Allele-Specific Gene Expression di polimorfismi associati al carcinoma tiroideo differenziato

Il carcinoma tiroideo rappresenta una delle neoplasie che interessano il sistema endocrino. La forma più comune di tumore della tiroide si manifesta a carico delle cellule follicolari che producono la maggioranza degli ormoni tiroidei, questa viene classificata come carcinoma tiroideo differenziato (DTC) di cui i sottotipi sono il papillare (75%), il follicolare (10%), il carcinoma a cellule di Hürtle (5%) ed i carcinomi scarsamente differenziati (1-6%). Vi sono poi forme tumorali a carico delle cellule parafollicolari dette carcinoma midollare della tiroide (meno del 5% dei casi). Il DTC è una patologia complessa, causata da fattori di rischio genetici ed ambientali. Per individuare possibili polimorfismi a singolo nucleotide (SNP) coinvolti nell’eziologia del DTC, sono stati effettuati studi di associazione sull’intero genoma (GWAS) che hanno portato all’identificazione di una forte relazione con SNPs dei geni DIRC3, FOXE1, NRG1, NKX2-1, BATF ed DHX3. Inoltre gli SNPs dei geni ARSB e SPATA13 (ASEF2) sono stati associati al rischio della malattia solo nella popolazione italiana. Gli studi di GWAS si basano sull’analisi degli SNPs presenti nei blocchi di Linkage Disequilibrium (LD) e mirano a determinare la loro associazione con la patologia, senza però svelare il loro ruolo funzionale. Questo studio si pone quindi lo scopo di analizzare il possibile ruolo funzionale degli SNPs rs13184587 e rs1220597. In particolare, è stato valutato se lo SNP rs13184587 altera l’espressione dei geni ARSB e BHMT2 e se lo SNP rs1220597 influenza i livelli di mRNA del gene SPATA13. La prima parte di questo lavoro si basa sulla raccolta di campioni di sangue periferico, estrazione di DNA ed RNA e genotipizzazione tramite TaqMan Real Time PCR. Nella seconda parte è stata valutata la variazione di espressione dei geni selezionati tramite la tecnica di Allele-Specific Gene Expression (ASGE) nei campioni eterozigoti. Infine è stata eseguita un’analisi statistica dei risultati ottenuti. Dalle analisi effettuate risulta che gli SNP rs13184587 e rs1220597 non influenzano l’espressione dei geni in esame. Ulteriori studi sono necessari per valutare il ruolo funzionale di queste variazioni geniche o di SNP nello stesso blocco di LD

The metastatic potential of grade I breast carcinoma of no special type: A deep insight into putative molecular mechanisms

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Mouse mammary tumour virus-like env nucleotide and p14 signal peptide are present in feline mammary carcinomas, but not in neoplastic or dysplastic canine mammary lesions

Mouse mammary tumour virus-like (MMTV-like) is suspected to be involved in human breast cancer and it has been hypothesized that companion animals might have a role in viral transmission. The aim of our study was to investigate the presence of MMTV-like nucleotide sequences and viral protein in a larger number of feline (FMCs) and canine mammary carcinomas (CMCs) by nested PCR and immunohistochemistry. Results showed that the presence of MMTV-like env sequence in FMCs was 7% (6/86), while all the CMCs and canine dysplastic lesions scored negative. All PCR-positive FMCs scored positive for the MMTV p14 signal peptide of the envelope precursor protein of the virus. In contrast, all PCR-negative FMCs and canine mammary lesions were also negative for immunohistochemistry analysis. Canine and feline normal mammary gland tissues scored negative for both PCR and MMTV-p14 protein. Multiple nucleotide alignment of MMTV-like env gene sequences isolated from cat showed 97% and 99% similarity with HMTV and MMTV, respectively, while the others two presented some polimorphisms. Particularly the sequences of one of these two tumors showed a polymorphism (c.7575 A> G), that causes a previously unreported amino acid substitution (Thr > Ala). In conclusion, the results of our study showed the presence of MMTV-like sequences and viral protein in some FMCs. Further studies are needed to understand whether this virus does play a role in the development of FMCs, if MMTV-like is an exogenous virus as these data suggest and, in such a case, how and from whom this virus was acquired

Exposure to nature is associated with decreased functional connectivity within the distress network: A resting state EEG study

Introduction: despite the well-established evidence supporting the restorative potential of nature exposure, the neurophysiological underpinnings of the restorative cognitive/emotional effect of nature are not yet fully understood. the main purpose of the current study was to investigate the association between exposure to nature and electroencephalography (EEG) functional connectivity in the distress network. methods: fifty-three individuals (11 men and 42 women; mean age 21.38 +/- 1.54 years) were randomly assigned to two groups: (i) a green group and (ii) a gray group. a slideshow consisting of images depicting natural and urban scenarios were, respectively, presented to the green and the gray group. before and after the slideshow, 5 min resting state (RS) EEG recordings were performed. the exact low-resolution electromagnetic tomography (eLORETA) software was used to execute all EEG analyses. results: compared to the gray group, the green group showed a significant increase in positive emotions (F-1; 50 = 9.50 p = 0.003) and in the subjective experience of being full of energy and alive (F-1; 50 = 4.72 p = 0.035). furthermore, as compared to urban pictures, the exposure to natural images was associated with a decrease of delta functional connectivity in the distress network, specifically between the left insula and left subgenual anterior cingulate cortex (T = -3.70, p = 0.023). discussion: our results would seem to be in accordance with previous neurophysiological studies suggesting that experiencing natural environments is associated with brain functional dynamics linked to emotional restorative processes

Patients perception of ionising radiation risks in CT ionising exposure. Does dose bill works?

Communicating to patients the magnitude of risk related to ionizing radiation exposure is problematic because of the uncertainty in estimates derived principally from epidemiological studies of large populations [1-6]. Euratom directive 59/2013 requires that dose bill will be part of the radiological report in European Countries [7]. However, how a risk is framed has a profound effect on risk perception. To date, no previous studies evaluated which could be the best way to make patient friendly dose bill. Our aim was to evaluate patients' perception of radiation exposure related to routine CT and their understanding after dose bill

MicroRNAs distribution in different phenotypes of Aortic Stenosis

Aortic valve stenosis (AVS) represents a cluster of different phenotypes, considering gradient and flow pattern. Circulating micro RNAs may reflect specific pathophysiological processes and could be useful biomarkers to identify disease. We assessed 80 patients (81, 76.7-84 years; 46, 57.5%females) with severe AVS. We performed bio-humoral evaluation (including circulating miRNA-1, 21, 29, 133) and 2D-echocardiography. Patients were classified according to ACC/AHA groups (D1-D3) and flow-gradient classification, considering normal/low flow, (NF/LF) and normal/high gradient, (NG/HG). Patients with reduced ejection fractionwere characterized by higher levels of miRNA1 (p = 0.003) and miRNA 133 (p = 0.03). LF condition was associated with higher levels of miRNA1 (p = 0.02) and miRNA21 (p = 0.02). Levels of miRNA21 were increased in patients with reduced Global longitudinal strain (p = 0.03). LF-HG and LF-LG showed higher levels of miRNA1 expression (p = 0.005). At one-year follow-up miRNA21 and miRNA29 levels resulted significant independent predictors of reverse remodeling and systolic function increase, respectively. Different phenotypes of AVS may express differential levels and types of miRNAs, which may retain a pathophysiological role in pro-hypertrophic and pro-fibrotic processes

PATIENTS' KNOWLEDGE AND AWARENESS OF RADIATION DOSE AND RISKS FROM CT: DO PATIENTS NEED A PERSONALIZED COMMUNICATION OF DOSE BILL?

In the last decades exposure to ionizing radiations in computed tomography (CT) has constantly increased. Only a few years ago it was quite difficult to assess how much radiation had been delivered to a patient during a CT examination. Nowadays, the technical challenges of dose data reporting between CT scanners from different vendors have been met, making dose tracking a reality since these dose data are automatically stored in the picture achieving system of the radiology department. Most authors affirm communication of CT risk to patients should be personalized, but no studies investigate if a tailored communication is needed. Aim of our study is to understand how patients' characteristics may condition the comprehension of this information

ANKRd44 gene silencing: a putative role in trastuzumab resistance in HER2-like breast cancer

Trastuzumab is an effective therapeutic treatment for Her2-like breast cancer; despite this most of these tumors develop resistance to therapy due to specific gene mutations or alterations in gene expression. Understanding the mechanisms of resistance to Trastuzumab could be a useful tool in order to identify combinations of drugs that elude resistance and allow a better response for the treated patients. Twelve primary biopsies of Her2+/hormone receptor negative (ER-/PgR-) breast cancer patients were selected based on the specific response to neoadjuvant therapy with Trastuzumab and their whole exome was sequenced leading to the identification of 18 informative gene mutations that discriminate patients selectively based on response to treatment. Among these genes, we focused on the study of the ANKRD44 gene to understand its role in the mechanism of resistance to Trastuzumab. The ANKRD44 gene was silenced in Her2-like breast cancer cell line (BT474), obtaining a partially Trastuzumab-resistant breast cancer cell line that constitutively activates the NF-kb protein via the TAK1/AKT pathway. Following this activation an increase in the level of glycolysis in resistant cells is promoted, also confirmed by the up-regulation of the LDHB protein and by an increased TROP2 protein expression, found generally associated with aggressive tumors. These results allow us to consider the ANKRD44 gene as a potential gene involved in Trastuzumab resistance

Cancer astrocytes have a more conserved molecular status in long recurrence free survival (RFS) IDH1 wild-type glioblastoma patients: New emerging cancer players

Glioblastoma is a devastating disease that despite all the information gathered so far, its optimal management remains elusive due to the absence of validated targets from clinical studies. A better clarification of the molecular mechanisms is needed. In this study, having access to IDH1 wild-type glioblastoma of patients with exceptionally long recurrence free survival (RFS), we decided to compare their mutational and gene expression profile to groups of IDH1 wild-type glioblastoma of patients with shorter RFS, by using NGS technology. The exome analysis revealed that Long-RFS tumors have a lower mutational rate compared to the other groups. A total of 158 genes were found differentially expressed among the groups, 112 of which distinguished the two RFS extreme groups. Overall, the exome data suggests that shorter RFS tumors could be, chronologically, in a more advanced state in the muli-step tumor process of sequential accumulation of mutations. New players in this kind of cancer emerge from the analysis, confirmed at the RNA/DNA level, identifying, therefore, possible oncodrivers or tumor suppressor genes