Evaluation of early post-natal pig mammary gland development and human breast cancer gene expression.

Breast cancer is the second leading cause of death in women after lung cancer, and only 5% of patients with metastatic breast cancer survive beyond ten years of diagnosis. Considering the heterogeneous subclasses of breast cancer, current cancer models have shortfalls due to copy number variants, and genetic differences of humans and immunocompromised animal models. Preclinical studies indicate stem cell activity in early post-natal mammary development may be reactivated in the human adult as a trigger to initiate cell proliferation leading to breast cancer. The goal of the work reported herein was to compare genetic expression of early development, post-natal pig mammary glands to the literature reported genes implicated in different subclasses of human breast cancer. Differentially expressed genes associated with breast cancer and present in early developing pig samples include NUCB2, ANGPTL4 and ACE. Histological staining confirmed E-cadherin, Vimentin, N-cadherin, and Claudin-1, which are all implicated in malignant cancer. Due to the homology of gene expression patterns in the developing pig mammary gland and reported genes in human breast cancer profiles, this research is worthy of further study to address a potential model using mammary development cues to unravel breast cancer biology.

Effects of corticosteroids on COPD lung macrophage phenotype and function.

The numbers of macrophages are increased in the lungs of chronic obstructive pulmonary disease (COPD) patients. COPD lung macrophages have reduced ability to phagocytose microbes and efferocytose apoptotic cells. Inhaled corticosteroids (ICSs) are widely used anti-inflammatory drugs in COPD; however, their role beyond suppression of cytokine release has not been explored in COPD macrophages. We have examined the effects of corticosteroids on COPD lung macrophage phenotype and function. Lung macrophages from controls and COPD patients were treated with corticosteroids; effects on gene and protein expression of CD163, CD164, CD206, MERTK, CD64, CD80 and CD86 were studied. We also examined the effect of corticosteroids on the function of CD163, MERTK and cluster of differentiation 64 (CD64). Corticosteroid increased CD163, CD164, CD206 and MERTK expression and reduced CD64, CD80 and CD86 expression. We also observed an increase in the uptake of the haemoglobin-haptoglobin complex (CD163) from 59 up to 81% and an increase in efferocytosis of apoptotic neutrophils (MERTK) from 15 up to 28% following corticosteroid treatment. We observed no effect on bacterial phagocytosis. Corticosteroids alter the phenotype and function of COPD lung macrophages. Our findings suggest mechanisms by which corticosteroids exert therapeutic benefit in COPD, reducing iron available for bacterial growth and enhancing efferocytosis.

Methodology to analyze gene expression patterns of early mammary development in pig models.

In mammary gland development, normal stem cell activity occurs in the embryonic stage and postnatally. Research supports that certain breast cancers contain a small sub-population of cells that mimic stem-like activity. It is believed stem cell activation in the mutated mature human mammary tissue is what drives quiescent epithelial cells to convert to mesenchymal states initiating migration, invasion, and metastasis in breast cancer. The goal of the work reported herein was to investigate early mammary development gene expression in the postnatal pig using fine needle biopsy methods in order to establish a reliable model for human breast cancer detection. Tissue samples were collected from pig mammary glands beginning at Day 11 of age through Day 39 in order to capture early postnatal-growth gene expression. Based on the initial clustering analysis, two distinct clusters of gene expression profiles occurred before and after Day 25 of mammary development. Gene set enrichment analysis (GSEA) ontology indicated the cellular processes that changed after Day 25, and many of these processes were implicated in epithelial-mesenchymal transition (EMT) signaling events. Gene expression in the postnatal pig was compared with the Epithelial-Mesenchymal Transition gene database (dbEMT) confirming the presence of EMT activity in this early developmental program. Information from this study will provide insight into early postnatal mammary gland development. In addition, mechanisms exploited by mutated mammary epithelial cells leading to cancer initiation and growth may be detected considering that mutated mammary epithelial cells can reactivate early developmental signals.

Fluorophosphonate-functionalised titanium via a pre-adsorbed alkane phosphonic acid: a novel dual action surface finish for bone regenerative applications.

Enhancing vitamin D-induced human osteoblast (hOB) maturation at bone biomaterial surfaces is likely to improve prosthesis integration with resultant reductions in the need for revision arthroplasty consequent to aseptic loosening. Biomaterials that are less appealing to microorganisms implicated in implant failures through infection are also highly desirable. However, finding surfaces that enhance hOB maturation to active vitamin D yet deter bacteria remain elusive. In addressing this, we have sought to bio-functionalise titanium (Ti) with lysophosphatidic acid (LPA) and related, phosphatase-resistant, LPA analogues. The impetus for this follows our discovery that LPA co-operates with active vitamin D3 metabolites to secure hOB maturation in vitro including cells grown upon Ti. LPA has also been found, by others, to inhibit virulence factor production and biofilm formation of the human opportunistic pathogen Pseudomonas aeruginosa. Collectively, selected LPA species might offer potential dual-action surface finishes for contemporary bone biomaterials. In attaching a phosphatase-resistant LPA analogue to Ti we took advantage of the affinity of alkane phosphonic acids for TiO2. Herein, we provide evidence for the facile development of a dual-action Ti surface for potential orthopaedic and dental applications. Successful conjugation of an LPA analogue (3S)1-fluoro-3-hydroxy-4-(oleoyloxy)butyl-1-phosphonate (FHBP) to the Ti surface was supported through physiochemical characterisation using x-ray photoelectron spectroscopy and secondary ion mass spectrometry. hOB maturation to active vitamin D3 was enhanced for cells grown on FHBP-Ti whilst these same surfaces exhibited clear antiadherent properties towards a clinical isolate of Staphylococcus aureus.

Using near infrared transmittance to generate sorted fractions of Fusarium-infected wheat and the impact on broiler performance.

The objective of this study was to investigate the effects of naturally contaminated Fusarium wheat containing deoxynivalenol (DON) on growth and performance of broiler chickens from 0 to 35 d. The BoMill TriQ individual kernel sorting technology uses near infrared transmittance (NIT) spectra to separate Fusarium-damaged kernels (FDK) from healthy kernels based on individual kernel CP. Three Fusarium-contaminated wheat sources were individually sorted into 3 test fractions: outlier (10% of the source), high mycotoxin (20% of the source), and low mycotoxin (70% of the source). These fractions were reconstituted into 4 ratios-M0, M20, M40, and M60-relating to the proportion of the high mycotoxin fraction in the reconstituted diets. These 12 reconstituted wheat sources with varying levels of DON were incorporated at ∼70% (starter) or ∼75% (grower/finisher) into diets. The fractions of wheat used had FDK ranging from 0.1 to 25.8% and DON from 0.0 to 14.3 ppm. A total of 480 newly hatched Ross 308 male broilers were randomly divided into 96 cages. Each test diet was assigned to 8 replicates with 5 birds per replicate cage. At 21 d, 180 birds were transferred to 36 cages, allowing 3 replicates of 5 birds per diet until 35 d. A factorial arrangement analysis compared the 3 wheat sources and 4 ratios produced from each sorted wheat. Growth and performance were evaluated as BW (g), feed intake (FI; g/bird/day), feed conversion ratio (FCR; g:g), AME (kcal ME/kg diet), nitrogen retention (NR; %), and mortality (%) for 0 to 21 d and 21 to 35 d. Results indicate no significant difference in BW, FI, and FCR (P > 0.05). Significant differences were seen in AME and NR (P < 0.01). This study demonstrates the potential of this novel sorting technology to produce naturally contaminated diets with a large range of mycotoxin concentrations from a single wheat source to enable future investigations of mycotoxin exposure in any species.

Examining the residual radiological footprint of a former colliery: An industrial nuclear archaeology investigation.

Nuclear industrial archaeology utilises radiation mapping and characterisation technologies to gain an insight into the radiological footprint of industrial heritage sites. Increased concentrations of naturally occurring radioactive materials at legacy mine sites are the result of elemental enrichment during coal mining and subsequent combustion. Public safety is of concern around these sites, and therefore, an increased understanding of their associated hazard is essential. Using coincident laser scanning and gamma detection technologies, this study sought to assess the radiological legacy of a coal mine located in Bristol, UK. From this, we can increase our understanding of the residual footprints associated with the local coal mining industry. Samples taken from inside the site were characterised using high resolution gamma spectrometry, wherein the radionuclide content and activities of samples were then quantified. An area of elevated low-level radioactivity was observed at and around buildings believed to belong to the colliery, while Th, U, and K are confirmed at the site from photopeak's of daughter radionuclides. Activities of the radionuclides K-40, U-238, and Th-232 were further quantified during subsequent laboratory analysis. Results highlight an enrichment of naturally occurring radionuclides when compared with global averages for unburned coal. Employing these techniques at further legacy sites would enable an increased understanding of the lasting traces of the coal mining industry, with a focus on NORM enrichment in residual fly ash.

Quantifying impacts of titanium dioxide nanoparticles on natural assemblages of riverine phytobenthos and phytoplankton in an outdoor setting.

Impacts of widespread release of engineered titanium dioxide nanoparticles (nTiO2) on freshwater phytoplankton and phytobenthic assemblages in the field, represents a significant knowledge gap. Using outdoor experiments, we quantified impacts of nTiO2 on phytoplankton and periphyton from UK rivers, applied at levels representative of environmentally realistic concentrations (0.05 mg/L) and hot spots of accumulation (5.0 mg/L). Addition of nTiO2 to river water led to rapid temporal size changes in homoagglomerates and many heteroaggregates of nTiO2 with cells in the phytoplankton, including green algae, pennate and centric diatoms, increasing settlement of some cells. Changes in phytoplankton composition were evident after 72-h resulting from a significant decline in the relative abundance of very small phytoplankton cells (1-3 µm), often accompanied by increases in centric diatoms at both concentrations. Significant changes detected in the composition of the phytobenthos after 12 days, following nTiO2 treatments, were not evident when using benthic diatoms alone after 56 days. A lack of inhibition in the maximum quantum yield (Fv/Fm) in phytobenthos after 72-h exposures contrasted with a significant inhibition in Fv/Fm in 75% of phytoplankton samples, the highest recorded in Rutile nTiO2 exposures at both concentrations of nTiO2. After 12 days, strong positive stimulatory responses were recorded in the maximum relative electron transport rate (rETRmax) and the maximum non-photochemical coefficient (NPQmax), in phytoplankton and phytobenthos samples exposed to the higher Anatase nTiO2 concentration, were not measured in Rutile exposed biota. Collectively, these results indicate that the Rutile phase of nTiO2 has more negative impacts on freshwater algae than the Anatase form, at specific time scales, and phytoplankton may be more impacted by nTiO2 than phytobenthos. We caution that repeated release of nTiO2, could lead to significant changes in riverine algal biomass and species composition, dependent on the phase and concentration of nTiO2.

Arterial blood gas measurements during deep open-water breath-hold dives.

Arterial blood gas (ABG) measurements at both maximum depth and at resurfacing prior to breathing have not previously been measured during free dives conducted to extreme depth in cold open-water conditions. An elite free diver was instrumented with a left radial arterial cannula connected to two sampling syringes through a low-volume splitting device. He performed two open-water dives to a depth of 60 m (197', 7 atmospheres absolute pressure) in the constant weight with fins competition format. ABG samples were drawn at 60 m (by a mixed-gas scuba diver) and again on resurfacing before breathing. An immersed surface static apnea, of identical length to the dives and with ABG sampling at identical times, was also performed. Both dives lasted approximately 2 min. Arterial partial pressure of oxygen ([Formula: see text]) increased during descent from an indicative baseline of 15.8 kPa (after hyperventilation and glossopharyngeal insufflation) to 42.8 and 33.3 kPa (dives 1 and 2) and decreased precipitously (to 8.2 and 8.6 kPa) during ascent. Arterial partial pressure of carbon dioxide ([Formula: see text]) also increased from a low indicative baseline of 2.8 kPa to 6.3 and 5.1 kPa on dives 1 and 2; an increase not explained by metabolic production of CO2 alone since [Formula: see text] actually decreased during ascent (to 5.2 and 4.5 kPa). Surface static apnea caused a steady decrease in [Formula: see text] and increase in [Formula: see text] without the inflections provoked by depth changes. Lung compression and expansion provoke significant changes in both [Formula: see text] and [Formula: see text] during rapid descent and ascent on a deep free dive. These changes generally support predictive hypotheses and previous findings in less extreme settings.NEW & NOTEWORTHY Arterial blood gas measurements at both maximum depth and the surface before breathing on the same dive have not previously been obtained during deep breath-hold dives in cold open-water conditions and competition dive format. Such measurements were obtained in two dives to 60 m (197') of 2 min duration. Changes in arterial oxygen and carbon dioxide (an increase during descent, and a decrease during ascent) support previous observations in less extreme dives and environments.

Radiation Tolerance Testing Methodology of Robotic Manipulator Prior to Nuclear Waste Handling.

Dramatic cost savings, safety improvements and accelerated nuclear decommissioning are all possible through the application of robotic solutions. Remotely-controlled systems with modern sensing capabilities, actuators and cutting tools have the potential for use in extremely hazardous environments, but operation in facilities used for handling radioactive material presents complex challenges for electronic components. We present a methodology and results obtained from testing in a radiation cell in which we demonstrate the operation of a robotic arm controlled using modern electronics exposed at 10 Gy/h to simulate radioactive conditions in the most hazardous nuclear waste handling facilities.

Multifunctional, Room-Temperature Processable, Heterogeneous Organic Passivation Layer for Oxide Semiconductor Thin-Film Transistors.

In recent decades, oxide thin-film transistors (TFTs) have attracted a great deal of attention as a promising technology in terms of next-generation electronics due to their outstanding electrical performance. However, achieving robust electrical characteristics under various environments is a crucial challenge for successful realization of oxide-based electronic applications. To resolve the limitation, we propose a highly flexible and reliable heterogeneous organic passivation layer composed of stacked parylene-C and diketopyrrolopyrrole-polymer films for improving stability of oxide TFTs under various environments and mechanical stress. The presented multifunctional heterogeneous organic (MHO) passivation leads to high-performance oxide TFTs by: (1) improving their electrical characteristics, (2) protecting them from external reactive molecules, and (3) blocking light exposure to the oxide layer. As a result, oxide TFTs with MHO passivation exhibit outstanding stability in ambient air as well as under light illumination: the threshold voltage shift of the device is almost 0 V under severe negative bias illumination stress condition (white light of 5700 lx, gate voltage of -20 V, and drain voltage of 10.1 V for 20 000 s). Furthermore, since the MHO passivation layer exhibits high mechanical stability at a bending radius of ≤5 mm and can be deposited at room temperature, this technique is expected to be useful in the fabrication of flexible/wearable devices.

Thrombocytes respond to lipopolysaccharide through Toll-like receptor-4, and MAP kinase and NF-kappaB pathways leading to expression of interleukin-6 and cyclooxygenase-2 with production of prostaglandin E2.

Chicken thrombocytes are equivalent in hemostatic function to mammalian platelets. Platelets are enucleated components of mammalian blood, while thrombocytes are nucleated blood leukocytes of chickens. Platelets and thrombocytes share characteristics that contribute to innate immunity. Experiments were conducted to determine if thrombocytes could respond in vitro to lipopolysaccharide (LPS) of Salmonella minnesota through Toll-like receptor-4 (TLR4). The aim was to activate the signal pathways leading to expression of interleukin-6 (IL-6) and inducible cyclooxygenase (COX-2) and to production of prostaglandin E2 (PGE2). Chicken thrombocytes were found to express TLR4, and LPS-induced an increase in thrombocyte mRNA expression of IL-6 and COX-2 with release of PGE2 into culture media. An increase of COX-2 and PGE2 due to LPS stimulation was inhibited by MEK1 inhibitor PD98059, but IL-6 expression was unaffected by PD98059. The IKK-2 inhibitor BMS345541 inhibited IL-6 and COX-2 with reduction of PGE2 concentrations. Therefore, the MAP kinase (MAPK) pathway activates expression of COX-2 and ultimately PGE2 production, but this pathway has little or no influence on IL-6 expression in thrombocytes. The NF-kappaB pathway also influences COX-2 expression and PGE2 production, and it is a primary activation signaling cascade for IL-6 gene expression in chicken thrombocytes. Thrombocytes represent a major component of the innate immune system of chickens in response to LPS and possibly other microbial products.

Spatial pattern of plutonium and radiocaesium contamination released during the Fukushima Daiichi nuclear power plant disaster.

Plutonium and radiocaesium are hazardous contaminants released by the Fukushima Daiichi nuclear power plant (FDNPP) disaster and their distribution in the environment requires careful characterisation using isotopic information. Comprehensive spatial survey of 134Cs and 137Cs has been conducted on a regular basis since the accident, but the dataset for 135Cs/137Cs atom ratios and trace isotopic analysis of Pu remains limited because of analytical challenges. We have developed a combined chemical procedure to separate Pu and Cs for isotopic analysis of environmental samples from contaminated catchments. Ultra-trace analyses reveal a FDNPP Pu signature in environmental samples, some from further afield than previously reported. For two samples, we attribute the dominant source of Pu to Reactor Unit 3. We review the mechanisms responsible for an emergent spatial pattern in 134,135Cs/137Cs in areas northwest (high 134Cs/137Cs, low 135Cs/137Cs) and southwest (low 134Cs/137Cs, high 135Cs/137Cs) of FDNPP. Several samples exhibit consistent 134,135Cs/137Cs values that are significantly different from those deposited on plant specimens collected in previous works. A complex spatial pattern of Pu and Cs isotopic signature is apparent. To confidently attribute the sources of mixed fallout material, future studies must focus on analysis of individual FDNPP-derived particles.

Molecularly selective nanoporous membrane-based wearable organic electrochemical device for noninvasive cortisol sensing.

Wearable biosensors have emerged as an alternative evolutionary development in the field of healthcare technology due to their potential to change conventional medical diagnostics and health monitoring. However, a number of critical technological challenges including selectivity, stability of (bio)recognition, efficient sample handling, invasiveness, and mechanical compliance to increase user comfort must still be overcome to successfully bring devices closer to commercial applications. We introduce the integration of an electrochemical transistor and a tailor-made synthetic and biomimetic polymeric membrane, which acts as a molecular memory layer facilitating the stable and selective molecular recognition of the human stress hormone cortisol. The sensor and a laser-patterned microcapillary channel array are integrated in a wearable sweat diagnostics platform, providing accurate sweat acquisition and precise sample delivery to the sensor interface. The integrated devices were successfully used with both ex situ methods using skin-like microfluidics and on human subjects with on-body real-sample analysis using a wearable sensor assembly.

Sequential adhesion of platelets and leukocytes from flowing whole blood onto a collagen-coated surface: requirement for a GpVI-binding site in collagen.

The adhesion of leukocytes to immobilised platelets may contribute to inflammatory and thrombotic responses in damaged tissue. To investigate the conditions under which platelets and leukocytes might be deposited together in vessels, we perfused fluorescently-labelled whole blood through glass capillaries coated with various collagen preparations. Video-microscopic observations of the surface showed that platelets formed numerous, individual, rolling and stationary attachments to surfaces coated with acid-soluble, monomeric collagen. However, leukocyte interactions with the deposited platelets were rare. If the blood was washed out, the adherent platelets became more activated, and many rolling adherent leukocytes were observed if a second bolus of blood was perfused over them. This suggested that platelet activation had initially been inadequate to support leukocyte capture. Next, fibrillar collagen was adsorbed to the capillaries to present an ordered array of peptide motifs to platelet receptor glycoprotein (Gp)VI and transduce an activating signal. In this case, platelets were deposited in discrete, stable aggregates and the bound platelets captured many flowing leukocytes. Alternatively, acid-soluble collagen was seeded with collagen-related peptide (CRP) known to contain a GpVI-binding motif. Again, platelet adhesion became stable, and numerous flowing leukocytes were captured. Addition of antibody against GpVI or against P-selectin greatly reduced leukocyte adhesion to the platelets. Thus, in whole blood, platelets binding to exposed collagen need to be activated through GpVI in order to expose sufficient P-selectin to allow efficient capture of flowing leukocytes to take place.

Enhanced Cell-Chip Coupling by Rapid Femtosecond Laser Patterning of Soft PEDOT:PSS Biointerfaces.

Interfacing soft materials with biological systems holds considerable promise for both biosensors and recording live cells. However, the interface between cells and organic substrates is not well studied, despite its crucial role in the effectiveness of the device. Furthermore, well-known cell adhesion enhancers, such as microgrooves, have not been implemented on these surfaces. Here, we present a nanoscale characterization of the cell-substrate interface for 3D laser-patterned organic electrodes by combining electrochemical impedance spectroscopy (EIS) and scanning electron microscopy/focused ion beam (SEM/FIB). We demonstrate that introducing 3D micropatterned grooves on organic surfaces enhances the cell adhesion of electrogenic cells.

The use of near infrared transmittance kernel sorting technology to salvage high quality grain from grain downgraded due to Fusarium damage.

The mycotoxins associated with specific Fusarium fungal infections of grains are a threat to global food and feed security. These fungal infestations are referred to as Fusarium Head Blight (FHB) and lead to Fusarium Damaged Kernels (FDK). Incidence of FDK >0.25% will lower the grade, with a tolerance of 5% FDK for export feed grain. During infestation, the fungi can produce a variety of mycotoxins, the most common being deoxynivalenol (DON). Fusarium Damaged Kernels have been associated with reduced crude protein (CP), lowering nutritional, functional and grade value. New technology has been developed using Near Infrared Transmittance (NIT) spectra that estimate CP of individual kernels of wheat, barley and durum. Our objective is to evaluate the technology's capability to reduce FDK and DON of downgraded wheat and ability to salvage high quality safe kernels. In five FDK downgraded sources of wheat, the lowest 20% CP kernels had significantly increased FDK and DON with the high CP fractions having decreased FDK and DON, thousand kernel weights (TKW) and bushel weight (Bu). Strong positive correlations were observed between FDK and DON (r = 0.90); FDK and grade (r = 0.62) and DON and grade (r = 0.62). Negative correlations were observed between FDK and DON with CP (r = -0.27 and -0.32); TKW (r = -0.45 and -0.54) and Bu (r = -0.79 and -0.74). Results show improved quality and value of Fusarium downgraded grain using this technology.

Phase Stability and Stoichiometry in Thin Film Iron Pyrite: Impact on Electronic Transport Properties.

The use of pyrite FeS2 as an earth-abundant, low-cost, nontoxic thin film photovoltaic hinges on improved understanding and control of certain physical and chemical properties. Phase stability, phase purity, stoichiometry, and defects, are central in this respect, as they are frequently implicated in poor solar cell performance. Here, phase-pure polycrystalline pyrite FeS2 films, synthesized by ex situ sulfidation, are subject to systematic reduction by vacuum annealing (to 550 °C) to assess phase stability, stoichiometry evolution, and their impact on transport. Bulk probes reveal the onset of pyrrhotite (Fe(1-δ)S) around 400 °C, rapidly evolving into the majority phase by 425 °C. This is supported by X-ray photoelectron spectroscopy on (001) crystals, revealing surface Fe(1-δ)S formation as low as 160 °C, with rapid growth near 400 °C. The impact on transport is dramatic, with Fe(1-δ)S minority phases leading to a crossover from diffusive transport to hopping (due to conductive Fe(1-δ)S nanoregions in an FeS2 matrix), followed by metallicity when Fe(1-δ)S dominates. Notably, the crossover to hopping leads to an inversion of the sign, and a large decrease in magnitude of the Hall coefficient. By tracking resistivity, magnetotransport, magnetization, and structural/chemical parameters vs annealing, we provide a detailed picture of the evolution in properties with stoichiometry. A strong propensity for S-deficient minority phase formation is found, with no wide window where S vacancies control the FeS2 carrier density. These findings have important implications for FeS2 solar cell development, emphasizing the need for (a) nanoscale chemical homogeneity, and (b) caution in interpreting carrier types and densities.