Microenvironment and tumor cells: two targets for new molecular therapies of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC), is one of the most frequent human cancer and is characterized by a high mortality rate. The aggressiveness appears strictly related to the liver pathological background on which cancer develops. Inflammation and the consequent fibro/cirrhosis, derived from chronic injuries of several origins (viral, toxic and metabolic) and observable in almost all oncological patients, represents the most powerful risk factor for HCC and, at the same time, an important obstacle to the efficacy of systemic therapy. Multiple microenvironmental cues, indeed, play a pivotal role in the pathogenesis, evolution and recurrence of HCC as well as in the resistance to standard therapies observed in most of patients. The identification of altered pathways in cancer cells and of microenvironmental changes, strictly connected in pathogenic feedback loop, may permit to plan new therapeutic approaches targeting tumor cells and their permissive microenvironment, simultaneousl

Integrating Survey and Administrative Data on Local Social Protection

Welfare systems can be observed according to two different perspectives. The former deals with the supply of social protection, i.e. with the funding and provision of social benefits and the production of social services and goods. The latter focuses on the demand of social protection, and particularly on the characteristics of people benefiting from social protection or asking for it. Typically, data on the supply of social benefits have an administrative nature (registers and budgets data) whereas data on beneficiaries come from sample surveys. In theory, administrative data, being census data, can be detailed by territory. On the contrary, sample surveys are usually planned to provide accurate estimates at the national level or for large sub-national areas. This chapter provides an example on the use of different data sets for the Old age and Family/children functions at the province level (LAU 1 in the EU nomenclature). Data on the supply of benefits derive from the SISSIM (Istat Survey on Interventions and Social Services of Individual and associated Municipalities) and from municipalities' budgets. Data on the demand of social protection come from EU-SILC (European Union - Statistics on Income and Living Conditions), a survey that is annually conducted by Istat in a comparable European framework. Earned benefits are estimated applying small area estimation methods, given that the sample size of the EU-SILC survey at the province level is small, so the traditional design-based estimators usually are unreliable. Results are analysed to understand whether administrative and sample survey data can be used to to compose a coherent picture of social protection delivered at the provincial level

The effect of robenacoxib on the concentration of C-reactive protein in synovial fluid from dogs with osteoarthritis

Background: Robenacoxib is a novel and highly selective inhibitor of COX-2 in dogs and cats and because of its acidic nature is regarded as being tissue-selective. Thirty four dogs with stifle osteoarthritis secondary to failure of the cranial cruciate ligament were recruited into this study. Lameness, radiographic features, synovial cytology and C-reactive protein concentrations in serum and synovial fluid were assessed before and 28 days after commencing a course of Robenacoxib at a dose of 1 mg/kg SID.<p></p> Results: There was a significant reduction in the lameness score (P <0.01) and an increase in the radiographic score (P < 0.05) between pre- and post-treatment assessments. There was no difference between pre- (median 1.49 mg/l; Q1-Q3 0.56-4.24 mg/L) and post – (1.10 mg/L; 0.31-1.78 mg/L) treatment serum C-reactive protein levels although synovial fluid levels were significantly reduced (pre- : 0.44 mg/L; 0.23-1.62 mg/L; post- : 0.17 mg/L; 0.05-0.49 mg/L) (P < 0.05). There was no correlation between C-reactive protein concentrations in serum and matched synovial fluid samples.<p></p> Conclusions: Robenacoxib proved effective in reducing lameness in dogs with failure of the cranial cruciate ligament and osteoarthritis of the stifle joint. The drug also reduced levels of C-reactive protein in the synovial fluid taken from the affected stifle joint. Robenacoxib appears to reduce articular inflammation as assessed by C-reactive protein which supports the concept that Robenacoxib is a tissue-selective non-steroidal anti-inflammatory drug.<p></p&gt

Influence of different drying parameters on the composition of volatile compounds of thyme and rosemary cultivated in Sardinia

The shelf life of spices is traditionally extended by drying. Fresh herbs, due to their high water content, undergo microorganism growth and adverse biochemical reactions. On the other hand drying may result in a lot of physical and chemical alterations. Air and oven-dehydration are the main methods used to stabilize spices. During oven drying, in general, losses of volatile compounds are directly dependent on the temperature and time used. This paper deals with the effect of different drying temperatures and air fluxes on the volatiles in rosemary (Rosmarinus officinalis L.) and thyme (Thymus officinalis L.) cultivated in Sardinia. Fresh leaves were collected and soon divided in two batches, which were subjected to hydro distillation and GC-MS analysis, the first batch as fresh, the second one after drying in a laboratory pilot dryer. Three drying temperatures were used, 30, 38 and 45°C, and for each one two airflow rates were set. The fresh and dried plant material were hydro distilled for 4 hours using a Clevenger-type apparatus (Italian Official Pharmacopeias X). The oils (liquid and light yellow) were recovered directly from above the distillate without adding any solvent and stored at –20°C before analyses, which were carried out on two replicates of each sample by gas chromatography, using a flame ionization detector. The diluted samples were injected using a split/splitless automatic injector (using 2,6-dimethylphenol as internal standard). Qualitative analysis was done by GC/Mass and mass units were monitored from 10 to 450 at 70 eV. Results of the influence of the different drying conditions on volatile compounds of the two herbs will be reported

An epistatic mini-circuitry between the transcription factors Snail and HNF4a controls liver stem cell and hepatocyte features exhorting opposite regulation on stemness-inhibiting microRNAs

Preservation of the epithelial state involves the stable repression of EMT program while maintenance of the stem compartment requires the inhibition of differentiation processes. A simple and direct molecular mini-circuitry between master elements of these biological processes, may provide the best device to keep balanced such complex phenomena. In this work, we show that in hepatic stem cell Snail, a transcriptional repressor of the hepatocyte differentiation master gene HNF4, directly represses the expression of the epithelial microRNAs-200c and -34a, which in turn target several stem cell genes. Notably, in differentiated hepatocytes HNF4, previously identified as a transcriptional repressor of Snail, induces the microRNAs-34a and -200a, b, c that, when silenced, causes epithelial dedifferentiation and reacquisition of stem traits. Altogether these data unveiled Snail, HNF4 and microRNAs -200a, b, c and -34a as epistatic elements controlling hepatic stem cell maintenance/differentiation

TGFβ impairs HNF1α functional activity in Epithelial-to-Mesenchymal Transition interfering with the recruitment of CBP/p300 acetyltransferases

The cytokine transforming growth factor β (TGFβ) plays a crucial role in the induction of both epithelial-to-mesenchymal transition (EMT) program and fibro-cirrhotic process in the liver, where it contributes also to organ inflammation following several chronic injuries. All these pathological situations greatly increase the risk of hepatocellular carcinoma (HCC) and contribute to tumor progression. In particular, late-stage HCCs are characterized by constitutive activation of TGFβ pathway and by an EMT molecular signature leading to the acquisition of invasive and metastatic properties. In these pathological conditions, the cytokine has been shown to induce the transcriptional downregulation of HNF1α, a master regulator of the epithelial/hepatocyte differentiation and of the EMT reverse process, the mesenchymal-to-epithelial transition (MET). Therefore, the restoration of HNF1α expression/activity has been proposed as targeted therapeutic strategy for liver fibro-cirrhosis and late-stage HCCs. In this study, TGFβ is found to trigger an early functional inactivation of HNF1α during EMT process that anticipates the effects of the transcriptional downregulation of its own gene. Mechanistically, the cytokine, while not affecting the HNF1α DNA-binding capacity, impaired its ability to recruit CBP/p300 acetyltransferases on target gene promoters and, consequently, its transactivating function. The loss of HNF1α capacity to bind to CBP/p300 and HNF1α functional inactivation have been found to correlate with a change of its posttranslational modification profile. Collectively, the results obtained in this work unveil a new level of HNF1α functional inactivation by TGFβ and contribute to shed light on the early events triggering EMT in hepatocytes. Moreover, these data suggest that the use of HNF1α as anti-EMT tool in a TGFβ-containing microenvironment may require the design of new therapeutic strategies overcoming the TGFβ-induced HNF1α inactivation

YAP integrates the regulatory Snail/HNF4α circuitry controlling epithelial/hepatocyte differentiation

Yes-associated protein (YAP) is a transcriptional co-factor involved in many cell processes, including development, proliferation, stemness, differentiation, and tumorigenesis. It has been described as a sensor of mechanical and biochemical stimuli that enables cells to integrate environmental signals. Although in the liver the correlation between extracellular matrix elasticity (greatly increased in the most of chronic hepatic diseases), differentiation/functional state of parenchymal cells and subcellular localization/activation of YAP has been previously reported, its role as regulator of the hepatocyte differentiation remains to be clarified. The aim of this study was to evaluate the role of YAP in the regulation of epithelial/hepatocyte differentiation and to clarify how a transducer of general stimuli can integrate tissue-specific molecular mechanisms determining specific cell outcomes. By means of YAP silencing and overexpression we demonstrated that YAP has a functional role in the repression of epithelial/hepatocyte differentiation by inversely modulating the expression of Snail (master regulator of the epithelial-to-mesenchymal transition and liver stemness) and HNF4α (master regulator of hepatocyte differentiation) at transcriptional level, through the direct occupancy of their promoters. Furthermore, we found that Snail, in turn, is able to positively control YAP expression influencing protein level and subcellular localization and that HNF4α stably represses YAP transcription in differentiated hepatocytes both in cell culture and in adult liver. Overall, our data indicate YAP as a new member of the HNF4/Snail epistatic molecular circuitry previously demonstrated to control liver cell state. In this model, the dynamic balance between three main transcriptional regulators, that are able to control reciprocally their expression/activity, is responsible for the induction/maintenance of different liver cell differentiation states and its modulation could be the aim of therapeutic protocols for several chronic liver diseases

Memory-processor co-scheduling in fixed priority systems

A major obstacle towards the adoption of multi-core platforms for real-time systems is given by the difficulties in characterizing the interference due to memory contention. The simple fact that multiple cores may simultaneously access shared memory and communication resources introduces a significant pessimism in the timing and schedulability analysis. To counter this problem, predictable execution models have been proposed splitting task executions into two consecutive phases: a memory phase in which the required instruction and data are pre-fetched to local memory (M-phase), and an execution phase in which the task is executed with no memory contention (C-phase). Decoupling memory and execution phases not only simplifies the timing analysis, but it also allows a more efficient (and predictable) pipelining of memory and execution phases through proper co-scheduling algorithms. In this paper, we take a further step towards the design of smart co-scheduling algorithms for sporadic real-time tasks complying with the M/C (memory-computation) model. We provide a theoretical framework that aims at tightly characterizing the schedulability improvement obtainable with the adopted M/C task model on a single-core systems. We identify a tight critical instant for M/C tasks scheduled with fixed priority, providing an exact response-time analysis with pseudo-polynomial complexity. We show in our experiments that a significant schedulability improvement may be obtained with respect to classic execution models, placing an important building block towards the design of more efficient partitioned multi-core systems

Response-Time Analysis of Conditional DAG Tasks in Multiprocessor Systems

Different task models have been proposed to represent the parallel structure of real-time tasks executing on manycore platforms: fork/join, synchronous parallel, DAG-based, etc. Despite different schedulability tests and resource augmentation bounds are available for these task systems, we experience difficulties in applying such results to real application scenarios, where the execution flow of parallel tasks is characterized by multiple (and nested) conditional structures. When a conditional branch drives the number and size of sub-jobs to spawn, it is hard to decide which execution path to select for modeling the worst-case scenario. To circumvent this problem, we integrate control flow information in the task model, considering conditional parallel tasks (cp-tasks) represented by DAGs composed of both precedence and conditional edges. For this task model, we identify meaningful parameters that characterize the schedulability of the system, and derive efficient algorithms to compute them. A response time analysis based on these parameters is then presented for different scheduling policies. A set of simulations shows that the proposed approach allows efficiently checking the schedulability of the addressed systems, and that it significantly tightens the schedulability analysis of non-conditional (e.g., Classic DAG) tasks over existing approaches