The effect of dietary soy genistein on pig growth and viral replication during a viral challenge.
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Twelve replications of four littermate pigs from a herd naive for porcine reproductive and respiratory syndrome (PRRS) were weaned (10 +/- 2 d of age) and penned individually in isolation rooms. Pigs were randomly allotted within litter to one of four dietary soy genistein concentrations (0, 200, 400, and 800 ppm) to quantify the effect of soy genistein on pig growth and virus replication during a viral challenge. Genistein was provided as the soy glycoside, genistin. At 29 +/- 2 d of age (4.9 +/- 1.4 kg BW), pigs were oronasally inoculated with 10(4.3) PRRS virus/mL from strain JA142 in a 2-mL dose. Blood was collected every 4 d from d 0 to 24 postinoculation and analyzed for serum PPRS virus, interferon activity, and alpha1-acylglycoprotein (AGP) concentrations. Serum virus and interferon peaked at 10(5) virus/mL and 57% protection, respectively, at 4 d postinoculation and then declined steadily. Serum AGP concentration peaked at 12 d postinoculation. Each log increase in serum virus was associated with a reduction of daily gain of 0.034 kg in 5.3-kg pigs and 0.004 kg in 11-kg pigs. As dietary genistein concentration increased, serum concentrations of PRRS virus decreased linearly (10(2.46), 10(2.26), 10(2.05), and 10(2.14) virus per milliliter of serum, P < 0.07) and interferon responded quadratically (28.4, 25.7, 22.8, and 30.9% protection, P < 0.06) independent of days postinoculation. The AGP concentrations increased (P < 0.01) quadratically with the magnitude of the response to dietary genistein maximized at 12 to 16 d postinoculation. Effects of dietary genistein on daily pig gain and feed intake were dependent on dietary genistein concentration and stage of viremia. Daily pig gains from d 0 to 24 postinoculation were improved as dietary genistein increased, but the magnitude of the response to dietary genistein concentration lessened as the serum virus concentrations were minimized resulting in a linear genistein x period interaction (P < 0.07). Daily feed intakes also were increased quadratically as genistein concentration increased. These data indicate that soy genistein at dietary concentrations of 200 to 400 ppm is an orally active immune modulator that enhances systemic serum virus elimination and body growth in virally challenged pigs. (+info)
Palmitate uptake by neonatal rat myocytes and hepatocytes. Role of extracellular protein.
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The role of extracellular binding proteins in the rate of [3H]palmitate uptake by neonatal cardiac myocytes and hepatocytes was investigated using a model-independent approach. Binding proteins used in this study included alpha1-acid glycoprotein [isoelectric point (pI) approximately 2.7], conalbumin (pI approximately 6.4), lysozyme (pI approximately 11.0), albumin (pI approximately 4.9), and albumin which had been modified to yield proteins with pI values of 3.5, 4.7, 7.5 and 8.6. All uptake studies were conducted at similar unbound ligand fractions. There was a linear relationship between the rate of neonatal hepatocyte [3H]palmitate clearance and protein pI (r2 = 0.98). In contrast, there was an overall poor relationship between neonatal cardiac myocyte [3H]palmitate-clearance rate and protein pI (r2 = 0.48). However, the relationship improved when the data on [3H]palmitate-clearance were analyzed using only the modified albumins. The study indicates that an ionic interaction between extracellular proteins and the hepatocyte surface enhances the overall uptake of [3H]palmitate. This interaction may be limited to albumin for neonatal cardiac myocytes. (+info)
Acute phase protein alpha 1-acid glycoprotein interacts with plasminogen activator inhibitor type 1 and stabilizes its inhibitory activity.
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alpha(1)-Acid glycoprotein, one of the major acute phase proteins, was found to interact with plasminogen activator inhibitor type 1 (PAI-1) and to stabilize its inhibitory activity toward plasminogen activators. This conclusion is based on the following observations: (a) alpha(1)-acid glycoprotein was identified to bind PAI-1 by a yeast two-hybrid system. Three of 10 positive clones identified by this method to interact with PAI-1 contained almost the entire sequence of alpha(1)-acid glycoprotein; (b) this protein formed complexes with PAI-1 that could be immunoprecipitated from both the incubation mixtures and blood plasma by specific antibodies to either PAI-1 or alpha(1)-acid glycoprotein. Such complexes could be also detected by a solid phase binding assay; and (c) the real-time bimolecular interactions monitored by surface plasmon resonance indicated that the complex of alpha(1)-acid glycoprotein with PAI-1 is less stable than that formed by vitronectin with PAI-1, but in both cases, the apparent K(D) values were in the range of strong interactions (4.51 + 1.33 and 0.58 + 0.07 nm, respectively). The on rate for binding of PAI-1 to alpha(1)-glycoprotein or vitronectin differed by 2-fold, indicating much faster complex formation by vitronectin than by alpha(1)-acid glycoprotein. On the other hand, dissociation of PAI-1 bound to vitronectin was much slower than that from the alpha(1)-acid glycoprotein, as indicated by 4-fold lower k(off) values. Furthermore, the PAI-1 activity toward urokinase-type plasminogen activator and tissue-type plasminogen activator was significantly prolonged in the presence of alpha(1)-acid glycoprotein. These observations suggest that the complex of PAI-1 with alpha(1)-acid glycoprotein can play a role as an alternative reservoir of the physiologically active form of the inhibitor, particularly during inflammation or other acute phase reactions. (+info)
In vitro and in vivo properties of novel nucleoside transport inhibitors with improved pharmacological properties that potentiate antifolate activity.
(52/703)
The activity of antimetabolite inhibitors of de novo deoxyribonucleotide biosynthesis can be compromised by the salvage of extracellular preformed nucleosides and nucleobases. Dipyridamole (DP) is a nucleoside transport inhibitor that has been used clinically in an attempt to increase antimetabolite activity; however, DP binds tightly to the serum protein alpha1-acid glycoprotein (AGP) thereby rendering this therapeutic strategy largely ineffective. Four novel DP analogues (NU3076, NU3084, NU3108, and NU3121) have been developed with substitutions at the 2,6- and 4,8-positions of the pyrimidopyrimidine ring. The novel DP analogues inhibit thymidine (dThd) uptake into L1210 cells in vitro (NU3076 IC(50), 0.25 microM; NU3084 IC(50), 0.27 microM; NU3108 IC(50), 0.31 microM; NU3121 IC(50), 0.26 microM; and DP IC(50), 0.37 microM), but, unlike DP, their activity remains largely unaffected in the presence of 5 mg/ml AGP. The four DP analogues inhibit dThd and hypoxanthine rescue from Alimta (multitargeted antifolate)-induced growth inhibition in A549 and COR L23 human lung carcinoma cell lines in the presence of 2.5 mg/ml AGP, whereas the activity of DP is completely abolished. i.p. administration of 10 mg/kg NU3108, NU3121, and DP produced peak plasma concentrations of 4.4, 2.1, and 6.7 microM, respectively, and levels were sustained above 1 microM for approximately 45 min (DP) and 120 min (NU3108 and NU3121). [3H]thymidine incorporation into COR L23 xenografts grown in CD1 nude mice was reduced by 64% (NU3108), 44% (NU3121), and 65% (DP) 2 h after administration of the nucleoside transport inhibitors. In conclusion, two novel DP analogues (NU3108 and NU3121) have been identified that do not bind to AGP and that display superior pharmacokinetic profiles in comparison to DP and inhibit [3H]thymidine incorporation into human tumor xenografts in vivo. (+info)
Specific glycosylation of alpha(1)-acid glycoprotein characterises patients with familial Mediterranean fever and obligatory carriers of MEFV.
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BACKGROUND: Familial Mediterranean fever (FMF) is a periodic febrile disorder, characterised by fever and serositis. The acute phase response during attacks of FMF results from the release of cytokines, which in turn induce increased expression and changed glycosylation of acute phase proteins. A recent study indicated that attacks in FMF are accompanied by a rise of plasma concentrations of serum amyloid A (SAA) and C reactive protein (CRP), which remain significantly raised during remission relative to healthy controls. Another study suggested that obligatory heterozygotes also display an inflammatory acute phase response. OBJECTIVE: To determine the state of inflammation in homozygotic and heterozygotic MEFV genotypes. METHODS: CRP and SAA were studied by enzyme linked immunosorbent assay (ELISA). The glycosylation of the acute phase protein, alpha(1)-acid glycoprotein (AGP), was visualised with crossed affinoimmunoelectrophoresis with concanavalin A as diantennary glycan-specific component and Aleuria aurantia lectin as fucose-specific affinity component. RESULTS: FMF attacks were associated with an increase (p<0.05) in the serum inflammation parameters CRP, SAA, and AGP. The glycosylation of AGP showed an increase (p<0.05) in fucosylated AGP glycoforms, whereas the branching of the glycans remained unaffected. The glycosylation of AGP in the MEFV carrier group, compared with that in a healthy control group, was characterised by a significant increase (p<0.05) in branching of the glycans, whereas the fucosylation remained unaffected. CONCLUSION: The findings suggest an FMF-specific release of cytokines, resulting in a different glycosylation of AGP between a homozygotic and heterozygotic MEFV genotype. (+info)
Association of a alpha1 acidic glycoprotein and squamous cell carcinoma of the head and neck.
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Serum from patients with different malignancies contain an abnormal concentration of a a1-acidic-glycoprotein (AAG) and also, increased levels of AAG are associated with the presence of tumor mass. In the present report, serum levels of AAG were measured by radial immunodiffusion in squamous cell carcinoma of the head and neck (SCCHN) patients taking into account disease status parameters such as tumor localization, stage and extension of disease. Immunohistochemical methods, SDS-PAGE and Western-blotting were employed to study the expression of AAG and a carbohydrate related antigen (sialyl Lewis x) in tumor tissues and derived fractions. AAG showed abnormal levels in 7/15 oral cavity tumor patients sera, 2/5 oropharynx and 5/10 larynx tumors; increased AAG serum levels belonged to patients with disseminated disease. On the other hand, the presence of AAG and sialyl Lewis x were demonstrated in carcinoma cells and in derived fractions from tumor tissues belonging to patients with elevated AAG serum levels. In the present study, we have found elevated levels of AAG in serum samples from SCCHN patients; these neoplastic cells are capable to express AAG. (+info)
Acute phase response in naturally occurring coliform mastitis.
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Changes in the activities of serum cytokines and in acute phase response were observed in dairy cows with naturally occurring coliform mastitis. Seven cows with severe mastitis showed systemic and mammary inflammatory response throughout the observation period, and 11 cows with mild mastitis recovered and were able to be milked within 3 days of onset of mastitis. Serum interleukin (IL)-I and tumor necrosis factor (TNF) activities were higher in the severe group than in the mild group at the first appearance of symptoms. Elevated IL-1 activity was evident in the severe group throughout the observation period. Serum alpha-1-acidglycoprotein (alpha1AG) concentration began to rise with the beginning of mastitis in the severe group, and peaked at 9 days. Serum haptoglobin (Hp) concentrations peaked at 3 days, and decreased gradually after 3 days in the severe group. These results showed that there are dynamic changes in serum IL-1 activity and in serum alpha1AG and Hp concentrations in cows with severe coliform mastitis. (+info)
Effect of alpha1-acid glycoprotein expressed in cancer cells on malignant characteristics.
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The alpha1-acid glycoprotein (AAG) is a prototypical serum acute phase reactant in most mammalian species; it is synthesized mainly in liver parenchymal cells. Recently, we found that mRNAs of AAG were expressed in non-hepatic cancer cells, and the expression levels were regulated by the cytokines--IL-1, IL-6, and TNF-alpha. The functional role of AAG in non-hepatic cancer cells has not yet been established. In order to understand the functional role of the AAG expressed in HT-29 cells, the cancer cells were transfected with cloned cDNA for AAG, or exposed to antisense oligodeoxynucleotide (ODN) for AAG. The colony-forming capacity, invasion, and adhesion to laminin of these transformed cancer cells were measured. Overexpression of AAG by transfection, and inhibition of the AAG expression by antisense ODNs were identified by Western blot as well as nested reverse transcriptase-polymerase chain reaction (nested RT-PCR), respectively. Results showed that the overexpression of AAG by transfection reduced colony-forming capacities, invasion, and adhesion to laminin of the cancer cells; on the other hand, the antisense ODN for AAG elevated colony-forming capacities, invasion, and adhesion to laminin of the cancer cells. These results suggest that AAG, expressed in cancer cells inhibited proliferation, invasion, and metastasis of the cancer cells. (+info)