CARATTERISTICHE CLINICO-PATOLOGICHE, PROFILO MOLECOLARE E BERSAGLI PER L\u2019IMMUNOTERAPIA NEI CARCINOMI INDIFFERENZIATI DEL PANCREAS CON CELLULE GIGANTI SIMIL-OSTEOCLASTICHE

Pancreatic carcinoma is a malignant neoplasm with a high mortality rate. The study of conventional pancreatic ductal adenocarcinoma (PDAC) and its variants can improve the comprehension of its biology. The aim of this thesis is a comprehensive study of a PDAC variant, the undifferentiated carcinoma of the pancreas with osteoclast-like giant cells (UCOGC). After collecting a significant number of UCOGC (23 pancreatic and 5 extra-pancreatic), we recorded clinic-pathological data. Then we performed whole-exome sequencing for comprehensive molecular analysis. Tumor samples were also tested with immunohistochemistry (IHC) for immunotherapy biomarkers PD-1, PD-L1 and CD163. The prognostic role of clinic-pathological, molecular and IHC results was evaluated with ad-hoc survival analysis. We found that UCOGC could present in a \u201cpure\u201d form or associated with a differentiated neoplasia (usually PDAC). The \u201cpure\u201d form of UCOGC has a better prognosis. At molecular level, UCOGC shares the same somatic mutations with PDAC, involving the four most important driver genes KRAS, TP53, SMAD4 and CDKN2A. This demonstrates definitively that UCOGC is a real PDAC variant. No typical UCOGC genes were found but, interestingly, SERPINA3, a driver gene in some gynecological malignancies never found in association with pancreatic carcinoma, presented the same mutation in two distinct UCOGC. The IHC for the immunological targets revealed that PD-L1 is more expressed in UCOGC associated with PDAC; its expression has a negative prognostic role. PD-1, if present, was expressed by peri-tumor lymphocytes. CD163 was significantly and diffusely expressed in all the histiocytes of all UCOGC. We demonstrated an important role of morphology, having the \u201cpure\u201d form a better prognosis. Molecular results have clarified that UCOGC is a PDAC variant. IHC of PD-1, PD-L1 and CD163 showed that UCOGC may represent a potential target for new immunotherapy strategies. In particular, data concerning PD-L1 appear very promising, demonstrating its expression in above all cases associated with PDAC and its negative prognostic value. Furthermore, CD163 is expressed diffusely and strongly in all cases, with possible implications for future immunotherapies

Streamwise oscillation of spanwise velocity at the wall of a channel for turbulent drag reduction

Steady forcing at the wall of a channel flow is studied via DNS to assess its ability of yielding reductions of turbulent friction drag. The wall forcing consists of a stationary distribution of spanwise velocity that alternates in the streamwise direction. The idea behind the forcing builds upon the existing technique of the spanwise wall oscillation, and exploits the convective nature of the flow to achieve an unsteady interaction with turbulence. The analysis takes advantage of the equivalent laminar flow, that is solved analytically to show that the energetic cost of the forcing is unaffected by turbulence. In a turbulent flow, the alternate forcing is found to behave similarly to the oscillating wall; in particular an optimal wavelength is found that yields a maximal reduction of turbulent drag. The energetic performance is significantly improved, with more than 50% of maximum friction saving at large intensities of the forcing, and a net energetic saving of 23% for smaller intensities. Such a steady, wall-based forcing may pave the way to passively interacting with the turbulent flow to achieve drag reduction through a suitable distribution of roughness, designed to excite a selected streamwise wavelength

RASSF1 tumor suppressor gene in pancreatic ductal adenocarcinoma: correlation of expression, chromosomal status and epigenetic changes

Background: The Ras Association Domain Family Member 1 (RASSF1) is one of the most frequently reported methylation-inactivated tumor suppressor genes in primary pancreatic ductal adenocarcinomas (PDAC). Limited information is still available about the impact of RASSF1 gene silencing on the expression of its different isoforms in neoplastic cells. Methods: A series of 96 primary PDAC, with known clinico-pathological parameters, was tested for RASSF1 methylation status by methylation-specific PCR, RASSF1 locus copy number alterations by fluorescence in situ hybridization, and Rassf1a protein expression by immunohistochemistry. A further series of 14 xenografted primary PDAC and 8 PDAC-derived cell lines were tested to obtain a detailed methylation mapping of CpG islands A and C of the RASSF1 locus by pyrosequencing and to evaluate the expression of Rassf1 variants by qRT-PCR. Results: Methylation of CpG island A of the RASSF1 gene was observed in 35% of the tumors and allelic loss of RASSF1 locus was seen in 30 disomic and in 20 polysomic cases (52%). Rassf1a immunohistochemical expression was downregulated in half of primary PDAC, and this downregulation was neither correlated with methylation of RASSF1 promoter nor with RASSF1 copy number alterations. RASSF1 status did not influence patients' prognosis. The expression of the seven RASSF1 isoforms in xenografts and cell lines showed that RASSF1A, RASSF1B, and RASSF1C isoforms were present in all xenografts and cell lines, whereas RASSF1D, RASSF1E, and RASSF1F isoforms were variably expressed among samples. RASSF1G was never expressed in either xenografts or cell lines. The variable expression of RASSF1 isoforms in PDAC xenografts and cell lines was not dependent on RASSF1 methylation status of CpG islands A and C. Conclusions:RASSF1 alterations occurring in PDAC mainly consist in variations of expression of the different isoforms. Different genetic mechanisms seem to contribute to RASSF1 deregulation in this setting, but RASSF1 methylation does not seem to substantially affect RASSF1 isoforms expression

Relationship between Mast Cells and the Colitis with Relapse Induced by Trinitrobenzesulphonic Acid in Wistar Rats

The present study aimed to clarify the role of mast cells in colitis with relapse induced in Wistar rats by trinitrobenzenosulphonic acid. Colitis induction increased the histamine concentration in the colon, which peaked on day 26. The number of mast cells, probably immature, was ten times higher on day 8. Different from animals infected with intestinal parasites, after colitis remission, mast cells do not migrate to the spleen, showing that mast cell proliferation presents different characteristics depending on the inflammation stimuli. Treatment with sulfasalazine, doxantrazole, quercetin, or nedocromil did not increase the histamine concentration or the mast cell number in the colon on day 26, thereby showing absence of degranulation of these cells. In conclusion, although mast cell proliferation is associated with colitis, these cells and their mediators appear to play no clear role in the colitis with relapses

Oncofetal gene SALL4 and prognosis in cancer: A systematic review with meta-analysis

The Spalt-Like Transcription Factor 4 (SALL4) oncogene plays a central function in embryo-fetal development and is absent in differentiated tissues. Evidence suggests that it can be reactivated in several cancers worsening the prognosis. We aimed at investigating the risk associated with SALL4 reactivation for all-cause mortality and recurrence in cancer using the current literature. A PubMed and SCOPUS search until 1st September 2016 was performed, focusing on perspective studies reporting prognostic parameters in cancer data. In addition, 17 datasets of different cancer types from The Cancer Genome Atlas were considered. A total of 9,947 participants across 40 cohorts, followed-up for about 5 years on average, were analyzed comparing patients showing SALL4 presence (SALL4+, n = 1,811) or absence (SALL4-, n = 8,136). All data were summarised using risk ratios (RRs) for the number of deaths/recurrences and hazard ratios (HRs) for the time-dependent risk related to SALL4+, adjusted for potential confounders. SALL4+ significantly increased overall mortality (RR = 1.34, 95% confidence intervals (CI)=1.21-1.48, p<0.0001, I2=66%; HR=1.4; 95%CI: 1.19-1.65; p<0.0001; I2=63%) and recurrence of disease (RR = 1.25, 95% CI = 1.1-1.42, p=0.0006, I2=62%); HR=1.52; 95% CI: 1.22-1.89, p=0.0002; I2=69%) compared to SALL4-. Moreover, SALL4 remained significantly associated with poor prognosis even using HRs adjusted for potential confounders (overall mortality: HR=1.4; 95%CI: 1.19-1.65; p<0.0001; I2=63%; recurrence of disease: HR=1.52; 95% CI: 1.22-1.89, p=0.0002; I2=69%). These results suggest that SALL4 expression increases both mortality and recurrence of cancer, confirming this gene as an important prognostic marker and a potential target for personalized medicine

The association between smoking prevalence and eating disorders: a systematic review and meta-analysis

Background and Aims: Cigarette smoking is associated with severe mental illness, including schizophrenia and bipolar disorder, and with morbidity and mortality, but the association with anorexia (AN), bulimia nervosa (BN) and binge eating disorder (BED) is unclear. This meta-analysis compared the odds of smoking in eating disorders (ED) (ED = AN or BN or BED) versus healthy controls (HC) and calculated the prevalence of smokers in people with ED.&nbsp; Methods: Three independent authors searched PubMed, MEDLINE and Scopus from database inception until 31 December 2015 for studies reporting data on life-time or current smoking prevalence in BED, BN and AN with or without control group. Meta-analyses were undertaken, calculating odds ratios (ORs) of life-time smoking in BED, BN, AN versus healthy controls (HCs) or prevalence of smoking in BED, BN and AN with 95% confidence intervals (CI).&nbsp; Results: Thirty-one studies (ED = 8517, controls = 68 335) were meta-analysed. Compared with HCs, there were significantly more smokers among people with BN (life-time OR = 2.165) and BED (life-time OR = 1.792) but not AN (life-time OR = 0.927). BED was associated with smoking the most (life-time prevalence = 47.73%) followed by BN (life-time prevalence = 39.4%) and AN (life-time prevalence = 30.8%). In BN, life-time smoking prevalence was highest in Europe. In AN, higher age moderated both life-time and current smoking prevalence, and body mass index moderated higher life-time smoking prevalence. In BN, female sex moderated higher life-time smoking prevalence.&nbsp; Conclusions: People with binge eating disorder and bulimia nervosa are significantly more likely to be life-time smokers than healthy controls, which is not the case for anorexia nervosa

MiR-21 up-regulation in ampullary adenocarcinoma and its pre-invasive lesions

Poor information is available on the molecular landscape characterizing the carcinogenetic process leading to ampullary carcinoma. MiR-21 is one of the most frequently up-regulated miRNAs in pancreatic adenocarcinoma, a tumor sharing similar molecular features with ampullary adenocarcinomas (AVCs), above all with the pancreatic-biliary type. We profiled, by in situ hybridization (ISH), miR-21 expression in a series of 26 AVCs, 50 ampullary dysplastic lesions (35 low-grade [LG-IEN] and 15 high-grade [HG-IEN]) and 10 normal duodenal mucosa samples. The same series was investigated by immunohistochemistry for β-catenin, p53 and HER2 expression. HER2 gene amplification was evaluated by chromogenic in situ hybridization. To validate miR-21 ISH results we performed miR-21 qRT-PCR analysis in a series of 10 AVCs and their matched normal samples. All the normal control samples showed a negative or faint miR-21 expression, whereas a significant miR-21 up-regulation was observed during the carcinogenetic cascade (p &lt; 0.001), with 21/26 (80.8%) of cancer samples showing a miR-21 overexpression. In comparison to control samples, a significant overexpression was found in samples of LG-IEN (p = .0003), HG-IEN (p = .0001), and AVCs (p &lt; 0.0001). No significant difference in miR-21 overexpression was observed between LG-IEN, HG-IEN and AVCs. By qRT-PCR analysis, AVCs showed a 1.7-fold increase over the controls (p = .003). P53 was frequently dysregulated in both dysplastic and carcinoma samples (44 out of 76; 57.9%). A 20% (10/50) of dysplastic lesions and 11% (3/26) of carcinomas were characterized by a nuclear localization of β-catenin. Only 2 AVCs (7.7%; both intestinal-type) showed a HER2 overexpression (both 2+), which corresponded to a HER2 gene amplification at CISH analysis. This is the first study demonstrating a miRNA dysregulation in the whole spectrum of ampullary carcinogenesis. MiR-21 overexpression is an early molecular event during ampullary carcinogenesis and its levels increase with the neoplastic progression

Rare histotypes of epithelial biliary tract tumors: A literature review

Adenocarcinoma represents the most frequent biliary tract cancer. However, other rare histotypes can be found in the biliary tract, such as cholangiolocellular carcinoma, cholangiocarcinoma with ductal plate malformation pattern, adenosquamous carcinoma, mucinous carcinoma, signet ring cell carcinoma, clear cell carcinoma, mucoepidermoid carcinoma, lymphoepithelioma-like carcinoma, and sarcomatous cholangiocarcinoma. These cancer types account for less than 10 % of all the already rare biliary tract tumors. Yet, they represent a relevant issue in everyday clinical practice, given the lack of therapeutic recommendations and the overall scarcity of data, mainly deriving from isolated small center-specific cohorts of patients.The shifts of such histotypes from the most common ones reflect genetic and molecular differences, determine changes in clinical aggressiveness, and suggest a possible variability in sensitivity to the standard treatments of biliary adenocarcinomas. The consistency and degree of these variables are still to be solidly demonstrated and investigated. Therefore, this paper aims to review the current literature concerning very infrequent and rare epithelial biliary tract cancers, focusing our attention on the clinical, molecular, and immunohistochemical features of these tumors

Inflammation and frailty in the elderly: A systematic review and meta-analysis

The pathogenesis of frailty and the role of inflammation is poorly understood. We examined the evidence considering the relationship between inflammation and frailty through a systematic review and meta-analysis. A systematic literature search of papers providing data on inflammatory biomarkers and frailty was carried out in major electronic databases from inception until May 2016. From 1856 initial hits, 35 studies (32 cross-sectional studies n=3232 frail, n=11,483 pre-frail and n=8522 robust, and 563 pre-frail+robust; 3 longitudinal studies n=3402 participants without frailty at baseline) were meta-analyzed. Cross-sectional studies reported that compared to 6757 robust participants, both 1698 frail (SMD=1.00, 95%CI: 0.40-1.61) and 8568 pre-frail (SMD=0.33, 95%CI: 0.04-0.62) participants had significantly higher levels of C-reactive protein (CRP). Frailty (n=1057; SMD=1.12, 95%CI: 0.27-2.13) and pre-frailty (n=4467; SMD=0.56, 95%CI: 0.00-1.11) were associated with higher serum levels of interleukin-6 compared to people who were robust (n=2392). Frailty and pre-frailty were also significantly associated with elevated white blood cell and fibrinogen levels. In three longitudinal studies, higher serum CRP (OR=1.06, 95%CI: 0.78-1.44,) and IL-6 (OR=1.19, 95%CI: 0.87-1.62) were not associated with frailty. In conclusion, frailty and pre-frailty are associated with higher inflammatory parameters and in particular CRP and IL-6. Further longitudinal studies are needed

SORPTION OF S-TRIAZINES IN BRAZILIAN RAINFOREST SOILS

This research was conducted to evaluate the sorption of Ametryn, Atrazine, Simazine, Prometrine and Metamitron to soils from "Mata Atlântica" at Ubatuba region (Atlantic rainforest soils), employing the batch equilibrium approach. The herbicides were weakly retained in soils with low soil organic matter (SOM) content and thus presenting high potential to water contamination. All herbicides have shown high Koc at Typic Humaquepts soil, the higher in SOM content. The sorption isotherms for the herbicides at Typic Humaquepts soil suggested specific interactions between herbicides and SOM probably with partial protonation of herbicides followed by ion-exchange processes and/or hydrogen bonding formation of hydroxyl groups on the SOM surface.Esta pesquisa foi conduzida para avaliar a sorção de ametrina, atrazina, simazina, prometrina e metamitron em solos de Mata Atlântica na região de Ubatuba, empregando-se o método em batelada. Os herbicidas foram fracamente retidos em solos com baixo teor de matéria orgânica (MO) e, portanto, apresentaram elevado potencial de contaminação da água. Todos os herbicidas mostraram alto valor de Koc em solos da classe Gleissolo Melânico Distrófico, que contém o teor mais elevado de MO. As isotermas de sorção dos herbicidas no Gleissolo Melânico Distrófico sugerem interações específicas entre os herbicidas e a MO, provavelmente com protonação parcial dos herbicidas seguida por processos de troca-iônica e/ou formação de pontes de hidrogênio dos grupos hidroxila sobre a superfície da MO