Per-pixel unmixing of spectrally overlapping fluorophores using intra-exposure excitation modulation.

Multilabel fluorescence imaging is essential for the visualization of complex systems, though a major challenge is the limited width of the useable spectral window. Here, we present a new method, exNEEMO, that enables per-pixel quantification of spectrally-overlapping fluorophores based on their light-induced dynamics, in a way that is compatible with a very broad range of timescales over which these dynamics may occur. Our approach makes use of intra-exposure modulation of the excitation light to distinguish the different emitters given their reference responses to this modulation.

Latest trends in bioimaging and building a proactive network of early-career young scientists around bioimaging in Europe.

Biological research is in constant need of new methodological developments to assess organization and functions at various scales ranging from whole organisms to interactions between proteins. One of the main ways to evidence and quantify biological phenomena is imaging. Fluorescence microscopy and label-free microscopy are in particular highly active fields of research due to their compatibility with living samples as well as their versatility.

Emerging applications of nanobodies in cancer therapy.

Cancer is a heterogeneous disease, requiring treatment tailored to the unique phenotype of the patient's tumor. Monoclonal antibodies (mAbs) and variants thereof have enabled targeted therapies to selectively target cancer cells. Cancer cell-specific mAbs have been used for image-guided surgery and targeted delivery of radionuclides or toxic agents, improving classical treatment strategies.

Consequences of the constitutive NOX2 activity in living cells: Cytosol acidification, apoptosis, and localized lipid peroxidation.

The phagocyte NADPH oxidase (NOX2) is a key enzyme of the innate immune system generating superoxide anions (O), precursors of reactive oxygen species. The NOX2 protein complex is composed of six subunits: two membrane proteins (gp91 and p22) forming the catalytic core, three cytosolic proteins (p67, p47 and p40) and a small GTPase Rac. The sophisticated activation mechanism of the NADPH oxidase relies on the assembly of cytosolic subunits with the membrane-bound components.

tdLanYFP, a Yellow, Bright, Photostable, and pH-Insensitive Fluorescent Protein for Live-Cell Imaging and Förster Resonance Energy Transfer-Based Sensing Strategies.

Yellow fluorescent proteins (YFPs) are widely used as optical reporters in Förster resonance energy transfer (FRET)-based biosensors. Although great improvements have been done, the sensitivity of the biosensors is still limited by the low photostability and the poor fluorescence performances of YFPs at acidic pH values. Here, we characterize the yellow fluorescent protein tdLanYFP, derived from the tetrameric protein from the cephalochordate , LanYFP.

Separation of spectrally overlapping fluorophores using intra-exposure excitation modulation.

Multicolor fluorescence imaging is an excellent method for the simultaneous visualization of multiple structures, although it is limited by the available spectral window. More labels can be measured by distinguishing these on properties, such as their fluorescence dynamics, but usually these dynamics must be directly resolvable by the instrument. We propose an approach to distinguish emitters over a much broader range of light-induced dynamics by combining fast modulation of the light source with the detection of the time-integrated fluorescence.

Occurrence of type A, B and D trichothecenes, zearalenone and stachybotrylactam in straw.

Straw is the main by-product of grain production, used as bedding material and animal feed. If produced or stored under adverse hygienic conditions, straw is prone to the growth of filamentous fungi. Some of them, e.

The NADPH Oxidase and the Phagosome.

The key purpose of phagocytosis is the destruction of pathogenic microorganisms. The phagocytes exert a wide array of killing mechanisms that allow mastering the vast majority of pathogens. One of these mechanisms consists in the production of reactive oxygen species inside the phagosome by a specific enzyme, the phagocyte NADPH oxidase.

Plasma kinetics and matrix residues of deoxynivalenol (DON) and zearalenone (ZEN) are altered in endotoxaemic pigs independent of LPS entry site.

This study aimed to investigate a potential modulatory effect of E. coli lipopolysaccharide (LPS) on the kinetics of deoxynivalenol (DON) and zearalenone (ZEN) after pre- or post-hepatic LPS administration to unravel the putative role of the liver. Fifteen barrows were fed a diet containing mycotoxin-contaminated maize (4.

Determination of T-2 toxin, HT-2 toxin, and three other type A trichothecenes in layer feed by high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS)--comparison of two sample preparation methods.

A sensitive method for the simultaneous determination of T-2 toxin, HT-2 toxin, neosolaniol, T-2 triol, and T-2 tetraol in layer feed using high-performance liquid chromatography coupled to triple quadrupole mass spectrometry in the positive ionization mode (LC-ESI-MS/MS) is described. Two fast and easy clean-up methods-with BondElut Mycotoxin and MycoSep 227 columns, respectively-were tested. The separation of the toxins was conducted on a Pursuit XRs Ultra 2.

Haematological and immunological adaptations of non-pregnant, non-lactating dairy cows to a high-energetic diet containing mycotoxins.

Diet change and fatness are supposed to challenge the immune system of the cow. Therefore, immunological and haematological consequences of adaptation to and continued feeding of a high-energy diet were studied in eight non-pregnant, non-lactating Holstein cows over 16 weeks. Blood haptoglobin concentration remained unaltered, suggesting that an acute phase reaction was not induced.

Effects of Dietary Exposure to Zearalenone (ZEN) on Carp (Cyprinus carpio L.).

The mycotoxin zearalenone (ZEN) is frequently contaminating animal feeds including feed used in aquaculture. In the present study, the effects of dietary exposure to ZEN on carp (Cyprinus carpio L.) were investigated.

Development of a multi-toxin method for investigating the carryover of zearalenone, deoxynivalenol and their metabolites into milk of dairy cows.

A dose-response study was carried out to examine the carryover of zearalenone (ZEN), deoxynivalenol (DON) and their metabolites into bovine milk. Therefore, a feeding trial with 30 dairy cows fed with three different levels of Fusarium (FUS) toxin-contaminated maize was performed. A control group (0.

Diagnosis of intoxications of piglets fed with Fusarium toxin-contaminated maize by the analysis of mycotoxin residues in serum, liquor and urine with LC-MS/MS.

Concentrations of zearalenone (ZEN), deoxynivalenol (DON) and their metabolites α-zearalenol (α-ZEL), β-zearalenol (β-ZEL), zearalanone (ZAN), α-zearalanol (α-ZAL), β-zearalanol (β-ZAL) and de-epoxy-deoxynivalenol (de-DON) in serum, liquor and urine of female piglets fed diets containing 0.01, 0.05, 0.

Residues of deoxynivalenol (DON) and its metabolite de-epoxy-DON in eggs, plasma and bile of laying hens of different genetic backgrounds.

In the present study, the potential for carry-over of deoxynivalenol (DON) into eggs and DON residues in plasma and bile of laying hens of different genetic backgrounds after long-term feeding trial was investigated. A total of 80, 23-week-old laying hens were assigned to a feeding trial with two diets, a control diet and a Fusarium toxin-contaminated diet (FUS) (0.4 and 9.

Lipopolysaccharides (LPS) modulate the metabolism of deoxynivalenol (DON) in the pig.

Pigs might be exposed to lipopolysaccharides (LPS) and deoxynivalenol (DON) at the same time, and both toxins are thought to interactively affect the intestinal barrier, the innate immune system, and the xenobiotics metabolism. Hence, we aimed at examining the single and combined effects of both toxins on nutrient digestibility and DON metabolism. For this purpose, barrows (26 ± 4 kg) were fed restrictedly either a control diet (CON) or a diet contaminated with 3.

Development of a liquid chromatography tandem mass spectrometry method for the simultaneous determination of zearalenone, deoxynivalenol and their metabolites in pig serum.

A sensitive and selective liquid chromatography tandem mass spectrometry method using negative electrospray ionisation (LC-ESI-MS/MS) was developed for the simultaneous determination of zearalenone (ZEN), deoxynivalenol (DON) and their metabolites α-zearalenol, β-zearalenol, zearalanone, α-zearalanol, β-zearalanol and de-epoxy-deoxynivalenol in pig serum. For method development, different sample preparation columns were tested for their suitability for extraction and clean up. Finally, preparation of serum samples was carried out using Oasis™ HLB solid-phase extraction (SPE) columns.

Effects of feeding deoxynivalenol (DON)-contaminated wheat to laying hens and roosters of different genetic background on the reproductive performance and health of the newly hatched chicks.

A total of 216 23-week-old laying hens from two different genetic backgrounds (half of the birds were Lohmann brown [LB] and [LSL] hens, respectively) and 24 adult roosters were assigned to a feeding trial to study the effect of increasing concentrations of deoxynivalenol (DON) in the diet (0, 5, 10 mg/kg) on the reproductive performance of hens and roosters, and the health of the newly hatched chicks. Hatchability was adversely affected by the presence of DON in LB hens' diet, while the hatchability of the LSL chicks was significantly higher than LB chicks. An interaction effect between DON in the hens' diet and the breed was noticed on fertility, as the fertility was decreased in the eggs of LB hens receiving 10 mg/kg DON in their diet and increased in the eggs of LSL hens fed 10 mg/kg DON.

In vivo effects of deoxynivalenol (DON) on innate immune responses of carp (Cyprinus carpio L.).

Deoxynivalenol (DON) is one of the most important members of Fusarium toxins since it often can be found in relevant concentrations in animal feeds. The effects of this group of toxins on fish are mostly unknown. The present study shows results from a feeding trial with carp (Cyprinus carpio L.

Ascaridia galli infection affects pullets differently when feed is contaminated with the Fusarium toxin deoxynivalenol (DON).

The Fusarium mycotoxin deoxynivalenol (DON) is a common contaminant of cereal grains used as animal feed. DON is known for its cytotoxic and anti-proliferative properties and might adversely affect the health of poultry. The prevalence of the intestinal parasitizing roundworm Ascaridia galli is higher in outdoor housing systems and has been associated with maldigestion and malabsorption.

UV-induced cis-trans isomerization of zearalenone in contaminated maize.

In the literature, it has been shown that the naturally occurring trans-zearalenone (ZEN) is transformed by ultraviolet irradiation to cis-ZEN. However, the practical relevance of this transformation in animal feeding remains unclear. The aim of the present preliminary investigation was to examine the effect of UV-irradiation on the concentration of trans-ZEN in a natural feed matrix at different dry matter contents to simulate the dry and wet feeding techniques usually applied in pig feeding.

Humic substances failed to prevent the systemic absorption of deoxynivalenol (DON) and its adverse effects on piglets.

The aim of this study was to examine the effects of a control diet (CON, 0.25 mg DON/kg diet) or a Fusarium toxin-contaminated diet (FUS, 4.49 mg DON/kg diet) without and with humic substances (HS) (CON-HS and FUS-HS, 0.

Inactivation of deoxynivalenol-contaminated cereal grains with sodium metabisulfite: a review of procedures and toxicological aspects.

Both hydrothermal treatment and wet preservation of mainly deoxynivalenol (DON)-containing, Fusarium toxin (FUS)-contaminated cereal grains with sodium metabisulfite (Na2S2O5 [SBS]) were successfully demonstrated to reduce the DON contamination through formation of the sulfonated derivative of DON, termed as DON sulfonate (DONS). The wet preservation is particularly interesting from a practical viewpoint as it can be easily performed at the farm level where the cereal grains are harvested and utilized in pig feeding. This review compiles the literature with regard to the chemical characterization and the detection of DONS, technical procedures and their efficacies, toxicological aspects and toxic effects of DON, DONS and SBS, and detection of DONS, DON and further metabolites in physiological specimens of pigs.

Hydrothermal treatment of naturally contaminated maize in the presence of sodium metabisulfite, methylamine and calcium hydroxide; effects on the concentration of zearalenone and deoxynivalenol.

Fusarium toxin-contaminated ground maize was hydrothermally treated in the presence of different combinations of chemicals in order to simultaneously reduce zearalenone (ZEA) and deoxynivalenol (DON) concentrations. Treatments were carried out in a laboratory conditioner at 80 °C and 17 % moisture. Six different treatments were performed, consisting of 3 doses of methylamine (MMA; 2.

Bioavailability of the Fusarium toxin deoxynivalenol (DON) from wheat straw and chaff in pigs.

Fusarium infections do not only affect the grain, but also the rest of the plant, which result in contamination of plants with the mycotoxin deoxynivalenol (DON). The bioavailability of DON may be influenced by the matrix due to the differences in nutrient composition between grain and straw, particularly the high fibre component in straw. The experiment was carried out by exposing 18 male castrated pigs (30-40 kg live weight) with a single dose of DON from wheat grain, straw and chaff in the diet.