Summary Immunohistological Determination of Chlamydia psittaci/pecorum and C. trachomatis in the Piglet Gut The jejunum, ileum, caecum and colon of 200 piglets were investigated immunohistochemically for the presence of Chlamydia psittaci and C. trachomatis using a vitelline IgY. Positive samples were later labelled using a commercial C. trachomatis polyclonal antiserum. Chlamydia were present in 33 (16.4%) of the animals, and 30 out of 33 were labelled by C. trachomatis polyclonal antiserum. Inclusions occurred predominantly (67%) in the large intestine. The serological results (CFT, ELISA) did not correlate well with immunohistochemical labelling in the gut. The incidence of Chlamydia rose from 6.9% in animals up to 4 weeks, to 41.8% in those over 4 weeks of age. A correlation between chlamydia and enteric disease was not obvious. Besides chlamydia, most of the diseased animals harboured other additional agents. In conclusion, intestinal chlamydiae in piglets, predominantly C. trachomatis, exist in Switzerland, although their pathogenic potential seems to be low. Zusammenfassung Därme (Jejunum, Ileum, Zaekum, Kolon) von insgesamt 200 Ferkeln wurden immunhistologisch auf Chlamydien untersucht. Der Chlamydien‐Nachweis erfolgte mittels der Avidin‐Biotin‐Methode. Als primärer Antikörper wurde ein aus dem Eidotter von mit C. psittaci und C. trachomatis immunisierten Hühnern gewonnenes Vitellinimmunglobulin IgY und ein kommerzieller polyklonaler Antiserum gegen C. trachomatis verwendet. Insgesamt 33 (16,4%) Schweine erwiesen sich als Chlamydienträger, 30 von 33 beherbergten C. trachomatis. Chlamydien‐Einschlüsse fanden sich überwiegend im Dickdarm (67%). Bei Tieren über 4 Wochen wurden Chlamydien häufiger nachgewiesen (41,8%) als bei Tieren bis zu 4 Wochen (6,9%). Die Ergebnisse der serologischen Untersuchung korrelieren nicht eindeutig mit den immunhistologischen Befunden im Darm. Ein eindeutiger Zusammenhang zum Durchfallgeschehen konnte nicht festgestellt werden, da mit Ausnahme von zwei Fällen bei den an Durchfall erkrankten Ferkeln neben Chlamydien auch andere darmpathogene Erreger gefunden wurden. Die vorliegenden Untersuchungen zeigen, daß auch in der Schweiz latente Darminfektionen mit Chlamydien bei Ferkeln und Läufern vorkommen, und dass Chlamydien als alleinige Durchfallerreger für Schweine vermutlich nur eine geringe Pathogenität aufweisen.

Background: Untreated disorders of the adrenocortical system, such as Cushing's or Addison's disease, can be fatal, and accurate quantification of a patient's cortisol levels is vital for diagnosis. The objective of this study was to assess the analytical performance of a new fullyautomated Elecsys ® Cortisol II assay (second generation) to measure cortisol levels in serum and saliva. Methods: Four European investigational sites assessed the intermediate precision and reproducibility of the Cortisol II assay (Roche Diagnostics) under routine conditions. Method comparisons of the Cortisol II assay vs. liquid chromatography-tandem mass spectrometry (LC-MS/MS), the gold standard for cortisol measurement, were performed. Cortisol reference ranges from three US sites were determined using samples from self-reported healthy individuals.

Highlights FDA-EUA approved anti-SARS-CoV-2 tests compare favorably with others. longitudinal patterns of antibody development are subject to high individual variability. comparing the assays of leading manufacturers, there are considerable differences in terms of achieved and stated sensitivity/specificity. Immune response relies on disease severity, thus need to be considered for validation.

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Anterior cruciate ligament (ACL) cell sheets combined with biomechanically competent scaffolds might facilitate ACL tissue engineering. Since thermoresponsive polymers allow a rapid enzyme-free detachment of cell sheets, we evaluated the applicability of a thermoresponsive poly(glycidyl ether) (PGE) coating for cruciate ligamentocyte sheet formation and its influence on ligamentocyte phenotype during sheet-mediated colonization of embroidered scaffolds. Ligamentocytes were seeded on surfaces either coated with PGE or without coating. Detached ligamentocyte sheets were cultured separately or wrapped around an embroidered scaffold made of polylactide acid (PLA) and poly(lactic-co-ε-caprolactone) (P(LA-CL)) threads functionalized by gas-phase fluorination and with collagen foam. Ligamentocyte viability, protein and gene expression were determined in sheets detached from surfaces with or without PGE coating, scaffolds seeded with sheets from PGE-coated plates and the respective monolayers. Stable and vital ligamentocyte sheets could be produced within 24 h with both surfaces, but more rapidly with PGE coating. PGE did not affect ligamentocyte phenotype. Scaffolds could be colonized with sheets associated with high cell survival, stable gene expression of ligament-related type I collagen, decorin, tenascin C and Mohawk after 14 d and extracellular matrix (ECM) deposition. PGE coating facilitates ligamentocyte sheet formation, and sheets colonizing the scaffolds displayed a ligament-related phenotype.

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Spheroid culture might stabilize the ligamentocyte phenotype. Therefore, the phenotype of lapine cruciate ligamentocyte (L-CLs) minispheroids prepared either by hanging drop (HD) method or by using a novel spheroid plate (SP) and the option of methyl cellulose (MC) for tuning spheroid formation was tested. A total of 250 and 1000 L-CLs per spheroid were seeded as HDs or on an SP before performing cell viability assay, morphometry, gene expression (qRT-PCR) and protein immunolocalization after 7 (HD/SP) and 14 (SP) days. Stable and viable spheroids of both sizes could be produced with both methods, but more rapidly with SP. MC accelerated the formation of round spheroids (HD). Their circular areas decreased significantly during culturing. After 7 days, the diameters of HD-derived spheroids were significantly larger compared to those harvested from the SP, with a tendency of lower circularity suggesting an ellipsoid shape. Gene expression of decorin increased significantly after 7 days (HD, similar trend in SP), tenascin C tended to increase after 7 (HD/SP) and 14 days (SP), whereas collagen type 1 decreased (HD/SP) compared to the monolayer control. The cruciate ligament extracellular matrix components could be localized in all mini-spheroids, confirming their conserved expression profile and their suitability for ligament tissue engineering.

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