Causes and consequences of natural hybridisation among coral reef butterflyfishes (Chaetodon: Chaetodontidae)

Natural hybridisation is the successful interbreeding of individuals from different populations, distinguishable through one or more heritable characters, and is a widespread phenomenon in the plant and animal kingdoms. The development of hybridisation theories has largely been based on studies in terrestrial and freshwater ecosystems. Hybridisation was traditionally considered rare and unimportant in marine systems and therefore received little attention. Recently however, there has been a surge of reported marine hybrids, particularly in corals and reef fishes. The ecological promoters and evolutionary and adaptive consequences of reef fish hybridisation are yet to be thoroughly evaluated. Butterflyfishes (f. Chaetodontidae) form a disproportionate number of hybrids and therefore represent an appropriate model group to investigate hybridisation in reef fishes. This thesis examines the causes and consequences of hybridisation in reef fishes and focuses on butterflyfishes (genus Chaetodon) at Christmas Island (Indian Ocean), a global hotspot for reef fish hybridisation. The aims of this thesis were to i) review the incidence and ecological/behavioural precursors of hybridisation in reef fishes, while providing a tentative framework for conducting studies within hybrid zones; ii) develop a microsatellite toolkit for species of the Chaetodon genus; iii) compare the ecology, behaviour and population genetics of hybridising sister species of butterflyfishes in order to, not only provide a snapshot of the evolutionary consequences of hybridisation in this group, but also determine which processes are likely to promote it; iv) use a comparative life history approach to determine the fitness of butterflyfish hybrids relative to their parental species. Chapter 1 reviews the current knowledge of hybridisation with a focus on marine fishes. Hybridisation was found to be highly prevalent in marine fish, despite previous assertions of rarity, and showed a taxonomic as well as latitudinal bias. Further, the current marine fish hybridisation literature was found to be largely lacking ecological and behavioural data, in contrast with freshwater counterparts, therefore highlighting the need for a framework to fill the data gap in order to better accompany the wealth of genetic data produced in the assessment of hybridisation. The development of a molecular toolkit, necessary for the rest of our investigations, is presented in Chapter 2. Twenty microsatellite loci were developed using 454 sequencing, to apply to the population genetic analysis of the Chaetodon guttatissimus × C. punctatofasciatus complex. This was done to facilitate direct comparison of the genetic underpinnings of hybridisation in this group to those of another previously studied group (C. trifasciatus and C. lunulatus), for which species-specific microsatellite loci had been designed and used. Chapter 3 uses the molecular toolkit and experimental framework outlined in the previous chapters to examine hybridisation between two butterflyfish sister species, Chaetodon guttatissimus and C. punctatofasciatus. The largely overlapping spatial and dietary ecologies of these species favour heterospecific encounters. Lack of assortative mating and local rarity of C. punctatofasciatus promote the formation of heterospecific breeding pairs. Analyses of mtDNA and microsatellite DNA were consistent with the hybrid status of the intermediately coloured hybrids. Maternal contribution to hybridisation in this complex was bidirectional, and introgression by C. punctatofasciatus mtDNA was detected in C. guttatissimus individuals within and beyond the hybrid zone (almost 1000 km to the west), potentially indicating a Pacific invasion of an Indian Ocean species genome. The comparisons drawn with previous work on hybrdising Chaetodon trifasciatus and C. lunulatus showed that, despite being driven by similar factors, hybridisation in reef fishes can have varying evolutionary consequences, possibly due to the magnitude of the genetic distance between hybridising species. Chapter 4 evaluates hybrid fitness in both Chaetodon hybridising groups presented in the previous study. Histology confirmed the reproductive viability of hybrids, and liver lipid analyses showed that hybrid condition was not different from parental species. Further, otolith data highlighted no difference in growth rate and maximum length between hybrids and parents. According to the fitness-related traits measured here, Chaetodon hybrids are as fit as their parents, and unlikely to promote the formation of a hybrid swarm. However, sufficient fitness and rapid transfer of genetic material between species allow persistence of hybrids within the suture zone, where they positively contribute to genetic diversity. The cases of hybridisation studied here appear to be initiated by similar ecological and behavioural settings, albeit showing different genetic consequences. Determining this was possible through the use of a comprehensive approach, which combined molecular analyses and extensive field observations. Further, the apparent lack of differences in fitness between hybrids and parental species points at the persistence of hybrid individuals within the Indo- Pacific suture zone, where they may continue to contribute positively to genetic diversity. The role of hybridisation in evolution and adaptability had been appreciated in terrestrial and freshwater systems, and this thesis shows that hybridisation can have a role in maintaining reef fish diversity. The studies presented here constitute a comprehensive overview of the relevance of hybridisation for reef fishes and may be a stepping stone toward ascertaining its role in the evolution and adaptation of new species in such a diverse group

Soft Concurrent Constraint Programming

Soft constraints extend classical constraints to represent multiple consistency levels, and thus provide a way to express preferences, fuzziness, and uncertainty. While there are many soft constraint solving formalisms, even distributed ones, by now there seems to be no concurrent programming framework where soft constraints can be handled. In this paper we show how the classical concurrent constraint (cc) programming framework can work with soft constraints, and we also propose an extension of cc languages which can use soft constraints to prune and direct the search for a solution. We believe that this new programming paradigm, called soft cc (scc), can be also very useful in many web-related scenarios. In fact, the language level allows web agents to express their interaction and negotiation protocols, and also to post their requests in terms of preferences, and the underlying soft constraint solver can find an agreement among the agents even if their requests are incompatible.Comment: 25 pages, 4 figures, submitted to the ACM Transactions on Computational Logic (TOCL), zipped file

Deregulation of MiR-34b/Sox2 Predicts Prostate Cancer Progression.

Most men diagnosed with prostate cancer will have an indolent and curable disease, whereas approximately 15% of these patients will rapidly progress to a castrate-resistant and metastatic stage with high morbidity and mortality. Therefore, the identification of molecular signature(s) that detect men at risk of progressing disease remains a pressing and still unmet need for these patients. Here, we used an integrated discovery platform combining prostate cancer cell lines, a Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP) model and clinically-annotated human tissue samples to identify loss of expression of microRNA-34b as consistently associated with prostate cancer relapse. Mechanistically, this was associated with epigenetics silencing of the MIR34B/C locus and increased DNA copy number loss, selectively in androgen-dependent prostate cancer. In turn, loss of miR-34b resulted in downstream deregulation and overexpression of the stemness marker, Sox2. These findings identify loss of miR-34b as a robust biomarker for prostate cancer progression in androgen-sensitive tumors, and anticipate a potential role of progenitor/stem cell signaling in this stage of disease

Naturally occurring hybrids of coral reef butterflyfishes have similar fitness compared to parental species.

Hybridisation can produce evolutionary novelty by increasing fitness and adaptive capacity. Heterosis, or hybrid vigour, has been documented in many plant and animal taxa, and is a notable consequence of hybridisation that has been exploited for decades in agriculture and aquaculture. On the contrary, loss of fitness in naturally occurring hybrid taxa has been observed in many cases. This can have negative consequences for the parental species involved (wasted reproductive effort), and has raised concerns for species conservation. This study evaluates the relative fitness of previously documented butterflyfish hybrids of the genus Chaetodon from the Indo-Pacific suture zone at Christmas Island. Histological examination confirmed the reproductive viability of Chaetodon hybrids. Examination of liver lipid content showed that hybrid body condition was not significantly different from parent species body condition. Lastly, size at age data revealed no difference in growth rates and asymptotic length between hybrids and parent species. Based on the traits measured in this study, naturally occurring hybrids of Chaetodon butterflyfishes have similar fitness to their parental species, and are unlikely to supplant parental species under current environmental conditions at the suture zone. However, given sufficient fitness and ongoing genetic exchange between the respective parental species, hybrids are likely to persist within the suture zone

miRNAs Expression Analysis in Paired Fresh/Frozen and Dissected Formalin Fixed and Paraffin Embedded Glioblastoma Using Real-Time PCR

miRNAs are small molecules involved in gene regulation. Each tissue shows a characteristic miRNAs epression profile that could be altered during neoplastic transformation. Glioblastoma is the most aggressive brain tumour of the adult with a high rate of mortality. Recognizing a specific pattern of miRNAs for GBM could provide further boost for target therapy. The availability of fresh tissue for brain specimens is often limited and for this reason the possibility of starting from formalin fixed and paraffin embedded tissue (FFPE) could very helpful even in miRNAs expression analysis. We analysed a panel of 19 miRNAs in 30 paired samples starting both from FFPE and Fresh/Frozen material. Our data revealed that there is a good correlation in results obtained from FFPE in comparison with those obtained analysing miRNAs extracted from Fresh/Frozen specimen. In the few cases with a not good correlation value we noticed that the discrepancy could be due to dissection performed in FFPE samples. To the best of our knowledge this is the first paper demonstrating that the results obtained in miRNAs analysis using Real-Time PCR starting from FFPE specimens of glioblastoma are comparable with those obtained in Fresh/Frozen samples

Impact of gastrointestinal side effects on patients’ reported quality of life trajectories after radiotherapy for prostate cancer: Data from the prospective, observational pros-it CNR study

Radiotherapy (RT) represents an important therapeutic option for the treatment of localized prostate cancer. The aim of the current study is to examine trajectories in patients’ reported quality of life (QoL) aspects related to bowel function and bother, considering data from the PROState cancer monitoring in ITaly from the National Research Council (Pros-IT CNR) study, analyzed with growth mixture models. Data for patients who underwent RT, either associated or not associated with androgen deprivation therapy, were considered. QoL outcomes were assessed over a 2-year period from the diagnosis, using the Italian version of the University of California Los Angeles-Prostate Cancer Index (Italian-UCLA-PCI). Three trajectories were identified for the bowel function; having three or more comorbidities and the use of 3D-CRT technique for RT were associated with the worst trajectory (OR = 3.80, 95% CI 2.04–7.08; OR = 2.17, 95% CI 1.22–3.87, respectively). Two trajectories were identified for the bowel bother scores; diabetes and the non-Image guided RT method were associated with being in the worst bowel bother trajectory group (OR = 1.69, 95% CI 1.06–2.67; OR = 2.57, 95% CI 1.70–3.86, respectively). The findings from this study suggest that the absence of comorbidities and the use of intensity modulated RT techniques with image guidance are related with a better tolerance to RT in terms of bowel side effects

Disease-specific and general health-related quality of life in newly diagnosed prostate cancer patients: The Pros-IT CNR study

Background: The National Research Council (CNR) prostate cancer monitoring project in Italy (Pros-IT CNR) is an observational, prospective, ongoing, multicentre study aiming to monitor a sample of Italian males diagnosed as new cases of prostate cancer. The present study aims to present data on the quality of life at time prostate cancer is diagnosed. Methods: One thousand seven hundred five patients were enrolled. Quality of life is evaluated at the time cancer was diagnosed and at subsequent assessments via the Italian version of the University of California Los Angeles-Prostate Cancer Index (UCLA-PCI) and the Short Form Health Survey (SF-12). Results: At diagnosis, lower scores on the physical component of the SF-12 were associated to older ages, obesity and the presence of 3+ moderate/severe comorbidities. Lower scores on the mental component were associated to younger ages, the presence of 3+ moderate/severe comorbidities and a T-score higher than one. Urinary and bowel functions according to UCLA-PCI were generally good. Almost 5% of the sample reported using at least one safety pad daily to control urinary loss; less than 3% reported moderate/severe problems attributable to bowel functions, and sexual function was a moderate/severe problem for 26.7%. Diabetes, 3+ moderate/severe comorbidities, T2 or T3-T4 categories and a Gleason score of eight or more were significantly associated with lower sexual function scores at diagnosis. Conclusions: Data collected by the Pros-IT CNR study have clarified the baseline status of newly diagnosed prostate cancer patients. A comprehensive assessment of quality of life will allow to objectively evaluate outcomes of different profile of care

"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