Search

Acrocomia spp., a genus of wild-growing palms in the neotropics, is rapidly gaining interest as a promising multipurpose crop. Diverse products can be derived from various components of the palm, the oils being of highest interest. Acrocomia shows similar oil yield and fatty acid composition to the African oil palm (Elaeis guineensis). It is, however, able to cope with a wider range of environmental conditions, including temporary water scarcity and lower temperatures, thus potentially a more sustainable alternative to its tropical counterpart. Acrocomia’s research history is recent compared to other traditional crops and thus knowledge gaps, uncertainty, and challenges need to be addressed. This review attempts to assess the acrocomia’s preparedness for cultivation by highlighting the state-of-the-art in research and identifying research gaps. Based on a systematic literature search following a value web approach, it (a) provides a comprehensive overview of research topics, (b) shows the development of publication activities over time and the drivers of this development, and (c) compiles main findings to assess the acrocomia’s preparedness for commercial cultivation. Our results confirm its multipurpose characteristic as a potential feedstock for manifold sectors. Research has continued to increase over the last decade, especially on A. aculeata and is driven by the interest in bioenergy. Increasing knowledge on botany has contributed to understanding the genetic diversity and genus-specific biology. This has enabled applied research on seed germination and propagation toward domestication and initial plantation activities, mostly in Brazil. Main research gaps are associated with genotype–environment interaction, planting material, crop management, and sustainable cropping systems. Overall, we conclude that acrocomia is at an early phase of development as an alternative and multipurpose crop and its up-scaling requires the integration of sustainability strategies tailored to location-based social-ecological conditions.

Electron microscopy (EM) is widely used for nanoparticle (NP) sizing. Following an initial assessment of two sample preparation protocols described in the current literature as “unperturbed”, we found that neither could accurately measure the size of NPs featuring a broad size distribution, e.g., aggregates. Because many real-world NP samples consist of aggregates, this finding was of considerable concern. The data showed that the protocols introduced errors into the measurement by either inducing agglomeration artefacts or providing a skewed size distribution towards small particles (skewing artefact). The focus of this work was to develop and apply a mathematical refinement to correct the skewing artefact. This refinement provided a much improved agreement between EM and a reference methodology, when applied to the measurement of synthetic amorphous silica NPs. Further investigation, highlighted the influence of NP chemistry on the refinement. This study emphasised the urgent need for greater and more detailed consideration regarding the sample preparation of NP aggregates to routinely achieve accurate measurements by EM. This study also provided a novel refinement solution applicable to the size characterisation of silica and citrate-coated gold NPs featuring broad size distributions. With further research, this approach could be extended to other NP types.

Novel applications of nanotechnology may lead to the release of engineered nanoparticles (ENPs), which result in concerns over their potential environmental hazardous impact. It is essential for the research workers to be able to quantitatively characterise ENPs in the environment and subsequently to assist the risk assessment of the ENPs. This study hence explored the application of nanoparticle tracking system (NTA) to quantitatively describe the behaviour of the ENPs in natural sediment-water systems. The NTA allows the measurement of both particle number concentration (PNC) and particle size distribution (PSD) of the ENPs. The developed NTA method was applied to a range of gold and magnetite ENPs with a selection of surface properties. The results showed that the positively-charged ENPs interacted more strongly with the sediment than neutral and negatively-charged ENPs. It was also found that the citrate coated Au ENPs had a higher distribution percentage (53%) than 11-mercaptoundecanoic acid coated Au ENPs (20%) and citrate coated magnetite ENPs (21%). The principles of the electrostatic interactions between hard (and soft) acids and bases (HSAB) are used to explain such behaviours; the hard base coating (i.e. citrate ions) will interact more strongly with hard acid (i.e. magnetite) than soft acid (i.e. gold). The results indicate that NTA is a complementary method to existing approaches to characterise the fate and behaviour of ENPs in natural sediment.

Available measurement methods for nanomaterials are based on very different measurement principles and hence produce different values when used on aggregated nanoparticle dispersions. This paper provides a solution for relating measurements of nanomaterials comprised of nanoparticle aggregates determined by different techniques using a uniform expression of a mass equivalent diameter (MED). The obtained solution is used to transform into MED the size distributions of the same sample of synthetic amorphous silica (nanomaterial comprising aggregated nanoparticles) measured by six different techniques: scanning electron microscopy in both high vacuum (SEM) and liquid cell setup (Wet-SEM); gas-phase electrophoretic mobility molecular analyzer (GEMMA); centrifugal liquid sedimentation (CLS); nanoparticle tracking analysis (NTA); and asymmetric flow field flow fractionation with inductively coupled plasma mass spectrometry detection (AF4-ICP-MS). Transformed size distributions are then compared between the methods and conclusions drawn on methods' measurement accuracy, limits of detection and quantification related to the synthetic amorphous silca's size. Two out of the six tested methods (GEMMA and AF4-ICP-MS) cross validate the MED distributions between each other, providing a true measurement. The measurement accuracy of other four techniques is shown to be compromised either by the high limit of detection and quantification (CLS, NTA, Wet-SEM) or the sample preparation that is biased by increased retention of smaller nanomaterials (SEM). This study thereby presents a successful and conclusive cross-method comparison of size distribution measurements of aggregated nanomaterials. The authors recommend the uniform MED size expression for application in nanomaterial risk assessment studies and clarifications in current regulations and definitions concerning nanomaterials.

The potential impact of nanomaterials on the environment and on human health has already triggered legislation requiring labelling of products containing nanoparticles. However, so far, no validated analytical methods for the implementation of this legislation exist. This paper outlines a generic approach for the validation of methods for detection and quantification of nanoparticles in food samples. It proposes validation of identity, selectivity, precision, working range, limit of detection and robustness, bearing in mind that each “result” must include information about the chemical identity, particle size and mass or particle number concentration. This has an impact on testing for selectivity and trueness, which also must take these aspects into consideration. Selectivity must not only be tested against matrix constituents and other nanoparticles, but it shall also be tested whether the methods apply equally well to particles of different suppliers. In trueness testing, information whether the particle size distribution has changed during analysis is required. Results are largely expected to follow normal distributions due to the expected high number of particles. An approach of estimating measurement uncertainties from the validation data is given.

Imaging and characterization of engineered nanoparticles (ENPs) in water, soils, sediment and food matrices is very important for research into the risks of ENPs to consumers and the environment. However, these analyses pose a significant challenge as most existing techniques require some form of sample manipulation prior to imaging and characterization, which can result in changes in the ENPs in a sample and in the introduction of analytical artefacts. This study therefore explored the application of a newly designed instrument, the atmospheric scanning electron microscope (ASEM), which allows the direct characterization of ENPs in liquid matrices and which therefore overcomes some of the limitations associated with existing imaging methods. ASEM was used to characterize the size distribution of a range of ENPs in a selection of environmental and food matrices, including supernatant of natural sediment, test medium used in ecotoxicology studies, bovine serum albumin and tomato soup under atmospheric conditions. The obtained imaging results were compared to results obtained using conventional imaging by transmission electron microscope (TEM) and SEM as well as to size distribution data derived from nanoparticle tracking analysis (NTA). ASEM analysis was found to be a complementary technique to existing methods that is able to visualize ENPs in complex liquid matrices and to provide ENP size information without extensive sample preparation. ASEM images can detect ENPs in liquids down to 30 nm and to a level of 1 mg L(-1) (9×10(8) particles mL(-1) , 50 nm Au ENPs). The results indicate ASEM is a highly complementary method to existing approaches for analyzing ENPs in complex media and that its use will allow those studying to study ENP behavior in situ, something that is currently extremely challenging to do.

Published literature has been reviewed in order to (a) explore the (potential) applications of nanotechnology in pesticide formulation, (b) identify possible impacts on environmental fate, and (c) analyze the suitability of current exposure assessment procedures to account for the novel properties of nanopesticides within the EU regulatory context. The term nanopesticide covers a wide variety of products and cannot be considered to represent a single category. Many nanoformulations combine several surfactants, polymers, and metal nanoparticles in the nanometer size range. The aims of nanoformulations are generally common to other pesticide formulations, these being to increase the apparent solubility of poorly soluble active ingredients, to release the active ingredient in a slow/targeted manner and/or to protect against premature degradation. Nanoformulations are thus expected to (a) have significant impacts on the fate of active ingredients and/or (b) introduce new ingredients for which the environmenta...

Engineered nanomaterials (ENMs) are increasingly being used in the food industry. In order to assess the efficacy and the risks of these materials, it is essential to have access to methods that not only detect the nanomaterials, but also provide information on the characteristics of the materials (e.g., size and shape). This review presents an overview of electron microscopy (EM)-based methods that have been, or have the potential to be, applied to imaging ENMs in foodstuffs. We provide an overview of approaches to sample preparation, including drying, chemical treatment, fixation and cryogenic methods. We then describe standard and non-standard EM-based approaches that are available for imaging prepared samples. Finally, we present a strategy for selecting the most appropriate method for a particular foodstuff.

The increasing industrial production of engineered nanoparticles (ENPs) raises concern over their safety to humans and the environment. There is a lack of knowledge regarding the environmental fate and impact of ENPs and in situ methods are needed to investigate e.g. nanoparticle aggregation and adsorption in the media of concern such as water, sediment and soil. In this study, the application of wet scanning electron microscopy (WetSEM) technology in combination with energy dispersive x-ray spectroscopy (EDS) to visualise and elementally identify metal and metal oxide nanoparticles (Au, TiO(2), ZnO and Fe(2)O(3)) under fully liquid conditions in distilled and lake water as well as in a soil suspension has been investigated. WetSEM capsules comprise an electron transparent membrane enabling the imaging and EDS analysis of liquid samples. Results are compared with conventional SEM images and show that WetSEM/EDS is a promising complementary tool for the in situ investigation of ENPs and their aggregates in natural matrices. In combination with other analytical tools (e.g. HDC- or FFF-ICP-MS, DLS), WetSEM could help to provide a better understanding of the fate and behaviour of ENPs in the environment.

There is an increasing concern over the safety of engineered nanoparticles (ENPs) to humans and the environment and it is likely that the environmental risks of these particles will have to be tested under regulatory schemes such as REACH. Due to their unique properties and the fact that their detection and characterisation in complex matrices is challenging, existing analytical methods and test approaches for assessing environmental risk may not be appropriate for ENPs. In this article we discuss the challenges associated with the testing of ENPs to generate data on persistence, mobility, bioavailability and ecotoxicity in the environment. It is essential that careful consideration is given to the selection of the test material, the test system (including test vessels and study media) and the test exposure conditions. During a study it is critical that not only the concentration of the ENP is determined but also its characteristics (e.g. size, shape, degree of aggregation and dissolution). A range of analytical techniques is available including microscopy-based approaches (e.g transmission and scanning electron microscopy), dynamic light scattering, and size separation approaches (e.g. field flow fractionation and hydrodynamic chromatography) coupled to detection methods such as inductively coupled plasma MS. All of these have their disadvantages: some are unable to distinguish between ENPs and natural interferences; some techniques require sample preparation approaches that can introduce artefacts; and others are complex and time-consuming. A combination of techniques is therefore needed. Our knowledge in this area is still limited, and co-ordinated research is required to gain a better understanding of the factors and processes affecting ENP fate and effects in the environment as well as to develop more usable, robust and sensitive methods for characterisation and detection of ENPs in environmental systems.

This study explored the potential for engineered nanoparticles (ENPs) to contaminate the UK drinking water supplies and established the significance of the drinking water exposure route compared to other routes of human exposure. A review of the occurrence and quantities of ENPs in different product types on the UK market as well as release scenarios, their possible fate and behaviour in raw water and during drinking water treatment was performed. Based on the available data, all the ENPs which are likely to reach water sources were identified and categorized. Worst case concentrations of ENPs in raw water and treated drinking water, using a simple exposure model, were estimated and then qualitatively compared to available estimates for human exposure through other routes. A range of metal, metal oxide and organic-based ENPs were identified that have the potential to contaminate drinking waters. Worst case predicted concentrations in drinking waters were in the low- to sub-µg/l range and more realistic estimates were tens of ng/l or less. For the majority of product types, human exposure via drinking water was predicted to be less important than exposure via other routes. The exceptions were some clothing materials, paints and coatings and cleaning products containing Ag, Al, TiO2, Fe2O3 ENPs and carbon-based materials.

In addition to factors such as consumer acceptance and the development of cost-effective applications of the developed products, the advancement of nanotechnology in food and food-related goods is dependent on comprehensive regulation being in place. However, for legislators to produce meaningful legislation, they will require access to high quality toxicological data, underpinned by accurate and robust analytical science. The opportunities presented by the unique properties of materials produced on a ‘nano’ scale also present a significant challenge to the scientists charged with their characterization and measurement. This is because most analytical techniques and procedures have been developed for use with materials in their ‘bulk’ state, and have been found lacking when applied to materials in a ‘nano’ form. Hence, one of the key goals of the analytical community is to develop and validate robust, analytical approaches for the detection, characterization and quantification of nanomaterials (NMs) in complex matrices, i.e. foodstuffs. This chapter therefore focuses on the characterization of engineered NMs in foodstuffs and food contact materials, and will cover approaches for sample preparation prior to analysis; the analytical techniques available for NM detection and characterization in food matrices; and the appropriateness of various QA and QC methodologies. Specific challenges associated with NM detection and analysis in food, beverages and nutraceuticals are also discussed.

Electron microscopy is a recognized standard tool for nanomaterial characterization, and recommended by the European Food Safety Authority for the size measurement of nanomaterials in food. Despite this, little data have been published assessing the reliability of the method, especially for size measurement of nanomaterials characterized by a broad size distribution and/or added to food matrices. This study is a thorough investigation of the measurement uncertainty when applying electron microscopy for size measurement of engineered nanomaterials in foods. Our results show that the number of measured particles was only a minor source of measurement uncertainty for nanomaterials in food, compared to the combined influence of sampling, sample preparation prior to imaging and the image analysis. The main conclusion is that to improve the measurement reliability, care should be taken to consider replications and matrix removal prior to sample preparation.

PolyHIPEs of ethylene glycol dimethacrylate (EGDMA) and styrene/divinylbenzene were prepared by polymerization of water-in-oil high internal phase emulsions (HIPEs) within high pressure liquid chromatography (HPLC) columns. The columns were incorporated into a HPLC system affixed to an inductively-coupled plasma mass spectrometer, and their potential for the separation of engineered nanoparticles was investigated. Triplicate injections of 5 and 10 nm gold particles injected onto a poly(styrene-co-divinylbenzene) polyHIPE column produced an average difference in retention time of 135 s. On a poly(EGDMA) column, triplicate injections of dysprosium containing polystyrene particles of 52 and 155 nm produced a difference in retention time of 8 s. In both cases the smaller particles eluted from the column first. Comparison, using scanning electron microscopy, of the polyHIPE columns after the separations, against freestanding monoliths produced from the same HIPEs, revealed no apparent change in the internal porous structure of the polyHIPEs. © 2015 The Authors Journal of Applied Polymer Science Published by Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014, 132, 41229.

Nanopesticides or nano plant protection products represent an emerging technological development that, in relation to pesticide use, could offer a range of benefits including increased efficacy, durability, and a reduction in the amounts of active ingredients that need to be used. A number of formulation types have been suggested including emulsions (e.g., nanoemulsions), nanocapsules (e.g., with polymers), and products containing pristine engineered nanoparticles, such as metals, metal oxides, and nanoclays. The increasing interest in the use of nanopesticides raises questions as to how to assess the environmental risk of these materials for regulatory purposes. Here, the current approaches for environmental risk assessment of pesticides are reviewed and the question of whether these approaches are fit for purpose for use on nanopesticides is addressed. Potential adaptations to existing environmental risk assessment tests and procedures for use with nanopesticides are discussed, addressing aspects such as analysis and characterization, environmental fate and exposure assessment, uptake by biota, ecotoxicity, and risk assessment of nanopesticides in aquatic and terrestrial ecosystems. Throughout, the main focus is on assessing whether the presence of the nanoformulation introduces potential differences relative to the conventional active ingredients. The proposed changes in the test methodology, research priorities, and recommendations would facilitate the development of regulatory approaches and a regulatory framework for nanopesticides.

For the first time, regulatory protocols defined in the OECD guidelines were applied to determine the fate properties of a nanopesticide in two agricultural soils with contrasting characteristics. The nanoformulation studied had no effect on the degradation kinetics of atrazine indicating that (1) the release of atrazine from the polymer nanocarriers occurred rapidly relative to the degradation kinetics (half-lives 36–53 days) and/or that (2) atrazine associated with the nanocarriers was subject to biotic or abiotic degradation. Sorption coefficients, derived from a batch and a centrifugation technique at a realistic soil-to-solution ratio, were higher for the nanoformulated atrazine than for the pure active ingredient. Results indicate that the nanoformulation had an effect on the fate of atrazine. However, since the protocols applied were designed to assess solutes, conclusions about the transport of atrazine loaded onto the nanocarriers should be made extremely cautiously. The centrifugation method applied over time (here over 7 days) appears to be a useful tool to indirectly assess the durability of nanopesticides under realistic soil-to-solution ratios and estimate the period of time during which an influence on the fate of the active ingredient may be expected. More detailed investigations into the bioavailability and durability of nanopesticides are necessary and will require the development of novel methods suitable to address both the “nano” and “organic” characteristics of polymer-based nanopesticides.

A systematic literature search identified publications relevant to repeated (94 papers) and mixture exposure (152 papers) of aquatic organisms. Limited data are available on pulsed exposure of pesticides and reports on repeated pulse or fluctuating exposures are even rarer. Although there has been recognition of the need for more realistic testing scenarios these lack a systematic approach. Among the reports on pesticide mixtures most of the plant studies evaluated interactions between mixtures of herbicides or fungicides / algicides. A small number of reports evaluated the effects on invertebrates of mixtures of pesticides with the same mode of action and these showed additive toxicity. More studies in invertebrates assessed interactions between pesticides with different modes of action including herbicide/ insecticide combinations some of which showed toxicity greater than additive. The majority of studies in fish have investigated the toxicity of insecticide mixtures. The toxicity of most combinations was at most additive, with a small number antagonistic. A small number of papers reported mixture effects of pesticides in amphibians; more than additive effects were observed with atrazine combinations. Simple models can be used to determine the additive toxicity of pesticide mixtures based on concentration addition or independent action. More complex models to determine the scale of synergy are speciesand dose-dependent and rely on an understanding of the interactions involved. Further work is required to develop a greater understanding of the species, dose and time dependence of exposure to pesticide mixtures in which more complex interactions occur. In the interim, in the majority of cases the increase in toxicity is no greater than 3 fold when compared to additive, the deviation from additivity decreases with the increasing number of components in the mixture, and where no known interactions have been reported this may be an appropriate factor to include in risk assessments.

While nanotechnology offers a number of benefits to the food sector, the use of nanoparticles (NPs) in food may also cause risks. In order to understand the risks of NPs, it is essential to have access to good quality analytical methods that are able to detect and characterize NPs in a wide range of media. This chapter provides an overview of the characteristics of NPs that will need to be measured in food materials and describes methods that are available to derive these characteristics. The advantages and drawbacks of each method are described and the challenges for the development of new methods for NP analysis in food materials are highlighted.

There is increasing concern over the risks of nanoparticles to humans and the environment, but little is known about the properties of the nanoparticulate mineral filters, such as titanium dioxide and zinc oxide, in sunscreens. There is an urgent need to develop methods for characterizing nanoparticles in (NPs) such products to provide data for human and environmental risk assessments. This study explored three methods (transmission electron microscopy [TEM], conventional scanning electron microscopy [SEM], and wet-scanning electron microscopy [WetSEM]) for characterizing NPs in sunscreens. Our results showed that these products contained titanium dioxide and zinc oxide particles in the nanometer range; thus, it is likely that consumers and the environment are exposed to engineered NPs through the use of these products. Further, we found that the combination of all three microscopy methods provided the most comprehensive information on size-related properties, which are crucial parameters for risk assessment of NPs in wet matrices.

Biglycan, a small leucine-rich proteoglycan, is essential for scar formation and preservation of hemodynamic function after myocardial infarction, as shown in biglycan-knockout mice. Because of this important role in cardiac pathophysiology, we aimed to identify regulators of biglycan expression and posttranslational modifications in cardiac fibroblasts. Cardiac fibroblasts were isolated from neonatal Wistar-Kyoto rats and used in the first passage. Expression of biglycan was analyzed after metabolic labeling with [(35)S]-sulfate by SDS-polyacrylamide gel electrophoresis and molecular sieve chromatography; mRNA expression was examined by Northern analysis and real-time RT-PCR. Serum, thrombin, transforming growth factor beta1 (TGFbeta 1) and platelet-derived growth factor BB (PDGF-BB) strongly increased [(35)S]-labeled proteoglycan levels. Tumor necrosis factor alpha further increased the stimulatory effect of PDGF-BB. PDGF-BB increased glycosaminoglycan (GAG) chain length as shown by molecular sieve chromatography after beta-elimination to release GAG chains. Nitric oxide was the only negative regulator of biglycan as evidenced by marked downregulation in response to DETA-NO ((Z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate), a long acting nitric oxide donor and SNAP (S-nitroso-N-acetyl-l,l-penicillamine), which completely inhibited PDGF-BB-induced secretion of total [(35)S]-labeled proteoglycans and biglycan mRNA expression. Of note, the molecular weight of biglycan GAG chains was even further increased by NO donors compared to control and PDGF-BB stimulation. The current results suggest that in cardiac fibroblasts, biglycan is induced by a variety of stimuli including serum, thrombin and growth factors such as PDGF-BB and TGFbeta1. This response is counteracted by NO and enhanced by TNFalpha. Interestingly, both up- and downregulation were associated with posttranslational increase of GAG chain length.

Nanotechnology is developing rapidly and, in the future, it is expected that increasingly more products will contain some sort of nanomaterial. However, to date, little is known about the occurrence, fate and toxicity of nanoparticles. The limitations in our knowledge are partly due to the lack of methodology for the detection and characterisation of engineered nanoparticles in complex matrices, i.e. water, soil or food. This review provides an overview of the characteristics of nanoparticles that could affect their behaviour and toxicity, as well as techniques available for their determination. Important properties include size, shape, surface properties, aggregation state, solubility, structure and chemical composition. Methods have been developed for natural or engineered nanomaterials in simple matrices, which could be optimized to provide the necessary information, including microscopy, chromatography, spectroscopy, centrifugation, as well as filtration and related techniques. A combination of these is often required. A number of challenges will arise when analysing environmental and food materials, including extraction challenges, the presence of analytical artifacts caused by sample preparation, problems of distinction between natural and engineered nanoparticles and lack of reference materials. Future work should focus on addressing these challenges.

Environmental risk assessments of engineered nanoparticles require thorough characterization of nanoparticles and their aggregates. Furthermore, quantitative analytical methods are required to determine environmental concentrations and enable both effect and exposure assessments. Many methods still need optimization and development, especially for new types of nanoparticles in water, but extensive experience can be gained from the fields of environmental chemistry of natural nanomaterials and from fundamental colloid chemistry. This review briefly describes most methods that are being exploited in nanoecotoxicology for analysis and characterization of nanomaterials. Methodological aspects are discussed in relation to the fields of nanometrology, particle size analysis and analytical chemistry. Differences in both the type of size measures (length, radius, aspect ratio, etc.), and the type of average or distributions afforded by the specific measures are compared. The strengths of single particle methods, such as electron microscopy and atomic force microscopy, with respect to imaging, shape determinations and application to particle process studies are discussed, together with their limitations in terms of counting statistics and sample preparation. Methods based on the measurement of particle populations are discussed in terms of their quantitative analyses, but the necessity of knowing their limitations in size range and concentration range is also considered. The advantage of combining complementary methods is highlighted.

It is inevitable that, during their use, engineered nanoparticles will be released into soils and waters. There is therefore increasing concern over the potential impacts of engineered nanoparticles in the environment on aquatic and terrestrial organisms and on human health. Once released into the environment, engineered nanoparticles will aggregate to some degree; they might also associate with suspended solids, sediment, be accumulated by organisms and enter drinking water sources and food materials. These fate processes are dependent on the characteristics of the particle and the characteristics of the environmental system. A range of ecotoxicological effects have also been reported, including effects on microbes, plants, invertebrates and fish. Although available data indicate that current risks of engineered nanoparticles in the environment to environmental and human health are probably low, our knowledge of the potential impacts of engineered nanoparticles in the environment on human health is still limited. There is therefore a need for continued work to develop an understanding of the exposure levels for engineered nanoparticles in environmental systems and to begin to explore the implications of these levels in terms of the ecosystem and human health. This will require research in a range of areas, including detection and characterization, environmental fate and transport, ecotoxicology and toxicology.

This study compares the effect of two different strategies to inhibit the renin-angiotensin system in the setting of acute myocardial infarction (MI). Male Wistar rats were treated with placebo, the angiotensin-converting enzyme (ACE) inhibitor ramipril (1 mg/kg/day), or the AT1 receptor antagonist, olmesartan (1 mg/kg/day), both initiated 1 week before induction of MI and continued for 6 weeks after MI. The inflammatory reaction in the heart was investigated 7 days post-MI by determination of macrophage infiltration and the expression of tumor necrosis factor (TNF-alpha), interleukin (IL)-1beta and IL-6 at mRNA and protein levels. Six weeks post-MI, cardiac function was measured following chronic implantation of catheters in the LV and femoral artery, and cardiac morphology and coronary structure were investigated in picrosirius-red stained hearts. In placebo-treated rats, macrophage infiltration was accompanied by upregulation of IL-1beta and IL-6 mRNA in the peri-infarct zone. TNF-alpha and IL-1beta mRNA and protein were also upregulated in the non-infarcted myocardium. Whereas both treatment regimes significantly reduced IL-6 upregulation, olmesartan additionally reduced macrophage infiltration and IL-1beta expression. Six weeks post-MI, placebo-treated MI animals developed an impaired cardiac function with structural remodeling of the myocardium and coronaries. While olmesartan and ramipril both improved cardiac function and reduced infarct size and myocardial/coronary remodeling, olmesartan was more effective not only in increasing vascular perimeter, inner vascular diameter and septal thickness but also in lowering media thickness of coronary arteries, inner left ventricular diameter, left ventricular circumference and left ventricular end-diastolic pressure than ramipril. Thus, following MI the AT1 receptor blocker, olmesartan, attenuated cardiac inflammatory reactions and protected myocardial/coronary structure and function of the failing heart proving to be of similar, in some cases superior effectiveness in this respect than the ACE inhibitor, ramipril.

Although tightly regulated programmed cell death (apoptosis) possesses great importance for tissue homeostasis, several pathologic processes are associated with organ failure due to adversely activated cell apoptosis. Transient increase in apoptosis has been shown to cause organ damage during fulminant hepatitis B, autoimmune diseases, ischemia-reperfusion injury, sepsis, or allograft rejection. A defined and temporary inhibition of cell apoptosis may therefore be of high clinical relevance. Activation of death receptors results in caspase-8 recruitment to the death-inducing signaling complex, which initiates the apoptotic process through cleavage of caspase-8 and downstream substrates. This initial step may be inhibited by the caspase-8 inhibitor FLIP (FLICE inhibitory protein). To specifically inhibit the initiation of death receptor-mediated apoptosis we constructed a fusion protein containing FLIP fused N-terminally to the human immunodeficiency virus TAT domain. This TAT domain allows the fusion protein to cross the cell membrane and thus makes the FLIP domain able to interfere with the death-inducing signaling complex inside of the cell. We observed that incubation of lymphocytic Jurkat or BJAB cells with TAT-FLIPS proteins significantly inhibits Fas-induced activation of procaspase-8 and downstream caspases, preventing cells from undergoing apoptosis. Systemic application of TAT-FLIPS prolongs survival and reduces multi-organ failure due to Fas-receptor-mediated lethal apoptosis in mice. Therefore, application of cellular FLIPS in the form of a TAT fusion protein may open a promising, easily applicable new tool for providing protection against transient, pathologically increased apoptosis in various diseases.

Objective: After myocardial infarction, angiotensin II (AngII) promotes ventricular remodeling and deposition of extracellular matrix (ECM), e.g., collagen type 1 and 3. Whether AngII regulates the expression of small leucine-rich proteoglycans (SLRP) which are important modulators of collagen fibrillogenesis and are induced after experimental myocardial infarction in rats is not known. The aim of the present study was therefore to analyse in cultured cardiac fibroblasts the expression and secretion of the SLRP biglycan in response to AngII. Methods: Cardiac fibroblasts were isolated from neonatal Wistar Kyoto rats and used in the first passage. Expression of AT1- and AT2-receptors was verified by RT-PCR. Expression of protoeglycans was analyzed after metabolic labeling with [35S]-sulfate, by SDS-PAGE and Western analysis. In addition, mRNA expression was examined by means of RT-PCR and Northern analysis. The activity of the biglycan promoter was analyzed using three biglycan promoter–luciferase fusion constructs. Results: Biglycan was found to be the predominant proteoglycan produced by neonatal cardiac fibroblasts in vitro. In response to AngII (10−7 M), secretion of total [35S]-labeled proteoglycans and mRNA of biglycan were increased to 116±1.8% and 121±11% ( n = 5, mean±S.E.M.) of unstimulated controls, respectively. Biglycan induction in response to AngII was sensitive to Losartan (10−5 M) and unaffected by PD123177 (10−6 M), suggesting that the AT1-receptor mediates the induction of biglycan. Direct activation of the biglycan promoter downstream of the AT1-receptor was excluded by promoter activity assays. Instead, increased release of transforming growth factor beta 1 (TGFβ1) was detected by ELISA in response to AT1-receptor stimulation. Furthermore, neutralising antibodies to TGFβ1 inhibited biglycan induction in response to AngII. Conclusion: The results indicate that in cardiac fibroblasts AngII via the AT1-receptor causes autocrine release of TGFβ1, which in turn induces biglycan expression and secretion.

Overexpression of the small leucine-rich proteoglycan biglycan (BGN) in fibrosis and desmoplasia results from enhanced activity of transforming growth factor-beta (TGF-beta). In pancreatic adenocarcinoma, the tumor cells themselves may contribute to BGN synthesis in vivo, since 8 of 18 different pancreatic carcinoma cell lines constitutively expressed BGN mRNA, as shown by reverse transcription-PCR analysis. In PANC-1 cells, TGF-beta1 dramatically stimulated BGN mRNA accumulation through a BGN transcription-independent, cycloheximide-sensitive mechanism and strongly increased the synthesis and release of the proteoglycan form of BGN. The ability of TGF-beta1 to induce BGN mRNA was critically dependent on Smad signaling, since 1) the up-regulation of BGN mRNA was preceded by a marked increase in Smad2 phosphorylation in TGF-beta1-treated PANC-1 cells, 2) TGF-beta1 was unable to induce BGN mRNA in pancreatic carcinoma cell lines that carry homozygous deletions of the Smad4/DPC4 gene, 3) inhibition of the Smad pathway in PANC-1 cells by transfection with a dominant negative Smad4/DPC4 mutant significantly reduced TGF-beta1-induced BGN mRNA expression, 4) stable reintroduction of wild type Smad4/DPC4 into Smad4-null CFPAC-1 cells restored the TGF-beta1 effect, and 5) overexpression of Smad2 and Smad3 in PANC-1 cells augmented TGF-beta1 induction of BGN mRNA, whereas forced expression of Smad7, an inhibitory Smad, effectively blocked it. These results clearly show that a functional Smad pathway is crucial for TGF-beta regulation of BGN mRNA expression. Since BGN has been shown to inhibit growth of pancreatic cancer cells, the Smad4/DPC4 mediation of the TGF-beta effect may represent a novel tumor suppressor function for Smad4/DPC4: antiproliferation via expression of autoinhibitory BGN.

A hyphenated methodology has been developed and validated, which utilizes the extensive size separation range of hydrodynamic chromatography (here: 5–300 nm) combined with the multi-element selectivity of ICP-MS. This has been applied to the analysis of metal-based nanoparticles in environmental samples. The quality of the particle sizing data obtained from this exercise was enabled through the production of a range of gold nanoparticles (sterically stabilized to prevent aggregation in environmental matrices), which were validated for use as external size calibration standards as well as internal retention time markers, using TEM. The methodology was then successfully applied to a study where nanosilver was spiked into sewage sludge, preliminary data from which showed that a fraction of nanosilver survived as single nanoparticles in the sludge supernatant. The method was also tested on solutions containing other commonly used nanoparticles (TiO2, SiO2, Al2O3 and Fe2O3). Overall, the data showed that, by using ICP-MS with collision cell technology, the methodology would be helpful in investigating the fate of a significant range of nanoparticle types. Other characteristics of HDC-ICP-MS are: rapid analysis time ( 300 nm), and limited sample pre-treatment required.