Rapid identification and differentiation of Mycobacterium avium subspecies paratuberculosis types by use of real-time PCR and high-resolution melt analysis of the MAP1506 locus

High-resolution melt (HRM) analysis can identify sequence polymorphisms by comparing the melting curves of amplicons generated by real-time PCR amplification. We describe the application of this technique to identify Mycobacterium avium subspecies paratuberculosis types I, II, and III. The HRM approach was based on type-specific nucleotide sequences in MAP1506, a member of the PPE (proline-proline-glutamic acid) gene family.

Generation and screening of natural product-like compounds for antibiotic discovery

Avec l’apparition de plus en plus de souches de bactérie résistante aux antibiotiques, le développement de nouveaux antibiotiques est devenu une important problématique pour les agences de santé. C’est pour cela que la création de nouvelles plateformes pour accélérer la découverte de médicaments est devenu un besoin urgent. Dans les dernières décennies, la recherche était principalement orientée sur la modification de molécules préexistantes, la méta-analyse d’organismes produisant des molécules activent et l’analyse de librairies moléculaires pour trouver des molécules synthétiques activent, ce qui s’est avéré relativement inefficace. Notre but était donc de développer de nouvelles molécules avec des effets thérapeutiques de façon plus efficace à une fraction du prix et du temps comparé à ce qui se fait actuellement. Comme structure de base, nous avons utilisé des métabolites secondaires qui pouvaient altérer le fonctionnement des protéines ou l’interaction entre deux protéines. Pour générer ces molécules, j’ai concentré mes efforts sur les terpènes, une classe de métabolites secondaires qui possède un large éventail d’activités biologiques incluant des activités antibactériennes. Nous avons développé un système de chromosome artificiel de levure (YAC) qui permet à la fois l’assemblage directionnel et combinatoire de gènes qui permet la création de voies de biosynthèse artificielles. Comme preuve de concept, j’ai développé des YACs qui contiennent les gènes pour l’expression des enzymes impliquées dans la biosynthèse de la -carotène et de l’albaflavenone et produit ces molécules avec un haut rendement. Finalement, Des YACs produits à partir de librairies de gènes ont permis de créer une grande diversité de molécules.

Detectability of migrating raptors and its effect on bias and precision of trend estimates

Annual counts of migrating raptors at fixed observation points are a widespread practice, and changes in numbers counted over time, adjusted for survey effort, are commonly used as indices of trends in population size. Unmodeled year-to-year variation in detectability may introduce bias, reduce precision of trend estimates, and reduce power to detect trends. We conducted dependent double-observer surveys at the annual fall raptor migration count at Lucky Peak, Idaho, in 2009 and 2010 and applied Huggins closed-capture removal models and information-theoretic model selection to determine the relative importance of factors affecting detectability. The most parsimonious model included effects of observer team identity, distance, species, and day of the season. We then simulated 30 years of counts with heterogeneous individual detectability, a population decline (λ = 0.964), and unexplained random variation in the number of available birds. Imperfect detectability did not bias trend estimation, and increased the time required to achieve 80% power by less than 11%. Results suggested that availability is a greater source of variance in annual counts than detectability; thus, efforts to account for availability would improve the monitoring value of migration counts. According to our models, long-term trends in observer efficiency or migratory flight distance may introduce substantial bias to trend estimates. Estimating detectability with a novel count protocol like our double-observer method is just one potential means of controlling such effects. The traditional approach of modeling the effects of covariates and adjusting the index may also be effective if ancillary data is collected consistently.

Subject Ontogeny: Subject Access through Time and the Dimensionality of Classification

Classification schemes undergo revision. However, in a networked environment revisions can be used to add dimensionality to classification. This dimensionality can be used to help explain conceptual warrant, explain the shift from disciplinary to multidisciplinary knowledge production, and as a component method of domain analysis. Further, subject ontogeny might be used in cooperative networked projects like digital preservation, online access tools, and interoperability frameworks.

Tracing outsideness: young women's institutional journeys and the geographies of closed space

Understanding confinement and its complex workings between individuals and society has been the stated aim of carceral geography and wider studies on detention. This project contributes ethnographic insights from multiple sites of incarceration, working with an under-researched group within confined populations. Focussing on young female detainees in Scotland, this project seeks to understand their experiences of different types of ‘closed’ space. Secure care, prison and closed psychiatric facilities all impact on the complex geographies of these young women’s lives. The fluid but always situated relations of control and care provide the backdrop for their journeys in/out and beyond institutional spaces. Understanding institutional journeys with reference to age and gender allows an insight into the highly mobile, often precarious, and unfamiliar lives of these young women who live on the margins. This thesis employs a mixed-method qualitative approach and explores what Goffman calls the ‘tissue and fabric’ of detention as a complex multi-institutional practice. In order to be able to understand the young women’s gendered, emotional and often repetitive experiences of confinement, analysis of the constitution of ‘closed space’ represents a first step for inquiry. The underlying nature of inner regimes, rules and discipline in closed spaces, provide the background on which confinement is lived, perceived and processed. The second part of the analysis is the exploration of individual experiences ‘on the inside’, ranging from young women’s views on entering a closed institution, the ways in which they adapt or resist the regime, and how they cope with embodied aspects of detention. The third and final step considers the wider context of incarceration by recovering the young women’s journeys through different types of institutional spaces and beyond. The exploration of these journeys challenges and re-develops understandings of mobility and inertia by engaging the relative power of carceral archipelagos and the figure of femina sacra. This project sits comfortably within the field of carceral geography while also pushing at its boundaries. On a conceptual level, a re-engagement with Goffman’s micro-analysis challenges current carceral-geographic theory development. Perhaps more importantly, this project pushes for an engagement with different institutions under the umbrella of carceral geography, thus creating new dialogues on issues like ‘care’ and ‘control’. Finally, an engagement with young women addresses an under-represented population within carceral geography in ways that raise distinctly problematic concerns for academic research and penal policy. Overall, this project aims to show the value of fine grained micro-level research in institutional geographies for extending thinking and understanding about society’s responses to a group of people who live on the margins of social and legal norms.

Waiting Time Screening in Diagnostic Medical Imaging - A Case-Based View

Due to the high standards expected from diagnostic medical imaging, the analysis of information regarding waiting lists via different information systems is of utmost importance. Such analysis, on the one hand, may improve the diagnostic quality and, on the other hand, may lead to the reduction of waiting times, with the concomitant increase of the quality of services and the reduction of the inherent financial costs. Hence, the purpose of this study is to assess the waiting time in the delivery of diagnostic medical imaging services, like computed tomography and magnetic resonance imaging. Thereby, this work is focused on the development of a decision support system to assess waiting times in diagnostic medical imaging with recourse to operational data of selected attributes extracted from distinct information systems. The computational framework is built on top of a Logic Programming Case-base Reasoning approach to Knowledge Representation and Reasoning that caters for the handling of in-complete, unknown, or even self-contradictory information.

Fixing, storage time and DNA extraction applied to dT-RFLP for ecological and molecular studies of marine nematodes assemblages

The application of molecular methods offers an alternative faster than traditional methods based on morphology It is nearly impossible to process all the samples in short period using traditional methods, and the deterioration of marine sediments rapidly occurs The dT-RFLP (directed Terminal-Restriction Fragment Length Polymorphism) allows a rapid assessment of biodiversity changes of nematodes assemblages The use of a not suitable fixing, storage time and DNA extraction could be a limitation in molecular analysis like dT-RFLP and real time PCR.Objetives: the best fixative •the level of DNA degradation over the time •the best DNA extraction method for marine nematodes and suitable for dT-RFLP analysis

Real-Time Engine Diagnostic and Control Based on Acoustic Emissions Analysis

Engine developers are putting more and more emphasis on the research of maximum thermal and mechanical efficiency in the recent years. Research advances have proven the effectiveness of downsized, turbocharged and direct injection concepts, applied to gasoline combustion systems, to reduce the overall fuel consumption while respecting exhaust emissions limits. These new technologies require more complex engine control units. The sound emitted from a mechanical system encloses many information related to its operating condition and it can be used for control and diagnostic purposes. The thesis shows how the functions carried out from different and specific sensors usually present on-board, can be executed, at the same time, using only one multifunction sensor based on low-cost microphone technology. A theoretical background about sound and signal processing is provided in chapter 1. In modern turbocharged downsized GDI engines, the achievement of maximum thermal efficiency is precluded by the occurrence of knock. Knock emits an unmistakable sound perceived by the human ear like a clink. In chapter 2, the possibility of using this characteristic sound for knock control propose, starting from first experimental assessment tests, to the implementation in a real, production-type engine control unit will be shown. Chapter 3 focus is on misfire detection. Putting emphasis on the low frequency domain of the engine sound spectrum, features related to each combustion cycle of each cylinder can be identified and isolated. An innovative approach to misfire detection, which presents the advantage of not being affected by the road and driveline conditions is introduced. A preliminary study of air path leak detection techniques based on acoustic emissions analysis has been developed, and the first experimental results are shown in chapter 4. Finally, in chapter 5, an innovative detection methodology, based on engine vibration analysis, that can provide useful information about combustion phase is reported.

Study of Bone Biomineralization in 2D and 3D Cultures of a Human Osteosarcoma Cell Line As Osteoblast-like Model

This PhD project focuses on the study of the early stages of bone biomineralization in 2D and 3D cultures of osteoblast-like SaOS-2 osteosarcoma cells, exposed to an osteogenic cocktail. The efficacy of osteogenic treatment was assessed on 2D cell cultures after 7 days. A large calcium minerals production, an overexpression of osteogenic markers and of alkaline phosphatase activity occurred in treated samples. TEM microscopy and cryo-XANES micro-spectroscopy were performed for localizing and characterizing Ca-depositions. These techniques revealed a different localization and chemical composition of Ca-minerals over time and after treatment. Nevertheless, the Mito stress test showed in treated samples a significant increase in maximal respiration levels associated to an upregulation of mitochondrial biogenesis indicative of an ongoing differentiation process. The 3D cell cultures were realized using two different hydrogels: a commercial collagen type I and a mixture of agarose and lactose-modified chitosan (CTL). Both biomaterials showed good biocompatibility with SaOS-2 cells. The gene expression analysis of SaOS-2 cells on collagen scaffolds indicated an osteogenic commitment after treatment. and Alizarin red staining highlighted the presence of Ca-spots in the differentiated samples. In addition, the intracellular magnesium quantification, and the X-ray microscopy on mineral depositions, suggested the incorporation of Mg during the early stages of bone formation process., SaOS-2 cells treated with osteogenic cocktail produced Ca mineral deposits also on CTL/agarose scaffolds, as confirmed by alizarin red staining. Further studies are underway to evaluate the differentiation also at the genetic level. Thanks to the combination of conventional laboratory methods and synchrotron-based techniques, it has been demonstrated that SaOS-2 is a suitable model for the study of biomineralization in vitro. These results have contributed to a deeper knowledge of biomineralization process in osteosarcoma cells and could provide new evidences about a therapeutic strategy acting on the reversibility of tumorigenicity by osteogenic induction.

Comparison of different solutions for the hybridization of a motorcycle engine for Formula Student applications and evaluation of the effectiveness on the simulated lap time

Following the latest environmental concerns, the importance of minimising the detrimental effect of emissions of terrestrial vehicles has become a major goal for the whole automotive field. The key to achieve an emission-free long term future is the electrification of vehicle fleets; this huge step cannot be taken without intermediate technologies. In this context, hybrid vehicles are fundamental to reach this goal. Specifically, mild hybrid vehicles represent a trade-off between cost and emissions that could act now as a bridge towards electrification. Like the industry, also student engineering competitions are likely to take the same route: Combustion vehicles may well turn into hybrid vehicles. For this reason, a preliminary design overview is necessary to pinpoint the key performance indicators for the prototypes of the future.

Towards a Smart Campus Digital Twin: Promoting Awareness and Sustainability Through Wayfinding and Real-Time Environmental Data

The newly inaugurated Navile District of the University of Bologna is a complex created along the Navile canal, that now houses various teaching and research activities for the disciplines of Chemistry, Industrial Chemistry, Pharmacy, Biotechnology and Astronomy. A Building Information Modeling system (BIM) gives staff of the Navile campus several ways to monitor buildings in the complex throughout their life cycle, one of which is the ability to access real-time environmental data such as room temperature, humidity, air composition, and more, thereby simplifying operations like finding faults and optimizing environmental resource usage. But smart features at Navile are not only available to the staff: AlmaMap Navile is a web application, whose development is documented in this thesis, that powers the public touch kiosks available throughout the campus, offering maps of the district and indications on how to reach buildings and spaces. Even if these two systems, BIM and AlmaMap, don't seem to have many similarities, they share the common intent of promoting awareness for informed decision making in the campus, and they do it while relying on web standards for communication. This opens up interesting possibilities, and is the idea behind AlmaMap Navile 2.0, an app that interfaces with the BIM system and combines real-time sensor data with a comfort calculation algorithm, giving users the ability not just to ask for directions to a space, but also to see its comfort level in advance and, should they want to, check environmental measurements coming from each sensor in a granular manner. The end result is a first step towards building a smart campus Digital Twin, that can support all the people who are part of the campus life in their daily activities, improving their efficiency and satisfaction, giving them the ability to make informed decisions, and promoting awareness and sustainability.

The Value Of The 2005 International Society Of Urological Pathology (isup) Modified Gleason Grading System As A Predictor Of Biochemical Recurrence After Radical Prostatectomy.

To compare time and risk to biochemical recurrence (BR) after radical prostatectomy of two chronologically different groups of patients using the standard and the modified Gleason system (MGS). Cohort 1 comprised biopsies of 197 patients graded according to the standard Gleason system (SGS) in the period 1997/2004, and cohort 2, 176 biopsies graded according to the modified system in the period 2005/2011. Time to BR was analyzed with the Kaplan-Meier product-limit analysis and prediction of shorter time to recurrence using univariate and multivariate Cox proportional hazards model. Patients in cohort 2 reflected time-related changes: striking increase in clinical stage T1c, systematic use of extended biopsies, and lower percentage of total length of cancer in millimeter in all cores. The MGS used in cohort 2 showed fewer biopsies with Gleason score ≤ 6 and more biopsies of the intermediate Gleason score 7. Time to BR using the Kaplan-Meier curves showed statistical significance using the MGS in cohort 2, but not the SGS in cohort 1. Only the MGS predicted shorter time to BR on univariate analysis and on multivariate analysis was an independent predictor. The results favor that the 2005 International Society of Urological Pathology modified system is a refinement of the Gleason grading and valuable for contemporary clinical practice.

Minimal Invasive Ostheosintesis For Treatment Of Diaphiseal Transverse Humeral Shaft Fractures.

To evaluate patients with transverse fractures of the shaft of the humerus treated with indirect reduction and internal fixation with plate and screws through minimally invasive technique. Inclusion criteria were adult patients with transverse diaphyseal fractures of the humerus closed, isolated or not occurring within 15 days of the initial trauma. Exclusion criteria were patients with compound fractures. In two patients, proximal screw loosening occurred, however, the fractures consolidated in the same mean time as the rest of the series. Consolidation with up to 5 degrees of varus occurred in five cases and extension deficit was observed in the patient with olecranon fracture treated with tension band, which was not considered as a complication. There was no recurrence of infection or iatrogenic radial nerve injury. It can be concluded that minimally invasive osteosynthesis with bridge plate can be considered a safe and effective option for the treatment of transverse fractures of the humeral shaft. Level of Evidence III, Therapeutic Study.

Fractalkine (cx3cl1) Is Involved In The Early Activation Of Hypothalamic Inflammation In Experimental Obesity.

Hypothalamic inflammation is a common feature of experimental obesity. Dietary fats are important triggers of this process, inducing the activation of toll-like receptor-4 (TLR4) signaling and endoplasmic reticulum stress. Microglia cells, which are the cellular components of the innate immune system in the brain, are expected to play a role in the early activation of diet-induced hypothalamic inflammation. Here, we use bone marrow transplants to generate mice chimeras that express a functional TLR4 in the entire body except in bone marrow-derived cells or only in bone marrow-derived cells. We show that a functional TLR4 in bone marrow-derived cells is required for the complete expression of the diet-induced obese phenotype and for the perpetuation of inflammation in the hypothalamus. In an obesity-prone mouse strain, the chemokine CX3CL1 (fractalkine) is rapidly induced in the neurons of the hypothalamus after the introduction of a high-fat diet. The inhibition of hypothalamic fractalkine reduces diet-induced hypothalamic inflammation and the recruitment of bone marrow-derived monocytic cells to the hypothalamus; in addition, this inhibition reduces obesity and protects against diet-induced glucose intolerance. Thus, fractalkine is an important player in the early induction of diet-induced hypothalamic inflammation, and its inhibition impairs the induction of the obese and glucose intolerance phenotypes.

Growth Curves For Girls With Turner Syndrome.

The objective of this study was to review the growth curves for Turner syndrome, evaluate the methodological and statistical quality, and suggest potential growth curves for clinical practice guidelines. The search was carried out in the databases Medline and Embase. Of 1006 references identified, 15 were included. Studies constructed curves for weight, height, weight/height, body mass index, head circumference, height velocity, leg length, and sitting height. The sample ranged between 47 and 1,565 (total = 6,273) girls aged 0 to 24 y, born between 1950 and 2006. The number of measures ranged from 580 to 9,011 (total = 28,915). Most studies showed strengths such as sample size, exclusion of the use of growth hormone and androgen, and analysis of confounding variables. However, the growth curves were restricted to height, lack of information about selection bias, limited distributional properties, and smoothing aspects. In conclusion, we observe the need to construct an international growth reference for girls with Turner syndrome, in order to provide support for clinical practice guidelines.