Local Railway Tenders In Italy: The Impossible Competition

Institute of Transport and Logistics Studies. Faculty of Economics and Business. The University of Sydne

RNA Quantitative Analysis from Fixed and Paraffin-Embedded Tissues: Membrane Hybridization and Capillary Electrophoresis

Fixed and paraffin-embedded tissues from pathology department archives are available for RNA expression analysis. We describe a general method for quantitation of specific RNA sequence extracted fr..

A Scheme to Improve the Effectiveness of the Bus Services in Lombardy Through Tenders

In Lombardy public subsidies to bus companies have increased in the last ten years, while market share and quality of public transport have declined. In 1996, the Lombardy Region began a reform of public local transport. In line with EU regulations, the regional transport acts 13/1995 and 22/1998 aim to improve the effectiveness of the public transport through competition and economic incentives. This paper describes the scheme developed for the tenders. In the present concession system, the routes and the schedules for each service are decided by the Region. The reform calls for tenders for a network divided into areas. The areas must be homogeneous in terms of transport demand and attractive for the tenderisers. Their size is chosen to make the management of the services affordable for sufficient competitors. A methodology has been developed to support the grantor in setting the following service requirements for each area: the links to be included in the network and the minimum number of seats to be assured by each route. Further requirements concerning fares, environmental and quality standards, and the information provided to users and to the transport authority are also set. The effectiveness of the planned services is encouraged by economic incentives: part of the subsidy is paid only if specific objectives set by the public authorities are met. The criteria to compare the offers are complex, because of the need to evaluate not only the tendered price, but also the effectiveness of the network and schedule proposals, as well as the usefulness of the additional routes and seats served or the improved quality. A Decision Support System (DSS) software application based on multi-criteria analysis automatically chooses the best project, ensuring a transparent procedure. The reliability of this evaluation system has been tested in simulated tender involving representatives of Lombardy bus companies.Institute of Transport and Logistics Studies. Faculty of Economics and Business. The University of Sydne

The Feasibility of the Road and Park Pricing to Support the Transport Policies

Dissertação apresentada à Faculdade de Direito da Universidade de Coimbra no âmbito do 2.º Ciclo de Estudos em Direito (conducente ao grau de Mestre), especialização em Ciências Jurídico-Políticas / menção em Direito Constituciona

Tumour heterogeneity: principles and practical consequences

Two major reasons compel us to study tumour heterogeneity: firstly, it represents the basis of acquired therapy resistance, and secondly it may be one of the major sources of the low level of reproducibility in clinical cancer research. The present review focuses on the heterogeneity of neoplastic disease, both within the primary tumour, and between primary tumour and metastases. We discuss different levels of heterogeneity and the current understanding of the phenomenon, as well as imminent developments relevant for clinical research and diagnostic pathology. It is necessary to develop new tools to study heterogeneity and new biomarkers for heterogeneity. Established and new in situ methods will be very useful. In future studies, not only clonal heterogeneity needs to be addressed, but also non-clonal phenotypic heterogeneity which might be important for therapy resistance. We also review heterogeneity established in major tumour types, in order to explore potential similarities that might help to define new strategies for targeted therapy

In stage II/III lymph node positive breast cancer patients less than 55 years of age, keratin 8 expression in lymph node metastases but not in the primary tumor is an indicator of better survival

Axillary lymph node status is one of the most important prognostic variables for breast cancer (BC). To investigate and understand the clinical, histopathological and biological factors that affect prognosis in node positive young breast cancer patients, we compared the phenotype of 100 primary tumours with their corresponding loco- regional lymph node (LN) metastases using conventional immunohistochemistry (IHC) markers currently in use for molecular classification of breast cancer. By comparing the expression of ER, PR, HER-2, Ki67, K8, K5/6 and vimentin, we found that expression of HER-2, Ki67, K8 and vimentin is frequently lost in lymph node metastases. Between the primary tumour and corresponding lymph node metastases expression of keratins K8 and K5/6 significantly changed. Expression of K8 in lymph node metastases, but not in primary tumours, segregates patients in two sub-groups with different outcome. Survival of patients with K8 positive LN metastases at 5 years in comparison with patients with K8 negative LN metastases was 75% vs 48%, at 10 years 62% vs 22% and at 20 years 53% vs 14%(p<0.001). K8 immunostaining of tissue from the lymph node metastasis allows defining a sub-group of lymph node positive BC patients with a highly unfavourable outcome, for whom therapeutic options might have to be reconsidered

Impact of standardization in tissue processing: the performance of different fixatives

Most tissues in clinical practice are formalin-fixed and paraffin-embedded for histological as well as molecular analyses. The reproducibility and uniformity of molecular analyses is strictly dependent on the quality of the biomolecules, which is highly influenced by pre-analytical processes. In this study, the effect of different fixatives was compared, including formalin, Bouin's solution, RCL2® and TAG-1™ fixatives, by stringent application of ISO standards in mouse liver tissue processing, including formalin-free transport of tissues and tissue grossing in a refrigerated environment. The effect of fixatives was studied in terms of nucleic acid quality at the time of tissue processing and after one year of tissue storage at room temperature in the dark. Furthermore, a microcomputed tomography (CT) scan analysis was applied to investigate the paraffin embedding. The results show that the application of ISO standards in tissue processing allows analysis of 400 bases amplicons from RNA and 1000 bases from DNA, even in extracts from formalin-fixed and paraffin-embedded tissues. However, after one year storage at room temperature in the dark, a degradation of the nucleic acids was observed. Nevertheless, extracts can still be analyzed, but for metachronous tests it is highly recommended to repeat the quantitation of housekeeping genes in order to standardize the extent of nucleic acid degradation

Molecular characterisation of breast cancer patients at high and low recurrence risk

The ability to predict the recurrence risk in breast cancer patients is not available for the individual. It is commonly accepted that the different clinical course of tumours with identical histology and stage are the result of differences at the molecular level. This case study of 80 patients affected by breast cancer looked at the messenger ribonucleic acid expression level of 22 genes, by using quantitative reverse transcriptase-polymerase chain reaction. Our results showed that a panel of seven genes is associated to patients' survival. Moreover, the combination of two couples of genes is able to define short- and long-living cohorts of patients. In particular, our findings strongly demonstrate that retinoblastoma (RB) and cyclin-dependent kinase 2 (CDK2) on one side and cytokeratin 8 (CK8) and epidermal growth factor receptor 2 (HER2) on the other may affect the clinical course of the disease in 56% of patients. Groups characterised by low RB and high CDK2 as with low CK8 and high HER2 have a higher risk of recurrences and death in 5 years. The identification of these sub-groups of patients with higher risk of early relapse could have further involvement in the selection of the cases to submit to therapy against HER2 or CDK2 as a possible therapy target