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Background A coproantigen enzyme-linked immunosorbent assay (ELISA) has recently been proposed for detecting ascarid infections in chickens. The excretion pattern of ascarid antigens through chicken faeces and the consistency of measurements over the course of infections are currently unknown. This study evaluates the pattern and repeatability of worm antigen per gram of faeces (APG) and compares the diagnostic performance of the coproantigen ELISA with a plasma and egg yolk antibody ELISA and McMaster faecal egg counts (M-FEC) at different weeks post-infection (wpi). Methods Faecal, blood and egg yolk samples were collected from laying hens that were orally infected with a mix of Ascaridia galli and Heterakis gallinarum eggs (N = 108) or kept as uninfected controls (N = 71). Measurements including (a) APG using a coproantigen ELISA, (b) eggs per gram of faeces (EPG) using the McMaster technique and (c) ascaridspecific IgY in plasma and in egg yolks using an ascarid-specific antibody ELISA) were performed between wpi 2 and 18. Results Time-dependent significant differences in APG between infected and non-infected laying hens were quantified. At wpi 2 (t(164) = 0.66, P = 1.00) and 4 (t(164) = −3.09, P = 0.094) no significant differences were observed between the groups, whereas infected hens had significantly higher levels of APG than controls by wpi 6 (t(164) = −6.74, P < 0.001). As indicated by a high overall repeatability estimate of 0.91 (CI = 0.89–0.93), APG could be measured consistently from the same individual. Compared to McMaster and antibody ELISA, coproantigen ELISA showed the highest overall diagnostic performance (area under curve, AUC = 0.93), although the differences were time-dependent. From wpi 6 to 18 coproantigen ELISA had an AUC > 0.95, while plasma IgY ELISA showed the highest diagnostic&nbsp

Erik Őrfi,1,2 Tamás Mészáros,1,2 Mark Hennies,3 Tamás Fülöp,1,2 László Dézsi,1,2 Alexander Nardocci,1 László Rosivall,1,2 Péter Hamar,4,5 Barry W Neun,6 Marina A Dobrovolskaia,6 János Szebeni,1,2,7,* Gábor Szénási1,8,* 1Nanomedicine Research and Education Center, Institute of Pathophysiology, Semmelweis University, Budapest, Hungary; 2SeroScience LCC., Cambridge, MA, USA; 3TECOdevelopment GmbH, Rheinbach, Germany; 4Institute of Clinical Experimental Research, Semmelweis University, Budapest, Hungary; 5Institute for Translational Medicine, Medical School, University of Pécs, Pécs, Hungary; 6Nanotechnology Characterization Laboratory, Frederick National Laboratory for Cancer Research, National Cancer Institute, Frederick, MD, USA; 7Department of Nanobiotechnology and Regenerative Medicine, Faculty of Health, Miskolc University, Miskolc, Hungary; 8Institute of Pathophysiology, Semmelweis University, Budapest, Hungary *These authors contributed equally to this work Purpose: Undesirable complement (C) activation by nanomedicines can entail an adverse immune reaction known as C activation-related pseudoallergy (CARPA) in sensitive patients. The syndrome includes cardiopulmonary, hemodynamic, and a variety of other physiological changes that have been well described in man, pigs, dogs, and rats. However, the information on CARPA is scarce and ambiguous in mice, a species widely used in preclinical studies. The present study aimed to fill this gap by exploring signs of CARPA in mice following i.v. administration of AmBisome and Abelcet, which are nano-formulations of Amphotericin B with high risk to cause CARPA. Materials and methods: Anesthetized NMRI mice were intravenously injected with liposomal amphotericin B (Abelcet and AmBisome; 30–300 mg phospholipid/kg), drug-free high cholesterol multilamellar vesicles (HC-MLV), and positive controls, cobra venom factor (CVF) and zymosan, followed by the measurement of blood pressure (BP), heart rate, white blood cell, and platelet counts and plasma thromboxane B2 (TXB2) levels. C activation was assessed by C3a ELISA, a C3 consumption assay (PAN-C3) and a modified sheep red blood cell hemolytic assay. Results: All test agents, except HC-MLV, caused transient hypertension, thrombocytopenia, and elevation of plasma TXB2, which were paralleled by significant rises of plasma C3a in CVF and zymosan-treated animals, wherein the initial hypertension turned into hypotension and shock. Abelcet and AmBisome caused minor, delayed rise of C3a that was not associated with hypertension. The C3a receptor inhibitor SB-290157 attenuated the hypertension caused by Abelcet and decreased the BP thereafter. Conclusion: The parallelism between C3a anaphylatoxin production and severity of physiological changes caused by the different agents is consistent with CARPA underlying these changes. Although the reactive dose of liposomal phospholipids was substantially higher than that in other species (pigs, dogs), the mouse seems suitable for studying the mechanism of hypersensitivity reactions to liposomal formulations of amphotericin B, a frequent side effect of these drugs. Keywords: hypersensitivity, infusion reactions, zymosan, cobra venom factor, TXB2, cholesterol, anaphylatoxins, platelets

Background Classical faecal egg counts (FEC) provide less reliable diagnostic information for nematode infections in chickens. We developed an ELISA based on Ascaridia galli antigens and tested two hypotheses, as follows: (i) IgY antibodies developed against A. galli will also be useful to identify Heterakis gallinarum infections, and (ii) circulating antibodies stored in egg yolks are as good as plasma samples, so a non-invasive diagnosis is possible. The aim of this study, therefore, was to compare the diagnostic accuracy of the ELISA system with FEC, using both plasma and egg yolks from experimentally infected hens. In addition, naturally infected animals were evaluated to validate the assay. Results The assay quantified large differences (P 0.90 for both nematodes using either plasma or egg yolks. Sensitivity of the assay was 94 and 93% with plasma and egg yolk samples, respectively, whereas FEC yielded in a sensitivity of 84% in A. galli experiment. Total test accuracy of the assay with plasma samples (AUC = 0.99) tended to be higher (P = 0.0630) than FEC (AUC = 0.92) for A. galli, while the assay with either sample matrix performed similar to FEC (AUC ≥ 0.91) for H. gallinarum. Among the three tests, the FECs correlated better with A. galli burden than the ELISA. Although 90% of naturally infected hens were correctly identified by the ELISA, 45% of the infected hens tested negative with FEC, indicating the validity of the higher test accuracy of the ELISA. Conclusions Antigens of A. galli can be used successfully to identify H. gallinarum-infected animals, indicating that chickens develop cross-reactive antibodies against the two closely related species. Egg yolks are as informative as plasma samples, so that animal welfare-friendly sampling is possible. Although the assay with plasma samples reveals qualitative information of higher quality than FECs on the infection status of naturally infected birds, the latter is still a better tool to assess the intensity of A. galli but not of H. gallinarum infections. Electronic supplementary material The online version of this article (doi:10.1186/s13071-017-2121-9) contains supplementary material, which is available to authorized users.

Average (Mean ± SE) plasma and egg-yolk antibody concentrations of infected birds and their uninfected-control counterparts in A. galli and H. gallinarum experiments. (DOC 29 kb)

Anhand von 75 seitlichen Fernröntgenaufnahmen und korrespondierenden Gipsmodellen wurde untersucht, ob die konventionell am FRS gewonnenen Werte der Frontzahninklinationswinkel im Ober- und Unterkiefer und des Interinzisalwinkels aus am Modell bestimmten Werten berechnet werden können. Nach der Modellduplikation erfolgte die Entfernung der linken Seite durch Trimmen. Durch den höchsten Punkt der Inzisalkante und die Hälfte der labio-oralen Distanz, gemessen zwischen den tiefsten Punkten des Sulkus, wurde eine Gerade auf die zur Kauebene parallelen jeweiligen Sockelebenen gezogen, der dorsale Winkel gemessen und mit entsprechenden Messwerten aus dem FRS verglichen. In einem weiteren Schritt wurden in Analogie der Messungen am Modell die Frontzahninklinationen auf eine am FRS neu eingeführte Kauebene bezogen. In einer Vorauswertung wurden diese Messmethoden als reproduzierbar und wiederholbar bestätigt. Die mit Hilfe einer linearen Regression untersuchte Vorhersagbarkeit der am FRS ermittelten Werte für die Frontzahninklinationswinkel im Ober- und Unterkiefer (R² = 0,33 / 0,54) und den Interinzisalwinkel (R² = 0,73) aus am Modell bestimmten Werten erreichte nicht die klinisch erforderliche Präzision. Am FRS konnte im Vergleich zu den konventionell verwendeten Bezugsebenen durch die Messung der Frontzahninklinationswinkel zur Kauebene die Güte der Vorhersagbarkeit aus am Modell ermittelten Werten gesteigert werden (R² = 0,53 / 0,74). Eine Auswertung nach Angle Klasse, skelettaler Klasse, Wachstumsmuster, Overjet, Overbite und dem Verhältnis des Zahnmaterials beider Kiefer ergab keine wesentliche Verbesserung der Vorhersagbarkeit. Das Gipsmodell kann derzeit das FRS als diagnostische Grundlage zur Bestimmung betreffender Winkel nicht ersetzen. Methodische Unterschiede der am FRS und am Modell untersuchten Messtechniken und auf beiden Seiten bestehende Fehlerquellen erschweren einen Vergleich der Messungen. Zukünftig erscheint es denkbar, durch eine computergestützte Bestimmung der Winkel anhand von digitalisierten Gipsmodellen die Präzision der Methode zu erhöhen. A total of 75 lateral cephalometric radiographs and corresponding dental casts were evaluated to determine if radiographic measurements of the incisor inclination and the interincisal angle can be calculated from dental casts. The dental casts were duplicated and the left sides of the copies were trimmed off. A line was drawn from the highest point of the incisal edge to the half-way point between the labial and the oral gingival sulcus. The dorsal angle formed by this line and the base of the cast parallel to the occlusal plane was then compared to the corresponding values from cephalometric radiographs. The incisor inclinations were then compared to a newly introduced occlusion plane in the cephalometric radiographs as in the measurements of the dental casts. A preliminary evaluation had shown these measurement methods to be reproducible and repeatable. Linear regression analysis was used to establish the predictability of the cephalometric radiographs values from those based on dental casts. This analysis showed that the predictability of measurements of the upper and lower central incisor inclination (R² = 0.33 / 0.54) and of the interincisal angle (R² = 0.73) was not sufficiently accurate for clinical application. While predictability could be increased by determining the incisor inclination in relation to an occlusal plane in the cephalometric radiograph (R² = 0.53 / 0.74), it was not enhanced by evaluation of Angle class, skeletal class, growth pattern, overjet, overbite or the relation of the tooth size of both jaws. It can thus be concluded that dental casts cannot presently be used in place of lateral cephalometric radiograph as a diagnostic tool for the determination of the angles in question. Methodical differences in measurement techniques and other sources of error also complicate the comparison of the two methods. However, it is conceivable that the accuracy of this method can be enhanced in future by computer-aided evaluation of the angles in digitalized dental casts.