Degenerative encephalopathy in Nova Scotia Duck Tolling Retrievers presenting with a rapid eye movement sleep behavior disorder

BACKGROUND: Neurodegenerative diseases are a heterogeneous group of disorders characterized by loss of neurons and are commonly associated with a genetic mutation. HYPOTHESIS/OBJECTIVES: To characterize the clinical and histopathological features of a novel degenerative neurological disease affecting the brain of young adult Nova Scotia Duck Tolling Retrievers (NSDTRs). ANIMALS: Nine, young adult, related NSDTRs were evaluated for neurological dysfunction and rapid eye movement sleep behavior disorder. METHODS: Case series review. RESULTS: Clinical signs of neurological dysfunction began between 2 months and 5 years of age and were progressive in nature. They were characterized by episodes of marked movements during sleep, increased anxiety, noise phobia, and gait abnormalities. Magnetic resonance imaging documented symmetrical, progressively increasing, T2‐weighted image intensity, predominantly within the caudate nuclei, consistent with necrosis secondary to gray matter degeneration. Abnormalities were not detected on clinicopathological analysis of blood and cerebrospinal fluid, infectious disease screening or urine metabolite screening in most cases. Postmortem examination of brain tissue identified symmetrical malacia of the caudate nuclei and axonal dystrophy within the brainstem and spinal cord. Genealogical analysis supports an autosomal recessive mode of inheritance. CONCLUSIONS AND CLINICAL IMPORTANCE: A degenerative encephalopathy was identified in young adult NSDTRs consistent with a hereditary disease. The prognosis is guarded due to the progressive nature of the disease, which is minimally responsive to empirical treatment

Pig α₁-Acid Glycoprotein: Characterization and First Description in Any Species as a Negative Acute Phase Protein.

The serum protein α1-acid glycoprotein (AGP), also known as orosomucoid, is generally described as an archetypical positive acute phase protein. Here, porcine AGP was identified, purified and characterized from pooled pig serum. It was found to circulate as a single chain glycoprotein having an apparent molecular weight of 43 kDa by SDS-PAGE under reducing conditions, of which approximately 17 kDa were accounted for by N-bound oligosaccharides. Those data correspond well with the properties of the protein predicted from the single porcine AGP gene (ORM1, Q29014 (UniProt)), containing 5 putative glycosylation sites. A monoclonal antibody (MAb) was produced and shown to quantitatively and specifically react with all microheterogenous forms of pig AGP as analyzed by 2-D electrophoresis. This MAb was used to develop an immunoassay (ELISA) for quantification of AGP in pig serum samples. The adult serum concentrations of pig AGP were in the range of 1-3 mg/ml in a number of conventional pig breeds while it was lower in Göttingen and Ossabaw minipigs (in the 0.3 to 0.6 mg/ml range) and higher in young (2-5 days old) conventional pigs (mean: 6.6 mg/ml). Surprisingly, pig AGP was found to behave as a negative acute phase protein during a range of experimental infections and aseptic inflammation with significant decreases in serum concentration and in hepatic ORM1 expression during the acute phase response. To our knowledge this is the first description in any species of AGP being a negative acute phase protein

Serum concentrations of pigAGP during the acute phase response.

<p>Serum concentration of pig AGP (left) and pig haptoglobin (right) at different days post infection after experimental <i>Streptococcus suis</i> (A), <i>Actinobacillus pleuropneumoniae</i> (haptoglobin data not included) (B), and <i>Staphylococcus aureus</i> (C) infection and after aseptic inflammation (D). Note: In the <i>Staphylococcus aureus</i> experiment, only two infected pigs were sampled at 48 hours.</p

Characterization of pig AGP by 2D electrophoresis and 2D blotting.

<p>A: Pig AGP 2-D electrophoresis, influence of sample preparation conditions, from left to right: reduced sample, non-reduced sample, non-denatured sample. Close-up from gels with pH gradient 2.5–5, individual pig serum sample (B16/221). Mw markers: 30,43,67,94 kDa (from bottom). Arrow: Pig AGP isoforms. B: The reaction of MAb 1.62 with non-purified pig PAGP isoforms by 2-D electrophoresis. Individual pig serum sample (Aus), non-reducing, complete gel/blot with IPG pH 2.5–5 in the first dimension, left: silver-stained, right: blot probed with MAb 1.62. Mw markers: 14, 20, 30, 43, 67, 94 kDa (from bottom). Arrow: Pig AGP isoforms C: Close-up of 2-D electrophoresis of two individual pig sera (827 µg/ml (left), 1692 µg/ml (right)), silver-stained (top), blotted and stained by RuBPS (general protein stain)(middle), and the same blot subsequently probed with MAb 1.62 (bottom). Non-reducing, pH gradient 2.5–5. Arrow: Pig AGP isoforms.</p

Hepatic expression of pig AGP gene during acute infection.

<p>A: Relative expression levels of pig AGP (left) and pigMAP (right) (mean of controls (CTRL, N = 6) set to 1) at 24 hours after experimental infection with <i>Actinobacillus pleuropneumoniae</i> serotype 6 (Ap6) and serotype 2 (Ap2), respectively, as indicated. Values for all individual animals are shown. Error bars depict SEM. Analysis was done on liver tissue samples by qPCR (see text). P<0.01: **, not significant: NS. B: Relative expression levels (mean of controls (CTRL, N = 2) set to 1) in <i>Staphylococcus aureus</i> liver samples 30 (N = 3), 36 (N = 2) and 48 (N = 2) hours after i.v. infection with the bacterium as determined by qPCR, pig AGP (left), pig MAP (middle) and haptoglobin (right). Controls received sterile isotonic saline and were euthanized at 48 hours. Values for individual animals are shown. Error bars depict SEM.</p

Characterization of pig AGP by SDS PAGE and Western blotting.

<p>A: Left panel: Silver-stained SDS PAGE, from the left: Salted-out pooled pig serum supernatant; purified pig AGP; purified pig AGP after sialidase treatment. Right panel: Western blot with the same samples probed with rabbit anti human AGP (DAKO). Arrow: Position of pig AGP in salted-out serum supernatant. B: Western blot probed with anti human AGP, from the left: Purified pig AGP; purified AGP after sialidase treatment; purified AGP after sialidase and PNGase F treatment; buffer control for PNGase F treatment. C: Western blot of pooled pig serum probed with antiserum (1/500) from mouse immunized with purified pig AGP (see text)(representative example). D: Western blot probed with MAb 1.62, from the left: Pooled pig serum; purified pig AGP; purified pig AGP after sialidase treatment.</p

ELISA quantification of pigAGP.

<p>A: Titration of purified pig AGP, pig AGP standard (Saikin Kagaku Institute Ltd.), and two individual pig sera in competitive MAb 1.62 based ELISA. B: Serum concentrations of pig AGP in newborn piglets and in 1-month old piglets (Landrace, Duroc, Yorkshire crossbreds, N = 31). Bars indicate mean and SEM. C: Serum concentrations of pig AGP in different pig breeds and rearing conditions (individual samples), mean and SEM shown. DD: Duroc (2 months, herd) LL: Landrace (2 months, herd) YY: Yorkshire (2 months, herd) Ossabaw minipigs, Experimental stables (14–16 months of age) Göttingen minipigs, Experimental stables (41–47 months of age) L/Y: Landrace/Yorkshire crossbreds (experimental stables, 8–9 months of age) Conventional herd (5 months, D/L/Y cross bred production pigs) SPF herd (5 months, D/L/Y cross bred production pigs).</p

Multiple independent variants at the TERT locus are associated with telomere length and risks of breast and ovarian cancer

<p>TERT-locus SNPs and leukocyte telomere measures are reportedly associated with risks of multiple cancers. Using the Illumina custom genotyping array iCOG, we analyzed similar to 480 SNPs at the TERT locus in breast (n = 103,991), ovarian (n = 39,774) and BRCA1 mutation carrier (n = 11,705) cancer cases and controls. Leukocyte telomere measurements were also available for 53,724 participants. Most associations cluster into three independent peaks. The minor allele at the peak 1 SNP rs2736108 associates with longer telomeres (P = 5.8 x 10(-7)), lower risks for estrogen receptor (ER)-negative (P = 1.0 x 10(-8)) and BRCA1 mutation carrier (P = 1.1 x 10(-5)) breast cancers and altered promoter assay signal. The minor allele at the peak 2 SNP rs7705526 associates with longer telomeres (P = 2.3 x 10(-14)), higher risk of low-malignant-potential ovarian cancer (P = 1.3 x 10(-15)) and greater promoter activity. The minor alleles at the peak 3 SNPs rs10069690 and rs2242652 increase ER-negative (P = 1.2 x 10(-12)) and BRCA1 mutation carrier (P = 1.6 x 10-14) breast and invasive ovarian (P = 1.3 x 10(-11)) cancer risks but not via altered telomere length. The cancer risk alleles of rs2242652 and rs10069690, respectively, increase silencing and generate a truncated TERT splice variant.</p>