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Cool S.K.,Ghent University | Geers B.,Ghent University | Roels S.,Coda Research | Stremersch S.,Ghent University | And 5 more authors.
Journal of Controlled Release | Year: 2013

Local extravasation and triggered drug delivery by use of ultrasound and microbubbles is a promising strategy to target drugs to their sites of action. In the past we have developed drug loaded microbubbles by coupling drug containing liposomes to the surface of microbubbles. Until now the advantages of this drug loading strategy have only been demonstrated in vitro. Therefore, in this paper, microbubbles with indocyanine green (ICG) containing liposomes at their surface or a mixture of ICG-liposomes and microbubbles was injected intravenously in mice. Immediately after injection the left hind leg was exposed to 1 MHz ultrasound and the ICG deposition was monitored 1, 4 and 7 days post-treatment by in vivo fluorescence imaging. In mice that received the ICG-liposome loaded microbubbles the local ICG deposition was, at each time point, about 2-fold higher than in mice that received ICG-liposomes mixed with microbubbles. We also showed that the perforations in the blood vessels allow the passage of ICG-liposomes up to 5 h after microbubble and ultrasound treatment. An increase in tissue temperature to 41 C was observed in all ultrasound treated mice. However, ultrasound tissue heating was excluded to cause the local ICG deposition. We concluded that coupling of drug containing liposomes to microbubbles may increase ultrasound mediated drug delivery in vivo. © 2013 Elsevier B.V.

Schmeiser S.,Justus Liebig University | Schmeiser S.,Institute For Virologie | Mast J.,Coda Research | Thiel H.-J.,Justus Liebig University | Konig M.,Justus Liebig University
Journal of Virology | Year: 2014

Knowledge on the morphogenesis of pestiviruses is limited due to low virus production in infected cells. In order to localize virion morphogenesis and replication sites of pestiviruses and to examine intracellular virion transport, a cell culture model was established to facilitate ultrastructural studies. Based on results of virus growth kinetic analysis and quantification of viral RNA, pestivirus strain Giraffe-1 turned out to be a suitable candidate for studies on virion generation and export from culture cells. Using conventional transmission electron microscopy and single-tilt electron tomography, we found virions located predominately in the lumen of the endoplasmic reticulum (ER) in infected cells and were able to depict the budding process of virions at ER membranes. Colocalization of the viral core protein and the envelope glycoprotein E2 with the ER marker protein disulfide isomerase (PDI) was demonstrated by immunogold labeling of cryosections. Moreover, pestivirions could be shown in transport vesicles and the Golgi complex and during exocytosis. Interestingly, viral capsid protein and double-stranded RNA (dsRNA) were detected in multivesicular bodies (MVBs), which implies that the endosomal compartment plays a role in pestiviral replication. Significant cellular membrane alterations such as those described for members of the Flavivirus and Hepacivirus genera were not found. Based on the gained morphological data, we present a consistent model of pestivirus morphogenesis. © 2014, American Society for Microbiology.

Thiry C.,Coda Research | Ruttens A.,Coda Research | De Temmerman L.,Coda Research | Schneider Y.-J.,Institute Des Science Of La Vie | Pussemier L.,Coda Research
Food Chemistry | Year: 2012

Selenium is an essential trace element that has raised interest because of its antioxidant and anticancer properties. The beneficial or toxic effect of Se is not only dose-dependent, but also relates to the chemical form of the element and its bioavailability. In this review, recently published data is summarised concerning both Se speciation and Se relative bioavailability in various foodstuffs. In addition, Se bioavailability is discussed in relation to the species-dependent metabolism in humans. In this way, the understanding of the potential health impact of Se species in commonly consumed food is aimed to be improved. It is strongly suggested on the basis of a higher retention and a lower toxicity, that organic Se (especially SeMet, the major species in food) is more recommendable than inorganic Se in the frame of a balanced diet. Further research is however desirable concerning the characterisation of unidentified Se species and determination of their health effects. © 2011 Elsevier Ltd. All rights reserved.

Rosseel T.,Coda Research | Ozhelvaci O.,Coda Research | Freimanis G.,The Pirbright Institute | Van Borm S.,Coda Research
Journal of Virological Methods | Year: 2015

Viral metagenomic approaches are increasingly being used for viral discovery. Various strategies are applied to enrich viral sequences, but there is often a lack of knowledge about their effective influence on the viral discovery sensitivity. We evaluate some convenient and widely used approaches for RNA virus discovery in clinical samples in order to reveal their sensitivity and potential bias introduced by the enrichment or amplifications steps. An RNA virus was artificially spiked at a fixed titer in serum and lung tissue, respectively, low and high nucleic acid content matrices. For serum, a simple DNase treatment on the RNA extract gave the maximum gain in proportion of viral sequences (83×), and a subsequent ribosomal RNA removal nearly doubled once more the proportion of viral sequences. For lung tissue, a ribosomal RNA depletion step on the RNA extract had the biggest gain in proportion of viral sequences (32×). We show also that direct sequencing of cDNA is recommended above an extra random PCR amplification step, and a that the virion enrichment strategy (filtration and nuclease treatment) has a beneficial effect for sequencing-based virus discovery. Our findings provide sample-dependent guidelines for targeted virus discovery strategies. © 2015 Elsevier B.V.

Machiels B.,University of Liège | Lete C.,University of Liège | Guillaume A.,University of Liège | Mast J.,Coda Research | And 3 more authors.
PLoS Pathogens | Year: 2011

All gammaherpesviruses encode a major glycoprotein homologous to the Epstein-Barr virus gp350. These glycoproteins are often involved in cell binding, and some provide neutralization targets. However, the capacity of gammaherpesviruses for long-term transmission from immune hosts implies that in vivo neutralization is incomplete. In this study, we used Bovine Herpesvirus 4 (BoHV-4) to determine how its gp350 homolog - gp180 - contributes to virus replication and neutralization. A lack of gp180 had no impact on the establishment and maintenance of BoHV-4 latency, but markedly sensitized virions to neutralization by immune sera. Antibody had greater access to gB, gH and gL on gp180-deficient virions, including neutralization epitopes. Gp180 appears to be highly O-glycosylated, and removing O-linked glycans from virions also sensitized them to neutralization. It therefore appeared that gp180 provides part of a glycan shield for otherwise vulnerable viral epitopes. Interestingly, this O-glycan shield could be exploited for neutralization by lectins and carbohydrate-specific antibody. The conservation of O-glycosylation sites in all gp350 homologs suggests that this is a general evasion mechanism that may also provide a therapeutic target. © 2011 Machiels et al.

Van Der Zande M.,Wageningen University | Vandebriel R.J.,National Institute for Public Health and the Environment | Van Doren E.,Coda Research | Kramer E.,Wageningen University | And 10 more authors.
ACS Nano | Year: 2012

We report the results of a 28-day oral exposure study in rats, exposed to <20 nm noncoated, or <15 nm PVP-coated silver nanoparticles ([Ag] = 90 mg/kg body weight (bw)), or AgNO 3 ([Ag] = 9 mg/kg bw), or carrier solution only. Dissection was performed at day 29, and after a wash-out period of 1 or 8 weeks. Silver was present in all examined organs with the highest levels in the liver and spleen for all silver treatments. Silver concentrations in the organs were highly correlated to the amount of Ag + in the silver nanoparticle suspension, indicating that mainly Ag +, and to a much lesser extent silver nanoparticles, passed the intestines in the silver nanoparticle exposed rats. In all groups silver was cleared from most organs after 8 weeks postdosing, but remarkably not from the brain and testis. Using single particle inductively coupled plasma mass spectrometry, silver nanoparticles were detected in silver nanoparticle exposed rats, but, remarkably also in AgNO 3 exposed rats, hereby demonstrating the formation of nanoparticles from Ag +in vivo that are probably composed of silver salts. Biochemical markers and antibody levels in blood, lymphocyte proliferation and cytokine release, and NK-cell activity did not reveal hepatotoxicity or immunotoxicity of the silver exposure. In conclusion, oral exposure to silver nanoparticles appears to be very similar to exposure to silver salts. However, the consequences of in vivo formation of silver nanoparticles, and of the long retention of silver in brain and testis should be considered in a risk assessment of silver nanoparticles. © 2012 American Chemical Society.

Thiry C.,Institute Des Science Of La Vie And Uclouvain | Ruttens A.,Coda Research | Pussemier L.,Coda Research | Schneider Y.-J.,Institute Des Science Of La Vie And Uclouvain
British Journal of Nutrition | Year: 2013

A range of Se species has been shown to occur in a variety of different foodstuffs. Depending on its speciation, Se is more or less bioavailable to human subjects. In the present study, the role of speciation as a determinant of Se bioavailability was addressed with an investigation of species-specific mechanisms of transport at the intestinal level. The present work focused on four distinct Se compounds (selenate (Se(VI)), selenite (Se(IV)), selenomethionine (SeMet) and methylselenocysteine (MeSeCys)), whose intestinal transport was mimicked through an in vitro bicameral model of enterocyte-like differentiated Caco-2 cells. Efficiency of Se absorption was shown to be species dependent (SeMet>MeSeCys>Se(VI)>Se(IV)). In the case of SeMet, MeSeCys and Se(VI), the highly polarised passage from the apical to basolateral pole indicated that a substantial fraction of transport was transcellular, whilst results for Se(IV) indicated paracellular diffusion. Passage of the organic Se species (SeMet and MeSeCys) became saturated after 3 h, but no such effect was observed for the inorganic species. In addition, SeMet and MeSeCys transport was significantly inhibited by their respective S analogues methionine and methylcysteine, which suggests a common transport system for both kinds of compounds. © 2012 The Authors.

Waegeneers N.,Coda Research | Thiry C.,Coda Research | De Temmerman L.,Coda Research | Ruttens A.,Coda Research
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2013

The total selenium content of about 800 food products purchased in Belgium was determined and combined with food records to determine the nutritional selenium status of Belgian people. The largest selenium concentrations (&1 mg kg-1) were found in Brazil nuts and offal, of which the consumption is limited. Usually consumed food groups with the highest selenium concentrations were fish and shellfish (0.2-0.9 mg kg-1), eggs, poultry meat, cheese, mushrooms and pasta (approximately 0.2 mg kg-1). The mean dietary selenium intake was calculated to be 60 μg day-1, which is at the lower end but within the range recommended by the Superior Health Council in Belgium (60-70 μg day-1), and adequate according to the 55 μg day-1 recommended by the Scientific Committee on Food (SCF) of the European Commission. The major sources of selenium intake are meat and meat products (31%), fish and shellfish (20%), pasta and rice (12%), and bread and breakfast cereals (11%). © 2013 Copyright Taylor and Francis Group, LLC.

De Temmerman L.,Coda Research | Ruttens A.,Coda Research | Waegeneers N.,Coda Research
Environmental Pollution | Year: 2012

Root crops, carrot and celeriac, were exposed to atmospheric deposition in a polluted versus reference area. An effect was observed on the As, Cd and Pb concentrations of the leaves and the storage organs. The concentrations in the whole storage organs correlated well with atmospheric deposition, which shows that they even could be used for biomonitoring. Nevertheless, leaves remain much more appropriate. The results revealed also a significant increase of the As and Cd concentration in the consumable part of the storage organs as a function of their atmospheric deposition. As such the experiments allowed deriving regression equations, useful for modeling the atmospheric impact of trace elements on the edible parts of root crops. For Pb, however, there was hardly any significant impact on the inner parts of the storage organs and as such the transfer of Pb in the food chain through root crops can be considered to be negligible. © 2012 Elsevier Ltd. All rights reserved.

Tangni E.K.,Coda Research | Motte J.-C.,Coda Research | Callebaut A.,Coda Research | Pussemier L.,Coda Research
Journal of Agricultural and Food Chemistry | Year: 2010

Cross-reactivity of antibodies in AGRAQUANT, DON EIA, VERATOX, ROSA LF-DONQ, and MYCONTROLDON designed for deoxynivalenol (DON) determination in food and feedstuffs was evaluated against nivalenol, 3-acetylDON, 15-acetylDON, de-epoxy metabolite 1 of DON, DON-3β-glucoside, T2-toxin, HT2-toxin, fusarenone X, diacetoxyscirpenol, verrucarol, and zearalenone. Cross-reactivity measurements were run in water using the 50% reduction of absorbance of the blank for ELISA kits or through direct DON determination upon using the standards of mycotoxins via ROSA LF-DONQ or MYCONTROLDON. For the tested toxin concentrations, all DON kits have low cross-reactivity toward diacetoxyscirpenol, T2-toxin, HT2-toxin, verrucarol, and zearalenone and moderate cross-reactivity toward 15-AcetylDON and fusarenone X. AGRAQUANT, DON EIA, and VERATOX kits showed high cross-reactivity in various ranking orders against DON-3-Glc, DOM-1, and 3AcDON. DON EIA showed also high cross-reactivity against nivalenol and fusarenone X. These mycotoxins could coexist in food or feedstuffs, and analytical results can be wrongly interpreted. Cross-reactivity does not allow checking the compliance with the legal norms, but it does allow an overall risk assessment for the consumers. Updating regularly the cross-reactivity evaluation of the produced batches is recommended for 3-acetylDON, nivalenol, DON-3-Glc, de-epoxy metabolite 1, and fusarenone X. © 2010 American Chemical Society.

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