Assessment of Surfactant Protein A (SP-A) dependent agglutination

Background: Monomers of the collectin surfactant associated protein-A (SP-A) are arranged in trimers and higher oligomers. The state of oligomerization differs between individuals and likely affects SP-A's functional properties. SP-A can form aggregates together with other SP-A molecules. Here we report and assess a test system for the aggregate forming properties of SP-A in serum and broncho-alveolar lavage samples. Methods: Anti-SP-A antibodies fixed to latex beads bound SP-A at its N-terminal end and allowed the interaction with other SP-A molecules in a given sample by their C-terminal carbohydrate recognition domain (CRD) to agglutinate the beads to aggregates, which were quantified by light microscopy. Results: SP-A aggregation was dependent on its concentration, the presence of calcium, and was dose-dependently inhibited by mannose. Unaffected by the presence of SP-D no aggregation was observed in absence of SP-A. The more complex the oligomeric structure of SP-A present in a particular sample, the better was its capability to induce aggregation at a given total concentration of SP-A. SP-A in serum agglutinated independently of the pulmonary disease; in contrast SP-A in lung lavage fluid was clearly inferior in patients with chronic bronchitis and particularly with cystic fibrosis compared to controls. Conclusions: The functional status of SP-A with respect to its aggregating properties in serum and lavage samples can be easily assessed. SP-A in lung lavage fluid in patients with severe neutrophilic bronchitis was inferior

Surfactant proteins SP-A and SP-D modulate uterine contractile events in ULTR myometrial cell line

Pulmonary surfactant proteins SP-A and SP-D are pattern recognition innate immune molecules. However, there is extrapulmonary existence, especially in the amniotic fluid and at the feto-maternal interface. There is sufficient evidence to suggest that SP-A and SP-D are involved in the initiation of labour. This is of great importance given that preterm birth is associated with increased mortality and morbidity. In this study, we investigated the effects of recombinant forms of SP-A and SP-D (rhSP-A and rhSP-D, the comprising of trimeric lectin domain) on contractile events in vitro, using a human myometrial cell line (ULTR) as an experimental model. Treatment with rhSP-A or rhSP-D increased the cell velocity, distance travelled and displacement by ULTR cells. rhSP-A and rhSP-D also affected the contractile response of ULTRs when grown on collagen matrices showing reduced surface area. We investigated this effect further by measuring contractility-associated protein (CAP) genes. Treatment with rhSP-A and rhSP-D induced expression of oxytocin receptor (OXTR) and connexin 43 (CX43). In addition, rhSP-A and rhSP-D were able to induce secretion of GROα and IL-8. rhSP-D also induced the expression of IL-6 and IL-6 Ra. We provide evidence that SP-A and SP-D play a key role in modulating events prior to labour by reconditioning the human myometrium and in inducing CAP genes and pro-inflammatory cytokines thus shifting the uterus from a quiescent state to a contractile one

A mRNA landscape of bovine embryos after standard and MAPK-inhibited culture conditions: a comparative analysis.

BACKGROUND: Genes and signalling pathways involved in pluripotency have been studied extensively in mouse and human pre-implantation embryos and embryonic stem (ES) cells. The unsuccessful attempts to generate ES cell lines from other species including cattle suggests that other genes and pathways are involved in maintaining pluripotency in these species. To investigate which genes are involved in bovine pluripotency, expression profiles were generated from morula, blastocyst, trophectoderm and inner cell mass (ICM) samples using microarray analysis. As MAPK inhibition can increase the NANOG/GATA6 ratio in the inner cell mass, additionally blastocysts were cultured in the presence of a MAPK inhibitor and changes in gene expression in the inner cell mass were analysed. RESULTS: Between morula and blastocyst 3,774 genes were differentially expressed and the largest differences were found in blastocyst up-regulated genes. Gene ontology (GO) analysis shows lipid metabolic process as the term most enriched with genes expressed at higher levels in blastocysts. Genes with higher expression levels in morulae were enriched in the RNA processing GO term. Of the 497 differentially expressed genes comparing ICM and TE, the expression of NANOG, SOX2 and POU5F1 was increased in the ICM confirming their evolutionary preserved role in pluripotency. Several genes implicated to be involved in differentiation or fate determination were also expressed at higher levels in the ICM. Genes expressed at higher levels in the ICM were enriched in the RNA splicing and regulation of gene expression GO term. Although NANOG expression was elevated upon MAPK inhibition, SOX2 and POU5F1 expression showed little increase. Expression of other genes in the MAPK pathway including DUSP4 and SPRY4, or influenced by MAPK inhibition such as IFNT, was down-regulated. CONCLUSION: The data obtained from the microarray studies provide further insight in gene expression during bovine embryonic development. They show an expression profile in pluripotent cells that indicates a pluripotent, epiblast-like state. The inability to culture ICM cells as stem cells in the presence of an inhibitor of MAPK activity together with the reported data indicates that MAPK inhibition alone is not sufficient to maintain a pluripotent character in bovine cells

The potential for immunoglobulins and host defense peptides (HDPs) to reduce the use of antibiotics in animal production

Abstract Innate defense mechanisms are aimed at quickly containing and removing infectious microorganisms and involve local stromal and immune cell activation, neutrophil recruitment and activation and the induction of host defense peptides (defensins and cathelicidins), acute phase proteins and complement activation. As an alternative to antibiotics, innate immune mechanisms are highly relevant as they offer rapid general ways to, at least partially, protect against infections and enable the build-up of a sufficient adaptive immune response. This review describes two classes of promising alternatives to antibiotics based on components of the innate host defense. First we describe immunoglobulins applied to mimic the way in which they work in the newborn as locally acting broadly active defense molecules enforcing innate immunity barriers. Secondly, the potential of host defense peptides with different modes of action, used directly, induced in situ or used as vaccine adjuvants is described

Computed tomographic, radiographic, and endoscopic tracheal dimensions in english bulldogs with grade 1 clinical signs of brachycephalic airway syndrome

Tracheal hypoplasia is commonly seen in English Bulldogs affected with brachycephalic airway syndrome. Previously published diagnostic criteria for tracheal hypoplasia in this breed have been a radiographic tracheal diameter:tracheal inlet ratio (TD:TI) < 0.12 or a tracheal diameter:third rib diameter ratio (TD:3R) < 2.0. Computed tomography has become increasingly used for airway evaluation, however published information is lacking regarding CT tracheal dimensions in English Bulldogs. Objectives of this prospective cross-sectional study were to describe radiographic and CT tracheal dimensions in a sample of clinically normal English Bulldogs and compare these values with tracheoscopy scores. Computed tomography (n = 40), radiography (n = 38), and tracheoscopy (n = 40) studies were performed during a single general anesthesia session for each included dog. Tracheal measurements were recorded at three locations: cervical, thoracic inlet, and thorax. Tracheal diameters were narrowest at the thoracic inlet with all techniques. Computed tomographic measurements averaged 19% greater than radiographic measurements. All included dogs had radiographic tracheal measurements greater than the previously published criteria for tracheal hypoplasia. Mean CT TD:TI was 0.26 (± 0.03, 0.20-0.33), and mean CT TT:3R was 2.27 (± 0.24, 1.71-2.74). Radiographic TD:TI and CT TD:TI were significantly correlated (P = 0.00); however radiographic TT:3R and CT TT:3R were not significantly correlated (P = 0.25). Tracheoscopy identified hypoplastic changes in all dogs and tracheoscopy scores were not correlated with CT or radiography diameter measurements. In conclusion, findings indicated that some CT and radiographic tracheal diameter measurements were comparable in English Bulldogs however diameters for both imaging techniques were not comparable with tracheoscopy scores

COMPUTED TOMOGRAPHIC, RADIOGRAPHIC, AND ENDOSCOPIC TRACHEAL DIMENSIONS IN ENGLISH BULLDOGS WITH GRADE 1 CLINICAL SIGNS OF BRACHYCEPHALIC AIRWAY SYNDROME

Tracheal hypoplasia is commonly seen in English Bulldogs affected with brachycephalic airway syndrome. Previously published diagnostic criteria for tracheal hypoplasia in this breed have been a radiographic tracheal diameter:tracheal inlet ratio (TD:TI) < 0.12 or a tracheal diameter:third rib diameter ratio (TD:3R) < 2.0. Computed tomography has become increasingly used for airway evaluation, however published information is lacking regarding CT tracheal dimensions in English Bulldogs. Objectives of this prospective cross-sectional study were to describe radiographic and CT tracheal dimensions in a sample of clinically normal English Bulldogs and compare these values with tracheoscopy scores. Computed tomography (n = 40), radiography (n = 38), and tracheoscopy (n = 40) studies were performed during a single general anesthesia session for each included dog. Tracheal measurements were recorded at three locations: cervical, thoracic inlet, and thorax. Tracheal diameters were narrowest at the thoracic inlet with all techniques. Computed tomographic measurements averaged 19% greater than radiographic measurements. All included dogs had radiographic tracheal measurements greater than the previously published criteria for tracheal hypoplasia. Mean CT TD:TI was 0.26 (± 0.03, 0.20-0.33), and mean CT TT:3R was 2.27 (± 0.24, 1.71-2.74). Radiographic TD:TI and CT TD:TI were significantly correlated (P = 0.00); however radiographic TT:3R and CT TT:3R were not significantly correlated (P = 0.25). Tracheoscopy identified hypoplastic changes in all dogs and tracheoscopy scores were not correlated with CT or radiography diameter measurements. In conclusion, findings indicated that some CT and radiographic tracheal diameter measurements were comparable in English Bulldogs however diameters for both imaging techniques were not comparable with tracheoscopy scores

The effect of interleukin-2 on canine peripheral nerve sheath tumours after marginal surgical excision : a double-blind randomized study

BACKGROUND: The objective of this study was to evaluate the effect on outcomes of intraoperative recombinant human interleukin-2 injection after surgical resection of peripheral nerve sheath tumours. In this double-blind trial, 40 patients due to undergo surgical excision (<5 mm margins) of presumed peripheral nerve sheath tumours were randomized to receive intraoperative injection of interleukin-2 or placebo into the wound bed. RESULTS: There were no significant differences in any variable investigated or in median survival between the two groups. The median recurrence free interval was 874 days (range 48-2141 days), The recurrence-free interval and overall survival time were significantly longer in dogs that undergone the primary surgery by a specialist-certified surgeon compared to a referring veterinarian regardless of whether additional adjunct therapy was given. CONCLUSION: Overall, marginal excision of peripheral nerve sheath tumours in dogs resulted in a long survival time, but adjuvant treatment with recombinant human interleukin-2 (rhIL-2) did not provide a survival advantage