STUDIO DI ASSOCIAZIONE TRA POLIMORFISMI DEL GENE DELLA DNA METILTRANSFERASI-3B (DNMT3B) E LA MIASTENIA GRAVIS IN PAZIENTI CON E SENZA PATOLOGIE DEL TIMO

La Miastenia gravis (MG) è una patologia autoimmune determinata, nell’80% dei casi, dalla presenza di autoanticorpi diretti contro il recettore nicotinico dell’ Acetilcolina (AchR+); recentemente sono stati descritti anche altri target, come le proteine MuSK e LRP4. È ormai ampiamente accettato che il timo sia il primo sito di sviluppo e mantenimento dell'autoimmunità nei pazienti affetti da MG. Inoltre, una percentuale del 10-20% dei pazienti affetti dalla MG presenta timomi ed un 70% dei pazienti di MG presenta iperplasia timica. In letteratura sono presenti diversi studi che suggeriscono un contributo dato dai processi epigenetici, come la metilazione del DNA, nel promuovere la cancerogenesi e l’autoimmunità. Tuttavia, il ruolo dell’epigenetica nella MG non è stato ancora completamente chiarito. L’obiettivo del nostro studio è stato quello di valutare il possibile contributo dato dai polimorfismi della DNMT3B (-149C>T e -579G>T), che codifica per la DNA metiltransferasi 3B, nel rischio di sviluppare la Miastenia gravis e/o timomi ad essa associati. I polimorfismi in studio sono stati selezionati sulla base di recenti reports e sulla base di meta-analisi prese dalla letteratura che indicano una associazione tra questi polimorfismi ed un aumentato rischio di sviluppare diverse neoplasie, come il carcinoma del colon-retto ed il carcinoma polmonare e patologie autoimmuni, come la porpora trombocitopenia idiopatica. La popolazione in studio è composta da 324 pazienti MG AchR+ , di cui 140 maschi e 184 femmine, e 409 controlli, di cui 147 maschi e 262 femmine. All’interno del gruppo dei pazienti di MG, 94 di questi presentano un timoma. Dallo studio non emerge alcuna associazione statisticamente significativa tra l’intera popolazione di pazienti ed i controlli per entrambi i polimorfismi. Dopo aver suddiviso i pazienti MG in tre sottogruppi (timoma, iperplasia timica, timo normale) si individua una associazione statisticamente significativa dell’ allele DNMT3B -579T (OR=1.58; 95%CI= 1.14-2.18, P=0.0063) e del genotipo mutato TT (OR= 2.56; 95%CI=1.29-5.07, P=0.0071) con il rischio di sviluppare il timoma. Per il polimorfismo DNMT3B -149C>T si individua una associazione statisticamente significativa del genotipo eterozigote CT (OR= 1.71; 95%CI=1.04-2.82, P=0.00347) rispetto al wild-type CC con l’aumentato rischio di sviluppare il timoma. Questo risultato suggerisce che la presenza dell’allele DNMT3B-579T potrebbe rappresentare un fattore di rischio per lo sviluppo dei timomi associati alla Miastenia gravis, in particolare nel genotipo omozigote TT. Perciò l’analisi di questo polimorfismo potrebbe aiutare ad identificare quali pazienti di MG AchR+ hanno un aumentato rischio di sviluppare un timoma

MICRONUCLEI, GENETIC POLYMORPHISMS AND OTHER BIOLOGICAL PARAMETERS AS EARLY BIOMARKERS FOR THE RISK OF CANCER

SCOPO: Lo scopo del presente studio caso-controllo appaiato e` stato quello di validare l’uso del test del MN in PBL come biomarcatore precoce di rischio di cancro, con una speciale attenzione al possibile effetto dei polimorfismi genetici (GSTM/GSTT1 e MTHFR C677T) sulla frequenza basale di MN ed in relazione ai livelli plasmatici di acido folico, omocisteina e vitamina B12. METODI: Sono stati raccolti campioni di sangue da 1.650 soggetti sani selezionati da una popolazione generale tra giugno 1991 e novembre 1993, e sono stati preparati immediatamente vetrini per il test del MN. Lo stato di vita, o la causa di morte, e` stato monitorato per tutti i soggetti fino a gennaio 2005. Alla fine del campionamento, sono stati registrati 111 decessi: 52 casi di cancro e 59 per altre cause (soprattutto malattie cardiovascolari). Per lo studio presente, sono stati selezionati 101 individui sani di controllo dall’intero campione di popolazione, appaiandoli per età` (± 5 anni) e genere. 2.000 cellule binucleate (BN) per soggetto sono state lette per il test del MN. RISULTATI: La frequenza di MN era significativamente più` elevata (test di Kruskall-Wallis, p=1.4 x 10-8) nei casi (4.7 ± 3.4 MN/1000 BN cellule) che nel gruppo di controllo (1.5 ± 1.7 MN/1000 BN cellule). Nell’analisi di regressione logistica non e` stata osservata l’influenza di altri fattori, come età, genere, genotipo, abitudine al fumo e al bere. CONCLUSIONI: MN e` stato confermato essere un buon biomarcatore citogenetico che può potenzialmente essere usato come predittore di rischio di cancro con una sensibilità` veramente elevata (0.80; 95%C.I. 0.69-0.91). Questo studio contribuirà agli studi correnti in Europa sulla stima del valore predittivo del MN per il rischio di sviluppare il cancro

Shopify: the valuation case

We start the coverage of Shopify with a Sell recommendation given our target price of $391.00corresponding a downside of 66$1180.00(as of May 20th2021).The market is overestimating Shopify’s external and internal opportunities. The launch of vaccines and low barriers to entry in the industry are likely to slow down the surging value of Shopify. Considering unrealistic expectations in high-tech stocks, Shopify’s value is expected to slow its growth pace, thus the SELL recommendation

Context-Aware Performance Benchmarking of a Fleet of Industrial Assets

Industrial assets are instrumented with sensors, connected and continuously monitored. The collected data, generally in form of time-series, is used for corrective and preventive maintenance. More advanced exploitation of this data for very diverse purposes, e.g. identifying underperformance, operational optimization or predictive maintenance, is currently an active area of research. The general methods used to analyze the time-series lead to models that are either too simple to be used in complex operational contexts or too difficult to be generalized to the whole fleet due to their asset-specific nature. Therefore, we have conceived an alternative methodology allowing to better characterize the operational context of an asset and quantify the impact on its performance. The proposed methodology allows to benchmark and profile fleet assets in a context-aware fashion, is applicable in multiple domains (even without ground truth). The methodology is evaluated on real-world data coming from a fleet of wind turbines and compared to the standard approach used in the domain. We also illustrate how the asset performance (in terms of energy production) is influenced by the operational context (in terms of environmental conditions). Moreover, we investigate how the same operational context impacts the performance of the different assets in the fleet and how groups of similarly behaving assets can be determined

Reactive oxygen species, antioxidant mechanisms and serum cytokine levels in cancer patients: impact of an antioxidant treatment

Objective. So far, it is not well established whether oxidative stress found in cancer patients results from an increased production of oxidants in the body or from a failure of physiological antioxidant systems. To further investigate this question we have assessed the blood levels of reactive oxygen species as a marker of free radicals producing oxidative stress and the most relevant of the physiological body enzymes counteracting reactive oxygen species, namely glutathione peroxidase and superoxide dismutase. Serum levels of proinflammatory cytokines and IL-2 were also investigated. All these parameters were studied in relation to the clinically most important index of disease progression, namely Performance Status (ECOG PS). We also tested the reducing ability of different antioxidant agents on reactive oxygen species levels by measuring the increase in glutathione peroxidase activity, and the reduction of serum levels of IL-6 and TNF. Design, setting and subjects. We carried out an open non randomized study on 28 advanced stage cancer patients (stage III, 10.7%, and stage IV, 89.3%) with tumours at different (8) sites: all were hospitalized in the Medical Oncology Dept, University of Cagliari Interventions. The patients were divided into 5 groups and a different antioxidant treatment was administered to each group. The selected antioxidants were: alpha lipoic acid 200 mg/day orally, N-acetylcysteine 1800 mg/day i.v. or carboxycysteine-lysine salt 2.7 g/day orally, amifostine 375 mg/day i.v., reduced glutathione 600 mg/day i.v., vitamin A 30000 IU/day orally plus vitamin E 70 mg/day orally plus Vitamin C 500 mg/day orally. The antioxidant treatment was administered for 10 consecutive days. Results. Our results show that all but one of the antioxidants tested were effective in reducing reactive oxygen species levels and 2 of them (cysteine-containing compounds and amifostine) had the additional effect of increasing glutathione peroxidase activity. Comprehensively, the “antioxidant treatment” was found to have an effect both on reactive oxygen species levels and glutathione peroxidase activity. The antioxidant treatment also reduced serum levels of IL-6 and TNF. Patients in both ECOG PS 0-1 and ECOG PS 2-3 responded to antioxidant treatment

Transplanted murine long-term repopulating hematopoietic cells can differentiate to osteoblasts in the marrow stem cell niche

Bone marrow transplantation (BMT) can give rise to donor-derived osteopoiesis in mice and humans; however, the source of this activity, whether a primitive osteoprogenitor or a transplantable marrow cell with dual hematopoietic and osteogenic potential, has eluded detection. To address this issue, we fractionated whole BM from mice according to cell surface immunophenotype and assayed the hematopoietic and osteopoietic potentials of the transplanted cells. Here, we show that a donor marrow cell capable of robust osteopoiesis possesses a surface phenotype of c-Kit(+) Lin(-) Sca-1(+) CD34(-/lo), identical to that of the long-term repopulating hematopoietic stem cell (LTR-HSC). Secondary BMT studies demonstrated that a single marrow cell able to contribute to hematopoietic reconstitution in primary recipients also drives robust osteopoiesis and LT hematopoiesis in secondary recipients. These findings indicate that LTR-HSC can give rise to progeny that differentiate to osteoblasts after BMT, suggesting a mechanism for prompt restoration of the osteoblastic HSC niche following BM injury, such as that induced by clinical BMT preparative regimens. An understanding of the mechanisms that regulate this differentiation potential may lead to novel treatments for disorders of bone as well as methods for preserving the integrity of endosteal hematopoietic niches.Bone marrow transplantation (BMT) can give rise to donor-derived osteopoiesis in mice and humans; however, the source of this activity, whether a primitive osteoprogenitor or a transplantable marrow cell with dual hematopoietic and osteogenic potential, has eluded detection. To address this issue, we fractionated whole BM from mice according to cell surface immunophenotype and assayed the hematopoietic and osteopoietic potentials of the transplanted cells. Here, we show that a donor marrow cell capable of robust osteopoiesis possesses a surface phenotype of c-Kit + Lin - Sca-1 + CD34 -/lo, identical to that of the long-term repopulating hematopoietic stem cell (LTR-HSC). Secondary BMT studies demonstrated that a single marrow cell able to contribute to hematopoietic reconstitution in primary recipients also drives robust osteopoiesis and LT hematopoiesis in secondary recipients. These findings indicate that LTR-HSC can give rise to progeny that differentiate to osteoblasts after BMT, suggesting a mechanism for prompt restoration of the osteoblastic HSC niche following BM injury, such as that induced by clinical BMT preparative regimens. An understanding of the mechanisms that regulate this differentiation potential may lead to novel treatments for disorders of bone as well as methods for preserving the integrity of endosteal hematopoietic niches. © The American Society of Gene & Cell Therapy

Polyphosphoester-stabilized cubosomes encapsulating a Ru(II) complex for the photodynamic treatment of lung adenocarcinoma

The clinical translation of photosensitizers based on ruthenium(II) polypyridyl complexes (RPCs) in photodynamic therapy of cancer faces several challenges. To address these limitations, we conducted an investigation to assess the potential of a cubosome formulation stabilized in water against coalescence utilizing a polyphosphoester analog of Pluronic F127 as a stabilizer and loaded with newly synthesized RPC-based photosensitizer [Ru(dppn)2(bpy-morph)](PF6)2 (bpy-morph = 2,2'-bipyridine-4,4'-diylbis(morpholinomethanone)), PS-Ru. The photophysical characterization of PS-Ru revealed its robust capacity to induce the formation of singlet oxygen (1O2). Furthermore, the physicochemical analysis of the PS-Ru-loaded cubosomes dispersion demonstrated that the encapsulation of the photosensitizer within the nanoparticles did not disrupt the three-dimensional arrangement of the lipid bilayer. The biological tests showed that PS-Ru-loaded cubosomes exhibited significant phototoxic activity when exposed to the light source, in stark contrast to empty cubosomes and to the same formulation without irradiation. This promising outcome suggests the potential of the formulation in overcoming the drawbacks associated with the clinical use of RPCs in photodynamic therapy for anticancer treatments

Early Cerebral Hemodynamic, Metabolic, and Histological Changes in Hypoxic–Ischemic Fetal Lambs during Postnatal Life

The hemodynamic, metabolic, and biochemical changes produced during the transition from fetal to neonatal life may be aggravated if an episode of asphyxia occurs during fetal life. The aim of the study was to examine regional cerebral blood flow (RCBF), histological changes, and cerebral brain metabolism in preterm lambs, and to analyze the role of oxidative stress in the first hours of postnatal life following severe fetal asphyxia. Eighteen chronically instrumented newborn lambs were randomly assigned to either a control group or the hypoxic–ischemic (HI) group, in which case fetal asphyxia was induced just before delivery. All the animals were maintained on intermittent positive pressure ventilation for 3 h after delivery. During the HI insult, the injured group developed acidosis, hypoxia, hypercapnia, lactic acidosis, and tachycardia (relative to the control group), without hypotension. The intermittent positive pressure ventilation transiently improved gas exchange and cardiovascular parameters. After HI injury and during ventilatory support, there continued to be an increased RCBF in inner regions among the HI group, but no significant differences were detected in cortical flow compared to the control group. Also, the magnitude of the increase in TUNEL positive cells (apoptosis) and antioxidant enzymes, and decrease of ATP reserves was significantly greater in the brain regions where the RCBF was not higher. In conclusion, our findings identify early metabolic, histological, and hemodynamic changes involved in brain damage in premature asphyxiated lambs. Such changes have been described in human neonates, so our model could be useful to test the safety and the effectiveness of different neuroprotective or ventilation strategies applied in the first hours after fetal HI injury

constraints on the collins function from new e e h_1 h_2 x measurements in a leading order analysis

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