Mycoplasma synoviae lipoprotein MSPB, the N-terminal part of VlhA haemagglutinin, induces secretion of nitric oxide, IL-6 and IL-1beta in chicken macrophages

Mycoplasma synoviae is a major pathogen of chickens and turkeys, causing respiratory disease and infectious synovitis. M. synoviae haemagglutinin VlhA is an abundant surface-exposed lipoprotein and immunodominant antigen. Post-translational cleavage of VlhA generates two proteins, MSPB and MSPA. MSPB, the amino-terminal end of VlhA, is a lipoprotein of about 40-50 kDa but can appear in truncated forms (tMSPB) of about 20-30 kDa. The aim of this study was to determine whether MSPB and tMSPB can stimulate chicken macrophages to secrete NO and cytokines. Macrophages derived from chicken monocytes (MDM) or the MQ-NCSU macrophage cell line were stimulated with M. synoviae protein extracts containing MSPB or tMSPB, as well as with purified MSPB and tMSPB. Proteins from detergent extractions induced IL-6 secretion in MDM and NO secretion in MQ-NCSU. Both MSPB and tMSPB were capable of inducing NO secretion in MQ-NCSU, as well as IL-6 and IL-1beta in MDM. The activity of IL-6 induced by purified tMSPB was similar to the effect of 60 pg/ml of recombinant chicken IL-6. The effect of IL-1beta induced by tMSPB was comparable to the effect of 10 ng/ml of recombinant IL-1beta. Whereas all samples containing MSPB were able to induce NO, IL-6 and IL-1beta, it seemed that the purified tMSPB of about 20 kDa was the most potent in its ability to induce IL-6 and IL-1beta in MDM. Compared to MSPB, tMSPB lacks about 200 amino acids in its carboxyl-terminal part. Therefore, our results suggest that the major part of the stimulating activity is associated with the amino-terminal part of MSPB, most likely with its lipid moiety.     

A study of measurement of noninvasive blood pressure with the oscillometric device,Sentinel, in isoflurane‐anaesthetized horses

ObjectiveTo assess accuracy of noninvasive blood pressure (NIBP) measured by oscillometric device Sentinel compared to invasive blood pressure (IBP) in anaesthetized horses undergoing surgery. To assess if differences between the NIBP measured by the Sentinel and IBP are associated with recumbency, cuff placement, weight of the horse or acepromazine premedication and to describe usefulness of the Sentinel.Study designProspective study examining replicates of simultaneous NIBP and IBP measurements.AnimalsTwenty-nine horses.MethodsInvasive blood pressure was measured via a catheter in the facial artery, transverse facial artery or metatarsal artery. NIBP was measured using appropriate size cuffs placed on one of two metacarpal or metatarsal bones or the tail in random order. With both techniques systolic (SAP), mean (MAP), and diastolic (DAP) arterial blood pressures and heart rates (HR) were recorded. A mixed effects model compared the IBP to the NIBP values and assessed potential effects of catheter placement, localisation of the cuffs in combination with recumbency, weight of the horse or acepromazine premedication.ResultsNoninvasive blood pressure yielded higher measurements than IBP. Agreement varied with recumbency and cuff position. Estimated mean differences between the two methods decreased from SAP (lateral recumbency: range -5.3 to -56.0 mmHg; dorsal recumbency: range 0.8 to -20.7 mmHg), to MAP (lateral recumbency: range -1.8 to -19.0 mmHg; dorsal recumbency: range 13.9 to -16.4 mmHg) to DAP (lateral recumbency: range 0.5 to -6.6 mmHg; dorsal recumbency: range 21.0 to -15.5 mmHg). NIBP measurement was approximately two times more variable than IBP measurement. No significant difference between IBP and NIBP due to horse's weight or acepromazine premedication was found. In 227 of 1047 (21.7%) measurements the Sentinel did not deliver a result.Conclusion and clinical relevanceAccording to the high variability of NIBP compared to IBP, NIBP measurements as measured by the Sentinel in the manner described here are not considered as an appropriate alternative to IBP to measure blood pressure in anaesthetized horses.     

Identification of Dynamic Changes in Proteins Associated with the Cellular Cytoskeleton after Exposure to Okadaic Acid

Exposure of cells to the diarrhetic shellfish poison, okadaic acid, leads to a dramatic reorganization of cytoskeletal architecture and loss of cell-cell contact. When cells are exposed to high concentrations of okadaic acid (100–500 nM), the morphological rearrangement is followed by apoptotic cell death. Okadaic acid inhibits the broad acting Ser/Thr protein phosphatases 1 and 2A, which results in hyperphosphorylation of a large number of proteins. Some of these hyperphosphorylated proteins are most likely key players in the reorganization of the cell morphology induced by okadaic acid. We wanted to identify these phosphoproteins and searched for them in the cellular lipid rafts, which have been found to contain proteins that regulate cytoskeletal dynamics and cell adhesion. By using stable isotope labeling by amino acids in cell culture cells treated with okadaic acid (400 nM) could be combined with control cells before the isolation of lipid rafts. Protein phosphorylation events and translocations induced by okadaic acid were identified by mass spectrometry. Okadaic acid was shown to regulate the phosphorylation status and location of proteins associated with the actin cytoskeleton, microtubules and cell adhesion structures. A large number of these okadaic acid-regulated proteins have previously also been shown to be similarly regulated prior to cell proliferation and migration. Our results suggest that okadaic acid activates general cell signaling pathways that induce breakdown of the cortical actin cytoskeleton and cell detachment.     

Preparation and comparative release characteristics of three anthocyanin encapsulation systems

Bilberries (Vaccinium myrtillus L.) and their major polyphenolic constituents, anthocyanins, have preventive activities inter alia against colon cancer and inflammatory bowel diseases. However, anthocyanins are sensitive to environmental conditions; thus their bioavailability in the gastrointestinal tract is an important determinant of their in vivo activity. In the study reported here, the potential benefits of encapsulating an anthocyanin rich bilberry extract (BE) on anthocyanin stability were investigated. Nonencapsulated BE and three different BE loaded microcapsule systems were incubated in simulated gastric fluid (SGF) and fed state simulated intestinal fluid (FeSSIF). After exposure to these media, released anthocyanins were identified and quantified by HPLC with UV/Vis detection. Although a rapid release of anthocyanins was observed within the first 20 min, encapsulation of anthocyanins doubled the amount of available anthocyanins after 150 min of incubation. These results illustrate the ability of encapsulation to inhibit early degradation of anthocyanins in the intestinal system.     

Microbiological aspects of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis>) and its processing—relevance for final product quality: a review

The aquaculture production of common carp (Cyprinus carpio) is one of the biggest on a global scale, although European production represents a minor part. Thus, common carp is a little-exploited, low-cost, and highly nutritious food source. For development of new quality products that have customers’ appeal and are safe, a combination of traditional and novel technologies can be used. However, good quality raw material is of basic importance for further processing and final product quality. Fish microbiota have a major role in fish spoilage and as potential human pathogens. To diminish the negative impact of the microbiota on fish, different methods and technologies can be used. The important steps before the final product processing in the production of common carp products include purging, transport, pre-slaughter storage, slaughter method, bleeding, cleaning, desliming, descaling, and gutting. The most important factor in fish spoilage prevention is chilling, although to assure longer freshness and shelf-life, the concept of hurdle technology should be used. Many preservation and packaging techniques have been developed for fish products, but not all have been researched for common carp products. This review aims to identify the gaps in research, knowledge, and practice for the microbiological aspects that impact upon the production of high-quality common carp food products.