647 resultados para Embryo Culture Techniques


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The objective of this chapter is to provide an overview of traffic data collection that can and should be used for the calibration and validation of traffic simulation models. There are big differences in availability of data from different sources. Some types of data such as loop detector data are widely available and used. Some can be measured with additional effort, for example, travel time data from GPS probe vehicles. Some types such as trajectory data are available only in rare situations such as research projects.

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The routine cultivation of human corneal endothelial cells, with the view to treating patients with endothelial dysfunction, remains a challenging task. While progress in this field has been buoyed by the proposed existence of progenitor cells for the corneal endothelium at the corneal limbus, strategies for exploiting this concept remain unclear. In the course of evaluating methods for growing corneal endothelial cells, we have noted a case where remarkable growth was achieved using a serial explant culture technique. Over the course of 7 months, a single explant of corneal endothelium, acquired from cadaveric human tissue, was sequentially seeded into 7 culture plates and on each occasion produced a confluent cell monolayer. Sample cultures were confirmed as endothelial in origin by positive staining for glypican-4. On each occasion, small cells, closest to the tissue explant, developed into a highly compact layer with an almost homogenous structure. This layer was resistant to removal with trypsin and produced continuous cell outgrowth during multiple culture periods. The small cells gave rise to larger cells with phase-bright cell boundaries and prominent immunostaining for both nestin and telomerase. Nestin and telomerase were also strongly expressed in small cells immediately adjacent to the wound site, following transfer of the explant to another culture plate. These findings are consistent with the theory that progenitor cells for the corneal endothelium reside within the limbus and provide new insights into expected expression patterns for nestin and telomerase within the differentiation pathway.

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For the renewable energy sources whose outputs vary continuously, a Z-source current-type inverter has been proposed as a possible buck-boost alternative for grid-interfacing. With a unique X-shaped LC network connected between its dc power source and inverter topology, Z-source current-type inverter is however expected to suffer from compounded resonant complications in addition to those associated with its second-order output filter. To improve its damping performance, this paper proposes the careful integration of Posicast or three-step compensators before the inverter pulse-width modulator for damping triggered resonant oscillations. In total, two compensators are needed for wave-shaping the inverter boost factor and modulation ratio, and they can conveniently be implemented using first-in first-out stacks and embedded timers of modern digital signal processors widely used in motion control applications. Both techniques are found to damp resonance of ac filter well, but for cases of transiting from current-buck to boost state, three-step technique is less effective due to the sudden intermediate discharging interval introduced by its non-monotonic stepping (unlike the monotonic stepping of Posicast damping). These findings have been confirmed both in simulations and experiments using an implemented laboratory prototype.

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Molecular interactions that underlie pathophysiological states are being elucidated using techniques that profile proteomicend points in cellular systems. Within the field of cancer research, protein interaction networks play pivotal roles in the establishment and maintenance of the hallmarks of malignancy, including cell division, invasion, and migration. Multiple complementary tools enable a multifaceted view of how signal protein pathway alterations contribute to pathophysiological states.One pivotal technique is signal pathway profiling of patient tissue specimens. This microanalysis technology provides a proteomic snapshot at one point in time of cells directly procured from the native context of a tumor micro environment. To study the adaptive patterns of signal pathway events over time, before and after experimental therapy, it is necessary to obtain biopsies from patients before, during, and after therapy. A complementary approach is the profiling of cultured cell lines with and without treatment. Cultured cell models provide the opportunity to study short-term signal changes occurring over minutes to hours. Through this type of system, the effects of particular pharmacological agents may be used to test the effects of signal pathway inhibition or activation on multiple endpoints within a pathway. The complexity of the data generated has necessitated the development of mathematical models for optimal interpretation of interrelated signaling pathways. In combination,clinical proteomic biopsy profiling, tissue culture proteomic profiling, and mathematical modeling synergistically enable a deeper understanding of how protein associations lead to disease states and present new insights into the design of therapeutic regimens.

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This is a case study of a young university striving to generate and sustain a vibrant Research Training culture. The university’s research training framework is informed by a belief in a project management approach to achieving successful research candidature. This has led to the definition and reporting of key milestones during candidature. In turn, these milestones have generated a range of training programs to support Higher Degree Research (HDR) students to meet these milestones in a timely fashion. Each milestone focuses on a specific set of skills blended with supporting the development of different parts of the doctoral thesis. Data on student progress and completion has provided evidence in highlighting the role that the milestones and training are playing in supporting timely completion. A university-wide reporting cycle generated data on the range of workshops and training provided to Higher Degree Research students and supervisors. The report provided details of thesis topic and format, as well as participation in research training events and participant evaluation of those events. Analysis of the data led to recommendations and comments on the strengths and weaknesses of the current research training program. Discussion considered strategies and drivers for enhancements into the future. In particular, the paper reflects on the significant potential role of centrally curated knowledge systems to support HDR student and supervisor access, and engagement and success. The research training program was developed using blended learning as a model. It covered face-to-face workshops as well as online modules. These were supplemented by web portals that offered a range of services to inform and educate students and supervisors and included opportunities for students to interact with each other. Topics ranged from the research life cycle, writing and publication, ethics, managing research data, managing copyright, and project management to use of software and the University’s Code of Conduct for Research. The challenges discussed included: How to reach off campus students and those studying in external modes? How best to promote events to potential participants? How long and what format is best for face-to-face sessions? What online resources best supplement face-to-face offerings? Is there a place for peer-based learning and what form should this take? These questions are raised by a relatively young university seeking to build and sustain a vibrant research culture. The rapid growth in enrolments in recent years has challenged previous one-to-one models of support. This review of research training is timely in seeking strategies to address changing research training support capacity and student needs. Part of the discussion will focus on supervisory training, noting that good supervision is the one remaining place where one-to-one support is provided. Ensuring that supervisors are appropriately equipped to address student expectations is considered in the context of the research training provisions. The paper concludes with reflection on the challenges faced, and recommended ways forward as the number of research students grows into the future.

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Increasingly, the library and information science (LIS) practitioner is being challenged to incorporate research into the context of their professional work. This paper reports on the Researcher-Librarian Partnership, a research-mentoring program that was initiated by the International Federation of the Library Association and Institutions. Six new LIS practitioners within their first seven years of professional practice took part in the program. Each was partnered with an experienced LIS researcher who provided mentoring and support. During the 12-month program the new professionals designed and implemented a research project on a topic of interest. This paper outlines the details of the program providing observations on how research mentoring can be a powerful way to ensure all stakeholders – practitioners, educators and professional associations – can plan an active role in supporting the development of a research culture within the profession.

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This paper proposes a combination of source-normalized weighted linear discriminant analysis (SN-WLDA) and short utterance variance (SUV) PLDA modelling to improve the short utterance PLDA speaker verification. As short-length utterance i-vectors vary with the speaker, session variations and phonetic content of the utterance (utterance variation), a combined approach of SN-WLDA projection and SUV PLDA modelling is used to compensate the session and utterance variations. Experimental studies have found that a combination of SN-WLDA and SUV PLDA modelling approach shows an improvement over baseline system (WCCN[LDA]-projected Gaussian PLDA (GPLDA)) as this approach effectively compensates the session and utterance variations.

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The foliage of a plant performs vital functions. As such, leaf models are required to be developed for modelling the plant architecture from a set of scattered data captured using a scanning device. The leaf model can be used for purely visual purposes or as part of a further model, such as a fluid movement model or biological process. For these reasons, an accurate mathematical representation of the surface and boundary is required. This paper compares three approaches for fitting a continuously differentiable surface through a set of scanned data points from a leaf surface, with a technique already used for reconstructing leaf surfaces. The techniques which will be considered are discrete smoothing D2-splines [R. Arcangeli, M. C. Lopez de Silanes, and J. J. Torrens, Multidimensional Minimising Splines, Springer, 2004.], the thin plate spline finite element smoother [S. Roberts, M. Hegland, and I. Altas, Approximation of a Thin Plate Spline Smoother using Continuous Piecewise Polynomial Functions, SIAM, 1 (2003), pp. 208--234] and the radial basis function Clough-Tocher method [M. Oqielat, I. Turner, and J. Belward, A hybrid Clough-Tocher method for surface fitting with application to leaf data., Appl. Math. Modelling, 33 (2009), pp. 2582-2595]. Numerical results show that discrete smoothing D2-splines produce reconstructed leaf surfaces which better represent the original physical leaf.

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Background Obtaining single parasite clones is required for many techniques in malaria research. Cloning by limiting dilution using microscopy-based assessment for parasite growth is an arduous and labor-intensive process. An alternative method for the detection of parasite growth in limiting dilution assays is using a commercial ELISA histidine-rich protein II (HRP2) detection kit. Methods Detection of parasite growth was undertaken using HRP2 ELISA and compared to thick film microscopy. An HRP2 protein standard was used to determine the detection threshold of the HRP2 ELISA assay, and a HRP2 release model was used to extrapolate the amount of parasite growth required for a positive result. Results The HRP2 ELISA was more sensitive than microscopy for detecting parasite growth. The minimum level of HRP2 protein detection of the ELISA was 0.11ng/ml. Modeling of HRP2 release determined that 2,116 parasites are required to complete a full erythrocytic cycle to produce sufficient HRP2 to be detected by the ELISA. Under standard culture conditions this number of parasites is likely to be reached between 8 to 14 days of culture. Conclusions This method provides an accurate and simple way for the detection of parasite growth in limiting dilution assays, reducing time and resources required in traditional methods. Furthermore the method uses spent culture media instead of the parasite-infected red blood cells, enabling culture to continue.

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Purpose Corneal confocal microscopy (CCM) is a rapid non-invasive ophthalmic technique, which has been shown to diagnose and stratify the severity of diabetic neuropathy. Current morphometric techniques assess individual static images of the subbasal nerve plexus; this work explores the potential for non-invasive assessment of the wide-field morphology and dynamic changes of this plexus in vivo. Methods In this pilot study, laser scanning CCM was used to acquire maps (using a dynamic fixation target and semi-automated tiling software) of the central corneal sub-basal nerve plexus in 4 diabetic patients with and 6 without neuropathy and in 2 control subjects. Nerve migration was measured in an additional 7 diabetic patients with neuropathy, 4 without neuropathy and in 2 control subjects by repeating a modified version of the mapping procedure within 2-8 weeks, thus facilitating re-identification of distinctive nerve landmarks in the 2 montages. The rate of nerve movement was determined from these data and normalised to a weekly rate (µm/week), using customised software. Results Wide-field corneal nerve fibre length correlated significantly with the Neuropathy Disability Score (r = -0.58, p < 0.05), vibration perception (r = -0.66, p < 0.05) and peroneal conduction velocity (r = 0.67, p < 0.05). Central corneal nerve fibre length did not correlate with any of these measures of neuropathy (p > 0.05 for all). The rate of corneal nerve migration was 14.3 ± 1.1 µm/week in diabetic patients with neuropathy, 19.7 ± 13.3µm/week in diabetic patients without neuropathy, and 24.4 ± 9.8µm/week in control subjects; however, these differences were not significantly different (p = 0.543). Conclusions Our data demonstrate that it is possible to capture wide-field images of the corneal nerve plexus, and to quantify the rate of corneal nerve migration by repeating this procedure over a number of weeks. Further studies on larger sample sizes are required to determine the utility of this approach for the diagnosis and monitoring of diabetic neuropathy.

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The Weberian sense of work and life suggests that working is something around which the rest of life flows. Moreover, work life and domestic life have been defined as separate for most people based on physical structures. That is, being physically in a building at work limited your ability to interact with those who are not nearby – not part of work. As such, social conventions regarding the uses of media at work have become part of our cultural sensibilities – we “know” it is not proper to have romantic discourse over the office phone, much less romance during work! Doing so becomes news. Yet, despite the construction of such distinctions, these workspaces and places have always been difficult to render as such. For example, one might consider the relatively recent development of teleworking from the 1980s or the “putting out system”[1] which dates back to the 1400s – both requiring work in the home. The papers in this special issue draw our attention to some of the ethical issues raised by the growing pervasiveness of information and communications technologies (ICTs) in our everyday lives and the fact that it is becoming increasingly difficult to make distinctions between being somewhere (like work) and being away from some things (like one’s friends, social interests and other parts of life that are not integrated into this space or place [2] )...

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Grading is basic to the work of Landscape Architects concerned with design on the land. Gradients conducive to easy use, rainwater drained away, and land slope contributing to functional and aesthetic use are all essential to the amenity and pleasure of external environments. This workbook has been prepared specifically to support the program of landscape construction for students in Landscape Architecture. It is concerned primarily with the technical design of grading rather than with its aesthetic design. It must be stressed that the two aspects are rarely separate; what is designed should be technically correct and aesthetically pleasing - it needs to look good as well as to function effectively. This revised edition contains amended and new content which has evolved out of student classes and discussion with colleagues. I am pleased to have on record that every delivery of this workbook material has resulted in my own better understanding of grading and the techniques for its calculation and communication.

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Joy Fear and Poetry is an original performance work written, designed and directed by Natasha Budd in collaboration with 15 performers aged 7-12 years. It was performed in Brisbane as part of La Boite Theatre’s 2013 Indie Season. The production employs contemporary performance, postdramatic and constructivist methodologies to make an intervention into habituated patterns of positioning children in society. It embodies a model of practice that moves beyond participant empowerment toward a more nuanced process of co-artists creating intersubjective ‘composite texts’ (McCall 2011) for mainstream audiences. Joy Fear and Poetry experiments with techniques for performance making that create conditions conducive to authentic theatre making with children. These focus on dramaturgical, directorial and design strategies harnessed to maintain the performers’ focus, motivation and cognitive engagement within a reflexive, collaborative process.

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The Third Sector is experiencing unprecedented change as nonprofit organisations pursue their mission, organisational excellence and sustainability. This research collected data from employees of Australian nonprofit human service organisations. Instruments to measure organisational culture and change readiness were validated. The relationships between perceptions of organisational cultures, change readiness, job satisfaction and intentions to leave were explored. Findings indicate flexible organisational cultures influence employee attitudes more positively than control cultures. It identified that leaders have a significant role to play in developing and fostering change readiness and job satisfaction. This improves employees' adaptation to change, increases staff retention and organisational sustainability.