A new monoclonal antibody, P2A8(6), that specifically recognizes a novel epitope on the multidrug resistance-associated protein 1 (MRP1), but not on MRP2 nor MRP3

Multidrug resistance (NIDR) is a major problem in the chemotherapeutic treatment of cancer. Overexpression of the multidrug resistance-associated protein 1 (MRP1), is associated with NIDR in certain tumors. A number of MRP1-specific MAbs, which facilitate both clinical and experimental investigations of this protein, are available. To add to this panel of existing antibodies, we have now generated an additional MRP1-specific monoclonal antibody (MAb), P2A8(6), which detects a unique heat stable epitope on the MRP1 molecule. Female Wistar rats were immunized via footpad injections with a combination of two short synthetic peptides corresponding to amino acids 235-246 (peptide A) and 246-260 (peptide B) of the MRP1 protein. Immune reactive B cells were then isolated from the popliteal lymph nodes for fusion with SP2/O-Ag14 myeloma cells. Resultant hybridoma supernatants were screened for MRP1-specific antibody production. Antibody P2A8(6) was characterized by Western blotting and immunocytochemistry on paired multidrug resistant (MRP1 overexpressing) and sensitive parental cell lines. The antibody detects a protein of 190 kDa in MRP1-expressing cell lines but not in MRP2- or MRP3-transfected cell lines. P2A8(6) stains drug-selected and MRP1-transfected cell lines homogeneously by immunocytochemistry and recognizes MRP1 by immunohistochemistry on formalin-fixed paraffin wax-embedded tissue sections. Peptide inhibition studies confirm that P2AS(6) reacts with peptide B (amino acids 246-260), therefore recognizing a different epitope from that of all currently available MRP1 MAbs. This new MAb, chosen for its specificity to the MRP1 protein, may be a useful addition to the currently available range of MRP1-specific MAbs.

Characterization of a ligand-attenuated binding site on glycoprotein IIb/IIIa

We describe an epitope on the platelet integrin, GPIIb/IIIa, identified by the monoclonal antibody, 4F8, which is attenuated by small-molecule GPIIb/IIIa ligands. 4F8 did not bind to the ligand binding pocket as it did not compete with a radiolabelled antagonist, H-3-SC-52012. This indicates that the 4F8 epitope behaves as a ligand-attenuated binding site (LABS). Ligand-induced attenuation of 4178 was an active process as it was prevented by pretreating platelets with cytochalasin D and reduced by prostaglandin E-1 or inhibition of protein kinase C. Disappearance of the epitope was required for full platelet activation as 4F8 prevented platelet aggregation without inhibiting fibrinogen binding. These results suggest a model where disappearance of the 4F8 epitope is a secondary event required for full

A work-based research assessment of the impact of 'lean manufacturing' on health and safety education within an SME

Globalisation has had a major impact on the engineering industry as pacific Rim countries undercut manufacturing costs and provide a more cost-effective location for many businesses. Engineering in Nortehrn Ireland has mostly declined owing to increased competition from these countries. Engineering companies are now forced to streamline their production processes and employ cost-reducing practices in order to meet customer demands at reduced prices. This article aims to analyse the effects of one such streamlining endeavour which was first introduced after World War II in Japan- 'lean manufacturing' . 'Lean manufacturing' aims to reduce all wasteful activities within the production process in order to improve productivity, while reducing manufacturing costs. The work-based project under consideration was concerned with the impact 'lean manufacturing' may have on health and safety performance and education within an engineering company. The focus of the project was to determine through work-based research, and quantitative analysis, the employee perception on health and safety: has it changed (either positively or negatively), as a consequence of implementing 'lean manufacturing'.

The effect of temperature and strain rate on the deformation behaviour, structure development and properties of biaxially stretched PET-clay nanocomposites.

The inclusion of a synthetic fluoromica clay in PET affects its processability via biaxial stretching and stretching temperature (95 °C and 102 °C) and strain rate (1 s-1 and 2 s-1) influence the structuring and properties of the stretched material. The inclusion of clay has little effect on the temperature operating window for the PET–clay but it has a major effect on deformation behaviour which will necessitate the use of much higher forming forces during processing. The strain hardening behaviour of both the filled and unfilled materials is well correlated with tensile strength and tensile modulus. Increasing the stretching temperature to reduce stretching forces has a detrimental effect on clay exfoliation, mechanical and O2 barrier properties. Increasing strain rate has a lesser effect on the strain hardening behaviour of the PET–clay compared with the pure PET and this is attributed to possible adiabatic heating in the PET–clay sample at the higher strain rate. The Halpin–Tsai model is shown to accurately predict the modulus enhancement of the PET–clay materials when a modified particle modulus rather than nominal clay modulus is used.

Anthrax Lethal Factor as an Immune Target in Humans and Transgenic Mice and the Impact of HLA Polymorphism on CD4+ T Cell Immunity

Bacillus anthracis produces a binary toxin composed of protective antigen (PA) and one of two subunits, lethal factor (LF) or edema factor (EF). Most studies have concentrated on induction of toxin-specific antibodies as the correlate of protective immunity, in contrast to which understanding of cellular immunity to these toxins and its impact on infection is limited. We characterized CD4+ T cell immunity to LF in a panel of humanized HLA-DR and DQ transgenic mice and in naturally exposed patients. As the variation in antigen presentation governed by HLA polymorphism has a major impact on protective immunity to specific epitopes, we examined relative binding affinities of LF peptides to purified HLA class II molecules, identifying those regions likely to be of broad applicability to human immune studies through their ability to bind multiple alleles. Transgenics differing only in their expression of human HLA class II alleles showed a marked hierarchy of immunity to LF. Immunogenicity in HLA transgenics was primarily restricted to epitopes from domains II and IV of LF and promiscuous, dominant epitopes, common to all HLA types, were identified in domain II. The relevance of this model was further demonstrated by the fact that a number of the immunodominant epitopes identified in mice were recognized by T cells from humans previously infected with cutaneous anthrax and from vaccinated individuals. The ability of the identified epitopes to confer protective immunity was demonstrated by lethal anthrax challenge of HLA transgenic mice immunized with a peptide subunit vaccine comprising the immunodominant epitopes that we identified.

Genomics and the impact of new technologies on the management of colorectal cancer

High-throughput genomic technologies have the potential to have a major impact on preclinical and clinical drug development and the selection and stratification of patients in clinical trials. These technologies, which are at varying stages of commercialization, include array-based comparative genomic hybridization, single-nucleotide polymorphism arrays, and (the most mature example) expression-based arrays. One of the rate-limiting steps in the routine clinical application of expression array-based technology is the need for suitable clinical samples. One of the major challenges moving forward, therefore, relates to the ability to use formalin-fixed, paraffin-embedded--derived tissue in expression profiling-based approaches.

“Islands” in Holocene forests: implications for forest openness, landscape clearance and “culture-steppe” species

Human activity has undoubtedly had a major impact on Holocene forested ecosystems, with the concurrent expansion of plants and animals associated with cleared landscapes and pasture, also known as 'culture-steppe'. However, this anthropogenic perspective may have underestimated the contribution of autogenic disturbance (e.g. wind-throw, fire), or a mixture of autogenic and anthropogenic processes, within early Holocene forests. Entomologists have long argued that the north European primary forest was probably similar in structure to pasture woodland. This idea has received support from the conservation biologist Frans Vera, who has recently strongly argued that the role of large herbivores in maintaining open forests in the primeval landscapes of Europe has been seriously underestimated. This paper reviews this debate from a fossil invertebrate perspective and looks at several early Holocene insect assemblages. Although wood taxa are indeed important during this period, species typical of open areas and grassland and dung beetles, usually associated with the dung of grazing animals, are persistent presences in many early woodland faunas. We also suggest that fire and other natural disturbance agents appear to have played an important ecological role in some of these forests, maintaining open areas and creating open vegetation islands within these systems. More work, however, is required to ascertain the role of grazing animals, but we conclude that fossil insects have a significant contribution to make to this debate. This evidence has fundamental implications in terms of how the palaeoecological record is interpreted, particularly by environmental archaeologists and palaeoecologists who may be more interested in identifying human-environment interactions rather than the ecological processes which may be preserved within palaeoecological records.

Structural and chemical embrittlement of grain boundaries by impurities: A general theory and first-principles calculations for copper

First-principles calculations of the Sigma 5(310)[001] symmetric tilt grain boundary in Cu with Bi, Na, and Ag substitutional impurities provide evidence that in the phenomenon of Bi embrittlement of Cu grain boundaries electronic effects do not play a major role; on the contrary, the embrittlement is mostly a structural or "size" effect. Na is predicted to be nearly as good an embrittler as Bi, whereas Ag does not embrittle the boundary in agreement with experiment. While we reject the prevailing view that "electronic" effects (i.e., charge transfer) are responsible for embrittlement, we do not exclude the role of chemistry. However, numerical results show a striking equivalence between the alkali metal Na and the semimetal Bi, small differences being accounted for by their contrasting "size" and "softness" (defined here). In order to separate structural and chemical effects unambiguously if not uniquely, we model the embrittlement process by taking the system of grain boundary and free surfaces through a sequence of precisely defined gedanken processes; each of these representing a putative mechanism. We thereby identify three mechanisms of embrittlement by substitutional impurities, two of which survive in the case of embrittlement or cohesion enhancement by interstitials. Two of the three are purely structural and the third contains both structural and chemical elements that by their very nature cannot be further unraveled. We are able to take the systems we study through each of these stages by explicit computer simulations and assess the contribution of each to the net reduction in intergranular cohesion. The conclusion we reach is that embrittlement by both Bi and Na is almost exclusively structural in origin; that is, the embrittlement is a size effect.

Multi component coupling reactions of N-acetyl-2-azetine

N-Acetyl-2-azetine undergoes Lewis acid catalysed formal [4+2]-cycloaddition with imines derived from aromatic amines to initially give an approximately 1: 1 mixture of exo-endo-diastereoisomeric 1-(2a,3,4,8b-tetrahydro-2H-1,4-diaza-cyclobuta[a]naphthalen-1-yl)-ethanone cycloadducts which were detected by proton NMR spectroscopy. These products, which were too unstable to isolate, and characterise, reacted further with aromatic amines to give 2,3,4-trisubstituted tetrahydroquinolines in good to excellent yield, predominantly as a single diastereoisomer, with the minor diastereoisomer converting to the major diastereoisomer on silica. The cycloaddition was irreversible and a mechanism is presented for the formation of the major diastereoisomer from the mixture of diastereoisomeric intermediates. A range of conditions is described for converting the 2,3,4-trisubsitituted tetrahydroquinolines into 2,3-disubstituted quinolines.

Where are they now? Tracking the longitudinal evolution of environmental businesses from the 1990s

In 1991 Bennett published one of the first major publications on the concept of ecopreneurship, business opportunities resulting from the emerging environmental agenda of the late 1980s and early 1990s. Since then a body of literature has developed that explores the idea of the intersection of entrepreneurship with environmentally and socially responsible behaviour. Many of the business cases presented by Bennett represent early adopters of green products, services and emerging eco-markets. Given the current emphasis on the transformation of business practices towards a more sustainable paradigm it is timely to review these 94 early ecopreneurial examples and consider their status two decades on from the original publication. This paper explores the definitions of environmental and social enterprise, and considers the longitudinal survival of these companies and the emerging trends in consolidation and failure of the sampled companies.

Mid- to late-Holocene climate variability and anthropogenic impacts: multi-proxy evidence from Lake Bliden, Denmark

We conducted multi-proxy geochemical analyses (including measurements of organic carbon, nitrogen and sulphur stable isotope composition, and carbonate carbon and oxygen isotope composition) on a 13.5 m sediment core from Lake Bliden, Denmark, which provide a record of shifting hydrological conditions for the past 6,700 years. The early part of the stratigraphic record (6,700-5,740 cal year BP) was wet, based on delta O-18(carb) and lithology, and corresponds to the Holocene Thermal Maximum. Shifts in primarily delta O-18(carb) indicate dry conditions prevailed from 5,740 to 2,800 cal year BP, although this was interrupted by very wet conditions from 5,300 to 5,150, 4,300 to 4,050 and 3,700 to 3,450 cal year BP. The timing of the latter two moist intervals is consistent with other Scandinavian paleoclimatic records. Dry conditions at Lake Bliden between 3,450 and 2,800 cal year BP is consistent with other paleolimnological records from southern Sweden but contrasts with records in central Sweden, possibly suggesting a more northerly trajectory of prevailing westerlies carrying moisture from the North Atlantic at this time. Overall, fluctuating moisture conditions at Lake Bliden appear to be strongly linked to changing sea surface temperatures in the Greenland, Iceland and Norwegian seas. After 2,800 cal year BP, sedimentology, magnetic susceptibility, delta C-13(ORG), delta C-13(carb) and delta O-18(carb) indicate a major reduction on water level, which caused the depositional setting at the coring site to shift from the profundal to littoral zone. The Roman Warm Period (2,200-1,500 cal year BP) appears dry based on enriched delta O-18(carb) values. Possible effects of human disturbance in the watershed after 820 cal year BP complicate attempts to interpret the stratigraphic record although tentative interpretation of the delta O-18(carb), magnetic susceptibility, delta C-13(ORG), delta C-13(carb) and delta O-18(carb) records suggest that the Medieval Warm Period was dry and the Little Ice Age was wet.

The Perils of Moviegoing in America, 1896-1950

The Perils of Moviegoing in America is a film history that examines the various physical and (perceived) moral dangers facing audiences during the first fifty years of film exhibition.

Chapter 1: “Conflagration”
As early as 1897, a major fire broke out at a film exhibition in San Francisco, with flames burning the projectionist and nearby audience members. From that point until the widespread adoption of safety stock in 1950, fires were a very common movie-going experience. Hundreds of audience members lost their lives in literally thousands of theatre fires, ranging from early nickelodeons to the movie palaces of the thirties and forties.

Chapter 2: “Thieves Among Us”
Bandits robbed movie theatres on hundreds of occasions from the early days of film exhibition through the end of the Great Depression. They held up ticket booths, and they dynamited theatre safes. They also shot theatre managers, ushers, and audience members, as a great many of the robberies occurred while movies were playing on the screens inside.

Chapter 3: “Bombs Away”
Bombings at movie theatres became common in small towns and large cities on literally hundreds of occasions from 1914 to the start of World War II. Some were incendiary bombs, and some were stench bombs; both could be fatal, whether due to explosions or to the trampling of panicked moviegoers

Chapter 4: “It’s Catching”
Widespread movie-going in the early 20th century provoked an outcry from numerous doctors and optometrists who believed that viewing films could do irreparable harm to the vision of audience members. Medical publications (including the Journal of the American Medical Association) published major studies on this perceived problem, which then filtered into popular-audience magazines and newspapers.

Chapter 5: “The Devil’s Apothecary Shops”
Sitting in the dark with complete strangers proved worrisome for many early filmgoers, who had good reason to be concerned. Darkness meant that prostitutes could easily work in the balconies of some movie theatres, as could “mashers” who molested female patrons (and sometimes children) after the lights were dimmed. That was all in addition to the various murderers who used the cover of darkness to commit their crimes at movie theatres.

Chapter 6: “Blue Sundays”
Blue laws were those regulations that prohibited businesses from operating on Sundays. Most communities across the US had such legislation on their books, which by the nickelodeon era were at odds with the thousands of filmgoers who went to the movies every Sunday. Theatre managers were often arrested, making newspaper headlines over and over again. Police sometimes even arrested entire film audiences as accomplices in the Blue Law violations.

Chapter 7: “Something for Nothing”
In an effort to bolster ticket sales, many movie theatres in the 1910s began to hold lotteries in which lucky audience members won cash prizes; by the time of the Great Depression, lotteries like “Bank Night” became a common aspect of the theatre-going enterprise. However, reception studies have generally overlooked the intense (and sometimes coordinated) efforts by police, politicians, and preachers to end this practice, which they viewed as illegal and immoral gambling.

Unique genetic variation at a species' rear edge is under threat from global climate change

Global climate change is having a significant effect on the distributions of a wide variety of species, causing both range shifts and population extinctions. To date, however, no consensus has emerged on how these processes will affect the range-wide genetic diversity of impacted species. It has been suggested that species that recolonized from low-latitude refugia might harbour high levels of genetic variation in rear-edge populations, and that loss of these populations could cause a disproportionately large reduction in overall genetic diversity in such taxa. In the present study, we have examined the distribution of genetic diversity across the range of the seaweed Chondrus crispus, a species that has exhibited a northward shift in its southern limit in Europe over the last 40 years. Analysis of 19 populations from both sides of the North Atlantic using mitochondrial single nucleotide polymorphisms (SNPs), sequence data from two singlecopy nuclear regions and allelic variation at eight microsatellite loci revealed unique genetic variation for all marker classes in the rear-edge populations in Iberia, but not in the rear-edge populations in North America. Palaeodistribution modelling and statistical testing of alternative phylogeographic scenarios indicate that the unique genetic diversity in Iberian populations is a result not only of persistence in the region during the last glacial maximum, but also because this refugium did not contribute substantially to the recolonization of Europe after the retreat of the ice. Consequently, loss of these rear-edge populations as a result of ongoing climate change will have a major effect on the overall genetic diversity of the species, particularly in Europe, and this could compromise the adaptive potential of the species as a whole in the face of future global warming.

Electric Vehicles - a well to wheel analysis

The European Union has set a target for 10% renewable energy in transport by 2020, which will be met using both biofuels and electric vehicles. In the case of biofuels, for the purposes of meeting the target, the biofuel must achieve greenhouse gas savings of 35% relative to the fossil fuel replaced. For biofuels, greenhouse gas savings can be calculated using life cycle analysis, or the European Union default values. In contrast, all electricity used in transport is considered to be the same, regardless of the source or the type of electric vehicle. However, the choice of the electric vehicle and electricity source will have a major impact on the greenhouse gas savings. This paper examines different electric-vehicle scenarios in terms of greenhouse gas savings, using a well-to-wheel life cycle analysis.

Immunotoxicity of aflatoxin B1: Impairment of the cell-mediated response to vaccine antigen and modulation of cytokine expression

Aflatoxin B1 (AFB1), a mycotoxin produced by Aspergillus flavus or A. parasiticus, is a frequent contaminant of food and feed. This toxin is hepatotoxic and immunotoxic. The present study analyzed in pigs the influence of AFB1 on humoral and cellular responses, and investigated whether the immunomodulation observed is produced through interference with cytokine expression. For 28 days, pigs were fed a control diet or a diet contaminated with 385, 867 or 1807 mu g pure AFB1/kg feed. At days 4 and 15, pigs were vaccinated with ovalbumin. AFB1 exposure, confirmed by an observed dose-response in blood aflatoxin-albumin adduct, had no major effect on humoral immunity as measured by plasma concentrations of total IgA, IgG and IgM and of anti-ovalbumin IgG. Toxin exposure did not impair the mitogenic response of lymphocytes but delayed and decreased their specific proliferation in response to the vaccine antigen, suggesting impaired lymphocyte activation in pigs exposed to AFB1. The expression level of pro-inflammatory (TNF-alpha, IL-1 beta, IL-6, IFN-gamma) and regulatory (IL-10) cytokines was assessed by real-time PCR in spleen. A significant up-regulation of all 5 cytokines was observed in spleen from pigs exposed to the highest dose of AFB1. In pigs exposed to the medium dose, IL-6 expression was increased and a trend towards increased IFN-gamma and IL-10 was observed. In addition we demonstrate that IL-6 impaired in vitro the antigenic- but not the mitogenic-induced proliferation of lymphocytes from control pigs vaccinated with ovalbumin. These results indicate that AFB1 dietary exposure decreases cell-mediated immunity while inducing an inflammatory response. These impairments in the immune response could participate in failure of vaccination protocols and increased susceptibility to infections described in pigs exposed to AFB1. (C) 2008 Elsevier Inc. All rights reserved.