Cyclin-dependent kinase 1 targeting improves sensitivity to radiation in BRAF V600E colorectal carcinoma cells.

Preoperative chemoradiation is currently the standard of care in locally advanced rectal carcinoma, even though a subset of rectal tumors does not achieve major clinically meaningful responses upon neoadjuvant chemoradiation. At present, no molecular biomarkers are available to predict response to neoadjuvant chemoradiation and select resistant tumors willing more intense therapeutic strategies. Thus, BRAF mutational status was investigated for its role in favoring resistance to radiation in colorectal carcinoma cell lines and cyclin-dependent kinase 1 as a target to improve radiosensitivity in BRAF V600E colorectal tumor cells.Colony-forming assay and apoptotic rates were evaluated to compare the sensitivity of different colon carcinoma cell lines to ionizing radiation and their radiosensitivity upon exposure to BRAF and/or cyclin-dependent kinase 1 inhibitory/silencing strategies. Cyclin-dependent kinase 1 expression/subcellular distribution was studied by immunoblot analysis.Colon carcinoma BRAF V600E HT29 cells exhibited poor response to radiation compared to BRAF wild-type COLO320 and HCT116 cells. Interestingly, neither radiosensitizing doses of 5-fluoruracil nor BRAF inhibition/silencing significantly improved radiosensitivity in HT29 cells. Of note, poor response to radiation correlated with upregulation/relocation of cyclin-dependent kinase 1 in mitochondria. Consistently, cyclin-dependent kinase 1 inhibition/silencing as well as its targeting, through inhibition of HSP90 quality control pathway, significantly inhibited the clonogenic ability and increased apoptotic rates in HT29 cells upon exposure to radiation.These data suggest that BRAF V600E colorectal carcinoma cells are poorly responsive to radiation, and cyclin-dependent kinase 1 represents a target to improve radiosensitivity in BRAF V600E colorectal tumor cells

Evaluation of Glucose Uptake in Normal and Cancer Cell Lines by Positron Emission Tomography.

To date, there is no definitive demonstration of the utility of positron emission tomography (PET) in studying glucose metabolism in cultured cell lines. Thus, this study was designed to compare PET to more standardized methods for the quantitative assessment of glucose uptake in nontransformed and transformed living cells and to validate PET for metabolic studies in vitro. Human colon and breast carcinoma cell lines and mouse embryo fibroblasts were evaluated for [ 18 F]fluorodeoxyglucose ([ 18 F]FDG) uptake by PET and autoradiography and 2-deoxyglucose (2-DG) incorporation by colorimetric assay and analyzed for the radiotoxic effects of [ 18 F]FDG and the expression levels of glucose transporters. Indeed, [ 18 F]FDG incorporation on PET was comparable to [ 18 F]FDG uptake by autoradiography and 2-DG incorporation by colorimetric assay, although radiotracer-based methods exhibited more pronounced differences between individual cell lines. As expected, these data correlated with glucose transporters 1 to 4 and hexokinase II expression in tumor cell lines and mouse fibroblasts. Notably, [ 18 F]FDG incorporation resulted in low apoptotic rates, with fibroblasts being slightly more sensitive to radiotracer-induced cell death. The quantitative analysis of [ 18 F]FDG uptake in living cells by PET represents a valuable and reproducible method to study tumor cell metabolism in vitro, being representative of the differences in the molecular profile of normal and tumor cell lines

F-18 FDG PET/CT metabolic tumor volume predicts overall survival in patients with disseminated epithelial ovarian cancer

OBJECTIVE: We evaluated the prognostic impact of quantitative assessment by maximum standardized uptake value (SUVmax), metabolic tumour volume (MTV) and tumour lesion glycolysis (TLG) on [F-18] FDG PET/CT for patients with peritoneal carcinomatosis from epithelial ovarian cancer (EOC). METHODS: Thirty-one patients with EOC underwent PET/CT for an early restaging after cytoreductive surgery, having been diagnosed with carcinomatosis (before chemotherapy). The SUVmax, MTV (cm3; 42% threshold) and TLG (g) were registered on residual peritoneal lesions. The patients were followed up 20±12months thereafter. The PET/CT results were compared to overall survival (OS). RESULTS: The Kaplan-Meier survival analysis for the SUVmax did not reveal significant differences in OS (p=0.48). The MTV survival analysis showed a significant higher OS in patients presenting with a higher tumour burden than those with less tumour burden (p=0.01; 26 vs. 14 months), whereas TLG exhibited a similar trend though not significant (p=0.06). Apart from chemo-resistance, the higher the MTV, the better will be the response to chemotherapy. CONCLUSIONS: Quantitative assessment by MTV rather than by SUVmax and TLG on PET/CT may be helpful for stratifying patients who present with peritoneal carcinomatosis from EOC, in order to implement the appropriate therapeutic regimen

Mediterranean monitoring and forecasting operational system for Copernicus Marine Service

The MEDiterranean Monitoring and Forecasting Center (Med-MFC) is part of the Copernicus Marine Environment Monitoring Service (CMEMS, http://marine.copernicus.eu/), provided on an operational mode by Mercator Ocean in agreement with the European Commission. Specifically, Med MFC system provides regular and systematic information about the physical state of the ocean and marine ecosystems for the Mediterranean Sea. The Med-MFC service started in May 2015 from the pre-operational system developed during the MyOcean projects, consolidating the understanding of regional Mediterranean Sea dynamics, from currents to biogeochemistry to waves, interfacing with local data collection networks and guaranteeing an efficient link with other Centers in Copernicus network. The Med-MFC products include analyses, 10 days forecasts and reanalysis, describing currents, temperature, salinity, sea level and pelagic biogeochemistry. Waves products will be available in MED-MFC version in 2017. The consortium, composed of INGV (Italy), HCMR (Greece) and OGS (Italy) and coordinated by the Euro-Mediterranean Centre on Climate Change (CMCC, Italy), performs advanced R&D activities and manages the service delivery. The Med-MFC infrastructure consists of 3 Production Units (PU), for Physics, Biogechemistry and Waves, a unique Dissemination Unit (DU) and Archiving Unit (AU) and Backup Units (BU) for all principal components, guaranteeing a resilient configuration of the service and providing and efficient and robust solution for the maintenance of the service and delivery. The Med-MFC includes also an evolution plan, both in terms of research and operational activities, oriented to increase the spatial resolution of products, to start wave products dissemination, to increase temporal extent of the reanalysis products and improving ocean physical modeling for delivering new products. The scientific activities carried out in 2015 concerned some improvements in the physical, biogeochemical and wave components of the system. Regarding the currents, new grid-point EOFs have been implemented in the Med-MFC assimilation system; the climatological CMAP precipitation was replaced by the ECMWF daily precipitation; reanalysis time-series have been increased by one year. Regarding the biogeochemistry, the main scientific achievement is related to the implementation of the carbon system in the Med-MFC biogeochemistry model system already available. The new model is able to reproduce the principal spatial patterns of the carbonate system variables in the Mediterranean Sea. Further, a key result consists of the calibration of the new variables (DIC and alkalinity), which serves to the estimation of the accuracy of the new products to be released in the next version of the system (i.e. pH and pCO2 at surface). Regarding the waves, the system has been validated against in-situ and satellite observations. For example, a very good agreement between model output and in-situ observations has been obtained at offshore and/or well-exposed wave buoys in the Mediterranean Sea.PublishedVienna3SR. AMBIENTE - Servizi e ricerca per la Societ

Common and rare variants in TMEM175 gene concur to the pathogenesis of Parkinson’s Disease in Italian patients

Parkinson’s disease (PD) represents the most common neurodegenerative movement disorder. We recently identified 16 novel genes associated with PD. In this study, we focused the attention on the common and rare variants identified in the lysosomal K+ channel TMEM175. The study includes a detailed clinical and genetic analysis of 400 cases and 300 controls. Molecular studies were performed on patient-derived fibroblasts. The functional properties of the mutant channels were assessed by patch-clamp technique and co-immunoprecipitation. We have found that TMEM175 was highly expressed in dopaminergic neurons of the substantia nigra pars compacta and in microglia of the cerebral cortex of the human brain. Four common variants were associated with PD, including two novel variants rs2290402 (c.-10C > T) and rs80114247 (c.T1022C, p.M341T), located in the Kozak consensus sequence and TM3II domain, respectively. We also disclosed 13 novel highly penetrant detrimental mutations in the TMEM175 gene associated with PD. At least nine of these mutations (p.R35C, p. R183X, p.A270T, p.P308L, p.S348L, p. L405V, p.R414W, p.P427fs, p.R481W) may be sufficient to cause the disease, and the presence of mutations of other genes correlated with an earlier disease onset. In vitro functional analysis of the ion channel encoded by the mutated TMEM175 gene revealed a loss of the K+ conductance and a reduced channel affinity for Akt. Moreover, we observed an impaired autophagic/lysosomal proteolytic flux and an increase expression of unfolded protein response markers in patient-derived fibroblasts. These data suggest that mutations in TMEM175 gene may contribute to the pathophysiology of PD

"Delirium Day": A nationwide point prevalence study of delirium in older hospitalized patients using an easy standardized diagnostic tool

Background: To date, delirium prevalence in adult acute hospital populations has been estimated generally from pooled findings of single-center studies and/or among specific patient populations. Furthermore, the number of participants in these studies has not exceeded a few hundred. To overcome these limitations, we have determined, in a multicenter study, the prevalence of delirium over a single day among a large population of patients admitted to acute and rehabilitation hospital wards in Italy. Methods: This is a point prevalence study (called "Delirium Day") including 1867 older patients (aged 65 years or more) across 108 acute and 12 rehabilitation wards in Italian hospitals. Delirium was assessed on the same day in all patients using the 4AT, a validated and briefly administered tool which does not require training. We also collected data regarding motoric subtypes of delirium, functional and nutritional status, dementia, comorbidity, medications, feeding tubes, peripheral venous and urinary catheters, and physical restraints. Results: The mean sample age was 82.0 ± 7.5 years (58 % female). Overall, 429 patients (22.9 %) had delirium. Hypoactive was the commonest subtype (132/344 patients, 38.5 %), followed by mixed, hyperactive, and nonmotoric delirium. The prevalence was highest in Neurology (28.5 %) and Geriatrics (24.7 %), lowest in Rehabilitation (14.0 %), and intermediate in Orthopedic (20.6 %) and Internal Medicine wards (21.4 %). In a multivariable logistic regression, age (odds ratio [OR] 1.03, 95 % confidence interval [CI] 1.01-1.05), Activities of Daily Living dependence (OR 1.19, 95 % CI 1.12-1.27), dementia (OR 3.25, 95 % CI 2.41-4.38), malnutrition (OR 2.01, 95 % CI 1.29-3.14), and use of antipsychotics (OR 2.03, 95 % CI 1.45-2.82), feeding tubes (OR 2.51, 95 % CI 1.11-5.66), peripheral venous catheters (OR 1.41, 95 % CI 1.06-1.87), urinary catheters (OR 1.73, 95 % CI 1.30-2.29), and physical restraints (OR 1.84, 95 % CI 1.40-2.40) were associated with delirium. Admission to Neurology wards was also associated with delirium (OR 2.00, 95 % CI 1.29-3.14), while admission to other settings was not. Conclusions: Delirium occurred in more than one out of five patients in acute and rehabilitation hospital wards. Prevalence was highest in Neurology and lowest in Rehabilitation divisions. The "Delirium Day" project might become a useful method to assess delirium across hospital settings and a benchmarking platform for future surveys

"Delirium Day": A nationwide point prevalence study of delirium in older hospitalized patients using an easy standardized diagnostic tool

Background: To date, delirium prevalence in adult acute hospital populations has been estimated generally from pooled findings of single-center studies and/or among specific patient populations. Furthermore, the number of participants in these studies has not exceeded a few hundred. To overcome these limitations, we have determined, in a multicenter study, the prevalence of delirium over a single day among a large population of patients admitted to acute and rehabilitation hospital wards in Italy. Methods: This is a point prevalence study (called "Delirium Day") including 1867 older patients (aged 65 years or more) across 108 acute and 12 rehabilitation wards in Italian hospitals. Delirium was assessed on the same day in all patients using the 4AT, a validated and briefly administered tool which does not require training. We also collected data regarding motoric subtypes of delirium, functional and nutritional status, dementia, comorbidity, medications, feeding tubes, peripheral venous and urinary catheters, and physical restraints. Results: The mean sample age was 82.0 \ub1 7.5 years (58 % female). Overall, 429 patients (22.9 %) had delirium. Hypoactive was the commonest subtype (132/344 patients, 38.5 %), followed by mixed, hyperactive, and nonmotoric delirium. The prevalence was highest in Neurology (28.5 %) and Geriatrics (24.7 %), lowest in Rehabilitation (14.0 %), and intermediate in Orthopedic (20.6 %) and Internal Medicine wards (21.4 %). In a multivariable logistic regression, age (odds ratio [OR] 1.03, 95 % confidence interval [CI] 1.01-1.05), Activities of Daily Living dependence (OR 1.19, 95 % CI 1.12-1.27), dementia (OR 3.25, 95 % CI 2.41-4.38), malnutrition (OR 2.01, 95 % CI 1.29-3.14), and use of antipsychotics (OR 2.03, 95 % CI 1.45-2.82), feeding tubes (OR 2.51, 95 % CI 1.11-5.66), peripheral venous catheters (OR 1.41, 95 % CI 1.06-1.87), urinary catheters (OR 1.73, 95 % CI 1.30-2.29), and physical restraints (OR 1.84, 95 % CI 1.40-2.40) were associated with delirium. Admission to Neurology wards was also associated with delirium (OR 2.00, 95 % CI 1.29-3.14), while admission to other settings was not. Conclusions: Delirium occurred in more than one out of five patients in acute and rehabilitation hospital wards. Prevalence was highest in Neurology and lowest in Rehabilitation divisions. The "Delirium Day" project might become a useful method to assess delirium across hospital settings and a benchmarking platform for future surveys

The current and evolving role of FDG–PET/CT in personalized iodine‑131 therapy of differentiated thyroid cancer

Purpose Several approaches have been recommended for the selection of patients with differentiated thyroid cancer amenable for postoperative radioiodine remnant ablation or repeated radioiodine treatment, though with inadequate results. 2-[18F]- fluoro-2-deoxy-d-glucose positron emission tomography is gaining consideration for predicting disease-free or survival of differentiated thyroid carcinoma patients, in particular in the peri-or post-surgery setting and for the detection of recurrence in patients with elevated or rising thyroglobulin without evidence of disease on neck ultrasound or iodine scintigraphy. This paper aims to review the role of FDG–PET/CT in tailoring iodine-131 empiric therapy for radioiodine remnant ablation and the detection of recurrence in patients with elevated thyroglobulin and negative iodine-123 scan. Methods A literature search up to May 2017 of MEDLINE® and SCOPUS® with the Mesh terms: “PET/CT”, “iodine-131 therapy”, “differentiated thyroid cancer” and “prognosis” was performed. Thereafter, papers dealing with radioiodine remnant ablation and empiric therapy were selected. Results Ninety papers were retrieved from the initial search and 19 considered for the review. The percentage of positive FDG–PET/CT performed at radioiodine remnant ablation or shortly after ranged from 17 to 69%, with highest values in high- and intermediate-to-high risk patients. The response rate to radioiodine remnant ablation and survival were consistently higher in negative FDG–PET/CT patients. Besides, FDG–PET/CT imaging was found to be a very accurate diagnostic tool for the detection of recurrence in patients with elevated thyroglobulin and negative iodine-123 scan, discriminating patients needing further empirical iodine-131 therapy from those who could benefit from alternative approaches. Conclusions Although a meta-analysis was not possible due to the heterogeneity and the small population samples of the studies retrieved, the results of the present review support the use of FDG–PET/CT in tailoring iodine-131 therapy when used close to radioiodine remnant ablation and in patients amenable to iodine-131 empiric therapy