazienti alla diagnosi. Tramite sequenziamento abbiamo identificato un profilo peculiare nei pazienti rispetto ai donatori sani. Dopo integrazione dei dati di sequenziamento con le informazioni cliniche, abbiamo deciso di concentrare la nostra attenzione su miR-122-5p che è risultato essere più abbondantemente caricato nelle vescicole isolate dai pazienti rispetto ai donatori sani e anche nei pazienti ricaduti rispetto ai non ricaduti.
Dall’analisi delle cellule mononucleate del sangue periferico abbiamo identificato un caratteristico profilo in termini di cellule circolanti associato con più alti livelli di questo miRNA.
La valutazione dei livelli circolanti di miR-122-5p può rappresentare quindi un importante strumento che permette di identificare i pazienti ad alto rischio e quindi indirizzarli ad una terapia più mirata come quella che utilizza anticorpi anti-PD1.
I dati ottenuti hanno messo in luce la superiorità della biopsia liquida rispetto alla classica biopsia tissutale. I biomarcatori che abbiamo identificato rappresentano uno strumento che consente di identificare i pazienti ad alto rischio e il loro possibile ruolo biologico mette le basi per una migliore conoscenza di queste due patologie.Hodgkin lymphoma (HL) and Anaplastic Large Cell Lymphoma (ALCL) are both CD30-positive lymphomas accounting for ~10% and 2%–3% of all cases of lymphoid neoplasms, respectively. Current first line treatments can cure most of these patients but 10% to 30% of them are resistant or relapse after first-line therapy. Liquid biopsy has the potential to help clinicians to screen for disease and monitor treatment response and OMICS approaches have been used to investigate the role of small-Extracellular vesicles (s-EVs) in tumor spread and progression.
We have started by analyzing miRNAs expression on ALCL primary tumor biopsy. Microarray analysis identified 19 miRNAs upregulated in patients with better prognosis. Among these, we demonstrated that miR-939 overexpression can contribute to fine-tuning the JUNB-mediated oncogenic signaling by modulating its transcriptional target platelet derived growth factor receptor (PDGFRB), further explaining the favorable outcome of these cases.
While a bioptic enrichment of miR-939 is associated to a better prognosis, from small-RNA-sequencing analysis performed on ALCL plasma s-EVs we identified 7 miRNAs more represented in relapsed patients and 10 miRNAs that otherwise are less represented. MiR-146a-5p, miR-378a-3p and let-7g-5p resulted to have highly significative prognostic impact both in univariate and in multivariate analysis, thus acting as a possible panel for the identification of high-risk patients. We focused on miR-146a-5p since its reported role in modulating tumor supporting-M2 macrophages differentiation. We confirmed this function, and we demonstrated its potential to increase macrophages migration toward tumor site and to potentiate tumor aggressiveness.
Another interesting component of s-EVs cargo are proteins. We performed a comprehensive characterization of proteins circulating in ALCL patients’ plasma s-EVs. Our results also indicated that proteins of the PI3K/AKT pathway circulate in ALCL patients’ bloodstream within s-EVs, particularly HSP90. Abnormally high levels of osteopontin (OPN/SPP1) are present in s-EVs, with higher levels in two patients that subsequently relapsed. Moreover, we highlighted the presence of more than twice the amount of TNC in patients s-EVs with respect to HD samples. Overall, proteomic analysis of pediatric ALCL plasmatic s-EVs suggests TNC, OPN/SPP1 and HSP90 as potential prognostic biomarkers for pediatric ALCL disease.
As for ALCL, also for cHL we focused our attention on s-EVs miRNAs cargo. We performed s-RNA-seq analysis on s-EVs isolated from plasma samples of 36 pediatric cHL identifying a peculiar cargo of miRNAs in cHL compared to HD. Sequencing data were then analyzed by integrating with clinical information. We decided to focus on miR-122-5p that resulted to be more abundantly loaded in patients s-EVs compared to HD and in those of patients who experienced relapse compared to not relapsed ones. By collecting paired samples of plasma and peripheral blood mononuclear cells (PBMC) obtained at diagnosis we were able to identify a peculiar profile in circulating immune cells by stratifying patients based on miR-122-5p relative abundance. The evaluation of circulating miR-122-5p could represent an effective tool to uncover high risk cHL patients yet at diagnosis and to address them to a more targeted therapy such as anti-PD-1.
Overall, the data obtained during this PhD have put in light the power of liquid biopsy over classical bioptic investigation both as prognostic and diagnostic tool. The identified biomarkers represent effective and simple tools to identify high risk patients and their proposed possible biological role put the basis for a better knowledge of these pathologies
Religion and the Global City
No full textReligion and the Global City examines the new realities of religion in global cities, bringing together in-depth case studies to reveal the presence, visibility, and social, political and cultural roles of religion in contemporary global urban landscapes, and offering an overview of the main debates and developments in this growing field
Grgur Garbin († 1621), captain of artillerymen, and his house in Zadar
No full textU radu se na temelju arhivskih i terenskih istraživanja donose nove spoznaje o luneti portala s nekadašnje kuće obitelji Garbin u Zadru, danas ugrađenoj u kuću u Ulici Mate Karamana. Preciznije se blazonira grb u luneti portala te ga se dovodi u vezu s kapetanom zadarskih topnika Grgurom Garbinom. Plemićka obitelj Garbin podrijetlom je iz Paga, a jedna grana prešla je početkom 17. stoljeća u Zadar. Zaključuje se da je luneta portala izrađena malo nakon 1609. godine kada prema arhivskim dokumentima Grgur Garbin kupuje kuću u blizini samostana sv. Dimitrija u Zadru. Utvrđuje se položaj kuće te se donose novi arhivski podatci iz života Grgura Garbina i njegovih potomaka. Saznaje se da je njegov sin Ivan Grgur (Zan Gregorio) zanimanjem bio zlatar.On the basis of the conducted archival and field research, the author presents new knowledge regarding the portal lunette earlier home of the Garbin family in Zadar, today built into the front facade of the house in Mate Karaman Street. The author describes the coat of arms in the portal lunette, and consequently – since it shows a cannon and a male head with the characteristic hat – brings it into connection with Grgur Garbin, captain of the Zadar artillerymen. In the 17th century, this type of hat – decorated with long feathers and identical to the one on Garbin’s grave – made an integral part of the uniform of Venetian commanders holding the rank of captain; this counts for artillery units, too. The noble Garbin family originates from the island of Pag, and one family branch moved to Zadar at the beginning of the 17th century. It is concluded that the portal lunette was built just after the year 1609, when – according to the archival documents – Grgur Garbin had bought a house near the monastery of St. Demetrius in Zadar. The author sets the accurate location of Garbin’s house and brings fresh archival data from the life of Grgur Garbin and his descendants. Record on the death of Grgur Garbin, captain of the Zadar artillerymen, was entered into registry books under the date 25 January 1621, and he was buried in the church of St. Demetrius near his family home. Grgur Garbin had two sons, both born in Zadar – Marco Antonio, baptised on 29 April 1609, and Ivan Grgur, baptised on 21st March 1611. Based on recent archival research, the author further learned that Garbin’s son Ivan Grgur, who died in Zadar in 1646, was professional goldsmith
Endoplasmic Reticulum Stress, NRF2 Signalling and Cardiovascular Diseases in a Nutshell
No full textThis short review is intended primarily to summarize the understanding of the interrelated roles of endoplasmic reticulum (ER) stress, oxidative stress and inflammation in cardiovascular diseases
Lung ultrasound in internal medicine: training and clinical practice
No full textAbstract Background Lung ultrasound (LUS) represents an emerging technique for bedside chest imaging in different clinical settings. A standardized approach allows the diagnosis, the quantification, and the follow-up of different conditions for which acute respiratory failure is the main clinical presentation. The aim of this study was to test what skill targets could be achieved in LUS, with a short-training course offered to 19 Medical Doctors attending the certification board school in Internal Medicine at the University of Verona, Italy. Methods The training course (theoretical and practical) consisted of 9 h subdivided in 4 days. Each trainee examined three healthy volunteers during the first day that was also the day of the theoretical lessons. Moreover, they examined nine patients per day (a total of 27 patients). Trainees were tested in the recognition of the basic signs in LUS, the managing of the Bedside Lung Ultrasound Evaluation (the BLUE protocol), and the recognition of the broad clinical scenarios recognized by the LUS. Kappa statistic was used to calculate the inter-observer agreement (trainees/tutor). Results Twenty-seven patients were examined by the 19 trainees (ten trainees had previous limited experience in general ultrasound). The agreement among the trainees and the tutor in the recognition of the LUS basic signs and in the recognition of the BLUE protocol profiles ranged from “fair” to “excellent”. In particular, the agreement among the trainees and the tutor in the final LUS diagnosis was “excellent” for the recognition of the interstitial syndrome and the pleural effusion, “substantial” for the recognition of the normal lung, and “moderate” for the recognition of consolidation and pneumothorax. LUS outcome gave useful information and drove change in therapy in 16 patients. It affected immediate management in nine patients. The concordance between the previous X chest ray and LUS was observed in 21 patients. Conclusions A short training in LUS provided good proficiency in the recognition only of the main signs of the BLUE protocol, but allowed a correct LUS diagnosis in the Internal Medicine most frequent clinical settings of acute respiratory failure. This study supports incorporating LUS into Internal Medicine fellowship training programs
Inhibition of lectin-like oxidized low-density lipoprotein receptor-1 expression: is it right now a safe and promising therapeutic approach for atherosclerosis?
No full textLectin -like oxidized(ox) LDL receptor-1 (LOX-1) is the major receptor for oxLDL; several studies in vitro and in animal models have shown that LOX-1 is involved in almost all proatherogenic effects of oxLDL. Growing evidence also demonstrated that LOX-1 plays a role not only in the initiation and formation, but also in plaque destabilization and rupture and strengthen the role of this receptor in cardiovascular disease. As several studies gathered from animal models, as well as genetic studies, support the hypothesis that the binding and internalization of oxLDL by scavenger receptors such as LOX-1 triggers a series of mechanisms leading to endothelial dysfunction and foam cell formation, modulation of LOX-1 levels has been suggested to be one therapeutic route in attenuating early events in atherosclerosis. This is an editorial comment to a paper in which the authors designed a pyrrole–imidazole polyamide to bind to a proximal site of the AP-1 binding region in the LOX-1 gene promoter. This molecule interfered with AP-1 protein binding to the LOX-1 gene promoter and inhibited the phorbol myristate acetate-mediated enhancement of LOX-1 gene expression in human umbilical vein endothelial cells and decreased apoptosis induced by Angiotensin II and oxLDL in HUVECs. As pyrrole–imidazole polyamide was efficacious in inhibition of LOX-1 expression and apoptosis, which has been shown to be associated with the vulnerability of plaques, the authors propose the use of this agent for the study and treatment of atherosclerotic disease. Although silencing LOX-1 expression opens an appealing scenario on treatment of atherosclerosis, some important questions remain to be elucidated before translating fascinating results, obtained in vitro and in animal models, to patients. Further studies on inhibition of LOX-1 expression or drugs targeting LOX-1 may help to enlighten these open issue
Short training in focused cardiac ultrasound in an Internal Medicine department: what realistic skill targets could be achieved?
No full textThe importance of focused cardiac ultrasound (FCU) in Internal Medicine care has been recognized by the American Society of Echocardiography. The aim of this study was to test what realistic skill targets could be achieved in FCU, with a relatively short training (theoretical and practical) of 9 h offered to Internal Medicine certification board attending students, and if the addition of further 9 h of training could significantly improve the level of competence. Kappa statistic was used to calculate the inter-observer agreement (trainees/tutor). The agreement between the trainees (who completed the entire training) and the tutor was, respectively, "substantial" (k = 0.71) for the identification of pericardial effusion, "moderate" (k = 0.56-0.54) for the identification of marked right ventricular and left ventricular enlargement, "substantial" (k = 0.77) for the assessment of global cardiac systolic function by visual inspection and "fair" (k = 0.35) for the assessment of size and respiratory change in the diameter of the inferior cave vein (IVC). 18 h training in FCU provided proficiency in obtaining adequate images from the parasternal window without providing the ability to correctly master the apical and subcostal windows. As concerns the interpretative skills, only pericardial effusion and visual estimation of global systolic function could be correctly identified, while ventricular enlargement and IVC prove to be more difficult to evaluate. This study supports incorporating FCU into Internal Medicine fellowship training programs, and should facilitate the design of other similar training courses
An exploratory look at NETosis in atherosclerosis
No full textCurrent evidence suggests the likelihood of a link between venous thromboembolism (VTE) and atherosclerosis, although they have been traditionally considered as different pathological entities. The contribution of neutrophils to human atherogenesis has been underestimated, if compared to their contribution established in VTE. This is due to the major importance attributed to macrophages in plaque destabilization. Nevertheless, the role of neutrophils in atherogenesis deserves increasing attention. In particular, neutrophil extracellular traps (NETs) are net-like chromatin fibres that are released from dying neutrophils. The death of neutrophils with NETs formation is called NETosis. During activation, neutrophils produce reactive oxygen species (ROS), through the activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. The main function of NETs is trapping and killing pathogens. Nevertheless, NETs formation has been observed in various chronic inflammatory diseases, autoimmune diseases, vasculitis, lung diseases, cancer and VTE. Recent studies suggest that NETs formation might contribute also to atherosclerosis progression. New data report the presence of NETs in the luminal portion of human atherosclerotic vessels and coronary specimens obtained from patients after acute myocardial infarction. Programmed death mechanisms in atherosclerosis such as apoptosis, efferocytosis and also NETosis, share common features and triggers. If defective, they can lead the cells to a switch from programmed death to necrosis, resulting in the release of pro-atherogenic factors, accumulation of cell debris and progression of the disease. This review provides evidence on the emerging role of neutrophils focusing on NETosis and oxidative stress burden in orchestrating common mechanisms in atherosclerosis and thrombosis
- …
