Monitoring mesenchymal stem cell cultures using image processing and pattern recognition techniques

Stem cells have attracted tremendous interest in recent times due to their promise in providing innovative new treatments for a great range of currently debilitating diseases. This is due to their potential ability to regenerate and repair damaged tissue, and hence restore lost body function, in a manner beyond the body's usual healing process. Bone marrow-derived mesenchymal stem cells or bone marrow stromal cells are one type of adult stem cells that are of particular interest. Since they are derived from a living human adult donor, they do not have the ethical issues associated with the use of human embryonic stem cells. They are also able to be taken from a patient or other donors with relative ease and then grown readily in the laboratory for clinical application. Despite the attractive properties of bone marrow stromal cells, there is presently no quick and easy way to determine the quality of a sample of such cells. Presently, a sample must be grown for weeks and subject to various time-consuming assays, under the direction of an expert cell biologist, to determine whether it will be useful. Hence there is a great need for innovative new ways to assess the quality of cell cultures for research and potential clinical application. The research presented in this thesis investigates the use of computerised image processing and pattern recognition techniques to provide a quicker and simpler method for the quality assessment of bone marrow stromal cell cultures. In particular, aim of this work is to find out whether it is possible, through the use of image processing and pattern recognition techniques, to predict the growth potential of a culture of human bone marrow stromal cells at early stages, before it is readily apparent to a human observer. With the above aim in mind, a computerised system was developed to classify the quality of bone marrow stromal cell cultures based on phase contrast microscopy images. Our system was trained and tested on mixed images of both healthy and unhealthy bone marrow stromal cell samples taken from three different patients. This system, when presented with 44 previously unseen bone marrow stromal cell culture images, outperformed human experts in the ability to correctly classify healthy and unhealthy cultures. The system correctly classified the health status of an image 88% of the time compared to an average of 72% of the time for human experts. Extensive training and testing of the system on a set of 139 normal sized images and 567 smaller image tiles showed an average performance of 86% and 85% correct classifications, respectively. The contributions of this thesis include demonstrating the applicability and potential of computerised image processing and pattern recognition techniques to the task of quality assessment of bone marrow stromal cell cultures. As part of this system, an image normalisation method has been suggested and a new segmentation algorithm has been developed for locating cell regions of irregularly shaped cells in phase contrast images. Importantly, we have validated the efficacy of both the normalisation and segmentation method, by demonstrating that both methods quantitatively improve the classification performance of subsequent pattern recognition algorithms, in discriminating between cell cultures of differing health status. We have shown that the quality of a cell culture of bone marrow stromal cells may be assessed without the need to either segment individual cells or to use time-lapse imaging. Finally, we have proposed a set of features, that when extracted from the cell regions of segmented input images, can be used to train current state of the art pattern recognition systems to predict the quality of bone marrow stromal cell cultures earlier and more consistently than human experts.

Teaching porn

This article gives an account of my experiences as a student and teacher of pornography in the UK university context. From my time as a student at Glasgow University in the late 1970s, to my classes on sexual transgression at Strathclyde in the 2000s, I trace changing attitudes to the pornographic, against the background of changing political and technological environments. The article considers the pedagogy of porn against the backdrop of pro- and anti-porn feminism, the rise of gay rights, and the impact of the internet. Under these influences, and over a period of three decades, pornography was destigmatized and redefined in a variety of contexts, from the irony of lad culture to the postmodern humour of the Graham Norton Show and the pro-porn feminism of the post-Madonna era.

Mileage, car ownership, experience of punishment avoidance and the risky driving of young drivers

Objective: Young drivers are at greatest risk of injury or death from a car crash in the first six months of independent driving. In Queensland, the graduated driver licensing (GDL) program was extensively modified in July 2007 in order to reduce this risk. Increased mileage and car ownership have been found to play a role in risky driving, offences and crashes; however GDL programs typically do not consider these variables. In addition, young novice drivers’ experiences of punishment avoidance have not previously been examined. The paper explores the mileage (duration and distance), car ownership and punishment avoidance behaviour of young newly-licensed intermediate (Provisional) drivers and their relationship with risky driving, crashes and offences. Methods: Drivers (n = 1032) aged 17-19 years recruited from across Queensland for longitudinal research completed Survey 1 exploring pre-licence and Learner experiences and sociodemographic characteristics. Survey 2 explored the same variables with a subset of these drivers (n = 341) after they had completed their first six months of independent driving. Results: Most young drivers in Survey 2 reported owning a vehicle and paying attention to Police presence. Drivers who had their own car reported significantly greater mileage and more risky driving. Novices who drove more kilometres, spent more hours each week driving, or avoided actual and anticipated Police presence were more likely to report risky driving. These drivers were also more likely to report being detected by Police for a driving-related offence. The media, parents, friends and other drivers play a pivotal role in informing novices of on-road Police enforcement operations. Conclusions: GDL programs should incorporate education for the parent and novice driver regarding the increased risks associated with greater driving particularly where the novices own a vehicle. Parents should be encouraged to delay exclusive access to a vehicle for the novice driver. Parents should also consider whether their young novice will deliberately avoid Police if they tell them their location. This may reinforce not only the risky behaviour but also the young novice’s beliefs that their parents condone this behaviour.

Teen content creators : experiences of using information to learn

As access to networked digital communities increases, a growing number of teens participate in digital communities by creating and sharing a variety of content. The affordances of social media - ease of use, ubiquitous access, and communal nature - have made creating and sharing content an appealing process for teens. Teens primarily learn the practices of encountering and using information through social interaction and participation within digital communities. This article adopts the position that information literacy is the experience of using information to learn. It reports on an investigation into teens experiences in the United States, as they use information to learn how to create content and participate within the context of social media. Teens that participate in sharing art on sites such as DeiviantArt, website creation, blogging, and/or posting edited videos via YouTube and Vimeo, were interviewed. The interviews explored teens' information experiences within particular social and digital contexts. Teens discussed the information they used, how information was gathered and accessed, and explored the process of using that information to participate in the communities.

Exponential asymptotics of free surface flow due to a line source

The steady problem of free surface flow due to a submerged line source is revisited for the case in which the fluid depth is finite and there is a stagnation point on the free surface directly above the source. Both the strength of the source and the fluid speed in the far field are measured by a dimensionless parameter, the Froude number. By applying techniques in exponential asymptotics, it is shown that there is a train of periodic waves on the surface of the fluid with an amplitude which is exponentially small in the limit that the Froude number vanishes. This study clarifies that periodic waves do form for flows due to a source, contrary to a suggestion by Chapman & Vanden-Broeck (2006, J. Fluid Mech., 567, 299--326). The exponentially small nature of the waves means they appear beyond all orders of the original power series expansion; this result explains why attempts at describing these flows using a finite number of terms in an algebraic power series incorrectly predict a flat free surface in the far field.

Investigation of two Wnt signalling pathway single nucleotide polymorphisms in a breast cancer-affected Australian population

In the mammary gland, Wnt signals are strongly implicated in initial development of the mammary rudiments and in the ductal branching and alveolar morphogenesis that occurs during pregnancy. Previously, we identified two Wnt signaling pathway-implicated genes, PPP3CA and MARK4, as having a role in more aggressive and potentially metastatic breast tumors. In this study, we examined two SNPs within PPP3CA and MARK4 in an Australian case-control study population for a potential role in human breast cancers. 182 cases and 180 controls were successfully genotyped for the PPP3CA SNP (rs2850328) and 182 cases and 177 controls were successfully genotyped for the MARK4 SNP (rs2395) using High Resolution Melt (HRM) analysis. Genotypes of randomly selected samples for both SNPs were validated by dye terminator sequencing. Chi-square tests were performed to determine any significant differences in the genotype and allele frequencies between the cases and controls. Chi-square analysis showed no statistically significant difference (p > .05) for genotype frequencies between cases and controls for rs2850328 (χ2 = 1.2, p = .5476) or rs2395 (χ2 = .3, p = .8608). Similarly, no statistical difference was observed for allele frequencies for rs2850328 (χ2 = .68, p = .4108) or rs2395 (χ2 = .02, p = .893). Even though an association of the polymorphisms rs2850328 and rs2395 and breast cancer was not detected in our case-control study population, other variants within the PPP3CA and MARK4 genes may still be associated with breast cancer, as both genes are implicated with processes involved in the disease as well as their mutual partaking in the Wnt signaling pathway.

The role of prostacyclin synthase and thromboxane synthase signaling in the development and progression of cancer

Prostacyclin synthase and thromboxane synthase signaling via arachidonic acid metabolism affects a number of tumor cell survival pathways such as cell proliferation, apoptosis, tumor cell invasion and metastasis, and angiogenesis. However, the effects of these respective synthases differ considerably with respect to the pathways described. While prostacyclin synthase is generally believed to be anti-tumor, a pro-carcinogenic role for thromboxane synthase has been demonstrated in a variety of cancers. The balance of oppositely-acting COX-derived prostanoids influences many processes throughout the body, such as blood pressure regulation, clotting, and inflammation. The PGI2/TXA2 ratio is of particular interest in-vivo, with the corresponding synthases shown to be differentially regulated in a variety of disease states. Pharmacological inhibition of thromboxane synthase has been shown to significantly inhibit tumor cell growth, invasion, metastasis and angiogenesis in a range of experimental models. In direct contrast, prostacyclin synthase overexpression has been shown to be chemopreventive in a murine model of the disease, suggesting that the expression and activity of this enzyme may protect against tumor development. In this review, we discuss the aberrant expression and known functions of both prostacyclin synthase and thromboxane synthase in cancer. We discuss the effects of these enzymes on a range of tumor cell survival pathways, such as tumor cell proliferation, induction of apoptosis, invasion and metastasis, and tumor cell angiogenesis. As downstream signaling pathways of these enzymes have also been implicated in cancer states, we examine the role of downstream effectors of PGIS and TXS activity in tumor growth and progression. Finally, we discuss current therapeutic strategies aimed at targeting these enzymes for the prevention/treatment of cancer. © 2010 Elsevier B.V. All rights reserved.

A vibrational spectroscopic study of the phosphate mineral lulzacite Sr2Fe2+(Fe2+,Mg)2Al4(PO4)4(OH)10

The mineral lulzacite from Saint-Aubin des Chateaux mine, France, with theoretical formula Sr2Fe2+(Fe2+,Mg)2Al4(PO4)4(OH)10 has been studied using a combination of electron microscopy with EDX and vibrational spectroscopic techniques. Chemical analysis shows a Sr, Fe, Al phosphate with minor amounts of Ga, Ba and Mg. Raman spectroscopy identifies an intense band at 990 cm�1 with an additional band at 1011 cm�1. These bands are attributed to the PO3� 4 m1 symmetric stretching mode. The m3 antisymmetric stretching modes are observed by a large number of Raman bands. The Raman bands at 1034, 1051, 1058, 1069 and 1084 together with the Raman bands at 1098, 1116, 1133, 1155 and 1174 cm�1 are assigned to the m3 antisymmetric stretching vibrations of PO3� 4 and the HOPO2� 3 units. The observation of these multiple Raman bands in the symmetric and antisymmetric stretching region gives credence to the concept that both phosphate and hydrogen phosphate units exist in the structure of lulzacite. The series of Raman bands at 567, 582, 601, 644, 661, 673 and 687 cm�1 are assigned to the PO3� 4 m2 bending modes. The series of Raman bands at 437, 468, 478, 491, 503 cm�1 are attributed to the PO3� 4 and HOPO2� 3 m4 bending modes. No Raman bands of lulzacite which could be attributed to the hydroxyl stretching unit were observed. Infrared bands at 3511 and 3359 cm�1 are ascribed to the OH stretching vibration of the OH units. Very broad bands at 3022 and 3299 cm�1 are attributed to the OH stretching vibrations of water. Vibrational spectroscopy offers insights into the molecular structure of the phosphate mineral lulzacite.

The molecular structure of the phosphate mineral beraunite Fe2+Fe⅗+(PO4)4(OH)5⋅4H2O– A vibrational spectroscopic study

The mineral beraunite from Boca Rica pegmatite in Minas Gerais with theoretical formula Fe2+Fe5 3+(PO4)4(OH)5⋅4H2O has been studied using a combination of electron microscopy with EDX and vibrational spectroscopic techniques. Raman spectroscopy identifies an intense band at 990 cm-1 and 1011 cm-1. These bands are attributed to the PO4 3- v, symmetric stretching mode. The m3 antisymmetric stretching modes are observed by a large number of Raman bands. The Raman bands at 1034, 1051, 1058, 1069 and 1084 together with the Raman bands at 1098, 1116, 1133, 1155 and 1174 cm-1 are assigned to the m3 antisymmetric stretching vibrations of PO4 3- and the HOPO3 2- units. The observation of these multiple Raman bands in the symmetric and antisymmetric stretching region gives credence to the concept that both phosphate and hydrogen phosphate units exist in the structure of beraunite. The series of Raman bands at 567, 582,601, 644, 661, 673, and 687 cm-1 are assigned to the PO4 3- v2 bending modes. The series of Raman bands at 437, 468, 478, 491, 503 cm-1 are attributed to the PO4 3- and OPO3 2- v4 bending modes. No Raman bands of beraunite which could be attributed to the hydroxyl stretching unit were observed. Infrared bands at 3511 and 3359 cm-1 are ascribed to the OH stretching vibration of the OH units. Very broad bands at 3022 and 3299 cm-1 are attributed to the OH stretching vibrations of water. Vibrational spectroscopy offers insights into the molecular structure of the phosphate mineral beraunite.

Faking it : how to kill a business through astroturfing on social media

There is no doubt about it the practice of astroturfing is lazy, misleading and potentially illegal public relations (PR). But on social media, astroturfing is not just lazy and misleading, it can be irresponsible and damaging.

The SOS screen in Arabidopsis: a search for functions involved in DNA metabolism

The SOS screen, as originally described by Perkins et al. (1999), was setup with the aim of identifying Arabidopsis functions that might potentially be involved in the DNA metabolism. Such functions, when expressed in bacteria, are prone to disturb replication and thus trigger the SOS response. Consistently, expression of AtRAD51 and AtDMC1 induced the SOS response in bacteria, even affecting E. coli viability. 100 SOS-inducing cDNAs were isolated from a cDNA library constructed from an Arabidopsis cell suspension that was found to highly express meiotic genes. A large proportion of these SOS+ candidates are clearly related to the DNA metabolism, others could be involved in the RNA metabolism, while the remaining cDNAs encode either totally unknown proteins or proteins that were considered as irrelevant. Seven SOS+ candidate genes are induced following gamma irradiation. The in planta function of several of the SOS-inducing clones was investigated using T-DNA insertional mutants or RNA interference. Only one SOS+ candidate, among those examined, exhibited a defined phenotype: silenced plants for DUT1 were sensitive to 5-fluoro-uracil (5FU), as is the case of the leaky dut-1 mutant in E. coli that are affected in dUTPase activity. dUTPase is essential to prevent uracil incorporation in the course of DNA replication.

Sense of home, sense of place : a morpho-typological comparative study of Florida and Queensland

Cities in the 21st century have become layered and complex systems not only in terms of physical form, but also social and cultural structure. Consolidated tools to analyze the urban environment have today to be improved including a strong interdisciplinary perspective in order to understand and manage the unprecedented complexity our cities are facing. Redevelopments, new estates, internal and external migrations are all dynamics which are deeply modifying the built environment directly or indirectly also affecting local identity, culture and social structure. This paper investigates the relationship between urban form and social behaviors, with particular attention to the perception of the built environment and its use by long term residents, recent migrants as well as tourists. A comparative study is suggested between South East Queensland and Florida; this two regions share common features such as subtropical climate, similar lifestyle, leisure cities and canal estates. Neighborhoods on the Gold and Sunshine Coasts have been designed using the communities of Florida, such as Celebration or Seaside, as models. These regions share also significant migration processes, similar social problems and high crime rates, which directly affect the local economies. Comparing Florida and SEQ could provide an understanding of different strategies adopted and how urban development and lifestyle can be managed maintaining social equity and security. This study, investigates people’s perception of built form and how this affects the use of public space. The relationship between built environment and social behaviour has been previously investigated, for example by environmental psychology; the innovation proposed by this research is to study the perception of place in leisure cities at multiple levels. Locals, migrants and tourists have different understanding of the built form in the same location; this understanding affects the use of space and the attitude to visit or avoid some precincts. The research methodology integrates traditional morpho-typological investigations with qualitative methods; data are collected in the first phase through online surveys about perception of urban forms. Findings guide then the selection of neighbourhoods to be investigated in detail through questionnaires and Nolli maps, specifying morphological regions as well as recurrent building typologies. A final phase includes interviews with selected stakeholders. Major urban projects are discussed addressing how they are used and perceived by locals, migrants or tourists; the comparison between SEQ and Florida allows the identification of strategies to address migration issues in both regions with particular attention to urban form and placemaking dynamics.