Human cardiac progenitor cell grafts as unrestricted source of supernumerary cardiac cells in healthy murine hearts

Human heart harbors a population of resident progenitor cells that can be isolated by stem cell antigen-1 antibody and expanded in culture. These cells can differentiate into cardiomyocytes in vitro and contribute to cardiac regeneration in vivo. However, when directly injected as single cell suspension, less than 1%-5% survive and differentiate. Among the major causes of this failure are the distressing protocols used to culture in vitro and implant progenitor cells into damaged hearts. Human cardiac progenitors obtained from the auricles of patients were cultured as scaffoldless engineered tissues fabricated using temperature-responsive surfaces. In the engineered tissue, progenitor cells established proper three-dimensional intercellular relationships and were embedded in self-produced extracellular matrix preserving their phenotype and multipotency in the absence of significant apoptosis. After engineered tissues were leant on visceral pericardium, a number of cells migrated into the murine myocardium and in the vascular walls, where they integrated in the respective textures. The study demonstrates the suitability of such an approach to deliver stem cells to the myocardium. Interestingly, the successful delivery of cells in murine healthy hearts suggests that myocardium displays a continued cell cupidity that is strictly regulated by the limited release of progenitor cells by the adopted source. When an unregulated cell source is added to the system, cells are delivered to the myocardium. The exploitation of this novel concept may pave the way to the setup of new protocols in cardiac cell therapy. STEM CELLS 2011;29:2051-206

Minimally invasive mitral valve surgery: State-of-the-art and our experience

The minimally invasive approach is becoming the standard-of-care for surgery of the mitral valve. As any less invasive strategy, it entails an increased surgical complexity. Standard-of-care mitral repair using the totally videoscopic approach is indeed reproducible; however, few specific data on patients with complex mitral valve disease are available in the published literature. The purpose of the present paper is to provide an overview of the current state-of-the-art in minimally invasive cardiac surgery, and a summary of recent evidence on the topic, with particular regard to the surgical techniques and comparisons with conventional surgery. The experience of the GVM Care and Research network in the field is also briefly reported

The October 2000 flooding in Valle d'Aosta (Italy): Event description and land planning measures for the risk mitigation

On October 13-16th, 2000 heavy rainfalls in the Northwestern Italian Alps caused huge flooding and landslides with significant damages to houses and infrastructures and several life losses. In this paper a description of the main events that affected Valle d’Aosta’s region and the subsequent land planning measures adopted for the risk mitigation are presented. After a first meteorological and hydrological framing, based on the data of the regional monitoring system (that pointed out rainfalls up to 236 mm in 24 h also in high‐altitude zones, because of the rise of the isotherm 0°C around 3000 m above sea level), the main effects of the event (extensive flooding, landslides, soil slips and debris flows) in the regional catchment of the Dora Baltea river are described. Through aerial and direct surveys those effects have been transferred into a thematic cartography within two months from the event, in order to have detailed elements for the technical, administrative and political land planning decisions, and, on this basis, a new regional directive containing detailed measures for the hydro‐geological risk mitigation and land safety has been adopted. © 2003 by Taylor nad Francis Group, LLC

Interspecific differences in swimming performance, behavior and survival between native Italian gudgeon (Gobio benacensis Pollini, 1816) and non-native European gudgeon (Gobio gobio Linnaeus, 1758)

Introduction of non-native species is an important cause of biodiversity decline in rivers. Separated by mountains and salt water, the freshwater fish fauna in Italy has experienced a natural isolation from fish in continental Europe. As a consequence, several Italian fish species have diverged from their European sister species, likely with unique adaptations to the local environment. Relatedly, the region is also susceptible to the invasion of non-native fish, and today almost half the fish species present are of non-native origin. Several of these non-native species have Italian congeners susceptible to competition and hybridizations, and, in the long run, displacements and extinctions. One such example is the Italian gudgeon (Gobio benacensis) and its European congener European gudgeon (Gobio gobio). During the last few decades the European gudgeon was introduced in Italian waters and has since spread rapidly, causing progressive declines in the Italian species. As for several other similar species pairs, little is known about potential differences in ecology and behavior. Here we study differences between Italian and European gudgeons in a controlled laboratory environment, using a combined open field and provoked escape response test, as well as tracking their sympatric survival over time in the hatchery. The smaller Italian gudgeon displayed a lower maximum swimming speed compared to the larger European gudgeon. The Italian gudgeon also experienced substantially higher sympatric mortality, resulting in only European gudgeon surviving to the end of the experiment. Contrary to expectations, no difference was seen in boldness and the Italian gudgeon displayed a higher movement activity, moving a larger distance in an open field test, compared to its European sister species. The reported differences could play a role in the replacement process, and may also have impacts on the surrounding ecosystem, and the prey and predators that have coevolved with the Italian species

Artificial light at night affects fish passage rates in two small-sized Cypriniformes fish

An increasing presence of instream structures such as weirs, dams, culverts and reservoirs degrades habitats, fragments rivers and blocks fish movements worldwide. Longitudinal river movements are fundamental for many fish species and the most widespread solution to restore longitudinal connectivity is the implementation of different fish passage solutions. Fishway functionality, however, is highly variable. To design a functional fishway, several aspects of the fish's interaction with its environment need to be taken into consideration. Artificial light at night (ALAN) can affect a range of different behaviours in fish, from activity and movement to feeding and predator-prey relationships. In a fish passage setting, fish are exposed to artificial light at night (ALAN) in the form of light pollution, but, sometimes, also as part of the fish passage solution. Although likely highly species specific, the effect of artificial light on fish passage behaviour has been little explored. Here we study the passage behaviour of two small-sized fish species, European gudgeon (Gobio gobio) and Italian riffle dace (Telestes muticellus), over a scaled deep side notch weir in a hydraulic flume in three different light conditions: daylight, darkness and ALAN. Although both species passed the obstacle at high efficiencies under all light conditions, their passage behaviours were influenced by light, particularly at the higher levels. While ALAN reduced passage success and resulted in delayed passage for gudgeon, riffle dace passed at higher rates under the artificial light compared to night treatment. Both results indicate a risk of negative effects from ALAN on passage performance at real fishways or movement rates in lit areas of natural streams for both species. Independent of light conditions, individuals of both species also passed faster after repeated trials, demonstrating learning in a fish passage context

Individual activity levels and presence of conspecifics affect fish passage rates over an in-flume barrier

Dams and other in-stream obstacles disrupt longitudinal connectivity and hinder fish from moving between habitats. Fishways and other fish passage solutions are used to pass fish over these artificial migration barriers. Fish passage functionality, however, varies greatly with fish passage design and environmental conditions and depends on fish species and characteristics. In particular, swimming performance and fish behaviour are considered key characteristics to predict fish passage performance. It is also well known, but not well quantified, that the presence of conspecifics affects fish passage behaviour. In this study, we quantified individual passage rates of PIT-tagged gudgeons (Gobio gobio) over a scaled deep side notch weir in an hydraulic flume. We then quantified individual swimming capability (time to fatigue) and activity level (distance moved in an open field test) for the same individual fish and tested for potential effects on fish passage rate. To check for potential group effects, we then repeated the passage experiment for fish individually or in groups of five. More active fish displayed higher passage rates compared to less active fish, and fish passed the obstacle at higher rates in groups of five compared to alone. No effect of fish swimming capability on passage rates was detected. This result highlights the need to take both individual variation as well as the presence and behaviour of conspecifics into account in fish passage studies and evaluations. Doing so has the potential to improve the understanding of fish behaviour, and in the end, the design of fish passage solutions. Future studies should explore these results on free ranging fish and in relation to in-situ fish passage solutions

Analysis of the efficacy of EPIONE therapies to treat phantom limb pain

Indiana University-Purdue University Indianapolis (IUPUI)The primary objectives of this thesis are (1) to discuss the current understanding of phenomena associated with, proposed mechanisms of, and suggested treatments for amputation related pain, (2) to describe the software developed for analyzing results of a clinical study for the treatment of phantom limb pain (PLP), (3) to discuss the methods for a multi-center trial by the EPIONE consortium along with presenting preliminary results, and (4) to discuss the methods and results of a case study involving a new therapy modality for alleviating PLP. Each objective has been expanded into a chapter as described below. Chapter 1 serves as a literature review introducing the topic of amputation, associ- ated phenomena, and proposed mechanisms. The chapter also discusses the currently available treatments and the instruments used to measure PLP. Key topics include the definition of PLP, the prevalence of PLP, current treatment options for PLP, and experimental measurement of PLP. The final objective of this chapter is to introduce topics related to the investigation paradigm utilized for the studies following in Chapter 4 and Chapter 5. Therefore, a minor emphasis has been put on surface electrical stimulation (SES) and operant conditioning. As with any multi-center clinical study, coordination is key. Chapter 2 introduces the common clinical protocol (CCP) and methods of analysis for the clinical trials conducted by the EPIONE consortium. In order to analyze results in an automated fashion, a software tool was developed. This tool, the EPIONE Extraction Program (EEP) along with its extension the Group Analysis Module (GAM), is the focus of Chapter 3. A high-level overview of the requirements, process flow, and software testing are described. This chapter also discusses the methods of analysis for several self-report instruments used to determine effect size in Chapter 4 and Chapter 5. The outputs of the software tools make up the results presented and described in these chapters. In addition to the details included in Chapter 3, supplemental information is available in Appendix A and Appendix B, which are the detailed User Guides for the EEP and GAM. Chapter 4 reviews the pilot study data conducted by the EPIONE consortium. The primary and two secondary instruments used for analysis are discussed. This chapter provides a brief overview of results from the group. Each clinical site used slightly different variations of a common clinical protocol to better understand what effectively drives alleviation of PLP and to allow comparison of results. The work done at Indiana University - Purdue University Indianapolis (IUPUI) represents a small part of several other universities involved in the EPIONE consortium. Chapter 5 focuses on a case study at IUPUI with a more in-depth review of data collected throughout the study period. Using SES, we seek to reverse cortical reorganization by giving meaningful stimuli through existing circuitry. In this chapter the present work is discussed by introducing a case study in detail with an analysis of psychophysical data

Targeting the MET oncogene by concomitant inhibition of receptor and ligand via an antibody-“decoy” strategy

MET, a master gene sustaining "invasive growth," is a relevant target for cancer precision therapy. In the vast majority of tumors, wild-type MET behaves as a "stress-response" gene and relies on the ligand (HGF) to sustain cell "scattering," invasive growth and apoptosis protection (oncogene "expedience"). In this context, concomitant targeting of MET and HGF could be crucial to reach effective inhibition. To test this hypothesis, we combined an anti-MET antibody (MvDN30) inducing "shedding" (i.e., removal of MET from the cell surface), with a "decoy" (i.e., the soluble extracellular domain of the MET receptor) endowed with HGF-sequestering ability. To avoid antibody/decoy interaction-and subsequent neutralization-we identified a single aminoacid in the extracellular domain of MET-lysine 842-that is critical for MvDN30 binding and engineered the corresponding recombinant decoyMET (K842E). DecoyMET(K842E) retains the ability to bind HGF with high affinity and inhibits HGF-induced MET phosphorylation. In HGF-dependent cellular models, MvDN30 antibody and decoyMET(K842E) used in combination cooperate in restraining invasive growth, and synergize in blocking cancer cell "scattering." The antibody and the decoy unbridle apoptosis of colon cancer stem cells grown in vitro as spheroids. In a preclinical model, built by orthotopic transplantation of a human pancreatic carcinoma in SCID mice engineered to express human HGF, concomitant treatment with antibody and decoy significantly reduces metastatic spread. The data reported indicate that vertical targeting of the MET/HGF axis results in powerful inhibition of ligand-dependent MET activation, providing proof of concept in favor of combined target therapy of MET "expedience.