Stage-specific embryonic antigen-4 is not a marker for chondrogenic and osteogenic potential in cultured chondrocytes and mesenchymal progenitor cells

One important challenge for regenerative medicine is to produce a clinically relevant number of cells with consistent tissue-forming potential. Isolation and expansion of cells from skeletal tissues results in a heterogeneous population of cells with variable regenerative potential. A more consistent tissue formation could be achieved by identification and selection of potent progenitors based on cell surface molecules. In this study, we assessed the expression of stage-specific embryonic antigen-4 (SSEA-4), a classic marker of undifferentiated stem cells, and other surface markers in human articular chondrocytes (hACs), osteoblasts, and bone marrow-derived mesenchymal stromal cells (bmMSCs) and characterized their differentiation potential. Further, we sorted SSEA-4-expressing hACs and followed their potential to proliferate and to form cartilage in vitro. Cells isolated from cartilage and bone exhibited remarkably heterogeneous SSEA-4 expression profiles in expansion cultures. SSEA-4 expression levels increased up to approximately 5 population doublings, but decreased following further expansion and differentiation cultures; levels were not related to the proliferation state of the cells. Although SSEA-4-sorted chondrocytes showed a slightly better chondrogenic potential than their SSEA-4-negative counterparts, differences were insufficient to establish a link between SSEA-4 expression and chondrogenic potential. SSEA-4 levels in bmMSCs also did not correlate to the cells' chondrogenic and osteogenic potential in vitro. SSEA-4 is clearly expressed by subpopulations of proliferating somatic cells with a MSC-like phenotype. However, the predictive value of SSEA-4 as a specific marker of superior differentiation capacity in progenitor cell populations from adult human tissue and even its usefulness as a stem cell marker appears questionable.

Discrepancy between antennal and behavioral responses for enantiomers of a-Pinene : electrophysiology and behavior of helicoverpa armigera (Lepidoptera)

The ability of adult cotton bollworm, Helicoverpa armigera (Hübner), to distinguish and respond to enantiomers of α-pinene was investigated with electrophysiological and behavioral methods. Electroantennogram recordings using mixtures of the enantiomers at saturating dose levels, and single unit electrophysiology, indicated that the two forms were detected by the same receptor neurons. The relative size of the electroantennogram response was higher for the (−) compared to the (+) form, indicating greater affinity for the (−) form at the level of the dendrites. Behavioral assays investigated the ability of moths to discriminate between, and respond to the (+) and (−) forms of α-pinene. Moths with no odor conditioning showed an innate preference for (+)-α-pinene. This preference displayed by naïve moths was not significantly different from the preferences of moths conditioned on (+)-α-pinene. However, we found a significant difference in preference between moths conditioned on the (−) enantiomer compared to naïve moths and moths conditioned on (+)-α-pinene, showing that learning plays an important role in the behavioral response. Moths are less able to distinguish between enantiomers of α-pinene than different odors (e.g., phenylacetaldehyde versus (−)-α-pinene) in learning experiments. The relevance of receptor discrimination of enantiomers and learning ability of the moths in host plant choice is discussed.

Landscape meaning and design : a new case study methodology

Understanding the effects of design interventions on the meanings people associate with landscapes is important to critical and ethical practice in landscape architecture. Case study research has become a common way researchers evaluate design interventions and related issues, with a standardised method promoted by the Landscape Architecture Foundation (LAF). However, the method is somewhat undeveloped for interpreting landscape meanings – something most commonly undertaken as historic landscape studies, but not as studies of design effect. This research proposes a new method for such interpretation, using a case study of Richard Haag’s radical 1971 proposal for a new kind of park on the site of the former Seattle gas works.

The Norfolk Island Eye Study (NIES) : rationale, methodology and distribution of ocular biometry (Biometry of the Bounty)

Aim: To describe the recruitment, ophthalmic examination methods and distribution of ocular biometry of participants in the Norfolk Island Eye Study, who were individuals descended from the English Bounty mutineers and their Polynesian wives. Methods: All 1,275 permanent residents of Norfolk Island aged over 15 years were invited to participate, including 602 individuals involved in a 2001 cardiovascular disease study. Participants completed a detailed questionnaire and underwent a comprehensive eye assessment including stereo disc and retinal photography, ocular coherence topography and conjunctival autofluorescence assessment. Additionally, blood or saliva was taken for DNA testing. Results: 781 participants aged over 15 years were seen (54% female), comprising 61% of the permanent Island population. 343 people (43.9%) could trace their family history to the Pitcairn Islanders (Norfolk Island Pitcairn Pedigree). Mean anterior chamber depth was 3.32mm, mean axial length (AL) was 23.5mm, and mean central corneal thickness was 546 microns. There were no statistically significant differences in these characteristics between persons with and without Pitcairn Island ancestry. Mean intra-ocular pressure was lower in people with Pitcairn Island ancestry: 15.89mmHg compared to those without Pitcairn Island ancestry 16.49mmHg (P = .007). The mean keratometry value was lower in people with Pitcairn Island ancestry (43.22 vs. 43.52, P = .007). The corneas were flatter in people of Pitcairn ancestry but there was no corresponding difference in AL or refraction. Conclusion: Our study population is highly representative of the permanent population of Norfolk Island. Ocular biometry was similar to that of other white populations. Heritability estimates, linkage analysis and genome-wide studies will further elucidate the genetic determinants of chronic ocular diseases in this genetic isolate.

Cross-sectional study of a microsatellite marker in the low densitity lipoprotein receptor gene in obese normotensives

1. The low density lipoprotein receptor is an important regulator of serum cholesterol which may have implications for the development of both hypertension and obesity. In this study, genotypes for a low density lipoprotein receptor gene (LDLR) dinucleotide polymorphism were determined in both lean and obese normotensive populations. 2. In previous cross-sectional association studies an ApaLI and a HincII polymorphism for LDLR were shown to be associated with obesity in essential hypertensives. However, these polymorphisms did not show an association with obesity in normotensives. 3. In contrast, this study reports that preliminary results for an LDLR microsatellite marker, located more towards the 3' end of the gene, show a significant association with obesity in the normotensive population studied. These results indicate that LDLR could play an important role in the development of obesity, which might be independent of hypertension.

Exploring expedition research methodology : a personal reflection

Research into the human dynamics of expeditions is a potentially rewarding and fruitful area of study. However, the complex nature of expedition work presents the researcher with numerous challenges. This paper presents a personal reflection on the challenges linked to determining appropriate methodological processes for a study into expedition teamwork. Previous expedition research is outlined and reviewed for appropriateness. Some alternative methodological theories are described and limitations highlighted. Lastly the actual data gathering and analysis processes are detailed. The aim being to show that what happened in the field inevitably dictated how methodological processes were adapted. Essentially the paper describes a personal journey into research. A journey that sparked numerous personal insights in the science of human dynamics and expeditions and one that I hope will add to the development of expedition research in general.

The 4C spectrum of fundamental behavioral relations for concurrent systems

The design of concurrent software systems, in particular process-aware information systems, involves behavioral modeling at various stages. Recently, approaches to behavioral analysis of such systems have been based on declarative abstractions defined as sets of behavioral relations. However, these relations are typically defined in an ad-hoc manner. In this paper, we address the lack of a systematic exploration of the fundamental relations that can be used to capture the behavior of concurrent systems, i.e., co-occurrence, conflict, causality, and concurrency. Besides the definition of the spectrum of behavioral relations, which we refer to as the 4C spectrum, we also show that our relations give rise to implication lattices. We further provide operationalizations of the proposed relations, starting by proposing techniques for computing relations in unlabeled systems, which are then lifted to become applicable in the context of labeled systems, i.e., systems in which state transitions have semantic annotations. Finally, we report on experimental results on efficiency of the proposed computations.

Food in an Australian primary school curriculum : a critical sociological study

Food is a multidimensional construct. It has social, cultural, economic, psychological, emotional, biological, and political dimensions. It is both a material object and a catalyst for a range of social and cultural action. Richly implicated in the social and cultural milieu, food is a central marker of culture and society. Yet little is known about the messages and knowledges in the school curriculum about food. Popular debates around food in schools are largely connected with biomedical issues of obesity, exercise and nutrition. This is a study of the sociological dimensions of food-related messages, practices and knowledge formations in the primary school curriculum. It uses an exploratory, qualitative case study methodology to identify and examine the food activities of a Year 5 class in a Queensland school. Data was gathered over a twoyear period using observation, documentation and interviews methods. Food was found to be an integral part of the primary school's activity. It had economic, symbolic, pedagogic, and instrumental value. Messages about food were found in the official, enacted and hidden curricular which were framed by a food governance framework of legislation, procedures and norms. In the school studied, food knowledge was commodified as a part of a political economy that centred on an 'eat more' message. Certain foods were privileged over others while myths about energy, fruit, fruit juice and sugar shaped student dispositions, values, norms and action. There was little engagement with the cognitive and behavioural dimensions of food and nutrition. The thesis concludes with recommendations for a whole scale reconsideration of food in schools as curricular content and knowledge.

A methodology to develop an urban transport disadvantage framework : the case of Brisbane, Australia

Most individuals travel in order to participate in a network of activities which are important for attaining a good standard of living. Because such activities are commonly widely dispersed and not located locally, regular access to a vehicle is important to avoid exclusion. However, planning transport system provisions that can engage members of society in an acceptable degree of activity participation remains a great challenge. The main challenges in most cities of the world are due to significant population growth and rapid urbanisation which produces increased demand for transport. Keeping pace with these challenges in most urban areas is difficult due to the widening gap between supply and demand for transport systems which places the urban population at a transport disadvantage. The key element in mitigating the issue of urban transport disadvantage is to accurately identify the urban transport disadvantaged. Although wide-ranging variables and multi-dimensional methods have been used to identify this group, variables are commonly selected using ad-hoc techniques and unsound methods. This poses questions of whether the current variables used are accurately linked with urban transport disadvantage, and the effectiveness of the current policies. To fill these gaps, the research conducted for this thesis develops an operational urban transport disadvantage framework (UTDAF) based on key statistical urban transport disadvantage variables to accurately identify the urban transport disadvantaged. The thesis develops a methodology based on qualitative and quantitative statistical approaches to develop an urban transport disadvantage framework designed to accurately identify urban transport disadvantage. The reliability and the applicability of the methodology developed is the prime concern rather than the accuracy of the estimations. Relevant concepts that impact on urban transport disadvantage identification and measurement and a wide range of urban transport disadvantage variables were identified through a review of the existing literature. Based on the reviews, a conceptual urban transport disadvantage framework was developed based on the causal theory. Variables identified during the literature review were selected and consolidated based on the recommendations of international and local experts during the Delphi study. Following the literature review, the conceptual urban transport disadvantage framework was statistically assessed to identify key variables. Using the statistical outputs, the key variables were weighted and aggregated to form the UTDAF. Before the variable's weights were finalised, they were adjusted based on results of correlation analysis between elements forming the framework to improve the framework's accuracy. The UTDAF was then applied to three contextual conditions to determine the framework's effectiveness in identifying urban transport disadvantage. The development of the framework is likely to be a robust application measure for policy makers to justify infrastructure investments and to generate awareness about the issue of urban transport disadvantage.

Knowledge inquiry in the construction management domain through structured research process and methodology

Construction works are project-based and interdisciplinary. Many construction management (CM) problems are ill defined. The knowledge required to address such problems is not readily available and mostly tacit in nature. Moreover, the researchers, especially the students in the higher education, often face difficulty in defining the research problem, adopting an appropriate research process and methodology for designing and validating their research. This paper describes a ‘Horseshoe’ research process approach and its application to address a research problem of extracting construction-relevant information from a building information model (BIM). It describes the different steps of the process for understanding a problem, formulating appropriate research question/s, defining different research tasks, including a methodology for developing, implementing and validating the research. It is argued that a structure research approach and the use of mixed research methods would provide a sound basis for research design and validation in order to make contribution to existing knowledge.

Behavioral evaluation of eight rat lines selected for high and low anxiety-related responses

Anxiety traits can be stable and permanent characteristics of an individual across time that is less susceptible of influences by a particular situation. One way to study trait anxiety in an experimental context is through the use of rat lines, selected according to contrasting phenotypes of fear and anxiety. It is not clear whether the behavioral differences between two contrasting rat lines in one given anxiety test are also present in others paradigms of state anxiety. Here, we examine the extent to which multiple anxiety traits generalize across selected animal lines originally selected for a single anxiety trait. We review the behavioral results available in the literature of eight rat genetic models of trait anxiety - namely Maudsley Reactive and Non-reactive rats, Floripa H and L rats, Tsukuba High and Low Emotional rats, High and Low Anxiety-related rats, High and Low Ultrasonic Vocalization rats, Roman High and Low Avoidance rats, Syracuse High and Low Avoidance rats, and Carioca High and Low Conditioned Freezing rats - across 11 behavioral paradigms of innate anxiety or aversive learning frequently used in the experimental setting. We observed both convergence and divergence of behavioral responses in these selected lines across the 11 paradigms. We find that predisposition for specific anxiety traits will usually be generalized to other anxiety provoking stimuli. However this generalization is not observed across all genetic models indicating some unique trait and state interactions. Genetic models of enhanced-anxiety related responses are beginning to help define how anxiety can manifest differently depending on the underlying traits and the current environmentally induced state.

Rodent Models of Conditioned Fear: Behavioral Measures of Fear and Memory

Pavlovian fear conditioning is a robust technique for examining behavioral and cellular components of fear learning and memory. In fear conditioning, the subject learns to associate a previously neutral stimulus with an inherently noxious co-stimulus. The learned association is reflected in the subjects' behavior upon subsequent re-exposure to the previously neutral stimulus or the training environment. Using fear conditioning, investigators can obtain a large amount of data that describe multiple aspects of learning and memory. In a single test, researchers can evaluate functional integrity in fear circuitry, which is both well characterized and highly conserved across species. Additionally, the availability of sensitive and reliable automated scoring software makes fear conditioning amenable to high-throughput experimentation in the rodent model; thus, this model of learning and memory is particularly useful for pharmacological and toxicological screening. Due to the conserved nature of fear circuitry across species, data from Pavlovian fear conditioning are highly translatable to human models. We describe equipment and techniques needed to perform and analyze conditioned fear data. We provide two examples of fear conditioning experiments, one in rats and one in mice, and the types of data that can be collected in a single experiment. © 2012 Springer Science+Business Media, LLC.

Understanding chronic pain complicating disability: Finding meaning through focus group methodology

Over the past few decades a major paradigm shift has occurred in the conceptualisation of chronic pain as a complex multidimensional phenomenon. Yet, pain experienced by individuals with a primary disability continues to be understood largely from a traditional biomedical model, despite its inherent limitations. This is reflected in the body of literature on the topic that is primarily driven by positivist assumptions and the search for etiologic pain mechanisms. Conversely, little is known about the experiences of and meanings attributed to, disability-related pain. Thus the purpose of this paper is to discuss the use of focus group methodology in elucidating the meanings and experiences of this population. Here, a distinction is made between the method of the focus group and focus group research as methodology. Typically, the focus group is presented as a seemingly atheoretical method of research. Drawing on research undertaken on the impact of chronic pain in people with multiple sclerosis, this paper seeks to theorise the focus group in arguing the methodological congruence of focus group research and the study of pain experience. It is argued that the contributions of group interaction and shared experiences in focus group discussions produce data and insights less accessible through more structured research methods. It is concluded that a biopsychosocial perspective of chronic pain may only ever be appreciated when the person-in-context is the unit of investigation.

Methodology for measuring the vertical displacements of bridges using fibre bragg grating sensors

In many bridges, vertical displacements are one of the most relevant parameters for structural health monitoring in both the short- and long-terms. Bridge managers around the globe are always looking for a simple way to measure vertical displacements of bridges. However, it is difficult to carry out such measurements. On the other hand, in recent years, with the advancement of fibre-optic technologies, fibre Bragg grating (FBG) sensors are more commonly used in structural health monitoring due to their outstanding advantages including multiplexing capability, immunity of electromagnetic interference as well as high resolution and accuracy. For these reasons, a methodology for measuring the vertical displacements of bridges using FBG sensors is proposed. The methodology includes two approaches. One of which is based on curvature measurements while the other utilises inclination measurements from successfully developed FBG tilt sensors. A series of simulation tests of a full-scale bridge was conducted. It shows that both approaches can be implemented to measure the vertical displacements for bridges with various support conditions, varying stiffness along the spans and without any prior known loading. A static loading beam test with increasing loads at the mid-span and a beam test with different loading locations were conducted to measure vertical displacements using FBG strain sensors and tilt sensors. The results show that the approaches can successfully measure vertical displacements.

Dynamic contrast-enhanced magnetic resonance imaging as a biomarker for the pharmacological response of PTK787/ZK 222584, an inhibitor of the vascular endothelial growth factor receptor tyrosine kinases, in patients with advanced colorectal cancer and liver metastases: Results from two phase I studies

Purpose: PTK787/ZK 222584 (PTK/ZK), an orally active inhibitor of vascular endothelial growth factor (VEGF) receptor tyrosine kinases, inhibits VEGF-mediated angiogenesis. The pharmacodynamic effects of PTK/ZK were evaluated by assessing changes in contrast-enhancement parameters of metastatic liver lesions using dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) in patients with advanced colorectal cancer treated in two ongoing, dose-escalating phase I studies. Patients and Methods: Twenty-six patients had DCE-MRI performed at baseline, day 2, and at the end of each 28-day cycle. Doses of oral PTK/ZK ranged from 50 to 2000 mg once daily. Tumor permeability and vascularity were assessed by calculating the bidirectional transfer constant (Ki). The percentage of baseline Ki (% of baseline Ki) at each time point was compared with pharmacokinetic and clinical end points. Results: A significant negative correlation exists between the % of baseline Ki and increase in PTK/ZK oral dose and plasma levels (P = .01 for oral dose; P = .0001 for area under the plasma concentration curve at day 2). Patients with a best response of stable disease had a significantly greater reduction in Ki at both day 2 and at the end of cycle 1 compared with progressors (mean difference in % of baseline Ki, 47%, P = .004%; and 51%, P = .006; respectively). The difference in % of baseline Ki remained statistically significant after adjusting for baseline WHO performance status. Conclusion: These findings should help to define a biologically active dose of PTK/ZK. These results suggest that DCE-MRI may be a useful biomarker for defining the pharmacological response and dose of angiogenesis inhibitiors, such as PTK/ZK, for further clinical development. © 2003 by American Society of Clinical Oncology.