Molecular profiling of single circulating tumor cells with diagnostic intention

Several hundred clinical trials currently explore the role of circulating tumor cell (CTC) analysis for therapy decisions, but assays are lacking for comprehensive molecular characterization of CTCs with diagnostic precision. We therefore combined a workflow for enrichment and isolation of pure CTCs with a non-random whole genome amplification method for single cells and applied it to 510 single CTCs and 189 leukocytes of 66 CTC-positive breast cancer patients. We defined a genome integrity index (GII) to identify single cells suited for molecular characterization by different molecular assays, such as diagnostic profiling of point mutations, gene amplifications and whole genomes of single cells. The reliability of >90% for successful molecular analysis of high-quality clinical samples selected by the GII enabled assessing the molecular heterogeneity of single CTCs of metastatic breast cancer patients. We readily identified genomic disparity of potentially high relevance between primary tumors and CTCs. Microheterogeneity analysis among individual CTCs uncovered pre-existing cells resistant to ERBB2-targeted therapies suggesting ongoing microevolution at late-stage disease whose exploration may provide essential information for personalized treatment decisions and shed light into mechanisms of acquired drug resistance

When the baby cries at night. Inelastic buyers in non-competitive markets *

Abstract We investigate empirically how sellers react to changes in the population of their consumers, identifying the effects of demand composition and demand size with limited information on costs. We show how pharmacists in Italy selectively increase the price of some products when they observe in their cities an exogenous influx of parents of newborns, conceivably less elastic buyers as compared with other more experienced and less pressed consumers. Exploiting population based laws that fix the number of pharmacies in a city, we use RDD to measure the effect of competition on sellers' ability to extract surplus from less elastic buyers. JEL-Code: D43, D83 L13

Molecular profiling of single circulating tumor cells with diagnostic intention

Several hundred clinical trials currently explore the role of circulating tumor cell (CTC) analysis for therapy decisions, but assays are lacking for comprehensive molecular characterization of CTCs with diagnostic precision. We therefore combined a workflow for enrichment and isolation of pure CTCs with a non-random whole genome amplification method for single cells and applied it to 510 single CTCs and 189 leukocytes of 66 CTC-positive breast cancer patients. We defined a genome integrity index (GII) to identify single cells suited for molecular characterization by different molecular assays, such as diagnostic profiling of point mutations, gene amplifications and whole genomes of single cells. The reliability of >90% for successful molecular analysis of high-quality clinical samples selected by the GII enabled assessing the molecular heterogeneity of single CTCs of metastatic breast cancer patients. We readily identified genomic disparity of potentially high relevance between primary tumors and CTCs. Microheterogeneity analysis among individual CTCs uncovered pre-existing cells resistant to ERBB2-targeted therapies suggesting ongoing microevolution at late-stage disease whose exploration may provide essential information for personalized treatment decisions and shed light into mechanisms of acquired drug resistance

Nuovi strumenti di pianificazione comunale per la città storica

Dalla metà degli anni ‘60 del secolo scorso l’Amministrazione ha affrontato il tema della rigenerazione del centro storico attraverso piani urbanistici e altri programmi (il “Piano del centro storico” del 1969, un piano di edilizia sociale nel centro storico adottato nel 1973, il “Piano d’intervento pubblico per la riorganizzazione e ristrutturazione dei servizi” del 1973-75) che nel loro insieme hanno realizzato significativi successi per quanto riguarda il risanamento igienico, strutturale e ambientale degli edifici, mentre hanno raggiunto solo parzialmente gli obiettivi di carattere funzionale, sociale e tipologico. Nel 1989 la Variante generale al Piano Regolatore Generale, ha confermato sostanzialmente le scelte di tutela del centro storico contenute nella precedente pianificazione, riservando attenzione a tutti gli edifici, estendendo la tutela dei singoli manufatti anche fuori dalle vecchie mura. Nel 2000 la Regione Emilia-Romagna emana la nuova legge sul governo del territorio, che riforma in maniera significativa i modi di pianificare, considerando le nuove esigenze di sostenibilità e la nuova stagione dello sviluppo urbano, più orientato alla riqualificazione dell’esistente che alla realizzazione di nuove addizioni urbane. Tra il 2005 e il 2009 la città di Bologna progetta e approva i nuovi strumenti di pianificazione e regolazione urbanistica comunale: il Piano Strutturale Comunale (approvato nel 2008), il Regolamento Urbanistico Edilizio e il Piano Operativo Comunale (approvati nel 2009)

The Italian version of the Circle of Security Parenting: clinical application and preliminary empirical data from a research study

Introduction The Circle of Security Parenting (COS-P) protocol is an early intervention program designed to prevent insecure attachment, child psychopathological distress and to promote psychological well-being that can be used with groups, dyads, and individuals. It is a visually based approach (utilizing extensive use of both graphics and video clips). Created in USA (Cooper, Hoffman, Powell, Marvin, 2005), COS-P is now widespread in many countries and has been translated into seven languages. The final purpose of COS-P is to support parents to recognize and respond to their children’s alternating needs for attachment and exploration (Powell, Cooper, Hoffman, Marvin, 2009). It provides an opportunity for caregivers to reflect on their child’s needs and the challenges each faces in meeting those needs (Cooper, Hoffman, & Powell, 2009). Whereas COS is an evidenced based approach, there are only preliminary empirical data on the efficacy of COS-P. Using case studies, the presentation describes the Italian version of COS-P, produced with the collaboration of its creators, and its preliminary empirical data from a research study. Particularly attention will be paid to the setting up of a research study designed to evaluate the efficacy of the COS-P in Italy. Method Case studies will be presented and preliminary data will be discussed on a pre-intervention/post-intervention design used to examine the COS-P efficacy. Self-report questionnaires and a projective tool were administered to patients participating to the protocol. The areas of investigation of used tools can be grouped in: dyadic adjustment; parenting; caregiving; parents’ subjective appraisal of child problems; parent’s attachment style and model. Results/discussion Preliminary data of the application of COS-P on case studies showed interesting results in terms of enhancement of the attachment-caregiving relationships and of parents’ subjective appraisal of child needs. COS constituted a type of treatment to support parenting with great potential in the Italian context

The circle of security parenting and parental conflict: a single case study

The Circle of Security Parenting (COS-P) is an early attachment based intervention that can be used with groups, dyads, and individuals. Created in the USA and now used in many countries, COS-P is a visually based approach that demonstrates its central principles through videos of parent/child interactions. The core purpose of the COS-P is to provide an opportunity for caregivers to reflect on their child’s needs and on the challenges each parent faces in meeting those needs. Even though there is a wide range of clinical settings in which child/parent attachment is an important component of assessment there is limited empirical data on when and how attachment based interventions are appropriate for specific clinical profiles and contexts. The aim of this paper is to present a clinical application of COS-P in order to explore and reflect on some specific therapeutic tasks where it works and on some clinical indicators and contexts appropriate for its application. A single case study of a father, “M.” (43 years old) in conflict for the custody of his 5 years old daughter is reported. The Adult Attachment Projective Picture System (AAP), the Parenting Stress Index, the Strengths and Difficulties Questionnaire, and the Parental Alliance Measure, were administered pre- and post-intervention. The clinical significance analysis method revealed that numerous changes occurred in the father. The AAP showed improvements in the level of agency of self. M. made gains in his capacity to use internal resources and to increase his agency of self. M. was classified as recovered in his perception of the child’s functioning and as improved in his parenting stress and parenting alliance with the mother. Considerations on specific contexts and clinical indicators for the application of COS-P are proposed

Non-Permissive Parvovirus B19 Infection: A Reservoir and Questionable Safety Concern in Mesenchymal Stem Cells

Mesenchymal stromal/stem cells (MSCs) are multipotent cells with differentiation, immunoregulatory and regenerative properties. Because of these features, they represent an attractive tool for regenerative medicine and cell-based therapy. However, MSCs may act as a reservoir of persistent viruses increasing the risk of failure of MSCs-based therapies and of viral transmission, especially in immunocompromised patients. Parvovirus B19V (B19V) is a common human pathogen that infects bone marrow erythroid progenitor cells, leading to transient or persistent anemia. Characteristics of B19V include the ability to cross the placenta, infecting the fetus, and to persist in several tissues. We thus isolated MSCs from bone marrow (BM-MSCs) and fetal membrane (FM-MSCs) to investigate their permissiveness to B19V infection. The results suggest that both BM- and FM- MSCs can be infected by B19V and, while not able to support viral replication, allow persistence over time in the infected cultures. Future studies are needed to understand the potential role of MSCs in B19V transmission and the conditions that can favor a potential reactivation of the virus