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1.
Poult Sci ; 97(6): 2064-2070, 2018 Jun 01.
Article in English | MEDLINE | ID: mdl-29471412

ABSTRACT

Two broiler trials were designed to investigate the relationship between the concentration of non-starch polysaccharides (NSP) in wheat and 1) its nutritional value for broilers and 2) the efficacy of exogenous enzymes. In a balance trial, diets were formulated with 3 wheat cultivars (Rustic and Viscount-medium NSP, Centenaire-high NSP) and were tested with or without the addition of an exogenous enzyme mixture. The diets were fed to 144 male Ross 308 broiler chickens housed in digestibility cages. Total tract nutrient digestibilities and AMEn were measured from 18 to 22 d of age. In a performance trial, diets were formulated with wheat (medium NSP diet) or with wheat mixed with rye and barley (high NSP diet) and were tested with or without the addition of an exogenous enzyme mixture. The diets were fed to 960 male Ross 308 broilers housed in pens and broiler performance during starter, grower and finisher periods was measured.In the balance trial, wheat cultivar did not affect nutrient digestibility or AMEn. Enzyme addition caused a significant increase in nutrient digestibilities and AMEn for the diet formulated with the high NSP wheat Centenaire only. In the performance trial, feeding the high NSP diet resulted in a higher feed conversion ratio and lower final body weight compared to the medium NSP diet. The largest improvements by enzyme addition were observed in the high NSP diet.In conclusion, the study was not able to show a consistent relationship between the NSP concentration of wheat and its nutritional value, but did demonstrate that the effect of an enzyme mixture on nutrient digestibility or broiler performance depends upon the NSP concentration in the diet.


Subject(s)
Chickens/physiology , Digestion , Endo-1,4-beta Xylanases/metabolism , Nutritive Value , Polysaccharides/metabolism , Animal Feed/analysis , Animal Nutritional Physiological Phenomena , Animals , Chickens/growth & development , Diet/veterinary , Male , Random Allocation , Triticum/chemistry
2.
Gene ; 610: 9-14, 2017 Apr 30.
Article in English | MEDLINE | ID: mdl-28185859

ABSTRACT

Interferon regulatory factor 5 (IRF5) regulates type I interferon (IFN)-responsive genes, and has been one of the most consistently associated genes with systemic lupus erythematosus (SLE). We sought to investigate whether IRF5 haplotypes are associated with risk for SLE in the genetically homogeneous Greek population of the island of Crete, as well as whether these haplotypes are associated with increased type I IFN. 322 SLE patients and 247 healthy controls from Crete were genotyped for rs2004640, rs3807306, rs10488631 and rs2280714 SNPs of IRF5 gene by using Taqman primer-probe sets. Type I IFN levels were measured using a functional reporter cell assay. All IRF5 SNPs examined were found to be associated with SLE in univariate case-control analysis. The 4 SNPs formed 5 major haplotypes and the Neanderthal-derived TACA risk haplotype was present in Crete and enriched in the SLE cases (OR=2.01, P=0.0003). Serum IFN levels were measured in a subset of the SLE patients, and carriage of the TACA haplotype was associated with higher circulating type I IFN levels (P=0.037). This study demonstrates the association of IRF5 with an increased susceptibility for SLE in the population of Crete and emphasizes the association of the Neanderthal-derived IRF5 haplotype with SLE susceptibility. Patients carrying allele the Neanderthal allele C had greater type I IFN, supporting a functional consequence of this polymorphism.


Subject(s)
Genetic Predisposition to Disease , Interferon Regulatory Factors/genetics , Lupus Erythematosus, Systemic/genetics , Adult , Animals , Case-Control Studies , Female , Greece , Haplotypes , Humans , Interferon Type I/blood , Interferon Type I/immunology , Lupus Erythematosus, Systemic/immunology , Male , Middle Aged , Neanderthals/genetics
3.
Lupus ; 26(3): 240-247, 2017 Mar.
Article in English | MEDLINE | ID: mdl-27365370

ABSTRACT

In 2012, the Systemic Lupus International Collaborating Clinics (SLICC) group published a new set of classification criteria for systemic lupus erythematosus (SLE). Studies applying these criteria to real-life scenarios have found either equal or greater sensitivity and equal or lower specificity to the 1997 ACR classification criteria (ACR 97). Nonetheless, there are no studies that have used the SLICC 12 criteria to investigate the incidence of lupus. We used the resource of the Rochester Epidemiology Project to identify incident SLE patients in Olmsted County, Minnesota, from 1993 to 2005, who fulfilled the ACR 97 or SLICC 12 criteria. A total of 58 patients met criteria by SLICC 12 and 44 patients met criteria by ACR 97. The adjusted incidence of 4.9 per 100,000 person-years by SLICC 12 was higher than that by ACR 97 (3.7 per 100,000 person-years, p = 0.04). The median duration from the appearance of first criterion to fulfillment of the criteria was shorter for the SLICC 12 than for ACR 97 (3.9 months vs 8.1 months). The higher incidence by SLICC 12 criteria came primarily from the ability to classify patients with renal-limited disease, the expansion of the immunologic criteria and the expansion of neurologic criteria.


Subject(s)
Lupus Erythematosus, Systemic/classification , Lupus Erythematosus, Systemic/epidemiology , Rheumatology , Adolescent , Adult , Age Distribution , Aged , Aged, 80 and over , Female , Humans , Incidence , Male , Middle Aged , Minnesota/epidemiology , Regression Analysis , Severity of Illness Index , Societies, Medical , Young Adult
5.
Vet Immunol Immunopathol ; 167(1-2): 51-6, 2015 Sep 15.
Article in English | MEDLINE | ID: mdl-26187439

ABSTRACT

During the last years, an antimicrobial protein from the RegIII family has been consistently identified as one of the main up-regulated mRNA transcripts in the pig small intestinal mucosa during different infections such as enterotoxigenic Escherichia coli (ETEC). This transcript has been mainly referred to in the literature as pancreatitis-associated protein (PAP/RegIIIα). However, the identity of this transcript has not been confirmed, and no evidence of its expression at the protein level is available in the literature, because the absence of a specific antibody. In this study, we first unequivocally identified the PAP/RegIII family protein mainly expressed in ETEC infected pig intestine as RegIIIγ by 2D-DIGE and MALDI-TOF/TOF. This shows that the pig differs from species like human and mice in that RegIIIγ (and not RegIIIα) might be the major RegIII isotype during intestinal infection. Immunoblotting analysis with a specifically generated polyclonal rabbit antibody revealed that pig RegIIIγ is expressed throughout the intestinal tract, but most abundantly in the ileum. Although a higher abundance of mRNA was paralleled by higher protein abundance, a lack of linear relationship was found between RegIIIγ mRNA and protein abundances in the jejunal mucosa, the latter most pronounced in the case of natural infection. This may be related to the secretory nature of RegIIIγ. This would mean that the antimicrobial protein RegIIIγ is a good candidate as a non-invasive faecal intestinal health biomarker in swine.


Subject(s)
Antigens, Neoplasm/metabolism , Biomarkers, Tumor/metabolism , Intestinal Mucosa/immunology , Lectins, C-Type/metabolism , Sus scrofa/immunology , Amino Acid Sequence , Animals , Antigens, Neoplasm/genetics , Antimicrobial Cationic Peptides/genetics , Antimicrobial Cationic Peptides/metabolism , Biomarkers/metabolism , Biomarkers, Tumor/genetics , Consensus Sequence , Enterotoxigenic Escherichia coli , Escherichia coli Infections/immunology , Escherichia coli Infections/metabolism , Escherichia coli Infections/veterinary , Humans , Ileum/immunology , Ileum/metabolism , Intestinal Mucosa/metabolism , Lectins/genetics , Lectins/metabolism , Lectins, C-Type/genetics , Liver/immunology , Liver/metabolism , Mice , Molecular Sequence Data , Pancreatitis-Associated Proteins , Rabbits , Sequence Homology, Amino Acid , Species Specificity , Sus scrofa/genetics , Sus scrofa/metabolism , Swine/genetics , Swine/immunology , Swine/metabolism , Swine Diseases
6.
Poult Sci ; 94(7): 1584-91, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25971947

ABSTRACT

Three wheat cultivars (Orpheus, Rustic, and Viscount) were used to formulate 3 test feeds (62.4% wheat) in a broiler digestibility trial. The diets were fed to male Ross 308 broiler chickens. The wheat cultivars mainly differed in their amount of non-starch polysaccharides ( NSP: ). The cultivar Orpheus was chosen to represent a high amount of NSP (102 g/kg DM), whereas the cultivars Rustic and Viscount represented low amounts of NSP (83.4 g/kg DM and 73.9 g/kg DM, respectively). Furthermore, the cultivars Orpheus and Viscount were feed quality wheat, whereas Rustic was a milling quality wheat. Nutrient digestibilities and AMEn contents of the diets were measured from 18 to 22-days-old by total excreta collection, or with the use of the indigestible marker titanium dioxide. In addition, the ileal viscosity was measured when the broilers were 25-days-old. Wheat cultivar affected N retention, DM digestibility, and AMEn. In general, the feed formulated with the high NSP wheat cultivar Orpheus resulted in the least favorable nutrient digestibilities and AMEn, whereas the results were better when the feed was formulated with the low NSP cultivars Viscount and Rustic. Feeding the Rustic cultivar caused the highest intestinal viscosity, although this was not reflected in the animal responses. Nutrient digestibilities and AMEn content of the diets were lower when calculated with the titanium dioxide marker than with the total excreta collection procedure. Moreover, the P-values of the effect of wheat cultivar on DM digestibility, N retention, crude fat digestibility and AMEn were lower with the use of the titanium dioxide marker. It can be concluded that wheat cultivar affected nutrient digestibility and AMEn, and that the observed differences were related to the amount of NSP. Furthermore, both the titanium dioxide marker and the total excreta collection methods showed the same trends despite the different values obtained. The titanium dioxide marker method was the simplest to use and produced valid results.


Subject(s)
Chickens/physiology , Digestion , Food Analysis/methods , Nutritive Value , Titanium/metabolism , Triticum/chemistry , Animal Feed/analysis , Animal Nutritional Physiological Phenomena , Animals , Biomarkers/metabolism , Diet/veterinary , Feces/chemistry , Male , Polysaccharides , Random Allocation
7.
PLoS One ; 10(4): e0123111, 2015.
Article in English | MEDLINE | ID: mdl-25923725

ABSTRACT

Pig vocalisations convey information about their current state of health and welfare. Continuously monitoring these vocalisations can provide useful information for the farmer. For instance, pig screams can indicate stressful situations. When monitoring screams, other sounds can interfere with scream detection. Therefore, identifying screams from other sounds is essential. The objective of this study was to understand which sound features define a scream. Therefore, a method to detect screams based on sound features with physical meaning and explicit rules was developed. To achieve this, 7 hours of labelled data from 24 pigs was used. The developed detection method attained 72% sensitivity, 91% specificity and 83% precision. As a result, the detection method showed that screams contain the following features discerning them from other sounds: a formant structure, adequate power, high frequency content, sufficient variability and duration.


Subject(s)
Vocalization, Animal/classification , Animals , Area Under Curve , ROC Curve , Stress, Physiological , Swine , Tape Recording
8.
Genes Immun ; 16(2): 142-50, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25569266

ABSTRACT

A classic T-cell phenotype in systemic lupus erythematosus (SLE) is the downregulation and replacement of the CD3ζ chain that alters T-cell receptor signaling. However, genetic associations with SLE in the human CD247 locus that encodes CD3ζ are not well established and require replication in independent cohorts. Our aim was therefore to examine, localize and validate CD247-SLE association in a large multiethnic population. We typed 44 contiguous CD247 single-nucleotide polymorphisms (SNPs) in 8922 SLE patients and 8077 controls from four ethnically distinct populations. The strongest associations were found in the Asian population (11 SNPs in intron 1, 4.99 × 10(-4) < P < 4.15 × 10(-2)), where we further identified a five-marker haplotype (rs12141731-rs2949655-rs16859085-rs12144621-rs858554; G-G-A-G-A; P(hap) = 2.12 × 10(-5)) that exceeded the most associated single SNP rs858554 (minor allele frequency in controls = 13%; P = 4.99 × 10(-4), odds ratio = 1.32) in significance. Imputation and subsequent association analysis showed evidence of association (P < 0.05) at 27 additional SNPs within intron 1. Cross-ethnic meta-analysis, assuming an additive genetic model adjusted for population proportions, showed five SNPs with significant P-values (1.40 × 10(-3) < P< 3.97 × 10(-2)), with one (rs704848) remaining significant after Bonferroni correction (P(meta) = 2.66 × 10(-2)). Our study independently confirms and extends the association of SLE with CD247, which is shared by various autoimmune disorders and supports a common T-cell-mediated mechanism.


Subject(s)
CD3 Complex/genetics , Lupus Erythematosus, Systemic/ethnology , Lupus Erythematosus, Systemic/genetics , Adult , Asian People/genetics , Case-Control Studies , Female , Genetic Association Studies , Genetic Predisposition to Disease , Haplotypes , Humans , Male , Polymorphism, Single Nucleotide , T-Lymphocytes/immunology , White People/genetics
9.
Genes Immun ; 16(1): 15-23, 2015.
Article in English | MEDLINE | ID: mdl-25338677

ABSTRACT

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder characterized by inflammation of multiple organ systems and dysregulated interferon responses. SLE is both genetically and phenotypically heterogeneous, greatly reducing the power of case-control studies in SLE. Elevated circulating interferon-alpha (IFN-α) is a stable, heritable trait in SLE, which has been implicated in primary disease pathogenesis. About 40-50% of patients have high IFN-α, and high levels correspond with clinical differences. To study genetic heterogeneity in SLE, we performed a case-case study comparing patients with high vs low IFN-α in over 1550 SLE cases, including genome-wide association study and replication cohorts. In meta-analysis, the top associations in European ancestry were protein kinase, cyclic GMP-dependent, type I (PRKG1) rs7897633 (P(Meta) = 2.75 × 10(-8)) and purine nucleoside phosphorylase (PNP) rs1049564 (P(Meta) = 1.24 × 10(-7)). We also found evidence for cross-ancestral background associations with the ankyrin repeat domain 44 (ANKRD44) and pleckstrin homology domain containing, family F member 2 gene (PLEKHF2) loci. These loci have not been previously identified in case-control SLE genetic studies. Bioinformatic analyses implicated these loci functionally in dendritic cells and natural killer cells, both of which are involved in IFN-α production in SLE. As case-control studies of heterogeneous diseases reach a limit of feasibility with respect to subject number and detectable effect size, the study of informative pathogenic sub-phenotypes becomes an attractive strategy for genetic discovery in complex disease.


Subject(s)
Cyclic GMP-Dependent Protein Kinase Type I/genetics , Interferon-alpha/immunology , Lupus Erythematosus, Systemic/genetics , Lupus Erythematosus, Systemic/immunology , Polymorphism, Single Nucleotide , Purine-Nucleoside Phosphorylase/genetics , Case-Control Studies , Female , Gene Regulatory Networks , Humans , White People/genetics
10.
Res Vet Sci ; 97(1): 124-8, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24957408

ABSTRACT

Most commonly, salivary cortisol is used in pig stress assessment, alternative salivary biomarkers are scarcely studied. Here, salivary cortisol and two alternative salivary biomarkers, haptoglobin and chromogranin A were measured in a pig stress study. Treatment pigs (n = 24) were exposed to mixing and feed deprivation, in two trials, and compared to untreated controls (n = 24). Haptoglobin differed for feed deprivation vs control. Other differences were only found within treatment. Treatment pigs had higher salivary cortisol concentrations on the mixing day (P < 0.05). Chromogranin A concentrations were increased on the day of refeeding (P < 0.05). Haptoglobin showed a similar pattern to chromogranin A. Overall correlations between the salivary biomarkers were positive. Cortisol and chromogranin A were moderately correlated (r = 0.49, P < 0.0001), correlations between other markers were weaker. The present results indicate that different types of stressors elicited different physiological stress responses in the pigs, and therefore including various salivary biomarkers in stress evaluation seems useful.


Subject(s)
Chromogranin A/metabolism , Haptoglobins/metabolism , Hydrocortisone/metabolism , Saliva/metabolism , Stress, Physiological/physiology , Sus scrofa/physiology , Animals , Biomarkers/metabolism , Crowding , Fluoroimmunoassay/veterinary , Food Deprivation/physiology , Immunoassay/veterinary , Linear Models , Sus scrofa/metabolism , Swine
11.
Lupus ; 23(9): 881-8, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24786785

ABSTRACT

OBJECTIVE: Interleukin-6 (IL-6), interleukin-10 (IL-10), interferon-alpha (IFN-α), and free light chains (FLCs: lambda, kappa) have all been noted to be of importance in systemic lupus erythematosus (SLE). Herein, we quantified and explored the relationship between these inflammatory mediators and disease activity in SLE; and stratified by their current anti-dsDNA antibody status. METHODS: Seventy-seven SLE patients underwent assessment of disease activity using the SLE disease activity index (SLEDAI). Serum FLC (lambda, kappa, and total), IL-6, IL-10, and IFN-α were quantified. Demographics of disease characteristics were determined by chart reviews. Statistical analyses included Mann-Whitney test, chi square, and linear regression analyses. RESULTS: Mean (SD) age of the patients was 44.9 ± 12.7 years; SLEDAI (mean ± SD) was 3.4 ± 4.0. Serum lambda FLC levels had a moderate correlation (r = 0.46 with physician global assessment, 0.44 with SLEDAI) and the strongest correlation with disease activity as compared with other inflammatory mediators including current dsDNA antibody status. After adjusting for prednisone use, the correlation of lambda FLC with PGA (r = 0.48) and SLEDAI (r = 0.52) was better than of current dsDNA antibody status with PGA (r = 0.33) and adjusted SLEDAI (r = 0.24), respectively. IL-10 and IFN-α activity did not correlate with disease activity. Serum FLC and IL-6 levels could differentiate between active and inactive SLE patients. Serum lambda FLC and IL-6 levels differed significantly among patients with and without current dsDNA antibodies. Serum lambda FLC levels accounted for 31% of variance in SLEDAI scores. CONCLUSION: Serum FLC and IL-6 are potentially useful biomarkers of disease activity in SLE. Further studies, with larger study sample and longitudinal design, are indicated.


Subject(s)
Antibodies, Antinuclear/blood , Immunoglobulin kappa-Chains/blood , Immunoglobulin lambda-Chains/blood , Interferon-alpha/blood , Interleukin-10/blood , Interleukin-6/blood , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/immunology , Adult , Cross-Sectional Studies , Female , Humans , Male , Middle Aged
12.
Lupus ; 22(14): 1523-5, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24014569

ABSTRACT

We report the case of a 19-year-old woman with progressive proliferative lupus nephritis (LN) class III after induction and maintenance therapy with mycophenolate mofetil (MMF). Despite a satisfying clinical improvement proteinuria progressed under this medication. We treated the patient with additional belimumab after discussing other options. Following treatment with belimumab, proteinuria rapidly improved to almost normal levels and clinical remission lasted. Belimumab might hold promise for this indication.


Subject(s)
Antibodies, Monoclonal, Humanized/therapeutic use , Immunosuppressive Agents/therapeutic use , Lupus Nephritis/drug therapy , Disease Progression , Female , Humans , Lupus Nephritis/physiopathology , Mycophenolic Acid/analogs & derivatives , Mycophenolic Acid/therapeutic use , Proteinuria/drug therapy , Proteinuria/etiology , Remission Induction , Treatment Outcome , Young Adult
13.
Genes Immun ; 14(8): 471-8, 2013 Dec.
Article in English | MEDLINE | ID: mdl-23965942

ABSTRACT

Alleles of interferon (IFN) regulatory factor 8 (IRF8) are associated with susceptibility to both systemic lupus erythematosus (SLE) and multiple sclerosis (MS). Although high-type I IFN is thought to be causal in SLE, type I IFN is used as a therapy in MS. We investigated whether IRF8 alleles were associated with type I IFN levels or serologic profiles in SLE and MS. Alleles that have been previously associated with SLE or MS were genotyped in SLE and MS patients. The MS-associated rs17445836G allele was associated with anti-double-stranded DNA (dsDNA) autoantibodies in SLE patients (meta-analysis odds ratio=1.92). The same allele was associated with decreased serum IFN activity in SLE patients with anti-dsDNA antibodies, and with decreased type I IFN-induced gene expression in peripheral blood mononuclear cell from anti-dsDNA-negative SLE patients. In secondary progressive MS patients, rs17445836G was associated with decreased serum type I IFN. Rs17445836G was associated with increased IRF8 expression in SLE patient B cells. In summary, IRF8 rs17445836G is associated with human autoimmune disease characterized by low-type I IFN levels, and this may have pharmacogenetic relevance as type I IFN is modulated in SLE and MS. The association with autoantibodies and increased IRF8 expression in B cells supports a role for rs17445836G in humoral tolerance.


Subject(s)
Interferon Regulatory Factors/genetics , Interferon Type I/blood , Lupus Erythematosus, Systemic/genetics , Multiple Sclerosis/genetics , Polymorphism, Single Nucleotide , Autoantibodies/immunology , Case-Control Studies , DNA/immunology , Humans , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/immunology , Multiple Sclerosis/blood , Multiple Sclerosis/immunology
14.
Vet Microbiol ; 163(3-4): 319-24, 2013 May 03.
Article in English | MEDLINE | ID: mdl-23391440

ABSTRACT

Recently, polyphenol extracts were suggested to inhibit binding of Escherichia coli heat labile enterotoxin (LT) to its intestinal receptor GM1. Therefore, polyphenols are promising feed or food supplements to combat enterotoxigenic infections. Little is known of the precise mechanism, or the type of polyphenol required. Here, seven different polyphenols were tested in vitro (1) for inhibition of LT binding to GM1 (GM1-ELISA), (2) for LT inhibitory activity in the cAMP Vero-cell assay, and (3) by testing the aggregating properties of polyphenols with LT using molecular weight exclusion membrane filters, and by centrifugation techniques. Results showed only three out of seven polyphenols, pentagalloylglucose (PGG), epigallocatechingallate (EGCG) and gallocatechingallate (GCG), to effectively inhibit binding of LT to GM1, and to inhibit induction of cAMP in Vero cells, and that PGG is the most effective. Blocking of the GM1 receptor is unlikely as a mechanism because pre-incubation of GM1 with polyphenols had no effect. Co-incubation of polyphenols with forskolin did not interfere with cAMP production in Vero cells, showing that polyphenol activity is not directly related to cAMP. It is concluded that the inhibitory activities of these three polyphenols may coincide with the formation of large (>100 kDa) LT-polyphenol aggregates. Enterotoxin inactivation appears to require a minimum of two galloyl moieties in polyphenol structure and the pentagalloyl PGG is the most effective.


Subject(s)
Bacterial Toxins/metabolism , Enterotoxins/metabolism , Escherichia coli Proteins/metabolism , Polyphenols/pharmacology , Animals , CHO Cells , Chlorocebus aethiops , Cricetulus , Cyclic AMP/metabolism , Enzyme-Linked Immunosorbent Assay , Escherichia coli , G(M1) Ganglioside/metabolism , Hydrolyzable Tannins/metabolism , Polyphenols/chemistry , Protein Binding/drug effects , Vero Cells
15.
J Theor Biol ; 317: 119-25, 2013 Jan 21.
Article in English | MEDLINE | ID: mdl-23073471

ABSTRACT

Serum amyloid A (SAA) is a major acute phase protein in most species, and is widely employed as a health marker. Systemic SAA isoforms (SAA1, and SAA2) are apolipoproteins synthesized by the liver which associate with high density lipoproteins (HDL). Local SAA (SAA3) isoforms are synthesized in other tissues and are present in colostrums, mastitic milk and mammary dry secretions. Of systemic SAA the bulk is monomeric and bound to HDL, and a small proportion is found in serum in a multimeric form with a buried HDL binding site. In most species, systemic SAA could easily be studied by purifying it from serum of diseased individuals by hydrophobic interaction chromatography methods. For years, we were not able to isolate systemic pig SAA using the latter methods, and found that the bulk of pig SAA did not reside in the HDL-rich serum fractions but in the soluble protein fraction mainly as a multimeric protein. Based on these surprising results, we analysed in silico the theoretical properties and predicted the secondary structure of pig SAA by using the published pig primary SAA amino acid sequence. Results of the analysis confirmed that systemic pig SAA had the highest homology with local SAA3 which in other species is the isoform associated with non-hepatic production in tissues such as mammary gland and intestinal epithelium. Furthermore, the primary sequence of the pig SAA N-terminal HDL binding site did differ considerably from SAA1/2. Secondary structure analysis of the predicted alpha-helical structure of this HDL binding site showed a considerable reduction in hydrophobicity compared to SAA1/2. Based on these results, it is argued that systemic acute phase SAA in the pig has the structural properties of locally produced SAA (SAA3). It is proposed that in pig SAA multimers the charged N-terminal sequence is buried, which would explain their different properties. It is concluded that pig systemic SAA is unique compared to other species, which raises questions about the proposed importance of acute phase SAA in HDL metabolism during inflammation in this species.


Subject(s)
Serum Amyloid A Protein/metabolism , Sus scrofa/metabolism , Amino Acid Sequence , Animals , Blotting, Western , Chromatography , Consensus Sequence , Electrophoresis, Polyacrylamide Gel , Hydrophobic and Hydrophilic Interactions , Lipoproteins, HDL/blood , Molecular Sequence Data , Protein Structure, Secondary , Sequence Analysis, Protein , Serum Amyloid A Protein/chemistry , Serum Amyloid A Protein/isolation & purification , Sus scrofa/blood
16.
Vet Immunol Immunopathol ; 152(1-2): 87-92, 2013 Mar 15.
Article in English | MEDLINE | ID: mdl-23078902

ABSTRACT

Diarrhoea in neonatal and early-weaned piglets due to enterotoxigenic Escherichia coli-F4 (ETEC-F4) is an important problem in the pig farming industry. There is substantial evidence for a genetic basis for susceptibility to ETEC-F4 since not all pigs suffer from diarrhoea after an ETEC-F4 infection. A region on SSC13 has been found to be in close linkage to the susceptibility of piglets for ETEC-F4ab,ac. Potential candidate genes on SSC13 have been examined and although some polymorphisms were found to be in linkage disequilibrium with the phenotype, the causative mutation has not yet been found. In this study we are looking at the expression of porcine genes in relation to ETEC-F4ab,ac. With the aid of the Affymetrix GeneChip Porcine Genome Array we were able to find differentially expressed genes between ETEC-F4ab,ac receptor positive (Fab,acR(+)) piglets without diarrhoea and F4ab,acR(+) piglets with diarrhoea or F4ab,acR(-) animals. Since the susceptibility to ETEC-F4ab,ac was described as a Mendelian trait, it is not so surprisingly that only two differentially expressed genes, transferrin receptor (TFRC) and trefoil factor 1 (TFF1), came out of the analysis. Although both genes could pass for functional candidate genes only TFRC also mapped to the region on SSC13 associated with susceptibility for ETEC-F4, which makes TFRC a positional functional candidate gene. Validation by qRT-PCR confirmed the differential expression of TFRC and TFF1. In piglets without diarrhoea, the expression of both genes was higher in F4ab,acR(+) than in F4ab,acR(-) piglets. Similarly, TFRC and TFF1 expression in F4ab,acR(+) piglets without diarrhoea was also higher than in F4ab,acR(+) piglets with diarrhoea. Consequently, although both genes might not play a role as receptor for F4 fimbriae, they could be of great importance during an ETEC-F4 outbreak. An upregulation of TFRC can be a consequence of the piglets ability to raise an effective immune response. An elevation of TFF1, a protein involved in mucin formation, may also affect the piglet's capability to cope with ETEC bacteria, rather than being a receptor for its fimbriae.


Subject(s)
Antigens, Bacterial/genetics , Diarrhea/veterinary , Enterotoxigenic Escherichia coli/genetics , Escherichia coli Infections/veterinary , Escherichia coli Proteins/genetics , Fimbriae Proteins/genetics , Fimbriae, Bacterial/genetics , Swine Diseases/microbiology , Animals , Animals, Newborn , Antigens, Bacterial/biosynthesis , Antigens, Bacterial/immunology , Diarrhea/genetics , Diarrhea/immunology , Diarrhea/microbiology , Enterotoxigenic Escherichia coli/immunology , Escherichia coli Infections/genetics , Escherichia coli Infections/immunology , Escherichia coli Infections/microbiology , Escherichia coli Proteins/biosynthesis , Escherichia coli Proteins/immunology , Fimbriae Proteins/biosynthesis , Fimbriae Proteins/immunology , Fimbriae, Bacterial/immunology , Gene Expression Profiling/veterinary , Genetic Predisposition to Disease , Oligonucleotide Array Sequence Analysis/veterinary , Peptides/genetics , Peptides/immunology , RNA/chemistry , RNA/genetics , Receptors, Transferrin/genetics , Receptors, Transferrin/immunology , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Swine , Swine Diseases/genetics , Swine Diseases/immunology , Trefoil Factor-2
17.
Genes Immun ; 13(5): 380-7, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22476155

ABSTRACT

Systemic lupus erythematosus (SLE) is an autoimmune disease with diverse clinical manifestations characterized by the development of pathogenic autoantibodies manifesting in inflammation of target organs such as the kidneys, skin and joints. Genome-wide association studies have identified genetic variants in the UBE2L3 region that are associated with SLE in subjects of European and Asian ancestry. UBE2L3 encodes an ubiquitin-conjugating enzyme, UBCH7, involved in cell proliferation and immune function. In this study, we sought to further characterize the genetic association in the region of UBE2L3 and use molecular methods to determine the functional effect of the risk haplotype. We identified significant associations between variants in the region of UBE2L3 and SLE in individuals of European and Asian ancestry that exceeded a Bonferroni-corrected threshold (P<1 × 10(-4)). A single risk haplotype was observed in all associated populations. Individuals harboring the risk haplotype display a significant increase in both UBE2L3 mRNA expression (P=0.0004) and UBCH7 protein expression (P=0.0068). The results suggest that variants carried on the SLE-associated UBE2L3 risk haplotype influence autoimmunity by modulating UBCH7 expression.


Subject(s)
Genetic Predisposition to Disease , Haplotypes , Lupus Erythematosus, Systemic/genetics , Ubiquitin-Conjugating Enzymes/genetics , Black or African American/genetics , Alleles , Asian People/genetics , Female , Hispanic or Latino/genetics , Humans , Linkage Disequilibrium , Lupus Erythematosus, Systemic/ethnology , Male , Polymorphism, Single Nucleotide , Ubiquitin-Conjugating Enzymes/metabolism , White People/genetics
18.
Sci Total Environ ; 426: 90-9, 2012 Jun 01.
Article in English | MEDLINE | ID: mdl-22503671

ABSTRACT

The Free Ion Activity Model (FIAM) predicts that metal uptake in biota is related to the free ion activity in the external solution and that metal complexes do not contribute. However, studies with plants have shown that labile metal complexes enhance metal bioavailability when the uptake is rate-limited by transport of the free ion in solution to the uptake site. Here, the role of labile complexes of Cd on metal bioavailability was assessed using Caco-2 cells, the cell model for intestinal absorption. At low Cd(2+) concentration (1 nM), the CdCl(n)(2-n) complexes contributed to the uptake almost to the same extent as the free ion. At large Cd(2+) concentration (10 µM), the contribution of the complexes was much smaller. At constant Cd(2+) concentration, Cd intake in the cells from solutions containing synthetic ligands such as EDTA increased as the dissociation rate of the cadmium complexes increased, and correlated well with the Cd diffusion flux in solution measured with the Diffusive Gradient in Thin Films technique (DGT). The Cd intake fluxes in the cells were well predicted assuming that the specific uptake is limited by diffusion of the free Cd(2+) ion to the cell surface. Our results underline that speciation of Cd has a major effect on its uptake by intestinal cells, but the availability is not simply related to the free ion concentration. Labile complexes of Cd enhance metal bioavailability in these cells, likely by alleviating diffusive limitations.


Subject(s)
Cadmium/metabolism , Coordination Complexes/chemistry , Environmental Pollutants/metabolism , Caco-2 Cells , Cadmium/chemistry , Coordination Complexes/metabolism , Edetic Acid , Environmental Pollutants/chemistry , Humans
19.
Anim Genet ; 43(3): 324-7, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22486505

ABSTRACT

Enterotoxigenic Escherichia coli (ETEC) is one of the most frequently isolated enteropathogens in production animals, especially pigs and calves. Economically, the swine industry is by far the most affected by infections with ETEC because of mortality, morbidity and decreased growth rate of newborn and early-weaned piglets. After ingestion by the animal, these bacteria attach themselves to specific receptors on the small intestinal epithelium by means of proteinaceous surface appendages, the fimbriae. The F4 fimbriae, which attach to the F4 receptor, are the most studied. The aim of our study was to investigate gene expression in the small intestine of piglets of MUC13 and MUC20 in relation to animals with a different treatment towards or a different reaction on ETEC-F4ac by means of quantitative reverse transcription chain reaction (qRT/PCR). MUC13 and MUC20 are positional candidate genes for this F4ac receptor and are located in the region on SSC13q41 that segregates with the susceptibility to ETEC-F4ac. The condition of the small intestine is crucial when examining expression differences between different samples. Therefore, the expression of two genes, fatty-acid binding protein 2, intestinal (FABP2) and pancreatitis-associated protein (PAP), now known as regenerating islet-derived 3 alpha (REG3A) in the small intestine was simultaneously checked. FABP2, a standard for epithelial content, reflects the state of damage, whereas REG3A is a measure for inflammation in the small intestine. The four different substudies presented here suggest that expression of MUC13 and MUC20 is not related to the susceptibility of piglets to ETEC-F4ac.


Subject(s)
Enterotoxigenic Escherichia coli/physiology , Escherichia coli Infections/veterinary , Intestine, Small/metabolism , Mucins/metabolism , Swine Diseases/genetics , Animals , Antigens, Neoplasm/genetics , Antigens, Neoplasm/metabolism , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Disease Susceptibility/immunology , Disease Susceptibility/veterinary , Escherichia coli Infections/genetics , Escherichia coli Infections/immunology , Fatty Acid-Binding Proteins/genetics , Fatty Acid-Binding Proteins/metabolism , Gene Expression , Gene Expression Regulation , Intestinal Mucosa/immunology , Intestinal Mucosa/metabolism , Intestine, Small/immunology , Lectins, C-Type/genetics , Lectins, C-Type/metabolism , Mucins/genetics , Pancreatitis-Associated Proteins , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Swine , Swine Diseases/immunology
20.
Genes Immun ; 13(3): 232-8, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22189356

ABSTRACT

Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by autoantibody production and organ damage. Lupus nephritis (LN) is one of the most severe manifestations of SLE. Multiple studies reported associations between renal diseases and variants in the non-muscle myosin heavy chain 9 (MYH9) and the neighboring apolipoprotein L 1 (APOL1) genes. We evaluated 167 variants spanning MYH9 for association with LN in a multiethnic sample. The two previously identified risk variants in APOL1 were also tested for association with LN in European-Americans (EAs) (N = 579) and African-Americans (AAs) (N = 407). Multiple peaks of association exceeding a Bonferroni corrected P-value of P < 2.03 × 10(-3) were observed between LN and MYH9 in EAs (N = 4620), with the most pronounced association at rs2157257 (P = 4.7 × 10(-4), odds ratio (OR) = 1.205). A modest effect with MYH9 was also detected in Gullah (rs8136069, P = 0.0019, OR = 2.304). No association between LN and MYH9 was found in AAs, Asians, Amerindians or Hispanics. This study provides the first investigation of MYH9 in LN in non-Africans and of APOL1 in LN in any population, and presents novel insight into the potential role of MYH9 in LN in EAs.


Subject(s)
Apolipoproteins/genetics , Black or African American/genetics , Lipoproteins, HDL/genetics , Lupus Nephritis/ethnology , Lupus Nephritis/genetics , Molecular Motor Proteins/genetics , Myosin Heavy Chains/genetics , Apolipoprotein L1 , Genetic Predisposition to Disease , Humans , Linkage Disequilibrium , Polymorphism, Single Nucleotide , White People/genetics
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