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This work demonstrates that a sustained galvanic coating of silver can be deposited onto copper substrates from a solution of Ag + ions in an ionic liquid (IL) based on a choline chloride (ChCl) eutectic. The sustained growth of the silver deposit was facilitated by the porous nature of the film and is proven using acoustic impedance spectroscopy on a quartz crystal microbalance (QCM). The silver and copper surfaces were characterised using atomic force microscopy (AFM) and scanning electron microscopy (SEM/EDX). In the commercial process of dip coating silver from ions in aqueous media, deposition does not continue once surface coverage has been achieved, without the use of catalysts. The process reported here using ionic liquids produces silver deposits of several microns by dip coating without the use of catalysts or strong inorganic acids.

Soil microbes are central to many soil processes, but due to the structural complexity of soil organic matter, the accurate quantification of microbial biomass contributions continues to pose a significant analytical challenge. In this study, microbes from a range of soils were cultured such that their molecular profile could be compared to that of soil organic matter and native vegetation. With the use of modern NMR spectroscopy, the contributions from microbial species can be discerned in soil organic matter and quantified. On the basis of these studies, the contributions of microbial biomass to soil organic matter appear to be much higher than the 1-5% reported by other researchers. In some soils, microbial biomass was found to contribute >50% of the extractable soil organic matter fractions and approximately 45% of the humin fraction and accounted for >80% of the soil nitrogen. These findings are significant because organic matter is intimately linked to nutrient release and transport in soils, nitrogen turnover rates, contaminant fate, soil quality, and fertility. Therefore, if in some cases soil organic matter and soil organic nitrogen are predominately of microbial origin, it is likely that this fraction, whether in the form of preserved material or living cells, plays an underestimated role in several soil processes.

Acute sediment toxicity tests do not test key life stage events such as moulting and reproduction and therefore do not reveal the longer-term effects of contaminant exposure. A laboratory method is described for determining the chronic toxicity of contaminants associated with whole sediments. The test is conducted using neonates of the estuarine amphipod Corophium volutator at 15 degrees C, salinity 25 psu and a 12 h light:12 h dark photoperiod. The endpoints are survival and growth after 28 days and survival, growth and reproduction of amphipods upon termination of test i.e. reproduction within all control vessels (ca 75 days). The sediment chronic toxicity test was used to investigate the effects of sediments spiked with environmentally relevant preparations of slightly weathered Alaskan North Slope crude oil, including a water-accommodated-fraction (WAF) and a chemically-dispersed (Corexit 9527) WAF. Sediment oil concentrations were quantified using ultra-violet fluorescence. The amphipods exposed to chemically dispersed oil had higher mortality and lower growth rates than control-, Corexit 9527- and WAF-exposed organisms, resulting in reduced reproduction. The described method supplements the standard acute sediment test and would be particularly useful when long-term ecological effects are suspected but acute tests reveal no significant mortality. The sediment chronic test reported herein has shown that sediment that was not evidently toxic during 10-day acute tests could have population-level effects on sediment-dwelling amphipods.

Amphipods are widely used in both acute and chronic (sub-lethal) sediment tests. Acute sediment tests provide relatively rapid results, but may fail to detect moderately toxic contaminants that are bound to the sediment, whereas chronic life-cycle tests are rarely performed as they are time consuming and expensive. Observations during chronic testing of oil-contaminated sediment suggested that there may be a link between the behavior of the marine amphipod Corophium volutator and reduction in growth rate. Behavior tests were performed with six individual amphipods per treatment using sediment spiked with weathered Forties oil with burrowing time, re-emergence from sediment, and activity prior to burrowing as endpoints. Further behavior tests were used to predict the chronic toxicity of sediments spiked with three crude oils each with a dominant unresolved complex mixture of hydrocarbons (UCM). The effect of sediment type on behavior was also investigated. The results suggested that although the b...

Transforming growth factor-beta3 (TGF-β3) and 1α,25-dihydroxyvitamin D3 (1α,25 (OH) 2D3) are essential factors in chondrogenesis and osteogenesis respectively. These factors also play a fundamental role in the developmental processes and the maintenance of skeletal integrity, but their respective direct effects on these processes are not fully understood. Using an organotypic bone rudiment culture system the current study has examined the direct roles the osteotropic factors 1α,25 (OH)2D3 and TGF-β3 exert on the development and modulation of the three dimensional structure of the embryonic femur. Isolated embryonic chick femurs (E11) were organotypically cultured for 10 days in basal media, or basal media supplemented with either 1α,25 (OH) 2D3 (25 nM) or TGF-β3 (5 ng/mL & 15 ng/mL). Analyses of the femurs were undertaken using micro-computed tomography (μCT), histology and immunohistochemistry. 1α,25 (OH)2D3 supplemented cultures enhanced osteogenesis directly in the developing femurs with elevated levels of osteogenic markers such as type 1 collagen. In marked contrast organotypic femur cultures supplemented with TGF-β3 (5 ng/mL & 15 ng/mL) demonstrated enhanced chondrogenesis with a reduction in osteogenesis. These studies demonstrate the efficacy of the ex vivo organotypic embryonic femur culture employed to elucidate the direct roles of these molecules, 1α,25 (OH) 2D3 and TGF-β3 on the structural development of embryonic bone within a three dimensional framework. We conclude that 1α,25(OH)2D and TGF-β3 modify directly the various cell populations in bone rudiment organotypic cultures effecting tissue metabolism resulting in significant changes in embryonic bone growth and modulation. Understanding the roles of osteotropic agents in the process of skeletal development is integral to developing new strategies for the recapitulation of bone tissue in later life.

Humin is the most recalcitrant and least understood fraction of soil organic matter. By definition, humin is that fraction not extracted by traditional aqueous alkaline soil extractants. Here we show thator = 70% of the traditional humin fraction is solubilized when 0.1 M NaOH + 6 M urea and dimethyl sulfoxide (DMSO) + 6% H2SO4 are used in series after conventional extraction. Multidimensional solution-state NMR is applied in this study to gain an understanding of the major constituents present in these "solubilized humin fractions". The spectra indicated strong contributions from five main categories of components, namely, peptides, aliphatic species, carbohydrates, peptidoglycan, and lignin. Diffusion edited spectroscopy indicated that all species are present as macromolecules (or stable aggregate species). Although the distribution of the components is generally similar, peptidoglycan is present at significant levels supporting a higher microbial contribution to humin than to humic and fulvic fractions. The abundance of plant- and microbial-derived materials found does not exclude "humic" materials (e.g., oxidized lignin) or the presence of novel compounds at lower concentrations but suggests that a large proportion of humin is formed from classes of known compounds and parent biopolymers.

The monoaromatic and total aromatic hydrocarbon fractions of two pairs of undegraded and moderately biodegraded crude oils from the Santa Maria basin (California) and the Vienna basin (Austria), all dominated by unresolved complex mixtures, were studied regarding their composition and toxicity towards the feeding rate of the marine mussel Mytilus edulis. Total aromatic and monoaromatic hydrocarbon fractions from sulphur-rich Monterey Formation crude oils were slightly more toxic than the fractions isolated from sulphur-lean Vienna basin oils. The ecotoxicity tests did not show any significant differences in toxicity of aromatic compounds from undegraded or in-reservoir biodegraded crude oils from the same oilfield although some differences in composition were observed. Organic sulphur compounds are suspected to cause the slightly higher toxicity of the aromatic hydrocarbon fractions from the Monterey oils.

FOXP3 has been identified as a key regulator of immune homeostasis. Mutations within the FOXP3 gene result in dysregulated CD4+ T-cell function and elevated cytokine production, leading to lymphoproliferative disease. FOXP3 expression in CD4+ T cells is primarily detected with the CD4+ CD25+ regulatory T-cell population. In humans the protein is detected as a doublet following immunoblot analysis. The lower band of the doublet has been identified as a splice isoform lacking a region corresponding to exon 2. The aim of this study was to investigate whether the splice variant form lacking exon 2 and a new novel splice variant lacking both exons 2 and 7, were functional inhibitors of CD4+ T-cell activation. The data generated showed that full-length FOXP3 and both splice variant forms of the protein were functional repressors of CD4+ T-cell activation.

Background Hydrocoele surgery is the recommended treatment for the common clinical manifestation of lymphatic filariasis in men. This study determined the geographical differences in surgery accessibility, and improvements in the quality of life of patients in Chikwawa district, Malawi. Methods Surgery records from Chikwawa District Hospital (CDH), between 2008 and 2013, were used to map surgery rates by village, spatial dependence by census enumeration area and relationship of distance (kilometres) to CDH. A subset of patients were selected to quantify and compare their physical and socio-economic well-being and level of disability pre- and post-surgery using a standardised questionnaire. Results A total of 476 hydrocoele surgical cases were identified with 260 cases geo-referenced and mapped. A significant negative relationship between village-level surgery rates and distance to CDH (r=−0.137; 95% CI: −0.47 to −0.26) was found, and clusters of enumeration areas with high surgery rates identified around the CDH. Significant improvements in patients' ability to walk and work were found and the overall level of disability was reduced post-surgery. Conclusions Hydrocoele surgery positively impacted on patients, improving their physical and socio-economic output. Surgical services need to scale-up and expand to reach cases that have less access to the best treatment currently available.

The fabrication and assembly of printed circuit boards (PCBs) involves the layering of metals onto an insulating substrate prior to the addition of electronic components. The interfaces between these metal layers are often subject to metal-metal interdiffusion, formation of brittle intermetallics and corrosion, compromising the integrity of the PCB-component solder joints. We have recently developed environmentally sustainable, non-toxic methodologies for application of metal PCB finishes using Deep Eutectic Solvents (DESs) as a novel solvent medium.1-3 Here we focus on the characterisation of metal-metal interfaces in DES media quantifying interfacial structure, layer thickness and morphology using a range of electrochemical and microscopy techniques, including chronoamperometry, cyclic voltammetry (CV) and novel simultaneous thickness electrochemical potential (STEP) measurements coupled with neutron reflectivity (NR.) We have developed the STEP methodology to ascertain some, if not all, of the same information that NR provides. Deposition of Ag and Cu layers has been carried out in different configurations onto Au substrates and characteristic STEP traces are presented for each upon galvanostatic stripping. The STEP methodology facilitates the identification and quantification of each metal layer by galvanostatic dissolution, which is comparable with time resolved NR measurements. The characteristic features of the STEP measurements are discussed including the shape and potential of the dissolution trace. The degree of mixing between the layers is qualitatively determined from E vs. t traces, along with the layer composition and thickness for systems in which layers remain discrete. In summary, we present a new, accessible and inexpensive technique, which probes the thickness, composition and interfacial mixing between layers in PCB metal systems using DES electrolytes and galvanostatic stripping methodologies. [1] A. D. Ballantyne, G. Forrest, M. Goosey, A. Griguceviciene, J. Juodkazyte, R. Kellner, A. Kosenko, R. Ramanauskas, K. S. Ryder, A. Selskis, R. Tarozaite and E. Veninga, Circuit World, 2012, 38(1), 21-29. [2] K. S. Ryder, A. D. Ballantyne, D. Price and T. Perrett, The PCB Magazine, February 2012, 22-30. [3] A. P. Abbott, S. Nandhra, S. Postlethwaite, E. L. Smith, and K. S. Ryder, Phys. Chem. Chem. Phys., 2007, 9, 3735–3743. Figure 1

Electrolytic coatings of metals such as nickel, chromium, zinc, silver and gold are commonly applied as decorative and/or functional coatings. Some of the functional benefits which metal coatings provide are corrosion resistance, wear resistance, electrocatalysis and use in magnetic applications. A range of strategic manufacturing industries are dependent on such electrolytic coatings including electronics, aerospace, automotive and energy. The vast majority of electrolytic coatings are plated from aqueous solutions but stringent health and safety legislation governing transport, use and disposal of feedstock chemicals has driven the challenge for innovation in electrolyte technology that facilitates coatings from less toxic materials and in a manner that is more efficient and more consistent with environmental and process sustainability. We have been motivated by this challenge and have developed a range of novel solvent systems for deposition of metal coatings as alternatives to existing process (e.g. Ni, Zn, Cu, Ag, Ag) as well as novel electrolytes for deposition of reactive metals such as Al. These electrolytes have become known as Deep Eutectic Solvents and have shown utility and promise in a range of metal finishing processes. [1] Whilst the thermodynamics, kinetics, metal ion speciation and growth processes of electrolytic coatings from aqueous solutions have been studied for more than one hundred years, little is currently known about these fundamental aspects of electrochemical deposition from DES media. [2] Here we describe our studies into the electrochemical characterisation of a range of metals in DES electrolytes. In particular, we will focus on aspects of growth and dissolution processes and how these effect the morphology and composition of metal films. We will describe the characterisation of thin metal films in DES media using a range of time resolved techniques including cyclic voltammetry, chronopotentiometry, chronoampertometry, gravimetry (QCM), neutron reflectometry (NR) and surface metrology. The use of in-situ electrochemical NR methods is novel and we will present new data on the growth and dissolution of single metal and bi-layer metals films. The Figure shows time resolved NR data for the electrochemical dissolution of a Cu/Ag bi-layer on a Au substrate. We will also present complimentary data on the characterisation of the same systems using so-called Simultaneous Thickness Electrochemical Potential (STEP) chronopotentiometric methods. We will consider the effect of small-molecule additives on surface morphology and finish (roughness) of electrolytic coatings. [3] Organic and inorganic additives are often added to electroplating solutions to improve surface finish, reduce roughness and promote uniform surface morphology of the coatings. Such additives are usually small molecules and often referred to as brighteners or levellers. However, there have been limited investigations into the effect of such additives on electrodeposition from ionic liquids (ILs) and deep eutectic solvents (DESs). Here we will describe the effect of four additives on electrolytic plating of exemplar metals, nickel and zinc, from an ethyleneglycol based DES; these are nicotinic acid (NA), methylnicotinate (MN), 5,5-dimethyl- hydantoin (DMH) and boric acid (BA). The additives show limited influence on the bulk speciation but have significant influence on the electrochemical behaviour of metal deposition. Small concentrations (ca. 15 mM) of NA and MN show inhibition of metal ion reduction whereas high concentrations of DMH and BA are required for a modest difference in behaviour from the additive free system. NA and MN also show that they significantly alter the nucleation and growth mechanism when compared to the additive free system and those with DMH and BA. Each of the additive systems had the effect of producing brighter and flatter bulk electrodeposits and with increased coating hardness. “Deep Eutectic Solvents (DESs) and their Applications”, Emma L. Smith, Karl S. Ryder and Andrew P. Abbott, Chem. Rev., 2014, 114, 11060. “EXAFS Study into the Speciation of Metal Salts Dissolved in Ionic Liquids and Deep Eutectic Solvents”, Jennifer M. Hartley, Chung-Man Ip, Gregory C.H. Forrest, Kuldip Singh, Stephen J. Gurman, Karl S. Ryder, Andrew P. Abbott and Gero Frisch, Inorg. Chem., 2014, 53(12), 6280. “Bright Metal Coatings from Sustainable Electrolytes: The Effect of Molecular Additives on Electrodeposition of Nickel from a Deep Eutectic Solvent”, Andrew P. Abbott, Andrew Ballantyne, Robert C. Harris, Jamil A. Juma and Karl S. Ryder, Phys. Chem. Chem. Phys., 2017, 19. 3219. Figure 1

Background Managing lymphatic filariasis (LF) morbidity and reducing disability is one of the two primary goals of the Global Programme to Eliminate Lymphatic Filariasis. However, in order to achieve this, the geographical distribution of LF morbidity needs to be better estimated. Methods All cases of lymphoedema within a single health centre catchment area (pop. 42 000) in the southern region of Malawi were examined. Maps of lymphoedema burden were produced and trends in patient demographics, severity of lymphoedema (Dreyer staging) and health-seeking behaviour were explored. The number of lymphoedema cases was compared with records maintained by the Ministry of Health, Malawi. Results A total of 69 lymphoedema cases were identified (32 per 10 000 population), of which 48 (70%) were female and 21 (30%) male. The majority of cases (51/69) had Dreyer stage 2–3, and almost all (65/69) had experienced acute attacks as a result of their lymphoedema. This burden was much greater than that estimated by Ministry of Health (33 cases). Conclusions Current case detection methods underestimate the burden of lymphoedema in Malawi. There is a continued need to develop new LF morbidity identification and surveillance approaches to ensure that future morbidity management strategies are effectively targeted.

Eating disorders in young people with type 1 diabetes mellitus confer additional health risks beyond those conferred by the disease itself. Risk factors for developing eating disorders are poorly understood.The current study aimed to examine risk factors for eating disturbance in young people with type 1 diabetes mellitus. Both diabetes specific risk factors, such as body mass index (BMI), glycaemic control and diabetes-related conflict, and also more general risk factors such as dysfunctional perfectionism and low self-esteem were assessed.Fifty young people aged 14-16 and their primary caregiver were asked to complete interviews and questionnaires about their eating attitudes and behaviours, dysfunctional perfectionism, self-esteem, family conflict, and general mental health symptoms. Recent weight and height and glycaemic control were extracted from the medical file.Different factors distinguished those young people who displayed eating disorder attitudes from those who did not (higher BMI-z, poorer glycaemic control, and lower self-esteem) and those young people who displayed eating disorder behaviour from those who did not (lower self-esteem and higher diabetes-related family conflict).The results of the current study suggest that there might be different factors associated with eating disorders (ED) attitudes and ED behaviours, but that food/eating-related factors, family factors, and intra-personal factors are all important. Furthermore there are some gender differences in the presence of ED attitudes and behaviours and preliminary evidence that higher body mass indexes (BMIs) impact on girls more than they do on boys.

A recent study has shown that some monoaromatic hydrocarbon constituents of the so-called unresolved complex mixtures (UCMs) or gas chromatographic humps, which are widespread in the marine environment, are toxic to the mussel Mytilus edulis. Here we describe the synthesis and toxicological assessment of 6-cyclohexyltetralin, 7-cyclohexyl-1-methyltetralin, and 7-cyclohexyl-1-n-propyltetralin, which contain structural features consistent with some monoaromatic UCM hydrocarbons. The compounds were all toxic to M. edulis when measured in the assay used previously to determine the toxicity of a monoaromatic UCM. The aqueous solubilities of the hydrocarbons in fresh and seawater at different temperatures were determined and found to range from about 10 to 110 microg/L (10-60 microg/L in seawater at 15 degrees C). Further studies of the aromatic UCM composition of a wide range of oils and oil residues are required to determine whether such alkylated compounds as 7-cyclohexyl-1-methyltetralin and 7-cyclohexyl-1-n-propyltetralin or their analogues are widespread in oils. If these aromatic compounds prove to be important in UCMs, toxicity experiments should be conducted with other biological end points and monitoring studies of pollutant hydrocarbons should probably include measurement of aromatic UCM hydrocarbons.

Gas chromatographic analysis of the hydrocarbons of environmental samples often reveals that unresolved complex mixtures (UCMs) or gas chromatographic "humps" of aliphatic and aromatic hydrocarbons are most abundant--yet little consideration seems to have been given to the possible toxicological impacts of hydrocarbon "humps". Here we show, using a well-accepted bioassay, that monoaromatic components of a UCM of hydrocarbons from a crude oil elicit a sublethal toxic response in a typical marine pollution indicator organism (the mussel, Mytilus edulis). Furthermore, coastal U.K. mussels shown previously to have unexplained impaired health contained high concentrations of UCMs, including monoaromatic UCMs. These findings may have important implications for our understanding of the toxicological sublethal effects of oil residues in the environment. Given the relatively resistant nature of UCM hydrocarbons, the effects of both acute oil spills and more chronic discharges may need further consideration.

Objectives To assess the efficacy of a public health intervention to reduce blood mercury (Hg) concentration levels in pregnant Bermudian women. Methods In 2003, we conducted a study entitled “Prenatal exposure of the Bermudian Population to Environmental Contaminants” which provided Bermuda’s first baseline data on prenatal exposure to several environmental contaminants, including Hg. The mean Hg concentration from 42 healthy newborns measured in umbilical cord blood was 41.3 nmol/L, ranging from 5–160 nmol/L. This concentration was much higher than expected, being approximately 8 times the general levels found in Canada and the U.S. Furthermore, we estimated that 85% of total Hg measured was in the form of methylmercury (MeHg), indicating that seafood consumption was the primary source of Hg exposure during pregnancy in Bermuda. Locally sourced seafood was identified as the most significant possible contributory source of Hg exposure. In 2005 the authors began a complementary research programme to study the levels of Hg in local commercial fish species. Coming out of this research were specific local fish consumption guidelines issued by the Department of Health advising pregnant women to avoid those local fish species found to be high in Hg while still encouraging consumption of fish species having lower Hg levels. Results In 2010, under another research initiative, we returned to Bermuda to carry out another evaluation of Hg in human blood. Hg was measured in the blood of 49 pregnant women. The arithmetic mean Hg blood concentration was 6.6 nmol/L and the geometric mean 4.2 nmol/L. The maximum concentration found was 24 nmol/L. Conclusions Hg exposure of Bermudian pregnant women has dropped significantly by a factor of around 5 since the foetal cord blood study in 2003.

Enhancement and application of our understanding of skeletal developmental biology is critical to developing tissue engineering approaches to bone repair. We propose that use of the developing embryonic femur as a model to further understand skeletogenesis, and the effects of key differentiation agents, will aid our understanding of the developing bone niche and inform bone reparation. We have used a three-dimensional organotypic culture system of embryonic chick femora to investigate the effects of two key skeletal differentiation agents, parathyroid hormone (PTH) and parathyroid hormone-related protein (PTHrP), on bone and cartilage development, using a combination of microcomputed tomography and histological analysis to assess tissue formation and structure, and cellular behavior. Stimulation of embryonic day 11 (E11) organotypic femur cultures with PTH and PTHrP initiated osteogenesis. Bone formation was enhanced, with increased collagen I and STRO-1 expression, and cartilage was reduced, with decreased chondrocyte proliferation, collagen II expression, and glycosaminoglycan levels. This study demonstrates the successful use of organotypic chick femur cultures as a model for bone development, evidenced by the ability of exogenous bioactive molecules to differentially modulate bone and cartilage formation. The organotypic model outlined provides a tool for analyzing key temporal stages of bone and cartilage development, providing a paradigm for translation of bone development to improve scaffolds and skeletal stem cell treatments for skeletal regenerative medicine.

Understanding the structural development of embryonic bone in a three dimensional framework is fundamental to developing new strategies for the recapitulation of bone tissue in latter life. We present an innovative combined approach of an organotypic embryonic femur culture model, microcomputed tomography (μCT) and immunohistochemistry to examine the development and modulation of the three dimensional structures of the developing embryonic femur. Isolated embryonic chick femurs were organotypic (air/liquid interface) cultured for 10 days in either basal, chondrogenic, or osteogenic supplemented culture conditions. The growth development and modulating effects of basal, chondrogenic, or osteogenic culture media of the embryonic chick femurs was investigated using μCT, immunohistochemistry, and histology. The growth and development of noncultured embryonic chick femur stages E10, E11, E12, E13, E15, and E17 were very closely correlated with increased morphometric indices of bone formation as determined by μCT. After 10 days in the organotpyic culture set up, the early aged femurs (E10 and E11) demonstrated a dramatic response to the chondrogenic or osteogenic culture conditions compared to the basal cultured femurs as determined by a change in μCT morphometric indices and modified expression of chondrogenic and osteogenic markers. Although the later aged femurs (E12 and E13) increased in size and structure after 10 days organotpypic culture, the effects of the osteogenic and chondrogenic organotypic cultures on these femurs were not significantly altered compared to basal conditions. We have demonstrated that the embryonic chick femur organotpyic culture model combined with the μCT and immunohistochemical analysis can provide an integral methodology for investigating the modulation of bone development in an ex vivo culture setting. Hence, these interdisciplinary techniques of μCT and whole organ bone cultures will enable us to delineate some of the temporal, structural developmental paradigms and modulation of bone tissue formation to underpin innovative skeletal regenerative technology for clinical therapeutic strategies in musculoskeletal trauma and diseases.

Immunohistochemical detection of cross-linked fibrillar collagens in mineralized tissues is much desired for exploring the mechanisms of biomineralization in health and disease. Mineralized teeth are impossible to section when embedded in conventional media, thus limiting on-section characterization of matrix proteins by immunohistochemistry. We hypothesized that by using an especially formulated acrylic resin suitable for mineralized dental tissues, not only sectioning of teeth would be possible, but also our recently developed immunofluorescence labeling technique would be amenable to fully calcified tissues for characterization of dentinal fibrillar collagens, which remains elusive. The hypothesis was tested on fixed rodent teeth embedded in Technovit 9100 New (R). It was possible to cut thin (1 mu m) sections of mineralized teeth, and immunofluorescence characterization of cross-linked type I fibrillar collagen was selected due to its abundance in dentine. Decalcified samples of teeth embedded in paraffin wax were also used to compare immuno-labeling from either method using the same immunoreagents in equivalent concentrations. In the decalcified tissue sections, type I collagen labeling in the dentine along the tubules was "patchy" and the signal in the predentine was very weak. However, enhanced signal in mineralized samples with type I collagen was detected not only in the predentine but also at the limit between intertubular dentine, within the elements of the enamel organ and subgingival stroma. This report offers advances in sectioning mineralized dental tissues and allows the application of immunofluorescence not only for on-section protein detection but importantly for detecting cross-linked fibrous collagens in both soft and mineralized tissue sections. Microsc. Res. Tech. 73:741-745, 2010. (C) 2009 Wiley-Liss, Inc.

In this paper, we describe the first example of a sustained galvanic coating deposited on a surface from a non-aqueous liquid. We present the surface characterization of electroless silver deposits on copper substrates from a solution of Ag(+) ions in an ionic liquid based on a choline chloride (ChCl) eutectic. Through a study of these deposits and the mechanism of formation using acoustic impedance spectroscopy (QCM), probe microscopy (AFM) and electron microscopy (SEM/EDX), we demonstrate that sustained growth of the silver deposit is facilitated by the porous nature of the silver. This is in contrast to the dip-coating reaction of silver ions in aqueous media, where the reaction stops when surface coverage is reached. Electroless silver deposits of up to several microns have been obtained by dip coating in ionic liquids without the use of catalysts of strong inorganic acids.

A combination of XPS, reflectance infrared spectroscopy, and neutron reflectivity measurements has been used to probe the spatial and global extents to which carboxylic acid motifs in electrodeposited conducting polymer films can be functionalized by reaction with vapor phase reagents (a carbodiimide together with trifluoroethanol) with the goal of controlling hydrophobicity. Across a range of polymer deposition and reaction temperatures, neutron reflectivity showed that, surprisingly, functionalization of the polymer matrix at depths >5-10 nm into the polymer film was always significantly lower than at the exposed surface. The most efficient functionalization was found to occur when a low-density polymer matrix was prepared by elution of motifs cleaved from the polymer by base hydrolysis. Finally, when trifluoroethanol functionalization was performed, the macroscopic property of hydrophobicity was related to the surface, internal microstructure, and composition of the reacted films as elucidated by the above combination of probes.

Honey is used as a therapy to aid wound healing. Previous data indicate that honey can stimulate cytokine production from human monocytes. The present study further examines this phenomenon in manuka honey. As inflammatory cytokine production in innate immune cells is classically mediated by pattern recognition receptors in response to microorganisms, bacterial contamination of honey and the effect of blocking TLR2 and -4 on stimulatory activity were assessed. No vegetative bacteria were isolated from honey; however, bacterial spores were cultured from one-third of samples, and low levels of LPS were detected. Blocking TLR4 but not TLR2 inhibited honey-stimulated cytokine production significantly. Cytokine production did not correlate with LPS levels in honey and was not inhibited by polymyxin B. Further, the activity was reduced significantly following heat treatment, indicating that component(s) other than LPS are responsible for the stimulatory activity of manuka honey. To identify the component responsible for inducing cytokine production, honey was separated by molecular weight using microcon centrifugal filtration and fractions assessed for stimulatory activity. The active fraction was analyzed by MALDI-TOF mass spectroscopy, which demonstrated the presence of a number of components of varying molecular weights. Additional fractionation using miniaturized, reverse-phase solid-phase extraction resulted in the isolation of a 5.8-kDa component, which stimulated production of TNF-α via TLR4. These findings reveal mechanisms and components involved in honey stimulation of cytokine induction and could potentially lead to the development of novel therapeutics to improve wound healing for patients with acute and chronic wounds.

Exposure of marine mussels (Mytilus edulis) to an unresolved complex mixture (UCM) of aromatic hydrocarbons isolated from a crude oil has been shown to reduce their feeding rate by 40%. The present study was undertaken to determine whether UCMs bioaccumulated by mussels in the field are also toxic. The feeding rate of mussels derived from polluted sites increased when they were placed in clean water, pointing to a loss of toxic agents from the tissues. At the end of the depuration period, water in which mussels from an oil-polluted site had been held contained a UCM. Steam-distillation extracts of the tissues of mussels taken from several polluted sites were shown to be highly toxic to the feeding activity of juvenile mussels. The tissues of mussels from these sites contained UCMs. Nontoxic steam distillates from clean mussels did not. Steam-distillation extracts of mussels from an oil-polluted site were fractionated by normal-phase high-performance liquid chromatography. A fraction, largely comprising a "monoaromatic" UCM, reduced the feeding rate of juvenile mussels by 70%. Two later-eluting fractions containing aromatic UCMs also produced smaller depressions in feeding rate. These results support our contention that some aromatic UCM hydrocarbons constitute a forgotten pollutant burden in the marine environment.

A small group of patients with manifest Huntington's disease (HD) were followed longitudinally to assess cognitive decline in relation to time from disease diagnosis. This article looks at performance on a range of computerised and pencil and paper cognitive tasks in patients 5 years post diagnosis, who were assessed annually for a 5 year follow up period. The almost universal cognitive decline reported in other longitudinal studies of HD was not replicated in this study. It was proposed that longitudinal follow up in HD is complicated by the varying degree to which different tasks are able to withstand repeated administration; a finding which would have significant implications on study design in future trials of cognitive enhansing interventions.

The acute toxicity of the oil dispersant Corexit 9527 reported in the literature is highly variable. No peer-reviewed data exist for Superdispersant-25 (SD-25). This study compares the toxicity of the two dispersants to a range of marine species representing different phyla occupying a wide range of niches: The marine sediment-dwelling amphipod Corophium volutator (Pallas), the common mussel Mytilus edulis (L.), the symbiotic snakelocks anemone Anemonia viridis (Forskål), and the seagrass Zostera marina (L.). Organisms were exposed to static dispersant concentrations for 48-h and median lethal concentration (LC50), median effect concentration (EC50), and lowest-observable-effect concentration (LOEC) values obtained. The sublethal effects of 48-h exposures and the ability of species to recover for up to 72 h after exposure were quantified relative to the 48-h endpoints. Results indicated that the anemone lethality test was the most sensitive with LOECs of 20 ppm followed by mussel feeding rate, seagrass photosynthetic index and amphipod lethality, with mussel lethality being the least sensitive with LOECs of 250 ppm for both dispersants. The results were consistent with current theory that dispersants act physically and irreversibly on the respiratory organs and reversibly, depending on exposure time, on the nervous system. Superdispersant-25 was found overall to be less toxic than Corexit 9527 and its sublethal effects more likely to be reversible following short-term exposure.