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Context: Although several risk proteins for hypothyroidism have been reported in recent years, many more plasma proteins have not been tested. Objective: To determine potential mechanisms and novel causal plasma proteins for hypothyroidism using Mendelian randomization (MR). Methods: A large-scale plasma proteome MR analysis was conducted using protein quantitative trait loci (pQTLs) for 2297 plasma proteins. We classified pQTLs into 4 different groups. MR analyses were conducted within the 4 groups simultaneously. Significant proteins were discovered and validated in 2 different cohorts. Colocalization analysis and enrichment analysis were conducted using proteins found with MR. Results: Thirty-one proteins were identified in the discovery cohort. Among them, 13 were validated in the validation cohort. Nine of the 13 proteins are risk factors (ISG15, Fc receptor-like protein 2, tumor necrosis factor ligand superfamily member 14, Rab-2A, FcRL3, thrombomodulin, interferon [IFN]-lambda-1, platelet glycoprotein Ib alpha chain, IL-7RA) for hypothyroidism, whereas others are protective proteins (protein O-glucosyltransferase 1 [POGLUT1], tumor necrosis factor ligand superfamily, 3-hydroxyisobutyryl-CoA hydrolase, transferrin receptor protein 1). Among the significant proteins, POGLUT1 strongly colocalized with expression quantitative trait loci from whole blood (posterior probability of colocalization [PP4]=0.978) and the thyroid (PP4=0.978). Two different trans-pQTLs (rs2111485 PP4=0.998; rs35103715 PP4= 0.998) for IFN-lambda-1 strongly colocalized with hypothyroidism in different chromosomes. Conclusion: Thirteen various proteins were identified and validated to be associated with hypothyroidism using univariable MR. We reinforced and expanded the effect of IFN on hypothyroidism. Several proteins identified in this study could explain part of the association between the coagulation system and hypothyroidism. Our study broadens the causal proteins for hypothyroidism and provides the relationships between plasma proteins and hypothyroidism. The proteins identified in this study can be used as early screening biomarkers for hypothyroidism. [ABSTRACT FROM AUTHOR]

Aims Fibromuscular dysplasia (FMD) is a poorly understood disease that predominantly affects women during middle-life, with features that include stenosis, aneurysm, and dissection of medium-large arteries. Recently, plasma proteomics has emerged as an important means to understand cardiovascular diseases. Our objectives were: (i) to characterize plasma proteins and determine if any exhibit differential abundance in FMD subjects vs. matched healthy controls and (ii) to leverage these protein data to conduct systems analyses to provide biologic insights on FMD, and explore if this could be developed into a blood-based FMD test. Methods and results Females with 'multifocal' FMD and matched healthy controls underwent clinical phenotyping, dermal biopsy, and blood draw. Using dual-capture proximity extension assay and nuclear magnetic resonance-spectroscopy, we evaluated plasma levels of 981 proteins and 31 lipid sub-classes, respectively. In a discovery cohort (N cases = 90, N controls = 100), we identified 105 proteins and 16 lipid sub-classes (predominantly triglycerides and fatty acids) with differential plasma abundance in FMD cases vs. controls. In an independent cohort (N cases = 23, N controls = 28), we successfully validated 37 plasma proteins and 10 lipid sub-classes with differential abundance. Among these, 5/37 proteins exhibited genetic control and Bayesian analyses identified 3 of these as potential upstream drivers of FMD. In a 3rd cohort (N cases = 506, N controls = 876) the genetic locus of one of these upstream disease drivers, CD2-associated protein (CD2AP), was independently validated as being associated with risk of having FMD (odds ratios  = 1.36; P  = 0.0003). Immune-fluorescence staining identified that CD2AP is expressed by the endothelium of medium-large arteries. Finally, machine learning trained on the discovery cohort was used to develop a test for FMD. When independently applied to the validation cohort, the test showed a c-statistic of 0.73 and sensitivity of 78.3%. Conclusion FMD exhibits a plasma proteogenomic and lipid signature that includes potential causative disease drivers, and which holds promise for developing a blood-based test for this disease. [ABSTRACT FROM AUTHOR]

The present study was carried out to determine the effect of garlic on egg counts and secretion of Galectins (Gals) in goat blood. Garlic (Allium sativum) has been used as medicinal herb since time immemorial in almost every known civilization. It has antibiotic, anti-helminthic and anti-inflammatory properties. Galectins (Gals) are a family of ß-galactosidase binding proteins that have a conserved carbohydrate recognition domain. They are involved in cell adhesion, migration, activation, proliferation, apoptosis and modulate pathological processes such as inflammation. Twelve (N=12) adult female clinically healthy Boer x Spanish goats were divided into two treatment groups (N=6 each) used. Treatment group 1 was drenched with 10 ml of Garlic barrier as recommended by the manufacturer (Garlic Barrier, Glendale, CA) and Treatment group 2 was drenched with water over a four-week period. Fecal and blood samples were collected weekly. Eggs per gram was evaluated using Modified McMaster method and the total plasma protein concentration was assessed using the BCA assay (Pierce). All variables were assessed using a t-test (SAS analysis software version 9.0 at p=0.05). Garlic treatment affected the fecal egg count and plasma protein concentration when compared to untreated controls. The fecal egg count was reduced in animals treated with garlic. The effect varied over time. Garlic EPG in weeks 1 (p<0.004) and 3 (p<0.027) of treatment. A significant increase in total protein concentration (p<0.004) was observed. Garlic treatment has an effect on GIN levels and the inflammatory response. The time dependent effects may indicate parasite, infection stage specific targets and need further study. [ABSTRACT FROM AUTHOR]

Two experiments, each consisting of 2 trials, were conducted to determine the effect of salmon protein hydrolysate (SPH) and spray-dried plasma protein (SDPP) fed during the first week postweaning and their subsequent effect on the growth performance of weanling pigs. Pigs were fed in a 3-phase feeding program with durations of 7 d for phase 1 in both Ex. 1 and 2; 14 or 15 d for phase 2 in Exp. 1 and 2, respectively; and 7 or 8 d for phase 3 in Exp. 1 and 2, respectively. Dietary treatments were fed only during phase 1, whereas the same diet was fed to all pigs in phases 2 and 3. Pigs were blocked by initial BW and sex, and littermates were balanced across treatments. Data from the 2 trials within each experiment were combined and analyzed together; no treatment x trial interactions (P > 0.10) were observed. In Exp. 1, a total of 324 weanling pigs (10 replications of 5 or 6 pigs per pen) with an average initial BW of 6.4 + 1.3 kg were assigned to 1) a control diet with no SPH or SDPP, 2) 1.5% SPH, 3) 3.0% SPH, 4) 1.5% SDPP, 5) 3.0% SDPP, or 6) 1.5% SPH + 1.5% SDPP. Experiment 2 was similar to Exp. 1, but red blood cells were removed from all diets to reduce diet complexity. In Exp. 2, weanling pigs (n = 320, 14 replications of 5 or 6 pigs per pen) with an average initial BW of 5.4 ± 1.2 kg were assigned to 1) a control diet with no SPH orSI3PP, 2) 1.5% SPH, 3)1.5% SDPP, or 4)1.5% SPH + 1.5% SDPP. Three batches of SPH were used, and each batch was analyzed for AA composition. In Exp. 1, the inclusion of SDPP or SPH during phase 1 did not affect (P> 0.10) ADG, ADFI, or G:F compared with those of pigs fed the control diet. No carryover effects on growth performance were observed in any of the subsequent phases. Overall, G:F was greater (P = 0.08) in pigs fed the 1.5% diets compared with those fed the 3.0% diets. In Exp. 2, no differences (P > 0.10) were observed in ADG, ADFI, or G:F among pigs fed the SPH or SDPP diets compared with those of pigs fed the control diet. Pigs fed the combined diet had greater (P < 0.10) overall ADFI compared with that of pigs fed the control diet, but ADFI was similar to that of pigs fed the SPH and SDPP diets. These results indicate that inclusion of up to 3% SDPP or SPH in diets fed during the first week postweaning did not affect the growth performance of weanling pigs, and no subsequent carryover effects were observed. Salmon protein hydrolysate did not affect the growth performance of weanling pigs and may be considered an alternative protein source in diets for weanling pigs. [ABSTRACT FROM AUTHOR]

Four experiments were conducted using 671 nursery pigs to evaluate fermented soybean meal (FSBM) as a new vegetable protein source for nursery pigs. In Exp. 1, a total of 192 pigs weaned at 19.2 ± 0.3 d of age were fed 3 diets (8 pens per treatment) for 2 wk: a control diet (without FSBM) and 2 diets with 3 and 6% FSBM replacing soybean meal, followed by a common diet for the next 2 wk. In Exp. 2, a total of 160 pigs weaned at 21.6 ± 0.2 d of age were fed 4 diets (5 pens per treatment) for 2 wk: a control diet (without FSBM but with 25% dried skim milk; DSM) and 3 diets with 3, 6, and 9% FSBM replacing DSM based on CP. Concentrations of CP, Lys, Met, Thr, and Trp were kept consistent among diets in Exp. 1 and 2. In Exp. 3, a total of 144 pigs weaned at 22.1 + 0.2 d of age were fed 3 diets (6 pens per treatment) for 2 wk: a control diet (without FSBM but with 40% DSM) and 2 diets with 5 and 10% FSBM replacing DSM based on CP. Concentrations of CP, Lys, Met, Thr, Trp, and lactose were kept consistent among diets. In Exp. 4, a total of 175 pigs weaned at 20.7 ± 0.4 d of age were fed 5 diets (5 pens per treatment) for 3 wk: a basal diet [15.5% CP without plasma protein (PP) and FSBM], 2 diets (18.4% CP) with 3.7% PP or 4.9% FSBM, and 2 diets (21.2% CP) with 7.3% PP or 9.8% FSBM. Concentrations of Lys, Met, Thr, and Trp were kept consistent among diets with the same CP concentrations. Pigs had access to feed and water ad libitum and their BW and feed intake were measured weekly for all experiments. Use of up to 6% FSBM replacing soybean meal improved (P < 0.05) G:F and diarrhea scores of nursery pigs (Exp. 1). Use of up to 9% FSBM replacing DSM reduced (P < 0.05) ADG and G:F (Exp. 2). When lactose concentrations were equal, FSBM could replace up to 10% DSM without adverse effects on ADG and C:F (Exp. 3). Relative bioavailability of protein in FSBM to PP was 99.1% (Exp. 4). Collectively, FSBM can serve as an alternative protein source for nursery pigs at 3 to 7 wk of age, possibly replacing the use of DSM and PP but excluding the first week postweaning for PP when balancing for AA and lactose. [ABSTRACT FROM AUTHOR]

Five experiments were conducted to determine the effects of different wheat gluten (WG) sources (Source 1 = enzymatically hydrolyzed, Source 2 = nonmodified ring-dried, Source 3 = spray-dried, and Source 4 = flash-dried) on growth performance of nursery pigs compared with soybean meal (SBM), spray-dried animal plasma (SDAP), or other specialty protein sources. In Exp. 1, pigs (n = 220, initially 6.1 ± 2.5 kg) were fed a control diet containing (as-fed basis) 6% SDAP or WG Source 1 or 2. The WG and L-lysine·HCl replaced 50 or 100% of the SDAP. From d 0 to 21, increasing WG (either source) decreased ADG and ADFI (linear, P < 0.01), but improved (linear, P < 0.02) G:F. In Exp. 2, pigs (n = 252, initially 6.2 ± 3.0 kg) were fed a negative control diet containing no SDAP or WG, diets containing (as-fed basis) 9% WG Source 1 or 5% SDAP, or combinations of WG and SDAP where WG and L-lysine. HCl replaced 25, 50, or 75% of SDAP. From d 0 to 14, pigs fed increasing WG had decreased ADG (linear, P < 0.05). In Exp. 3, pigs (n = 240, initially 7.0 ± 2.5 kg) were fed a negative control diet, a diet containing (as-fed basis) either 3, 6, 9, or 12% WG Source 3, or a positive control diet containing 5% SDAP. The diets containing 9% WG and 5% SDAP had the same amount of SBM. From d 0 to 7, pigs fed 5% SDAP had greater (P < 0.04) ADG than pigs fed the diet containing 9% WG. From d 0 to 14, increasing WG had no effect on ADG, ADFI, or G:F. In Exp. 4, pigs (n = 200, initially 6.0 ± 2.4 kg) were fed a negative control diet, the control diet with (as-fed basis) 4.5 or 9.0% WG Source 1, or the control diet with 2.5 or 5.0% SDAP. Diets containing WG and SDAP had similar SBM levels. From d 0 to 7 and 0 to 14, increasing SDAP tended to improve (linear, P < 0.06) ADG, but increasing WG had no effect. In Exp. 5, 170 barrows and gilts (initially 7.5 ± 2.8 kg) were used to determine the effects of WG Source 1 and 4 compared with select Menhaden fish meal or spray-dried blood cells and a negative control diet (SBM) on the growth performance of nursery pigs from d 5 to 26 postweaning (d 0 to 21 of experiment). No differences were found in ADG or G:F, but pigs fed the diet containing (as-fed basis) 2.5% spray-dried blood cells had greater ADFI than pigs fed the negative control from d 0 to 21. Wheat gluten source had no effect on ADG, ADFI, or G:F. The results of these studies suggest that increasing WG in diets fed immediately after weaning did not improve growth performance relative to SBM or SDAP. [ABSTRACT FROM AUTHOR]

Galectins are part of a conserved family of ß-galactoside-binding proteins that contribute to critical biological events during mammalian gestation and increasingly recognized for a possible role in the immune response of the cow. The objective of this study was to evaluate the effect of Galectins on signal transduction and cell activation in cow blood. Whole blood was collected aseptically from the jugular vein of healthy Holstein Friesian dairy cows (N=3). Blood samples (2.5ml) in duplicate were treated with 150µl of the four different type of recombinant galectins (1, 3, 4, and 9) respectively and untreated samples were served as control. The concentration of total plasma protein was determined using the Pierce BCA kit. Protein expression profiling was performed using1,358 antibodies on the Full Moon BioSystems' Signaling Explorer antibody array covering 20 cell signaling pathways, as recommended by the manufacturer using an Agilent microarray scanner. Data normalization was performed using GeneSpring GX software to generate fold changes in gene expression and then filtered to obtain a list of significantly upregulated and downregulated genes. Features were extracted from protein array images of samples treated with Galectin 1, 3, 4, 9, and untreated sample as a control group. Treatment with all four Galectins increased the concentration of total plasma protein. Average increases due to treatment with Gal1, Gal3, Gal4, and Gal9 were 27%, 10%, 20%, and 14% respectively. ANOVA test showed significance difference among the groups (p < 0.05). Dunnett option was used to compare each of the treated samples to the control group as a baseline. The results also showed that there was significant difference between the control group and any of the treated group (p < 0.05). Distinct signaling pathways are activated in response to Galectin exposure. Further studies are needed to define their regulation and functional impact on cow health. [ABSTRACT FROM AUTHOR]

A novel hyaluronan-binding protein (PHBP) was purified from human plasma by affinity chromatography on hyaluronan-conjugated Sepharose. The contaminating IgM and albumin in the partially purified preparation were removed with anti-IgG antibody-conjugated Sepharose and anti-albumin antibody-conjugated Sepharose, respectively, and no other contaminant was observed. Finally, 800 μg of PHBP was isolated from 500 ml of human plasma. PHBP gave a single 70-kDa band on SDS-PAGE under non-reducing conditions, and 50-kDa and 17-kDa bands under reducing conditions. Thus, PHBP was a heterodimer composed of 50-kDa and 17-kDa subunits, bridged by a disulfide linkage. Both subunits had novel N-terminal ami no acid sequences, indicating that PHBP was a novel hyaluronan-binding protein in human plasma. The amino acid sequence deduced from the nucleotide sequence of the cloned PHBP cDNA exhibited significant homology to that of hepatocyte growth factor activator (HGFA). The results of Northern blot analysis indicated that liver, kidney, and pancreas expressed PHBP mRNA. The predicted structure of PHBP showed three epidermal growth factor (EGF) domains, a kringle domain and a serine protease domain, from its N-terminus, although HGFA has a fibronectin type H domain, an EGF domain, a fibronectin type I domain, an EGF domain, a kringle domain, and a serine protease domain, from its N-terminus. [ABSTRACT FROM AUTHOR]

The intracellular localization of plasma proteins in human tumors transplanted in nude mice was investigated by immuno-electron microscopic method. Human plasma proteins produced by malignant tumors were also confirmed by immunochemical analysis of blood plasma of tumor-bearing mice. In general, human plasma proteins of adult type such as albumin, α-antitrypsin and transferrin were localized in the membrane system of endoplasmic reticula and Golgi complex. Albumin and transferrin were demonstrated also on the surface of microvilli of certain tumor cells but not on the basal cytoplasmlc membrane. On the other hand, α-fetoprotcln was localized on ribosomes as well as in endoplasmic reticula and Golgi complex. [ABSTRACT FROM PUBLISHER]

This study was conducted to evaluate the effects of using plasma protein (PP) as a highly digestible, functional protein source in nursery pig diets on growth performance and intestinal health. Pigs (n = 64; 6.53 ± 0.12 kg BW) weaned at 21 d of age were blocked by litter and BW and randomly assigned within blocks to 1 of 2 dietary treatments in a 35-d study. Four pigs were housed per pen, using a total of 16 pens (8 replicate pens per treatment). Treatments were: 1) control diet (without growth-promoting minerals or antibiotics) without PP, and 2) control diet with 4% of PP replacing SBM on a 1:1 basis. Diets were fed in 3 feeding phases (7, 14, and 14 days, respectively) and PP was included in all phases. Supplementation of PP tended (P = 0.07) to increase ADG (204 vs. 150 g/d) and improved (P = 0.02) gain:feed (812 vs 572 g/kg) during wk 2 only. Individual diarrhea scores (scale from 0=very hard to 6=watery diarrhea) determined using fecal loops changed over time (P < 0.001; 2.1, 2.4, 4.1 and 3.5 for d 1, 3, 7, and 14 respectively), but were not affected by PP. No differences were detected for intestinal morphology measured on d 35 in duodenum, jejunum and ileum. Cell proliferation using Ki-67 staining was increased by PP (86.6 vs. 72.2 positive cells) only in the ileum (P = 0.03). No differences in mucosal concentrations of IL-10, TNF-α, and IFN-γ in the duodenum, jejunum or ileum were detected. Number of eosinophils in blood decreased (14.1 vs. 34.8 cells/µL; P = 0.004), while red blood cell counts (5.9 vs 6.2 x 106 cells/µL; P = 0.09) and neutrophil counts (529 vs. 695 cells/µL; P = 0.07) tended to decrease with PP. Results suggest that PP was not effective in improving overall growth performance and had minimal effects on intestinal health of weanling pigs. [ABSTRACT FROM AUTHOR]

The objective of this research was to evaluate the effects of salmon protein hydrolysate (SPH) and spray-dried porcine plasma (SDPP) on growth performance of nursery pigs (17 to 21 d weaning age; 5.86 kg initial BW; Phase 1, 7 or 8 d; Phase 2, 14 d; and Phase 3, 7 or 8 d). The SPH is a protein source enzymatically-produced from Atlantic salmon and Rainbow trout tissues; it consists of free AA, peptides, and protein. A total of 644 nursery pigs (10 to 14 replicates per treatment with 5 to 6 pigs per pen) were used in a series of 2 experiments with 2 trials each. In Exp. 1, pigs were fed a conventional complex Phase 1 nursery diet containing: 1) no SPH or SDPP; 2) 1.5% SPH; 3) 1.5% SDPP; 4) 3.0% SPH; 5) 3.0% SDPP; or 6) 1.5% SPH and 1.5% SDPP. Pigs were then fed the same Phase 2 and 3 diets with no experimental ingredient. In Exp. 2, pigs were fed a conventional complex Phase 1 nursery diet (no mammalian protein source) containing: 1) no SPH or SDPP; 2) 1.5% SPH; 3) 1.5% SDPP; or 4) 1.5% SPH and 1.5% SDPP. Pigs were then fed the same Phase 2 and 3 diets with no experimental ingredient. Dietary Lys levels were the same in all diets within each phase. In Exp. 1, ADG, ADFI, and G:F were not affected by source or level of protein source during any growth phase (P > 0.10). Overall G:F was greater (P = 0.08) for pigs fed the 1.5% level of protein source than for those fed the 3.0% level of protein source, but there was no difference between protein source. In Exp. 2 during Phase 1, G:F was greater (P < 0.05) for pigs fed SDPP than for those fed SPH. During Phase 2, ADfiwas greater (P < 0.05) for pigs fed SPH than for those fed the control diet. During Phase 3, ADG was greater (P = 0.08) for pigs fed SDPP than for those fed SPH. Overall, ADG, ADFI, and G:F were not affected by protein source (P > 0.10). The results of these experiments indicate that SPH or SDPP are excellent protein sources for nursery pigs. We acknowledge partial financial support and SPH from Green Earth Industries, Washington, DC. [ABSTRACT FROM AUTHOR]