Development of a multiplex flow cytometric microsphere immunoassay for mycotoxins and evaluation of its application in feed

A multi-mycotoxin immunoassay—using the MultiAnalyte Profiling (xMAP) technology—is developed and evaluated. This technology combines a unique color-coded microsphere suspension array, with a dedicated flow cytometer. We aimed for the combined detection of aflatoxins, ochratoxin A, deoxynivalenol, fumonisins, zearalenone and T-2-toxin in an inhibition immunoassay format. Sets of six mycotoxin-protein conjugates and six specific monoclonal antibodies were selected, and we observed good sensitivities and no cross-interactions between the assays in buffer. However, detrimental effects of the feed extract on the sensitivities and in some cases on the slopes of the curves were observed and different sample materials showed different effects. Therefore, for quantitative analysis, this assay depends on calibration curves in blank matrix extracts or on the use of a suitable multi-mycotoxin cleanup. To test if the method was suitable for the qualitative detection at EU guidance levels, we fortified rapeseed meal, a feed ingredient, with the six mycotoxins, and all extracts showed inhibited responses in comparison with the non-fortified sample extract. Contaminated FAPAS reference feed samples assigned for a single mycotoxin showed strong inhibitions in the corresponding assays but also often in other assays of the multiplex. In most cases, the presence of these other mycotoxins was confirmed by instrumental analysis. The multiplex immunoassay can be easily extended with other mycotoxins of interest, but finding a suitable multi-mycotoxin cleanup will improve its applicability

Effect analysis of transient scenarios for successful water management strategies

Recent scenario studies on water management focus on one or two projection years and the effects on the water system and functions. The future is however more complex and dynamic. Therefore, we analyse transient scenarios in order to evaluate the performance of water management strategies. Current available simulation tools are not suitable for this purpose. Therefore, we have developed and used a tool to simulate 50-100 year long time series and that is good and fast enough to simulate the effects of these scenarios and strategies on the water system and the interaction with the human system. We present the first step by means of a case study

QRTEngine: An easy solution for running online reaction time experiments using Qualtrics

Performing online behavioral research is gaining increased popularity among researchers in psychological and cognitive science. However, the currently available methods for conducting online reaction time experiments are often complicated and typically require advanced technical skills. In this article, we introduce the Qualtrics Reaction Time Engine (QRTEngine), an open-source JavaScript engine that can be embedded in the online survey development environment Qualtrics. The QRTEngine can be used to easily develop browser-based online reaction time experiments with accurate timing within current browser capabilities, and it requires only minimal programming skills. After introducing the QRTEngine, we briefly discuss how to create and distribute a Stroop task. Next, we describe a study in which we investigated the timing accuracy of the engine under different processor loads using external chronometry. Finally, we show that the QRTEngine can be used to reproduce classic behavioral effects in three reaction time paradigms: a Stroop task, an attentional blink task, and a masked-priming task. These findings demonstrate that QRTEngine can be used as a tool for conducting online behavioral research even when this requires accurate stimulus presentation times

Structure of trans-bis[4-amino-3,5-bis(pyridin-2-yl)-1,2,4-triazole-N1,N'[diaqua- manganese(II) dibromide

[Mn(C12H10N6)2(H2O)2]Br2, Mr = 727.28, orthorhombic, Pbca, a = 10.734 (6), b = 17.084 (0), c = 15.182 (6) angstrom, V = 2784 angstrom 3, Z = 4, D(x) = 1.734 g cm-3, lambda-(Mo K-alpha) = 0.71073 angstrom, mu = 33.23 cm-1, F(000) = 1450, T = 295 K, final R = 0.032 for 1493 reflections [I > 2-sigma(I)]. The title compound is the first reported mononuclear compound with the ligand 4-amino-3,5-bis(pyridin-2-yl)-1,2,4-triazole. The manganese ions, situated on an inversion centre, are coordinated by four nitrogen atoms with an N-Mn-N angle of 74.1 (1)-degrees and Mn-N distances of 2.188 (4) and 2.266 (4) angstrom. Two axial water molecules [Mn-O = 2.200 (4) angstrom] complete the coordination sphere of the metal, which is pseudo-octahedral. The two bromide ions are not coordinated but are involved in an extended hydrogen-bridging network with the water ligands and the amino group of the triazole

Prognostic Value of Colonic Tissue and Blood Eosinophils in Ulcerative Colitis.

BACKGROUND It has been suggested that eosinophils may be a prognostic marker of disease outcome in ulcerative colitis (UC), but conflicting data exist. The objective was to investigate the extent of mucosal eosinophils and peripheral blood eosinophil count in newly diagnosed UC patients and to investigate its predictive value in short- and long-term disease outcomes. METHODS The degree of eosinophilia in baseline colonic biopsies and blood of newly diagnosed UC patients was retrospectively analyzed. It was investigated if tissue and blood eosinophilia could be a marker of a severe phenotype of UC, defined as the need for corticosteroids or immunomodulators in the first year or treatment with therapeutic monoclonal antibodies or colectomy during follow-up. Time to therapeutic monoclonal antibodies and time to colectomy were also evaluated as outcomes. RESULTS There were 103 UC patients (median age 26 years) included. Median tissue peak eosinophil count (PEC) was 70.0 and median peripheral blood eosinophil count was 0.3 × 109/L at diagnosis. Tissue PEC (r = -0.161, P = .104) and blood eosinophil count (r = 0.022, P = .877) were not correlated with the severity of histologic inflammation. Logistic regression analyses did not identify PEC and blood eosinophil count as predictors of more severe disease outcomes. Tissue PEC and peripheral blood eosinophil count did not predict the time the initiation of therapeutic monoclonal antibodies or colectomy. CONCLUSION Baseline tissue or peripheral blood eosinophils are not markers of disease activity and cannot be used as a predictor of severe disease outcomes in both adults and children with UC

Literatuuronderzoek omtrent patuline

Dit verslag heeft tot doel een literatuuroverzicht te geven omtrent het voorkomen van patuline en methoden voor het aantonen en bepalen van patuline in verschillende produkten. Patuline is een toxische uitscheidingsstof dat door verschillende penicillium- en aspergillus-schimmelsoorten geproduceerd wordt. Patuline bezit ook carcinogene en mutagene eigenschappen, wanneer ratten gedurende vijftien maanden met patuline worden ingespoten (onder de huid). P.expansum kan patuline vormen in o .a.: druiven, perziken, peren, abrikozen, pruimen of rozijnen, kersen , komkommers, worteltjes , tomaten , bananen, ananas en appels die rotte plekken vertonen. Na enten met P.expansum, P.urticae of Byssochlamys niveau kan er ook patuline gevormd worden in reineclaude, aardbei, dauw meloenen, rode en groene paprika, maar er wordt geen patuline gevormd in selderie, koolrabie, bloemkool, rode kool, radijs, paarde radijs, uien, vruchtvlees van een kalebas (als groente), aardappels en pootaardappels. Patuline komt vooral voor in appelprodukten zoals: appelsap , appelmoes en appelstroop. Vroeger werd patuline het meest bepaald m.b.v. dunnelaagchromatografie met als detectiereagens MBTH hydrachloride. Het MBTH hydrachloride wordt na het ontwikkelen van de silicagel plaat in tolueenethylacetaat- 90% mierezuur (5+4+1), op de plaat gespoten. Het patuline derivaat is dan onder invloed van langgolvig UV-licht als geel-bruine fluorescerende vlek waarneembaar, met een detectiegrens van 20-25 ug/1 appelsap

Biosensor immunoassay for traces of hazelnut protein in olive oil

The fraudulent addition of hazelnut oil to more expensive olive oil not only causes economical loss but may also result in problems for allergic individuals as they may inadvertently be exposed to potentially allergenic hazelnut proteins. To improve consumer safety, a rapid and sensitive direct biosensor immunoassay, based on a highly specific monoclonal antibody, was developed to detect the presence of hazelnut proteins in olive oils. The sample preparation was easy (extraction with buffer); the assay time was fast (4.5 min only) and the limit of detection was low (0.08 μg/g of hazelnut proteins in olive oil). Recoveries obtained with an olive oil mixed with different amounts of a hazelnut protein containing hazelnut oil varied between 93% and 109%

Українська шляхта між польським та українським етносами

Appearance and existence of the Ukrainian gentry relates to the traditions of Polish political culture, so during the whole period of its life it was between the Ukrainian and the Polish ethnic groups. Polanisation of the Ukrainian gentry begins at the date when some of the Ukrainian territories become a part of Poland and strengthens after Cossack revolution in the middle and at the end of the 16th century. Especially this process becomes effective at the beginning of the 18th century when a great part of gentry from other Polish lands migrates to Pravoberezhia (right-banked Ukraine). Nevertheless, having captured upper class and partially middle class of the Ukrainian gentry, polanisation mainly influenced consciousness and less religion of the lower class of the Ukrainian gentry. As for ethnoculture and language local gentry was mostly Ukrainian and it assimilated numerous Polish gentlemen-immigrants

Association between CTL Precursor Frequency to HLA-C Mismatches and HLA-C Antigen Cell Surface Expression

Previous studies showed the relevance of the cytotoxic T-cell precursor (CTLp) frequency assay for prediction of the outcome of HLA mismatched hematopoietic cell transplantation (HCT). Recently, it has been shown that HLA-C cell surface expression is correlated with virus specific cytotoxic T-cell responses and viremia control in HIV patients. The aim of the current study was to investigate the association between HLA-C antigen expression and the CTLp frequency to the mismatched HLA-C antigen. In total 115 recipient–donor pairs, for whom a successful CTLp assay was performed, were evaluated for this pilot study. All donor–recipient pairs were matched at 9/10 alleles with a single mismatch at the HLA-C locus. Antigen expression level of the mismatched HLA-C allele for each recipient and donor was based on the mean fluorescence intensity (MFI) values as described by Apps et al. (1). The cell surface expression of recipient’s mismatched HLA-C antigen was significantly lower among CTLp negative (n = 59) compared to CTLp positive (n = 56) pairs (154 and 193 MFI units, respectively, p = 0.0031). This difference was more pronounced in donor–recipient pairs that were mismatched for amino-acid residue-116 located in the groove of the HLA-C antigen, suggesting that the importance of peptide binding in the allo-recognition. Furthermore, in the particular case of low expression of the recipient mismatched HLA-C antigen (MFI < 115), CTLp reactivity depended on HLA-C expression level in the donor, the median MFI of donor’s mismatched HLA-C antigen was 114 in CTLp negative cases (n = 26), while in CTLp positive cases (n = 15) the median MFI of donor’s HLA-C antigen was 193 (p = 0.0093). We conclude that the expression level of the donor and recipient mismatched HLA-C antigens affect CTLp outcome. HLA-C antigen expression levels in combination with the CTLp assay may prove useful for the prediction of the clinical outcome of HLA-C mismatched HCT