A system for studying epithelial-stromal interactions reveals distinct inductive abilities of stromal cells from benign prostatic hyperplasia and prostate cancer

The development of normal and abnormal glandular structures in the prostate is controlled at the endocrine and paracrine levels by reciprocal interactions between epithelium and stroma. To study these processes it is useful to have an efficient method of tissue acquisition for reproducible isolation of cells from defined histologies. Here we assessed the utility of a standardized system for acquisition and growth of prostatic cells from different regions of the prostate with different pathologies, and we compared the abilities of stromal cells from normal peripheral zone (PZ-S), benign prostatic hyperplasia (BPH-S), and cancer (CA-S) to induce the growth of a human prostatic epithelial cell line (BPH-1) in vivo. Using the tissue recombination method, we showed that grafting stromal cells (from any histology) alone, or BPH-1 epithelial cells alone produced no visible grafts. Recombining PZ-S with BPH-1 cells also produced no visible grafts (n = 15). Recombining BPH-S with BPH-1 cells generated small, well-organized and sharply demarcated grafts approximately 3-4 mm in diameter (n = 9), demonstrating a moderate inductive ability of BPH-S. Recombining CA-S with BPH-1 cells generated highly disorganized grafts that completely surrounded the host kidney and invaded into adjacent renal tissue, demonstrating induction of an aggressive phenotype. We conclude that acquisition of tissue from toluidine blue dye stained specimens is an efficient method to generate high quality epithelial and/or stromal cultures. Stromal cells derived by this method from areas of BPH and cancer induce epithelial cell growth in vivo which mimics the natural history of these diseases.

Studying the Human Gut Microbiota in the Trans-Omics Era - Focus on Metagenomics and Metabonomics

The human gut microbiota comprises a diverse microbial consortium closely co-evolved with the human genome and diet. The importance of the gut microbiota in regulating human health and disease has however been largely overlooked due to the inaccessibility of the intestinal habitat, the complexity of the gut microbiota itself and the fact that many of its members resist cultivation and are in fact new to science. However, with the emergence of 16S rRNA molecular tools and "post-genomics" high resolution technologies for examining microorganisms as they occur in nature without the need for prior laboratory culture, this limited view of the gut microbiota is rapidly changing. This review will discuss the application of molecular microbiological tools to study the human gut microbiota in a culture independent manner. Genomics or metagenomics approaches have a tremendous capability to generate compositional data and to measure the metabolic potential encoded by the combined genomes of the gut microbiota. Another post-genomics approach, metabonomics, has the capacity to measure the metabolic kinetic or flux of metabolites through an ecosystem at a particular point in time or over a time course. Metabonomics thus derives data on the function of the gut microbiota in situ and how it responds to different environmental stimuli e.g. substrates like prebiotics, antibiotics and other drugs and in response to disease. Recently these two culture independent, high resolution approaches have been combined into a single "transgenomic" approach which allows correlation of changes in metabolite profiles within human biofluids with microbiota compositional metagenomic data. Such approaches are providing novel insight into the composition, function and evolution of our gut microbiota.

Methodology for studying postprandial lipid metabolism

Background: Postprandial lipid metabolism in humans has deserved much attention during the last two decades. Although fasting lipid and lipoprotein parameters reflect body homeostasis to some extent, the transient lipid and lipoprotein accumulation that occurs in the circulation after a fat-containing meal highlights the individual capacity to handle an acute fat input. An exacerbated postprandial accumulation of triglyceride-rich lipoproteins in the circulation has been associated with an increased cardiovascular risk. Methods: The important number of studies published in this field raises the question of the methodology used for such postprandial studies, as reviewed. Results: Based on our experiences, the present review reports and discuss the numerous methodological issues involved to serve as a basis for further works. These aspects include aims of the postprandial tests, size and nutrient composition of the test meals and background diets, pre-test conditions, characteristics of subjects involved, timing of sampling, suitable markers of postprandial lipid metabolism and calculations. Conclusion: In conclusion, we stress the need for standardization of postprandial tests.

Studying oil absorption in restructured potato chips

The oil-absorption capacity of different restructured potato chips during deep-fat frying was investigated. Low-leach potato flake was chosen as the major ingredient, whereas native and pregelatinized potato starches were studied as complementary ingredients. Results showed that off absorption increased significantly when reducing product thickness in all products. Interestingly, it was found that the product containing native potato starch as an ingredient picked up the lowest amount of on when sheeted into a thick chip, whereas it absorbed the largest amount of oil when sheeted into a thin chip. Those findings were mainly attributed to crust microstructure development as revealed by electron microscopy and confocal microscopy.

Studying reactive processes with classical dynamics: rebinding dynamics in MbNO

A new surface-crossing algorithm suitable for describing bond-breaking and bond-forming processes in molecular dynamics simulations is presented. The method is formulated for two intersecting potential energy manifolds which dissociate to different adiabatic states. During simulations, crossings are detected by monitoring an energy criterion. If fulfilled, the two manifolds are mixed over a finite number of time steps, after which the system is propagated on the second adiabat and the crossing is carried out with probability one. The algorithm is extensively tested (almost 0.5 mu s of total simulation time) for the rebinding of NO to myoglobin. The unbound surface ((FeNO)-N-...) is represented using a standard force field, whereas the bound surface (Fe-NO) is described by an ab initio potential energy surface. The rebinding is found to be nonexponential in time, in agreement with experimental studies, and can be described using two time constants. Depending on the asymptotic energy separation between the manifolds, the short rebinding timescale is between 1 and 9 ps, whereas the longer timescale is about an order of magnitude larger. NO molecules which do not rebind within 1 ns are typically found in the Xenon-4 pocket, indicating the high affinity of NO to this region in the protein.

Application of molecular biological methods for studying probiotics and the gut flora

Increasingly, the microbiological scientific community is relying on molecular biology to define the complexity of the gut flora and to distinguish one organism from the next. This is particularly pertinent in the field of probiotics, and probiotic therapy, where identifying probiotics from the commensal flora is often warranted. Current techniques, including genetic fingerprinting, gene sequencing, oligonucleotide probes and specific primer selection, discriminate closely related bacteria with varying degrees of success. Additional molecular methods being employed to determine the constituents of complex microbiota in this area of research are community analysis, denaturing gradient gel electrophoresis (DGGE)/temperature gradient gel electrophoresis (TGGE), fluorescent in situ hybridisation (FISH) and probe grids. Certain approaches enable specific aetiological agents to be monitored, whereas others allow the effects of dietary intervention on bacterial populations to be studied. Other approaches demonstrate diversity, but may not always enable quantification of the population. At the heart of current molecular methods is sequence information gathered from culturable organisms. However, the diversity and novelty identified when applying these methods to the gut microflora demonstrates how little is known about this ecosystem. Of greater concern is the inherent bias associated with some molecular methods. As we understand more of the complexity and dynamics of this diverse microbiota we will be in a position to develop more robust molecular-based technologies to examine it. In addition to identification of the microbiota and discrimination of probiotic strains from commensal organisms, the future of molecular biology in the field of probiotics and the gut flora will, no doubt, stretch to investigations of functionality and activity of the microflora, and/or specific fractions. The quest will be to demonstrate the roles of probiotic strains in vivo and not simply their presence or absence.

Studying the effect of systemisation on design activity

It is considered that systemisation, the use of standard systems of components and design solutions, has an effect on the activities of the designer. This paper draws on many areas of knowledge; design movements, economic theory, quality evaluation and organisation theory to substantiate the view that systemisation will reduce the designer's discretion at the point of design. A methodology to test this hypothesis is described which will be of use to other researchers studying the social processes of construction organisations.

Consumer robotic products: studying the factors that influence market adoption

The aim of this article is to identify the key factors that are associated with the adoption of a commercial robot in the home. This article is based on the development of the robot product Cybot by the University of Reading in conjunction with a publisher (Eaglemoss International Ltd.). The robots were distributed through a new part-work magazine series (Ultimate Real Robots) that had long-term customer usage and retention. A part-work is a serial publication that is issued periodically (e.g., every two weeks), usually in magazine format, and builds into a complete collection. This magazine focused on robotics and was accompanied by cover-mounted component parts that could be assembled, with instructions, by the user to build a working robot over the series. In total, the product contributed over half a million operational domestic robots to the world market, selling over 20 million robot part-work magazines across 18 countries, thereby providing a unique breadth of insight. Gaining a better understanding of the overall attitudes that customers of this product had toward robots in the home, their perception of what such devices could deliver and how they would wish to interact with them should provide results applicable to the domestic appliance, assistance/care, entertainment, and educational markets.

Higher education as a portfolio investment: students’ choices about studying, term time employment, leisure, and loans

Recent UK changes in the number of students entering higher education, and in the nature of financial support, highlight the complexity of students’ choices about human capital investments. Today’s students have to focus not on the relatively narrow issue of how much academic effort to invest, but instead on the more complicated issue of how to invest effort in pursuit of ‘employability skills’, and how to signal such acquisitions in the context of a highly competitive graduate jobs market. We propose a framework aimed specifically at students’ investment decisions, which encompasses corner solutions for both borrowing and employment while studying.

Exposure to fine particulate matter in ten night clubs in Athens Greece: studying the effect of ventilation, cigarette smoking and resuspension

The aim of the present work is to study the occupants' exposure to fine particulate concentrations in ten nightclubs (NCs) in Athens, Greece. Measurements of PM1 and PM 2.5 were made in the outdoor and indoor environment of each NC. The average indoorPM1 andPM 2.5 concentrations were found to be 181.77 μgm−3 and 454.08 μg m−3 respectively, while the corresponding outdoor values were 11.04 μg m−3 and 32.19 μg m−3. Ventilation and resuspension rates were estimated through consecutive numerical experiments with an indoor air quality model and were found to be remarkably lower than the minimum values recommended by national standards. The relative effects of the ventilation and smoking on the occupants' exposures were examined using multiple regression techniques. Itwas found that given the low ventilation rates, the effect of smoking as well as the occupancy is of the highest importance. Numerical evaluations showed that if the ventilation rates were at the minimum values set by national standards, then the indoor exposures would be reduced at the 70% of the present exposure values.

Studying the impact of ubiquitous monitoring technology on office worker behaviours: the value of sharing research data

Pervasive computing is a continually, and rapidly, growing field, although still remains in relative infancy. The possible applications for the technology are numerous, and stand to fundamentally change the way users interact with technology. However, alongside these are equally numerous potential undesirable effects and risks. The lack of empirical naturalistic data in the real world makes studying the true impacts of this technology difficult. This paper describes how two independent research projects shared such valuable empirical data on the relationship between pervasive technologies and users. Each project had different aims and adopted different methods, but successfully used the same data and arrived at the same conclusions. This paper demonstrates the benefit of sharing research data in multidisciplinary pervasive computing research where real world implementations are not widely available.

An in-vitro model for studying the interaction of Escherichia coli O157:H7 and other enteropathogens with bovine primary cell cultures

Sections of kidney, trachea, ileum, colon, rectum and rumen were removed at post mortem from a neonatal calf and, with the exception of the rumen, primary cell lines were established for each of the cell types. The adherence of enterohaemorrhagic Escherichia coli (EHEC) serotype O157:H7, enteropathogenic E. coli (EPEC) serotype O111, E. coli K12 (a laboratory adapted non-pathogenic strain) and Salmonella enterica serotype Typhimurium was assayed on each cell type. For all adherence assays on all cell lines, EHEC O157:H7 adhered to a significantly greater extent than the other bacteria. S. Typhimurium and EPEC O111 adhered to a similar extent to one another, whereas E. coli K12 was significantly less adherent by 100-fold. In all cell types, > 10% of adherent S. Typhimurium bacteria invaded, whereas c. 0.01-0.1% of adherent EHEC O157:H7 and EPEC O111 bacteria invaded, although they are regarded as non-invasive. EHEC O157 generated actin re-arrangements in all cell types as demonstrated by fluorescent actin staining (FAS) under densely packed bacterial micro-colonies. EPEC O111 readily generated the localised adherent phenotype on bovine cells but generated only densely packed micro-colonies on HEp-2 cells. The intensity of actin re-arrangements induced in bovine cells by EPEC O111 was less than that induced by EHEC O157:H7. The intimate attachment on all cell types by both EHEC O157:H7 and EPEC O111 was clearly demonstrated by scanning electron microscopy.

Studying economics reduces overexploitation in a common resource experiment

This paper studies the economic behavior of agents, who make decisions regarding the sustainability of Common-Pool Resources (CPR). For this purpose, economic experiments are applied to simulate the yield of a CPR, taking into account the influence of economics training on the learning process of individuals, regarding their decisions for sustainability. Based on a non-cooperative game with simultaneous choices, the results of experiments show that after several rounds the existence of economics knowledge reflects a better learning process for making decisions regarding sustainability of CPR.