The molecular background of equine sarcoids

Equine sarcoids (ES) are the most common skin tumors in equids all over the world. No universal treatment is yet available and the aetiology of the disease is not yet fully understood. Though, it is believed to be a multifactorial disease in which both genetic and infectious factors have been implicated. Association between ES and certain serotypes in the Major Histocompatibility Complex (MHC) region has previously been demonstrated and DNA from Bovine Papillomavirus (BPV) type 1 and 2 in and around equine sarcoid tumours has also been detected. The aim of this study is to further characterize the molecular genetic background of the susceptibility of ES with the main focus on the MHC-region. The high resolution and accuracy of microsatellite analyses may complement earlier serological studies. A total number of 78 horses were included in the study. 26 of these were classified as ES-affected by histological analysis. 32 of the horses were Swedish Warmbloods and 16 of these were ES-affected. 12 microsatellites in the MHC-region, including all three classes of MHC, as well as two microsatellites on chromosome 23 were tested for differences in heterozygosity and allele frequencies, occurrence of haplotypes as well as length of consecutive homozygous regions between cases and controls. The majority of the markers with significant results are located in MHC class II. Significant results were also observed for one marker located on chromosome 23. Longer consecutive regions of homozygous markers were observed in cases than in controls indicating a disadvantageous effect. The genetic profiles of horses affected with Equine Sarcoids are different from that of non-affected horses. In addition, the results implicate breed differences in the genotypes of the horses included in the study. Interestingly, this study also indicates protective effects of certain alleles and haplotyps towards the development of ES

Study of the collagen- and fibronectin- binding protein FNE

Strangles is a severe equine respiratory infection, caused by the bacterium Streptococcus equi subspecies equi affecting horses of all ages and types worldwide. Infected horses suffer from fever, coughing and copious nasal discharge as well as abscesses in lymph nodes in the head and throat. S. equi invades the body through the mouth or nose of the horse, and previous studies have shown that the interaction is dependent on various extracellular proteins, several of which have been characterized. In this project one of these proteins, FNE, has been studied. FNE is a secreted fibronectin- and collagen- binding protein unique to S. equi. The main purpose of this project was to identify the collagen- and fibronectin-binding parts of the protein. This was performed by designing primers and truncating the protein from both N- and C-termini (Appendix I). The binding capacities of the truncated proteins were assayed by dot blotting. To further examine the properties of the truncated proteins inhibition assays with antibodies against FNE were performed. Results show that fibronectin binding is restricted to the C-terminal part of the protein and that the amino acids GLEGGSS (228 – 234) are required for the binding. By contrast, collagen binding regions seem to be present at both N- and C-termini, i.e. the collagen binding domain is fairly large. Antibodies seem to be directed to the central part of the protein, and consistent with the above results, interaction with antibodies inhibits fibronectin- and collagen- binding. The results will hopefully contribute to a deeper basic knowledge about S. equi and the FNE protein as well as be important in the progress towards the development of a safe and effective vaccine against strangles

Acute sleep loss results in tissue-specific alterations in genome-wide DNA methylation state and metabolic fuel utilization in humans

Curtailed sleep promotes weight gain and loss of lean mass in humans, although the underlying molecular mechanisms are poorly understood. We investigated the genomic and physiological impact of acute sleep loss in peripheral tissues by obtaining adipose tissue and skeletal muscle after one night of sleep loss and after one full night of sleep. We find that acute sleep loss alters genome-wide DNA methylation in adipose tissue, and unbiased transcriptome-, protein-, and metabolite-level analyses also reveal highly tissue-specific changes that are partially reflected by altered metabolite levels in blood. We observe transcriptomic signatures of inflammation in both tissues following acute sleep loss, but changes involving the circadian clock are evident only in skeletal muscle, and we uncover molecular signatures suggestive of muscle breakdown that contrast with an anabolic adipose tissue signature. Our findings provide insight into how disruption of sleep and circadian rhythms may promote weight gain and sarcopenia

Mass spectrometry data from a quantitative analysis of protein expression in gills of immuno-challenged blue mussels (Mytilus edulis)

Here, we provide the dataset associated with our research article on the potential effects of ocean acidification on antimicrobial peptide (AMP) activity in the gills of Mytilus edulis, "Impact of ocean acidification on antimicrobial activity in gills of the blue mussel (Mytilus edulis)" [1]. Blue mussels were stimulated with lipopolysaccharides and samples were collected at different time points post injection. Protein extracts were prepared from the gills, digested using trypsin and a full in-depth proteome investigation was performed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Protein identification and quantification was performed using the MaxQuant 1.5.1.2 software, "MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification" [2]

Impact of ocean acidification on antimicrobial activity in gills of the blue mussel (Mytilus edulis)

Here, we aimed to investigate potential effects of ocean acidification on antimicrobial peptide (AMP) activity in the gills of Mytilus edulis, as gills are directly facing seawater and the changing pH (predicted to be reduced from 8.1 to 7.7 by 2100). The AMP activity of gill and haemocyte extracts was compared at pH 6.0, 7.7 and 8.1, with a radial diffusion assay against Escherichia coli. The activity of the gill extracts was not affected by pH, while it was significantly reduced with increasing pH in the haemocyte extracts. Gill extracts were also tested against different species of Vibrio (V. parahaemolyticus Vibrio tubiashii, V. splendidus and V. alginoyticus) at pH 7.7 and 8.1. The metabolic activity of the bacteria decreased by 65-90%, depending on species of bacteria, but was, as in the radial diffusion assay, not affected by pH. The results indicated that AMPs from gills are efficient in a broad pH-range. However, when mussels were pre-exposed for pH 7.7 for four month the gill extracts presented significantly lower inhibit of bacterial growth. A full in-depth proteome investigation of gill extracts, using LC-Orbitrap MS/MS technique, showed that among previously described AMPs from haemocytes of Mytilus, myticin A was found up-regulated in response to lipopolysaccharide, 3 h post injection. Sporadic occurrence of other immune related peptides/proteins also pointed to a rapid response (0.5?3 h p.i.). Altogether, our results indicate that the gills of blue mussels constitute an important first line defence adapted to act at the pH of seawater. The antimicrobial activity of the gills is however modulated when mussels are under the pressure of ocean acidification, which may give future advantages for invading pathogens

The X proteins of bornaviruses interfere with type I interferon signalling

Borna disease virus (BDV) is a neurotropic, negative-stranded RNA virus causing persistent infection and progressive neurological disorders in a wide range of warm-blooded animals. The role of the small non-structural X protein in viral pathogenesis is not completely understood. Here we investigated whether the X protein of BDV and avian bornavirus (ABV) interferes with the type I interferon (IFN) system, similar to other non-structural proteins of negative-stranded RNA viruses. In luciferase reporter assays, we found that the X protein of various bornaviruses interfered with the type I IFN system at all checkpoints investigated, in contrast to previously reported findings, resulting in reduced type I IFN secretion

Proteins secreted during <i>Giardia</i>-host cell interactions.

<p>During <i>Giardia</i>-host cell interactions <i>in vitro</i> specific proteins are released into the growth medium. The same types of proteins, mainly enzymes, are released from both the host and parasite (black) but there are also parasite-specific proteins being released (blue).</p

The effects of ESPs on IEC cell signaling.

<p>(A) Western blot analyses of differentiated Caco-2 cells exposed to <i>G</i>. <i>intestinalis</i> ESPs through a Transwell insert for 2h or 6h in the absence of inflammatory stimuli (e.g. lipopolysaccharides or tumour necrosis factor-alpha). Blots were probed for total proteins (phosphorylated/non-phosphorylated forms) of mitogen activated proteins kinases (MAPKs, ERK1/2 and P38) and nuclear factor kappa beta (NFκB). (B) Western blot analyses after the induction of inflammation with LPS and TNF-alpha.</p

The binding and internalization of <i>Giardia intestinalis</i> excreted-secreted products (ESPs) to intestinal epithelial cells (IECs).

<p>Confocal microscopy images of differentiated human colon carcinoma cell line, Caco-2, incubated with Alexa Fluor488-labelled ESPs of <i>G</i>. <i>intestinalis</i> isolate WB or GS. Note that the colour of nucleus, stained with DAPI, was changed to red for better contrast.</p