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1

Protein Crystal Bovine Insulin  

NASA Technical Reports Server (NTRS)

The comparison of protein crystal, Bovine Insulin space-grown (left) and earth-grown (right). Facilitates the incorporation of glucose into cells. In diabetics, there is either a decrease in or complete lack of insulin, thereby leading to several harmful complications. Principal Investigator is Larry DeLucas.

1991-01-01

2

Fabrication of a surface imprinted hydrogel shell over silica microspheres using bovine serum albumin as a model protein template.  

PubMed

Surface imprinting is an effective approach to improve the template transfer efficiency in applications of molecularly imprinted polymers as biosensors and separation materials. In this paper, we tried to fabricate a surface imprinted hydrogel over silica microspheres for selective recognition of bovine serum albumin by covalent immobilization of a water-soluble UV sensitive initiator onto the surface of silica beads. The polymerization was initiated by UV radiation with N-[3-(dimethylamino)propyl]methacrylamide and N-isopropylacrylamide as the functional monomer and assistant monomer, respectively, and a thin coat of stimuli-responsive hydrogel yielded over the silica gels. The surface imprinted hydrogels exhibited specific affinity toward the template protein with an association constant (K(a)) of 2.2 x 10(5)L mol(-1) and a maximum binding capacity (Q(max)) of 27.3 mgg(-1) in Tris-HCl buffer (pH 7.0). The rebinding and desorption kinetics of the surface imprinted hydrogels were determined and proven to be extremely fast (about 1 min compared to 3h for the previously prepared bulk imprinted hydrogel). Besides, the hydrogel-silica core-shell particles inherit both the stimuli-responsive property of the hydrogel and the good mechanical strength of the silica beads based on the on-line evaluation with high-performance liquid chromatography. The above comprehensive merits of the obtained surface imprinted hydrogel suggest the presented approach an attractive and broadly applicable way of developing biosensors and high-performance protein separation materials. PMID:19230646

Hua, Zhendong; Zhou, Shuang; Zhao, Meiping

2009-11-15

3

Bovine bone implant with bovine bone morphogenetic protein in healing a canine ulnar defect  

Microsoft Academic Search

Xenograft is considered an alternative material for bone transplantation, but its bone healing capacity is inferior compared to that of autografts and allografts. Here, we tested whether bone morphogenetic protein (BMP) addition enhances the suitability of demineralized xenogeneic bovine bone for bone grafting in dogs, and whether xenogeneic bone is a suitable carrier material for BMPs. The capacity of demineralized

T. Tuominen; T. Jämsä; J. Tuukkanen; A. Marttinen; T. S. Lindholm; P. Jalovaara

2001-01-01

4

A comparative study of milk serum proteins in camel ( Camelus dromedarius) and bovine colostrum  

Microsoft Academic Search

Camel (Camelus dromedarius) whey proteins were detected and compared to bovine whey proteins using size exclusion chromatography columns on HPLC. Camel whey proteins such as serum albumin and ?-lactalbumin appear to possess molecular weights similar to the respective bovine whey proteins. Camel whey lacks ?-lactoglobulin and consists of large amount of serum albumin, compared to bovine whey. Camel colostrum is

U Merin; S Bernstein; A Bloch-Damti; R Yagil; C van Creveld; P Lindner; N Gollop

2001-01-01

5

Bovine chromaffin cell cultures as model to study organophosporus neurotoxicity.  

PubMed

Based on the high level of phenyl valerate esterase activities, and in particular of neuropathy target esterase (NTE) found in bovine adrenal medulla, chromaffin cells culture have been proposed as an alternative model for the study of organophosphorus neurotoxicity. Organophosphorus-induced polyneuropathy is a syndrome related to the inhibition and further modification by organophosphorus compounds of NTE (a protein that displays phenyl valerate esterase activity resistant to mipafox and sensitive to paraoxon). Total phenyl valerate esterase activities found in homogenate, particulate and soluble fractions of bovine adrenal medulla were 5200+/-35, 5000+/-280 and 1700+/-260 mU/g tissue, respectively. Cultured chromaffin cells displayed a total hydrolysing activity of 41+/-5 mU/10(6) cells. Homogenates of bovine adrenal medulla displayed only about 6% of activity sensitive to paraoxon. Most of the phenyl valerate esterase activity inhibited by mipafox (a neuropathy inducing compound) was found in particulate fraction. Cultured chromaffin cells displayed kinetics of inhibition by mipafox similar to the kinetics displayed by homogenates of bovine adrenal medulla. We conclude that NTE could be assayed in this system by only using one inhibitor (mipafox) instead of two (paraoxon and mipafox). Also, the proposal is supported of using chromaffin cells as in vitro model for the study of the role of NTE and related esterases in organophosphorus-induced polyneuropathy. PMID:15177651

Quesada, E; Sogorb, M A; Vilanova, E; Carrera, V

2004-06-15

6

Killing of juvenile Fasciola hepatica by purified bovine eosinophil proteins.  

PubMed

Eosinophils were isolated from the mammary gland of Fasciola hepatica-infected cattle by intramammary infusion with a crude extract from adult F. hepatica. Up to 5 x 10(9) eosinophils with a purity of over 90% could be obtained from a single quarter of the gland. The major contaminating cells were monocytes which reached their peak several days following the eosinophil peak. Two major proteins were isolated from bovine eosinophil granules, a high molecular weight peroxidase-active protein and a smaller molecular weight predominantly basic protein. This smaller protein was thought to be the bovine equivalent of guinea-pig and human major basic protein (MBP), although it possessed an unusually high concentration of cysteine. The bovine MBP had a profound effect on juvenile F. hepatica in vitro causing damage and death at concentrations down to 1 x 10(-6) M. The damage was detected by a 51Cr release assay and/or a viability assay involving microscopical examination of the flukes. Other cations, especially protamine sulphate, were also shown to kill flukes, although both lysozyme, found in neutrophils, and the peroxidase-positive peak from bovine eosinophils were unable to mediate any detectable damage. PMID:7438542

Duffus, W P; Thorne, K; Oliver, R

1980-05-01

7

Killing of juvenile Fasciola hepatica by purified bovine eosinophil proteins.  

PubMed Central

Eosinophils were isolated from the mammary gland of Fasciola hepatica-infected cattle by intramammary infusion with a crude extract from adult F. hepatica. Up to 5 x 10(9) eosinophils with a purity of over 90% could be obtained from a single quarter of the gland. The major contaminating cells were monocytes which reached their peak several days following the eosinophil peak. Two major proteins were isolated from bovine eosinophil granules, a high molecular weight peroxidase-active protein and a smaller molecular weight predominantly basic protein. This smaller protein was thought to be the bovine equivalent of guinea-pig and human major basic protein (MBP), although it possessed an unusually high concentration of cysteine. The bovine MBP had a profound effect on juvenile F. hepatica in vitro causing damage and death at concentrations down to 1 x 10(-6) M. The damage was detected by a 51Cr release assay and/or a viability assay involving microscopical examination of the flukes. Other cations, especially protamine sulphate, were also shown to kill flukes, although both lysozyme, found in neutrophils, and the peroxidase-positive peak from bovine eosinophils were unable to mediate any detectable damage. Images Fig. 4

Duffus, W P; Thorne, K; Oliver, R

1980-01-01

8

Cholinergic regulation of protein phosphorylation in bovine adrenal chromaffin cells  

SciTech Connect

Chromaffin cells were isolated from bovine adrenal medullae and maintained in primary culture. After prelabeling with /sup 32/PO/sub 4/, exposure of the chromaffin cells to acetylcholine increased the phosphorylation of a M/sub r/ approx. = 100,000 protein and a M/sub r/ approx. = 60,000 protein (tyrosine hydroxylase), visualized after separation of total cellular proteins in NaDodSO/sub 4//polyacrylamide gels. Immunoprecipitation with antibodies to three known phosphoproteins (100-kDa, 87-kDa, and protein III) revealed an acetylcholine-dependent phosphorylation of these proteins. These three proteins were also shown to be present in bovine adrenal chromaffin cells by immunolabeling techniques. 100-kDa is a M/sub r/ approx. = 100,000 protein selectively phosphorylated by calcium/calmodulin-dependent protein kinase III, 87-kDa is a M/sub r/ approx. = 87,000 protein selectively phosphorylated by protein kinase C, and protein III is a phosphoprotein doublet of M/sub r/ approx. = 74,000 (IIIa) and M/sub r/ approx. = 55,000 (IIIb) phosphorylated by cAMP-dependent protein kinase and calcium/calmodulin-dependent protein kinase I. The data demonstrate that cholinergic activation of chromaffin cells increases the phosphorylation of several proteins and that several protein kinase systems may be involved in these effects.

Haycock, J.W.; Browning, M.D.; Greengard, P.

1988-03-01

9

Direct measurement of inositol in bovine myelin basic protein.  

PubMed Central

Myelin basic protein has been isolated from bovine central-nervous-system myelin by four methods, none of which exposes the protein to acid. After purification the inositol content of both hydrolysed and unhydrolysed protein was quantified by g.c.-m.s. Basic protein prepared by all methods contained less than 4 mol % of inositol. It is concluded, contrary to a previous proposal, that covalent binding to phosphoinositides does not represent a general mechanism for attachment of this cytoplasmically-oriented protein to its membrane. Images Fig. 1.

Smith, R; Braun, P E; Ferguson, M A; Low, M G; Sherman, W R

1987-01-01

10

Bovine and canine acute phase proteins  

Microsoft Academic Search

Acute phase proteins are serum proteins which increase in concentration during the acute phase response to inflammation or infection. The response occurs in all animals, but in different species the response of individual proteins can be significantly different. Of the numerous acute phase proteins which have been identified in humans, a number have been examined in cattle and dogs but

P. D. Eckersall; J. G. Conner

1988-01-01

11

Localisation of bovine colostral odorant-binding protein (bcOBP) mRNAs in several tissues of bovine body.  

PubMed

Bovine colostral odorant-binding protein (bcOBP) is a novel protein found in bovine colostrum and belonging to the lipocalin superfamily. Most of them are secretory proteins. We have examined the localisation of bcOBP messenger RNA in several tissues. The expression of bcOBP messenger RNAs was followed in bovine principal organs and female reproductive tracts with in situ hybridisation, but the localisation of it was not detected. The expression levels of bcOBP mRNAs were also extremely low in those tissues. On the other hand, the expression of bcOBP messenger RNAs has not been found in the airway epithelia and the gallbladder. These results suggest that bcOBP messenger RNAs are expressed in bovine several tissues without its localisation. In conclusion, the localisation of bcOBP messenger RNAs in bovine several tissues was not found. PMID:24339397

Katayama, Shota; Japaridze, Tamar

2014-03-01

12

Rapid thermal equilibration of differentially heated protein and water in bovine corneal stroma  

NASA Astrophysics Data System (ADS)

We measure and simulate the thermal response of bovine corneal stroma to a picosecond IR heating pulse. A thermal diffusion model is developed for this tissue based on the spatial distribution and properties of protein and water constituents in the stroma. In this idealized model, differentially heated protein and water constituents thermally equilibrate with a thermalization time of 515 ps. Using transient absorption spectroscopy for picosecond protein thermometry, a significantly faster thermalization time of 165 ps is measured. The implications of this faster than expected thermalization for the energy-partition model of short-pulse mid-IR tissue ablation are discussed.

Marcus, George Alexander; Schwettman, H. Alan

2011-10-01

13

The bovine patella as a model of early osteoarthritis.  

PubMed

The bovine patella model has been used extensively for studying important structure-function aspects of articular cartilage, including its degeneration. However, the degeneration seen in this model has, to our knowledge, never been adequately compared with human osteoarthritis (OA). In this study, bovine patellae displaying normal to severely degenerate states were compared with human tissue displaying intact cartilage to severe OA. Comparisons of normal and OA features were made with histological scoring, morphometric measurements, and qualitative observations. Differential interference contrast microscopy was used to image early OA changes in the articular cartilage matrix and to investigate whether this method provided comparable quality of visualisation of key structural features with standard histology. The intact bovine cartilage was found to be similar to healthy human cartilage and the degenerate bovine cartilage resembled the human OA tissues with regard to structural disruption, cellularity changes, and staining loss. The extent of degeneration in the bovine tissues matched the mild to moderate range of human OA tissues; however, no bovine samples exhibited late-stage OA. Additionally, in both bovine and human tissues, cartilage degeneration was accompanied by calcified cartilage thickening, tidemark duplication, and the advancement of the cement line by protrusions of bony spicules into the calcified cartilage. This comparison of degeneration in the bovine and human tissues suggests a common pathway for the progression of OA and thus the bovine patella is proposed to be an appropriate model for investigating the structural changes associated with early OA. PMID:24111904

Hargrave-Thomas, E J; Thambyah, A; McGlashan, S R; Broom, N D

2013-12-01

14

Iron absorption in humans as influenced by bovine milk proteins.  

PubMed

The effect of the two major bovine milk protein fractions on the dialyzability of iron in vitro under simulated gastrointestinal conditions and on the absorption of Fe by humans was studied. Liquid-formula meals were prepared from hydrolyzed maize starch, corn oil, and either spray-dried egg white or a milk-protein product. In meals containing egg white, 3.32% of the Fe was dialyzable. The substitution of casein and whey protein products reduced the dialyzable fraction to 0.19-0.56% and 0.86-1.60%, respectively. Percentage Fe absorption was also reduced by the substitution of casein or whey protein for egg white. Mean absorption values fell from 6.67 to 3.65% and 2.53 to 0.98%, respectively. When the intact milk-protein products were replaced by enzyme- or acid-hydrolyzed preparations, the dialyzable fraction increased markedly and in proportion to the extent of hydrolysis. A similar but much smaller effect on absorption was observed. These studies suggest that bovine casein and whey proteins are responsible at least in part for the poor bioavailability of the Fe in some infant formulas. PMID:2923087

Hurrell, R F; Lynch, S R; Trinidad, T P; Dassenko, S A; Cook, J D

1989-03-01

15

Characterization of a new bioactive protein from bovine seminal fluid.  

PubMed

A new acidic seminal fluid protein (aSFP) was purified from bovine seminal fluid, using anion exchange chromatography and FPLC on MonoQ. The purified aSFP displays a pI of 4.8 and an apparent molecular weight of 14 kDa. Homogeneity of aSFP was demonstrated by FPLC and SDS-polyacrylamide gel electrophoresis. Monospecific anti-aSFP IgGs were employed to characterize aSFP in bovine seminal plasma and seminal vesicle secretion by immuno blot analysis. Proteinchemical characterization of aSFP included amino acid analysis as well as determination of 23 amino acid residues of the N-terminal sequence of aSFP. According to this sequence, aSFP appears to represent a hitherto unknown protein. aSFP stimulated cell division and progesterone secretion of bovine granulosa cells in vitro in a potent and dose dependent manner. aSFP appears to be a potent growth factor with effects on ovarian granulosa cells. PMID:1898381

Einspanier, R; Einspanier, A; Wempe, F; Scheit, K H

1991-09-16

16

A comparative study of the backbone dynamics of two closely related lipid binding proteins: Bovine heart fatty acid binding protein and porcine ileal lipid binding protein  

Microsoft Academic Search

The backbone dynamics of bovine heart fatty acid binding protein (H-FABP) and porcine ileal lipid binding protein (ILBP) were studied by 15N NMR relaxation (T1 and T2) and steady state heteronuclear 15N{1H} NOE measurements. The microdynamic parameters characterizing the backbone mobility were determined using the ‘model-free’ approach. For H-FABP, the non-terminal backbone amide groups display a rather compact protein structure

Christian Lücke; David Fushman; Christian Ludwig; James A. Hamilton; James C. Sacchettini; Heinz Rüterjans

1999-01-01

17

Photo selective protein immobilization using bovine serum albumin  

NASA Astrophysics Data System (ADS)

A simple and selective technique which immobilizes protein onto a solid substrate by using UV illumination has been developed. In protein immobilization, a Bovine serum albumin (BSA) performed bifunctional role as a cross-linker between substrate and proteins and as a blocker inhibiting a nonspecific protein adsorption. A new photo-induced protein immobilization process has been investigated at each step by fluorescence microscopy, ellipsometry, and Fourier transform infrared (FT-IR) spectroscopy. A UV photomask has been used to induce selective protein immobilization on target regions of the surface of the SiO2 substrates under UV illumination with negligible nonspecific binding. The UV illumination also showed improved photostability than the conventional methods which employed bifunctional photo-crosslinker molecules of photo-reactive diazirine. This new UV illumination-based photo-addressable protein immobilization provides a new approach for developing novel protein microarrays for multiplexed sensing as well as other types of bio-immobilization in biomedical devices and biotechnologies.

Kim, Wan-Joong; Kim, Ansoon; Huh, Chul; Park, Chan Woo; Ah, Chil Seong; Kim, Bong Kyu; Yang, Jong-Heon; Chung, Kwang Hyo; Choi, Yo Han; Hong, Jongcheol; Sung, Gun Yong

2012-11-01

18

[Recent development for purification of active proteins from bovine pancreas with liquid chromatography].  

PubMed

Many active proteins exist in bovine pancreas and some of them have become protein drugs for human heath. These protein drugs sourcing from bovine pancreas are also high-tech product having high economic benefit. In the modern biological technology, the preparation of most active protein products relies on various liquid chromatographic techniques. The recent development of extraction of the active proteins from bovine pancreas and their separations and purifications, mainly with chromatographic methods are reviewed in this paper. It would be expected to be helpful for the preparation and application of the active proteins from natural products. PMID:21657047

Yang, Xiaoming; Geng, Xindu

2011-03-01

19

Low-molecular-weight hydrophobic proteins from bovine pulmonary surfactant.  

PubMed

Pulmonary surfactant stabilizes the lung by reducing the surface tension in the terminal air spaces. Lipid extract surfactant contains approx. 1% (w/w) low-molecular-weight hydrophobic proteins SP-B (15 kDa: nonreduced) and SP-C (3.5 kDa) and with the remainder being mainly phospholipids. The hydrophobic proteins were purified from bovine lipid extract surfactant using delipidation by phospholipase C digestion followed by hydroxyapatite chromatography. The phospholipase C step removed most of phosphatidylcholine resulting in a 10-fold enrichment of hydrophobic proteins relative to phospholipid. Chromatography of this preparation on a hydroxyapatite column resulted in the elution of phospholipids followed by SP-C and then SP-B. The column chromatography was repeated to remove residual phospholipids and yield purified SP-B and SP-C. The final recovery of SP-B from the lipid extracts was about 15-20% and that of SP-C was 5-10%. The bovine surfactant proteins were reconstituted with phospholipids and examined for their ability to lower the surface tension with a pulsating bubble surfactometer. Reconstituted surfactant preparations containing SP-B and dipalmitoylphosphatidylcholine plus dioleoylphosphatidylglycerol were capable of reducing the surface tension to near zero values at minimum bubble radius while the reconstitutes with SP-C only lowered the surface tension to approx. 20 mN/m. A more rapid decrease in surface tension was observed with reconstituted samples containing both hydrophobic proteins. These results indicate that both SP-B and SP-C can promote the adsorption and spreading of surfactant lipids at the air/liquid interface. In addition, SP-B appears to facilitate the squeeze-out of unsaturated phospholipids leading to an enrichment of dipalmitoylphosphatidylcholine in the monolayer. PMID:2378907

Mathialagan, N; Possmayer, F

1990-07-16

20

Purification of bovine bone morphogenetic protein by hydroxyapatite chromatography.  

PubMed Central

Bovine bone morphogenetic protein (bBMP) induces differentiation of mesenchymal-type cells into cartilage and bone. bBMP has an apparent Mr of 18,500 +/- 500 and represents less than 0.001% of the wet weight of bone tissue. A Mr 34,000 protein resembling osteonectin is separated by extraction with Triton X-100. A Mr 24,000 protein and about half of a Mr 22,000 protein are disassociated from bBMP by precipitation in 1.5 M guanidine hydrochloride. Aggregates of bBMP and a Mr 14,000 protein are insoluble in aqueous media; the bBMP becomes soluble when the Mr 14,000 protein is disassociated in 6 M urea and removed from the solution by ultrafiltration. Three separate molecular species with apparent Mrs 18,500, 17,500, and 17,000 are eluted at 0.10, 0.15, and 0.20 M phosphate ion concentrations, respectively, from a hydroxy-apatite column. The Mr 18,500 protein has the amino acid composition of acidic polylpeptide and includes four half-cystine residues; the pI is 4.9-5.1. The Mr 22,000 component is a chromoprotein resembling ferritin. The NH2-terminal amino acid sequence of the Mr 17,500 protein simulates histone H2B. The Mr 17,000 protein may possess calmodulin activity. Aggregates of the Mr 18,500 and other proteins induce formation of large deposits of bone; the Mr 18,500 protein alone is rapidly absorbed and induces formation of small deposits. None of the other proteins induces bone formation. Images

Urist, M R; Huo, Y K; Brownell, A G; Hohl, W M; Buyske, J; Lietze, A; Tempst, P; Hunkapiller, M; DeLange, R J

1984-01-01

21

Bovine Peptidoglycan Recognition ProteinS: Antimicrobial Activity, Localization, Secretion, and Binding Properties1  

Microsoft Academic Search

Peptidoglycan (PGN) recognition proteins (PGRPs) are pattern recognition molecules of innate immunity that are conserved from insects to humans. Various PGRPs are reported to have diverse functions: they bind bacterial molecules, digest PGN, and are essential to the Toll pathway in Drosophila. One family member, bovine PGN recognition protein-S (bPGRP-S), has been found to bind and kill microorganisms in a

C. Chace Tydell; Jun Yuan; Patti Tran; Michael E. Selsted

22

Crystal structure of bovine coronavirus spike protein lectin domain.  

PubMed

The spike protein N-terminal domains (NTDs) of bovine coronavirus (BCoV) and mouse hepatitis coronavirus (MHV) recognize sugar and protein receptors, respectively, despite their significant sequence homology. We recently determined the crystal structure of MHV NTD complexed with its protein receptor murine carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1), which surprisingly revealed a human galectin (galactose-binding lectin) fold in MHV NTD. Here, we have determined at 1.55 ? resolution the crystal structure of BCoV NTD, which also has the human galectin fold. Using mutagenesis, we have located the sugar-binding site in BCoV NTD, which overlaps with the galactose-binding site in human galectins. Using a glycan array screen, we have identified 5-N-acetyl-9-O-acetylneuraminic acid as the preferred sugar substrate for BCoV NTD. Subtle structural differences between BCoV and MHV NTDs, primarily involving different conformations of receptor-binding loops, explain why BCoV NTD does not bind CEACAM1 and why MHV NTD does not bind sugar. These results suggest a successful viral evolution strategy in which coronaviruses stole a galectin from hosts, incorporated it into their spike protein, and evolved it into viral receptor-binding domains with altered sugar specificity in contemporary BCoV or novel protein specificity in contemporary MHV. PMID:23091051

Peng, Guiqing; Xu, Liqing; Lin, Yi-Lun; Chen, Lang; Pasquarella, Joseph R; Holmes, Kathryn V; Li, Fang

2012-12-01

23

A bovine model of vaccine enhanced respiratory syncytial virus pathophysiology  

Microsoft Academic Search

A critical issue has been the observation that vaccination of children with a formalin-inactivated respiratory syncytial virus (RSV) vaccine is associated with disease enhancement. We have taken advantage of bovine RSV and our experience with this disease in calves to develop a natural model that parallels human disease. Using formalin-inactivated bovine RSV vaccine calves were either sham-vaccinated\\/infected, vaccinated\\/infected, or vaccinated\\/sham-infected

Laurel J. Gershwin; Edward S. Schelegle; Robert A. Gunther; Mark L. Anderson; Amelia R. Woolums; Danielle R. Larochelle; Gabrielle A. Boyle; Kathleen E. Friebertshauser; Randall S. Singer

1998-01-01

24

Immunostimulation of Murine Spleen Cells by Materials Associated with Bovine Milk Protein Fractions  

Microsoft Academic Search

Purified bovine milk proteins that were added to cultures of murine spleen cells significantly increased cell proliferation and production of immunoglobulin M. Casein and a whey mixture consisting of a- lactalbumin, bovine serum albumin, bovine gamma globulin, and b-lactoglobulin ( b-LG) were stimula- tory. Of the three b-LG preparations that are com- mercially available ( b-LG containing variants A and

K. F. Wong; N. Middleton; M. Montgomery; M. Dey; R. I. Carr

1998-01-01

25

Recombinant adenoviruses expressing the E2 protein of bovine viral diarrhea virus induce humoral and cellular immune responses  

Microsoft Academic Search

The E2 protein of bovine viral diarrhea virus (BVDV) is a major viral glycoprotein and an attractive target for BVDV vaccines. Three replication defective recombinant adenoviruses expressing the BVDV\\/E2 protein (rAds\\/E2) were constructed. Two contain a constitutive promoter, and one an inducible promoter. All three recombinant adenoviruses induced very strong BVDV specific antibody responses in a mouse model as detected

Seyyed Mehdy Elahi; Shi-Hsiang Shen; Brian Geoffrey Talbot; Bernard Massie; Serge Harpin; Youssef Elazhary

1999-01-01

26

Characterization of proteins produced in vitro by periattachment bovine conceptuses  

SciTech Connect

Bovine conceptuses from Days 16 (n = 4), 19 (n = 6), 22 (n = 3), and 24 (n = 4), and chorion from Day 69 (estrus/mating = Day 0) were cultured for 24 h in modified minimum essential medium (MEM) in the presence of radioactive L-leucine ((/sup 3/H) leucine) to characterize de novo synthesis and release of proteins. Proteins released into MEM were identified by two-dimensional polyacrylamide gel electrophoresis, fluorography, and gel and ion exchange chromatography. Major polypeptides identified in MEM were different from those identified in conceptus and chorionic tissues. Both uptake of (/sup 3/H) leucine and quality of polypeptides produced de novo and released into MEM were related to stage of conceptus development. Percent retention of (/sup 3/H) leucine in MEM was lowest in Day 16 cultures, increased in Days 19 and 22 cultures, and decreased in Day 24 cultures. Complexity of polypeptides increased after Day 16. Days 16, 19, 22 and 24 conceptus culture MEM was enriched in low-Mr, acidic polypeptides (Mr/isoelectric point ranges: 22K-26K/6.5-5.6, 20K-26K/5.5-5.4, and 16K-20K/5.0-4.5), which were not prominent products of Day 29 and 69 tissues. A high-Mr (Mr +/- SEM; 735K +/- 22K) glycoprotein was produced by all conceptus and chorionic tissues. The transient nature of production of low-Mr polypeptides suggests that they may be required during the periattachment period.

Bartol, F.F.; Roberts, R.M.; Bazer, F.W.; Lewis, G.S.; Godkin, J.D.; Thatcher, W.W.

1985-04-01

27

Presence of a metallothionein-like protein in the bovine pineal gland  

Microsoft Academic Search

Summary The high concentration of zinc in the bovine pineal gland prompted us to investigate the existence of a zinc-binding protein in this organ. In this study, we report that the subcellular distribution of zinc in the bovine pineal gland is nonuniform, with the crude nuclear, mitochondrial, microsomal, and supernatant fractions having 0.264±0.038, 0.160±0.019, 0.130±0.016, and 0.287±0.010 ?g zinc\\/mg protein,

A. Awad; P. Govitrapong; Y. Hama; M. Hegazy; M. Ebadi

1989-01-01

28

Genes and Proteins Differentially Expressed during In Vitro Malignant Transformation of Bovine Pancreatic Duct Cells1  

PubMed Central

Abstract Pancreatic carcinoma has an extremely bad prognosis due to lack of early diagnostic markers and lack of effective therapeutic strategies. Recently, we have established an in vitro model recapitulating the first steps in the carcinogenesis of the pancreas. SV40 large T antigen-immortalized bovine pancreatic duct cells formed intrapancreatic adenocarcinoma tumors on k-rasmut transfection after orthotopic injection in the nude mouse pancreas. Here we identified genes and proteins differentially expressed in the course of malignant transformation using reciprocal suppression subtractive hybridization and 2D gel electrophoresis and mass spectrometry, respectively. We identified 34 differentially expressed genes, expressed sequence tags, and 15 unique proteins. Differential expression was verified for some of the genes or proteins in samples from pancreatic carcinoma. Among these genes and proteins, the majority had already been described either to be influenced by a mutated ras or to be differentially expressed in pancreatic adenocarcinoma, thus proving the feasibility of our model. Other genes and proteins (e.g., BBC1, GLTSCR2, and rhoGDI?), up to now, have not been implicated in pancreatic tumor development. Thus, we were able to establish an in vitro model of pancreatic carcinogenesis, which enabled us to identify genes and proteins differentially expressed during the early steps of malignant transformation.

Jesnowski, R; Zubakov, Dmitri; Faissner, Ralf; Ringel, Jorg; Hoheisel, Jorg D; Losel, Ralf; Schnolzer, Martina; Lohr, Matthias

2007-01-01

29

Computational modelling of bovine ovarian follicle development  

PubMed Central

Background The development of ovarian follicles hinges on the timely exposure to the appropriate combination of hormones. Follicle stimulating hormone (FSH) and luteinizing hormone (LH) are both produced in the pituitary gland and are transported via the blood circulation to the thecal layer surrounding the follicle. From there both hormones are transported into the follicle by diffusion. FSH-receptors are expressed mainly in the granulosa while LH-receptors are expressed in a gradient with highest expression in the theca. How this spatial organization is achieved is not known. Equally it is not understood whether LH and FSH trigger distinct signalling programs or whether the distinct spatial localization of their G-protein coupled receptors is sufficient to convey their distinct biological function. Results We have developed a data-based computational model of the spatio-temporal signalling processes within the follicle and (i) predict that FSH and LH form a gradient inside the follicle, (ii) show that the spatial distribution of FSH- and LH-receptors can arise from the well known regulatory interactions, and (iii) find that the differential activity of FSH and LH may well result from the distinct spatial localisation of their receptors, even when both receptors respond with the same intracellular signalling cascade to their ligand. Conclusion The model integrates the large amount of published data into a consistent framework that can now be used to better understand how observed defects translate into failed follicle maturation.

2013-01-01

30

Expression of Bovine Leukemia Virus Genome is Blocked by a Nonimmunoglobulin Protein in Plasma from Infected Cattle  

NASA Astrophysics Data System (ADS)

Plasma of cattle infected with bovine leukemia virus contains a soluble factor that blocks the expression of the viral genome in cultured lymphocytes. The blocking factor is not present in plasma of bovine leukemia virus-free cattle or of cattle infected with common bovine viruses. Blocking of bovine leukemia virus expression by the plasma factor is reversible, and seems to be mediated by a nonimmunoglobulin protein molecule.

Gupta, P.; Ferrer, J. F.

1982-01-01

31

Conditioned medium from irradiated bovine pulmonary artery endothelial cells stimulates increased protein synthesis by irradiated bovine lung fibroblasts in vitro  

SciTech Connect

Pulmonary fibrosis, a potentially fatal consequence of radiation exposure, occurs by unknown mechanisms. The hypothesis that endothelial cells, injured by radiation, could alter the biochemical function of lung fibroblasts, was tested by exposing cultures of bovine pulmonary artery endothelial cells to 0 or 5 Gy radiation and then incubating them in fresh medium for 48 h. This endothelial cell conditioned medium (ECCM) was then applied to irradiated or nonirradiated cultures of bovine lung fibroblasts. Forty-eight hours later the fibroblasts were analyzed for their ability to synthesize DNA and protein. The ECCM from injured cells stimulated fibroblast protein synthesis twofold to threefold in irradiated fibroblasts without increasing DNA synthesis. It also stimulated a significant but less marked increase in protein synthesis in nonirradiated fibroblasts. Two-dimensional gel electrophoresis revealed this increased synthesis to be expressed in less than 10% of the 1100 separable fibroblast proteins. This study shows that endothelial cells injured by radiation produce factors that stimulate injured fibroblasts to markedly increase their synthesis of certain intracellular proteins, while not stimulating fibroblast replication.

Flavin, M.P.; Parton, L.A.; Bowman, C.M. (Childrens Hospital of Los Angeles, CA (USA))

1990-09-01

32

Engineered Lactobacillus rhamnosus GG expressing IgG-binding domains of protein G: Capture of hyperimmune bovine colostrum antibodies and protection against diarrhea in a mouse pup rotavirus infection model.  

PubMed

Rotavirus-induced diarrhea causes more than 500,000 deaths annually in the world, and although vaccines are being made available, new effective treatment strategies should still be considered. Purified antibodies derived from hyperimmune bovine colostrum (HBC), from cows immunized with rotavirus, were previously used for treatment of rotavirus diarrhea in children. A combination of HBC antibodies and a probiotic strain of Lactobacillus (L. rhamnosus GG) was also found to be more effective than HBC alone in reducing diarrhea in a mouse model of rotavirus infection. In order to further improve this form of treatment, L. rhamnosus GG was engineered to display surface expressed IgG-binding domains of protein G (GB1, GB2, and GB3) which capture HBC-derived IgG antibodies (HBC-IgG) and thus target rotavirus. The expression of IgG-binding domains on the surface of the bacteria as well as their binding to HBC-IgG and to rotavirus (simian strain RRV) was demonstrated by Western blot, flow cytometry, and electron microscopy. The prophylactic effect of engineered L. rhamnosus GG and anti-rotaviral activity of HBC antibodies was evaluated in a mouse pup model of RRV infection. The combination therapy with engineered L. rhamnosus GG (PG3) and HBC was significantly more effective in reducing the prevalence, severity, and duration of diarrhea in comparison to HBC alone or a combination of wild-type L. rhamnosus GG and HBC. The new therapy reduces the effective dose of HBC between 10 to 100-fold and may thus decrease treatment costs. This antibody capturing platform, tested here for the first time in vivo, could potentially be used to target additional gastrointestinal pathogens. PMID:24291196

Günayd?n, Gökçe; Zhang, Ran; Hammarström, Lennart; Marcotte, Harold

2014-01-16

33

Determination of bovine lymphocyte responses to extracted proteins of Brucella abortus by using protein immunoblotting.  

PubMed Central

Isolation and identification of Brucella antigenic determinants important to cellular responses have been difficult. In this study, bovine peripheral blood mononuclear (PBM) cells from cattle vaccinated with Brucella abortus 19 proliferated to extracted bacterial proteins blotted onto nitrocellulose. Proteins were extracted from gamma-irradiated B. abortus 19 with a sodium dodecyl sulfate extraction buffer. The extracted proteins were separated electrophoretically by sodium dodecyl sulfate-polyacrylamide gel electrophoresis prior to electroblotting onto nitrocellulose. Nitrocellulose sections corresponding to individual lanes of the gel (containing all separated proteins) were then cultured with the PBM cells. Primary and secondary stimulation responses of the PBM cells with the whole protein blots were similar kinetically to the responses of the PBM cells stimulated with whole irradiated B. abortus 19 or with whole irradiated B. abortus 19 blotted onto nitrocellulose. Although lipopolysaccharide was determined to be associated with the extracted proteins and transferred onto the blots, the lipopolysaccharide did not stimulate cellular proliferation, as indicated by the antigen-specific secondary responses. Stimulating PBM cells with portions of the blot containing high (greater than 45,000)-, medium (25,000 to 45,000)- or low (25,000)-molecular-weight proteins demonstrated that the responding cells were specific only to the proteins of corresponding molecular weights. These results indicate that cellular responses to individual proteins can be studied without cloning the bacterial genes or purifying the individual proteins. Images

Brooks-Alder, B; Splitter, G A

1988-01-01

34

Comparison of Human VDAC1 with Streptococcal Streptokinase and Bovine Bactericidal Permeability Increasing Protein: Role of Structural Information in Identifying Functionally Significant Domains  

Microsoft Academic Search

Comparison of the primary amino acid sequence of the human X-linked voltage dependent anion channel, with other sequences in data base searches, identified regions of similarity in streptococcal streptokinase and bovine bactericidal permeability increasing protein. These regions of similarity were in different areas of the protein and were relatively short. However, examination of an empirically derived structural model of the

K. M. Mccabe; D. A. Wheeler; V. Adams; E. R. B. Mccabe

1995-01-01

35

Effect of different culture systems on adipocyte differentiation-related protein (ADRP) in bovine embryos.  

PubMed

Bovine embryos cultured in serum-containing media abnormally accumulate lipid droplets, compared to their in vivo counterparts. The objective of this study was to investigate the effect of different culture systems on the mRNA expression and on the quantification and localisation of adipocyte differentiation-related protein (ADRP), a protein associated with lipid accumulation in bovine blastocysts. Two experiments were independently performed for ADRP mRNA expression analysis. In experiment A, blastocysts were produced in modified synthetic oviduct fluid (mSOF)+10% foetal calf serum (FCS), in coculture (bovine oviduct epithelial cells, Boec) and in ewe oviducts, whereas in experiment B, they were produced in mSOF+10?M docosahexaenoic acid (DHA) and in vivo. Control groups were also performed. ADRP mRNA expression was downregulated in the Boec, ewe oviduct and in vivo groups compared to the 10% FCS or DHA groups, respectively. Moreover, the expression of this protein was downregulated in the Boec group compared to the control group (P<0.05). A third experiment (experiment C) was performed to quantify and localise ADRP protein. Boec, in vivo and control groups were tested. After immunofluorescence staining followed by confocal microscopy analysis, embryonic ADRP was clearly localised around lipid droplets, indicating that ADRP is also a lipid droplet coat protein in bovine embryos. In conclusion, our results demonstrate that bovine embryos at the blastocyst stage expressed ADRP mRNA and protein, and that the embryonic culture system modified this expression. PMID:24560670

Al Darwich, A; Perreau, C; Tsikis, G; Coudert, E; Touzé, J L; Briant, E; Beckers, J F; Mermillod, P; Guignot, F

2014-03-01

36

Expression of cementum-derived attachment protein in bovine tooth germ during cementogenesis  

Microsoft Academic Search

Cementum-derived attachment protein (CAP) is a 56 kDa collagenous protein that promotes attachment of mesenchymal cells. Previous studies have shown that the presence of CAP is restricted to cementum in adult human tissues. In this study, we report generation of a monoclonal antibody against CAP and its use for the investigation of CAP in developing bovine tooth germs. Mice were

M Saito; M Iwase; S Maslan; N Nozaki; M Yamauchi; K Handa; O Takahashi; S Sato; T Kawase; T Teranaka; A. S Narayanan

2001-01-01

37

A bovine model of vaccine enhanced respiratory syncytial virus pathophysiology.  

PubMed

A critical issue has been the observation that vaccination of children with a formalin-inactivated respiratory syncytial virus (RSV) vaccine is associated with disease enhancement. We have taken advantage of bovine RSV and our experience with this disease in calves to develop a natural model that parallels human disease. Using formalin-inactivated bovine RSV vaccine calves were either sham-vaccinated/infected, vaccinated/infected, or vaccinated/sham-infected and their clinical signs, pulmonary function, and histological lung lesions quantitatively scored. Interestingly there was significantly greater disease in vaccinated/infected calves and histological lesions in calves were similar to those of affected children. Finally, we note that vaccination did not induce neutralizing antibodies, but IgG antibodies were detected by ELISA. Our model of RSV enhanced disease is important because it provides quantifiable evidence of disease severity that can be applied to evaluate the mechanisms of immunopathology and the safety of candidate RSV vaccines. PMID:9682383

Gershwin, L J; Schelegle, E S; Gunther, R A; Anderson, M L; Woolums, A R; Larochelle, D R; Boyle, G A; Friebertshauser, K E; Singer, R S

1998-07-01

38

A 23-kDa protein as a substrate for protein kinase C in bovine neutrophils. Purification and partial characterization  

Microsoft Academic Search

In ³²P{sub i}-loaded bovine neutrophils stimulated with phorbol myristate acetate (PMA), radioactivity was preferentially incorporated into a protein of low molecular mass, suggesting a PKC-dependent phosphorylation. This protein, termed 23-kDa protein, was predominantly localized in the cytosol. The apparent molecular mass of the purified protein range between 20 and 23 kDa. In the absence of mercaptoethanol, a dimer accumulated. Homogeneity

Marie Jose Stasia; Anne Christine Dianoux; Pierre V. Vignais

1989-01-01

39

Determination of minor proteins of bovine milk and colostrum by optical biosensor analysis.  

PubMed

Automated, rapid, sensitive, and label-free biosensor-based immunoassays for immunoglobulin G (IgG), folate binding protein, lactoferrin, and lactoperoxidase in bovine milk using surface plasmon resonance optical detection with direct binding assay format are described. Samples are prepared for analysis by direct dilution into buffer. Analysis conditions, including ligand immobilization, flow rate, contact time, and regeneration are defined and nonspecific binding considerations evaluated. The technique has been applied to the measurement of these proteins in consumer milks, colostrum, milk products, and infant formulas, and their temporal change during early bovine lactation followed. PMID:16792092

Indyk, Harvey E; Filonzi, Enrico L; Gapper, Leyton W

2006-01-01

40

Interaction of virally coded protein and a cell cycle-regulated cellular protein with the bovine parvovirus left terminus ori.  

PubMed Central

Replication of parvoviruses requires cis signals located in terminal palindromes that function as origins of replication in conjunction with trans-acting viral and cellular proteins. A gel retardation assay was used to identify proteins in crude nuclear extracts of bovine parvovirus (BPV)-infected bovine fetal lung cells that interact with the hairpinned left end (3' OH terminus of the viral minus strand in the flop conformation) of BPV. Three specific DNA-protein complexes formed. One complex was shown to involve a BPV structural protein(s) by inhibiting its formation when antiserum specific for these BPV proteins was used. By specific competition with serum containing antibodies against the BPV nonstructural proteins, a second complex was shown to involve a BPV nonstructural protein. A third complex contained protein of cellular origin and was also formed with extracts of uninfected bovine fetal lung cells. DNA competition assays suggest that the viral proteins do not bind to the right hairpin, which differs in sequence and secondary structure from the left terminus, or to a BPV terminus that lacks the first 52 nucleotides, preventing formation of the stem of the hairpin. The cellular protein is regulated in a cell cycle-dependent fashion, with its binding activity increased in uninfected, actively dividing cells compared with contact-inhibited cells. Since autonomous parvovirus replication requires an S-phase factor for progeny formation, the terminal binding protein demonstrated here is a candidate for this factor. Images

Metcalf, J B; Bates, R C; Lederman, M

1990-01-01

41

Pregnancy and interferon-tau induce conjugation of bovine ubiquitin cross-reactive protein to cytosolic uterine proteins.  

PubMed

Conceptus-derived interferon-tau (IFN-tau) induces bovine endometrial ubiquitin cross-reactive protein (UCRP) mRNA and protein on Days 15-21 of pregnancy. Bovine UCRP retains the Leu-Arg-Gly-Gly C-terminal sequence of ubiquitin that ligates to and directs degradation of cytosolic proteins. The objectives of the present experiments were to determine whether UCRP became conjugated to endometrial cytosolic proteins during early pregnancy and in response to recombinant bovine (rbo) IFN-tau. Ubiquitin (8 kDa), UCRP (17 kDa), and conjugates thereof (> or = 30 kDa) were quantitated using Western blotting and densitometry. Endometrial ubiquitin and its conjugates did not differ between Day 18 pregnant and nonpregnant cows, or between control and rboIFN-tau-treated (25 nM) explant cultures (Day 14; nonpregnant). Bovine UCRP was induced in endometrium from pregnant as compared with nonpregnant cows. Conjugation of endometrial proteins to UCRP was induced in pregnant as compared to nonpregnant cows. Recombinant boIFN-tau induced UCRP and its conjugates in cultured endometrial explants from nonpregnant cows. It is concluded that UCRP, in response to rboIFN-tau, becomes conjugated to endometrial cytosolic proteins during early pregnancy. The regulation of uterine proteins by UCRP may be integral to the maintenance of early pregnancy in ruminants. PMID:9546718

Johnson, G A; Austin, K J; Van Kirk, E A; Hansen, T R

1998-04-01

42

THz spectroscopy and molecular modeling of bovine serum albumin under various hydration conditions  

NASA Astrophysics Data System (ADS)

Bovine serum albumin (BSA) is the most abundant protein in bovine plasma; its three dimensional structure is yet unknown. We investigated the structure and dynamics of BSA in lyophilized samples, in 10% w/w and 50% w/w BSA aqueous solutions using THz spectroscopy and molecular modeling. THz spectra were recorded with a spectral resolution of 7.4 GHz. Theoretical spectra were simulated using a structural model of BSA based on the homology with the known structure of human serum albumin (HSA). The agreement between simulated THz spectra and THz spectra recorded experimentally allowed us to validate the BSA model and the solution models. Based on these models we investigated the flexibility of dry BSA and of BSA with one hydration layer. The hydrated structure of BSA is less flexible than the structure free of water molecules, except for residues 54 - 104 that are more mobile in the hydrated structure. We also investigated the fluctuations of the water molecules within the first hydration layer and identified two groups of water molecules: one that exhibits small fluctuations and one of highly mobile water molecules. These molecules are associated to highly mobile regions from the proteins and move in positive correlation with the neighboring protein regions. We also propose a BSA dimerization model in which the molecules strongly interact. The fluctuations of the BSA monomers and of their first hydration layer were investigated. The two molecules display similar fluctuation patterns, but one of them is slightly more flexible.

Mernea, Maria; Calborean, Octavian; Petrescu, Livia; Zatreanu, Diana; Sandu, Oana; Dascalu, Traian; Mihailescu, Dan Florin

2011-09-01

43

Bovine Papilloma Virus (BPV)Encoded E2 Protein Enhances Binding of E1 Protein to the BPV Replication Origin  

Microsoft Academic Search

The replication of bovine papilloma virus (BPV) DNA in vivo requires two viral-encoded proteins, E1 and E2, while all other proteins are derived from the host. We described previously the isolation of the E1 protein and showed that it contains multiple functions required for BPV DNA replication. The BPV transcription factor E2 was shown by others to stimulate BPV DNA

Yeon-Soo Seo; Friedemann Muller; Monika Lusky; Emma Gibbs; Hae-Yeong Kim; Barbara Phillips; Jerard Hurwitz

1993-01-01

44

Changes in Specific Blood Serum Protein Levels Associated with Parturition in the Bovine1  

Microsoft Academic Search

Beginning about 14 weeks before parturition, the level of serum proteins in the blood of the bovine started increasing, to reach a maximum about four weeks before parturition; and then started decreasing, to reach a minimum at parturition. The drop at parturition was caused by a loss of immune B~- and ~-globulins and some a-globulins from the blood; the level

B. L. Larson; K. A. Kendall

1957-01-01

45

Protection of calves against cryptosporidiosis with immune bovine colostrum induced by a Cryptosporidium parvum recombinant protein  

Microsoft Academic Search

The purpose of the study was to determine if immunization with a recombinant protein (rC7) of Cryptosporidium parvum would induce immune bovine colostrum that protected calves against cryptosporidiosis following oral challenge with C. parvum oocysts. Late gestation Holstein cows with low titers of antibody to the p23 antigen of C. parvum were immunized three times with 300 ?g affinity purified

Lance E Perryman; Sushila J Kapil; Michael L Jones; Elaine L Hunt

1999-01-01

46

Bovine prion protein as a modulator of protein kinase CK2.  

PubMed Central

On the basis of far-Western blot and plasmon resonance (BIAcore) experiments, we show here that recombinant bovine prion protein (bPrP) (25-242) strongly interacts with the catalytic alpha/alpha' subunits of protein kinase CK2 (also termed 'casein kinase 2'). This association leads to increased phosphotransferase activity of CK2alpha, tested on calmodulin or specific peptides as substrate. We also show that bPrP counteracts the inhibition of calmodulin phosphorylation promoted by the regulatory beta subunits of CK2. A truncated form of bPrP encompassing the C-terminal domain (residues 105-242) interacts with CK2 but does not affect its catalytic activity. The opposite is found with the N-terminal fragment of bPrP (residues 25-116), although the stimulation of catalysis is less efficient than with full-size bPrP. These results disclose the potential of the PrP to modulate the activity of CK2, a pleiotropic protein kinase that is particularly abundant in the brain.

Meggio, F; Negro, A; Sarno, S; Ruzzene, M; Bertoli, A; Sorgato, M C; Pinna, L A

2000-01-01

47

Protein Hydrolysates from Non-bovine and Plant Sources Replaces Tryptone in Microbiological Media  

NASA Astrophysics Data System (ADS)

Tryptone (pancreatic digest of casein) is a common ingredient in laboratory and fermentation media for growing wild-type and genetically modified microorganisms. Many of the commercially manufactured products such as human growth hormone, antibiotics, insulin, etc. are produced by recombinant strains grown on materials derived from bovine sources. With the emergence of Bovine Spongiform Encephalopathy (BSE) and the consequent increase in Food and Drug Administration (FDA) regulations, elimination of materials of bovine origin from fermentation media is of paramount importance. To achieve this objective, a number of protein hydrolysates derived from non-bovine animal and plant sources were evaluated. Tryptone in Luria-Bertani (LB) broth was replaced with an equal quantity of alternate protein hydrolysates. Four of the six hydrolysates (one animal and three from plants) were found to efficiently replace the tryptone present in LB-medium as measured by growth rate and growth yield of a recombinant Escherichia coli strain. In addition, we have determined plasmid stability, inducibility and activity of the plasmid encoded ?-galactosidase in the recombinant strain grown in the presence of various protein hydrolysates.

Ranganathan, Yamini; Patel, Shifa; Pasupuleti, Vijai K.; Meganathan, R.

48

Utilization of bovine blood plasma proteins for the production of angiotensin I converting enzyme inhibitory peptides  

Microsoft Academic Search

Hydrolysates of whole bovine plasma and its separated proteins, albumin and globulins, which inhibit the angiotensin I converting enzyme (ACE) were prepared by enzymic hydrolysis with several proteases available for industrial use. Alcalase produced ACE inhibitory peptides from plasma proteins most efficiently and the Alcalase hydrolysate of albumin showed the most high activity (IC50=0.56 mg\\/ml). Sequential ultrafiltration of the hydrolysate

Chang-Kee Hyun; Heuyn-Kil Shin

2000-01-01

49

Unique quadruplex structure and interaction of an RNA aptamer against bovine prion protein  

Microsoft Academic Search

RNA aptamers against bovine prion protein (bPrP) were obtained, most of the obtained aptamers being found to contain the r(GGAGGAGGAGGA) (R12) sequence. Then, it was revealed that R12 binds to both bPrP and its b-isoform with high affinity. Here, we present the structure of R12. This is the first report on the structure of an RNA aptamer against prion protein.

Tsukasa Mashima; Akimasa Matsugami; Fumiko Nishikawa; Satoshi Nishikawa; Masato Katahira

2009-01-01

50

Multiple, Distinct Forms of Bovine and Human Protein Kinase C Suggest Diversity in Cellular Signaling Pathways  

Microsoft Academic Search

A new family of protein kinase C-related genes has been identified in bovine, human, and rat genomes. The alpha-, beta-, and gamma-type protein kinase sequences are highly homologous, include a kinase domain, and potential calcium-binding sites, and they contain interspersed variable regions. The corresponding genes are located on distinct human chromosomes; the possibility of even greater genetic complexity of this

Lisa Coussens; Peter J. Parker; Lucy Rhee; Teresa L. Yang-Feng; Ellson Chen; Michael D. Waterfield; Uta Francke; Axel Ullrich

1986-01-01

51

Guanosine 3 prime ,5 prime -cyclic nucleotide binding proteins of bovine retina identified by photoaffinity labeling  

SciTech Connect

Cyclic GMP-binding proteins present in membrane fractions of bovine retina and, in particular, rod outer segments (ROS) were identified by photoaffinity labeling with 8-azido-({sup 32}P)cGMP. Two soluble proteins and two membrane-associated proteins were specifically labeled. The soluble proteins, 93 and 72 kDa, corresponded respectively to the {alpha} subunit of ROS cGMP phosphodiesterase and cGMP-dependent protein kinase. One of the two membrane-associated proteins, 53 kDa, was present in all particulate retinal fractions. Its function is unknown. It is distinct from cAMP-dependent protein kinase or the 63-kDa cGMP-activated channel from ROS. The second membrane-associated protein, 37 kDa, was present only in fractions that did not contain ROS. The molecular mass of this protein was similar to that of cGMP-binding protein previously attributed to rod cells.

Thompson, D.A.; Khorana, H.G. (Massachusetts Institute of Technology, Cambridge (USA))

1990-03-01

52

A 23-kDa protein as a substrate for protein kinase C in bovine neutrophils. Purification and partial characterization  

SciTech Connect

In {sup 32}P{sub i}-loaded bovine neutrophils stimulated with phorbol myristate acetate (PMA), radioactivity was preferentially incorporated into a protein of low molecular mass, suggesting a PKC-dependent phosphorylation. This protein, termed 23-kDa protein, was predominantly localized in the cytosol. The apparent molecular mass of the purified protein range between 20 and 23 kDa. In the absence of mercaptoethanol, a dimer accumulated. Homogeneity of the 23-kDa protein was verified by 2D-PAGE analysis. Gel isoelectric focusing (IEF) of the purified 23-kDa protein followed by Coomassie blue staining allowed the visualization of our discrete protein bands with isoelectric points ranging between pH 6.3 and 6.7. Phosphorylation of the 23-kDa protein by ({gamma}-{sup 32}P)ATP in the presence of bovine neutrophil PKC supplemented with Ca{sup 2+}, phosphatidylserine, and diacylglycerol or with PMA occurred on serine and required the presence of mercaptoethanol. IEF of the {sup 32}P-labeled 23-kDa protein followed by autoradiography revealed for discrete bands with distinct isoelectric points similar to those of the bands stained by Coomassie blue after IEF on nonlabeled 23-kDa protein. The bands of the 23-kDa protein resolved by IEF and transfered to nitrocellulose showed ability to bind ({sup 35}S)GTP-{gamma}-S. The immunoreactivity of antibodies raised in rabbits against the bovine neutrophil 23-kDa protein was demonstrated on immunoblots after SDS-PAGE. The 23-kDa protein differed also from several other proteins of similar molecular mass that have been identified in neutrophils, namely, calmodulin, the small subunit of the low-potential cytochrome b, and a low molecular weight protein which is ADP-ribosylated by the botulinum toxin.

Stasia, M.J.; Dianoux, A.C.; Vignais, P.V. (Centre d'Etudes Nucleaires (France))

1989-12-12

53

Spectral studies on the cadmium-ion-binding properties of bovine brain S-100b protein  

PubMed Central

The effect of Cd2+ binding on bovine brain S-100b protein was studied using c.d. u.v. difference spectroscopy and fluorescence measurements. At pH 7.5, S-100b protein binds two Cd2+ ions per monomer with a Kd value of 3 x 10(-5) M. Addition of Cd2+ resulted in perturbing the single tyrosine residue (Tyr17) in the protein as indicated by u.v. difference spectroscopy and aromatic c.d. measurements. In the presence of Cd2+, the tyrosine residue moves to a more non-polar environment, since a red shift was observed in the u.v. difference spectrum. When the protein was excited at 278 nm, the tyrosine fluorescence emission maximum was centred at 306 nm. Cd2+ addition resulted in an increase in intrinsic fluorescence intensity. Fluorescence titration with Cd2+ indicated the protein binds Cd2+ with a Kd value of 3 x 10(-5) M. 2-p-Toluidinylnaphthalene-6-sulphonate-labelled protein, when excited at 345 nm, had a fluorescence emission maximum at 440 nm. Addition of Cd2+ to labelled protein resulted in a 5-fold increase in fluorescence intensity accompanied by a 5 nm blue shift in the emission maximum, suggesting that the probe, in the presence of Cd2+, moves to a hydrophobic domain. U.v. difference spectroscopic studies indicated a unique Cd2(+)-binding site on the protein, since Cd2+ addition yielded a large positive absorption band in the 240 nm region that is not found with either Ca2+ or Zn2- ions. Similar absorption bands have been observed in Cd-protein complexes such as Cd-metallothionein [Vasak, Kagi & Hill (1981) Biochemistry 20, 2852-2856] and also in model complexes of Cd2+ with 2-mercaptoethanol. This absorption band is believed to arise as a result of charge-transfer transitions between the thiolate and Cd2+. Of the two Cd2- -binding sites on the beta-chain, one must be located at the N-terminal end near the single tyrosine residue, since Cd2- and Zn2+ produced similar effects on the intrinsic protein fluorescence. The other Cd2+ site which is unique to Cd2+ must be Cys84, located at the C-terminal end.

Donato, H; Mani, R S; Kay, C M

1991-01-01

54

Functional reconstitution of prostaglandin E receptor from bovine adrenal medulla with guanine nucleotide binding proteins  

SciTech Connect

Prostaglandin E/sub 2/ (PEG/sub 2/) was found to bind specifically to a 100,000 x g pellet prepared from bovine adrenal medulla. The PGE receptor was associated with a GTP-binding protein (G-protein) and could be covalently cross-linked with this G-protein by dithiobis(succinimidyl propionate) in the 100,000 x g pellet. In order to characterize the G-protein associated with the PGE receptor and reconstitute these proteins in phospholipid vesicles, the authors purified the G-protein to apparent homogeneity from the 100,000 x g pellet. The G-protein served as a substrate of pertussis toxin but differed in its ..cap alpha.. subunit from two known pertussis toxin substrate G-proteins (G/sub i/ and G/sub 0/) purified from bovine brain. The molecular weight of the ..cap alpha.. subunit was 40,000, which is between those of G/sub i/ and G/sub 0/. The purified protein was also distinguished immunologically from G/sub i/ and G/sub 0/ and was referred to as G/sub am/. Reconstitution of the PGE receptor with pure C/sub am/, G/sub i/, or G/sub 0/ in phospholipid vesicles resulted in a remarkable restoration of (/sup 3/H)PGE/sub 2/ binding activity in a GTP-dependent manner. The efficiency of these three G-proteins in this capacity was roughly equal. When pertussis toxin- or N-ethylmaleimide-treated G-proteins, instead of the native ones, were reconstituted into vesicles, the restoration of binding activity was no longer observed. These results indicate that the PGE receptor can couple functionally with G/sub am/, G/sub i/, or G/sub 0/ in phospholipid vesicles and suggest that G/sub am/ may be involved in signal transduction of the PGE receptor in bovine adrenal medulla.

Negishi, M.; Ito, S.; Yokohama, H.; Hayashi, H.; Katada, T.; Ui, M.; Hayaishi, O.

1988-05-15

55

N- and o-glycosylation of a commercial bovine whey protein product.  

PubMed

Bovine whey protein products are used as a base ingredient in infant formulas to optimize the amino acid pattern to a more human-like composition. Although the protein composition of bovine milk has been studied in detail, glycosylation details of commercial whey protein products are missing. To this end, both the N- and O-glycans of such a protein concentrate were sequentially released, the N-glycans enzymatically and the O-glycans chemically (reducing and nonreducing conditions). For the structural analysis of the N- and O-glycans a combination of MALDI-TOF-MS, one-dimensional (1)H NMR spectroscopy, Wisteria floribunda agglutinin affinity chromatography, HPAEC-PAD profiling, and HPLC-FD profiling (2-aminobenzamide derivatives), together with exoglycosidase treatments, were used. A mixture of over 60 N-glycans and 10 O-glycans was characterized, giving a detailed insight into the glycosylation of a bovine whey protein product, Deminal 90, which is applied as an ingredient for infant formulas. PMID:23194161

van Leeuwen, Sander S; Schoemaker, Ruud J W; Timmer, Christel J A M; Kamerling, Johannis P; Dijkhuizen, Lubbert

2012-12-26

56

Gene expression profiles of novel caprine placental prolactin-related proteins similar to bovine placental prolactin-related proteins  

PubMed Central

Background This study reports the identification of a full-length cDNA sequence for two novel caprine prolactin-related proteins (cPRP1 and cPRP6), and their localization and quantitative expression in the placenta. Caprine PRPs are compared with known bovine PRPs. We examined their evolution and role in the ruminant placenta. Results Full-length cPRP1 and cPRP6 cDNA were cloned with a 717- and 720- nucleotide open-reading frame corresponding to proteins of 238 and 239 amino acids. The cPRP1 predicted amino acid sequence shares a 72% homology with bovine PRP1 (bPRP1). The cPRP6 predicted amino acid sequence shares a 74% homology with bovine PRP6 (bPRP6). The two cPRPs as well as bPRPs were detected only in the placentome by RT-PCR. Analysis by in situ hybridization revealed the presence of both cPRPs mRNA in the trophoblast binucleate cells. These mRNA were quantified by real-time RT-PCR analysis of the placentome at 30, 50, 90 and 140 days of pregnancy. Both new cPRP genes were able to translate a mature protein in a mammalian cell-expression system. Western blotting established the molecular sizes of 33 kDa for cPRP1 with FLAG-tag and 45 kDa for cPRP6 with FLAG-tag. The sequence properties and localized expression of cPRP1 and cPRP6 were similar to those of bovine. However, their expression profiles differed from those in bovine placenta. Although this study demonstrated possible roles of PRPs in caprine placenta, PRPs may regulate binucleate-cell functions like those in bovine, but their crucial roles are still unclear. Conclusion We have identified the novel PRPs in caprine placenta. Localization and quantitative expression of caprine PRPs were compared with bovine PRPs. The data indicate that PRP genes in caprine placenta have coordination functions for gestation, as they do in bovine. This is the first study of PRPs function in caprine placenta.

Ushizawa, Koichi; Takahashi, Toru; Hosoe, Misa; Kizaki, Keiichiro; Abe, Yasuyuki; Sasada, Hiroshi; Sato, Eimei; Hashizume, Kazuyoshi

2007-01-01

57

Evaluation of protein expression in bovine bronchoalveolar fluid following challenge with Mannheimia haemolytica.  

PubMed

Proteomics analysis of bovine bronchoalveolar fluid (BAF) following induction of pneumonia with Mannheimia haemolytica using nanoflow liquid chromatography coupled with tandem mass spectrometry (nanoLC-MS/MS) resulted in the identification of 88 unique proteins. Proteins detected in BAF included antimicrobial peptides (AMPs), complement factors, acute-phase proteins, protease inhibitors, and proteins involved in oxidation-reduction. Notwithstanding biological variation, differences in relative protein abundance, determined using normalized peptide counts, were detected for select proteins in BAF from genuinely infected versus sham-infected animals. To demonstrate the applicability of using normalized peptide counts to assess protein expression trends, LC-MS/MS data for the acute-phase protein haptoglobin (HPT) were compared with ELISA data, and statistical evaluation of the relationship between the data revealed a strong measure of association. Differences were detected between sham- and genuinely infected animals for haptoglobin, as well as the AMPs cathelicidin-1 and cathelicidin-4, and inter-?-trypsin inhibitor heavy chain-4, a fairly novel protein involved in the acute phase response. Though the small sample size limited the scope of the inferences, the results indicate the likely importance of AMPs and acute-phase proteins during respiratory infection, and provide additional information regarding potential mechanisms involved in the bovine mucosal barrier defense. PMID:21800424

Boehmer, Jamie L; Degrasse, Jeffrey A; Lancaster, Vicki A; McFarland, Melinda A; Callahan, John H; Ward, Jeffrey L

2011-09-01

58

Partial purification and characterization of the lck protein-tyrosine kinase from bovine thymus.  

PubMed

Soluble extracts prepared from bovine thymus contain an angiotensin-I-phosphorylating activity that is activated several-fold by high concentrations of NaCl. Fractionation of this protein-tyrosine kinase activity by chromatography on DEAE-cellulose yields a major diffuse peak of activity. The enzymes responsible for this activity are found at much higher levels in extracts from bovine thymus than from bovine spleen. The peak of activity from the DEAE-cellulose column can be further separated into two major peaks by chromatography on heparin-agarose. The second peak to elute from the heparin-agarose column was previously purified through several chromatographic steps to yield a 40 kDa protein-tyrosine kinase (p40). We have now partially purified the early-eluting peak of kinase activity by chromatography on columns of butyl-agarose, protamine-agarose and Sephacryl S200. The enzyme was identified following covalent modification with 5'-fluorosulphonylbenzoyladenosine (FSBA) by reactivity with anti-FSBA antibodies. This procedure labelled a series of 52-56 kDa proteins. These proteins were also recognized by polyclonal anti-peptide antibodies raised against the C-terminal 33 amino acids of p56lck, a major T lymphocyte protein-tyrosine kinase. Peptide maps of the partially purified enzyme were identical to maps generated from p56lck obtained from LSTRA cells. These data suggest that bovine thymus p56lck is responsible for the activity found in the early-eluting peak from heparin-agarose. Antibodies raised against a peptide corresponding to amino acids 39-64 of p56lck, a sequence found near the N-terminus, recognized the slower-migrating, but not the faster-migrating, form of the enzyme, indicating that a fraction of the protein had been proteolysed near the N-terminus during purification. The partially purified bovine enzyme exhibited a restricted substrate specificity in vitro and did not readily phosphorylate human erythrocyte band 3, casein or histone, but was able to phosphorylate acid-treated enolase. The dilute enzyme present in fractions eluting from chromatography columns was unable to catalyse an autophosphorylation reaction. Autophosphorylation could be detected in more concentrated enzyme samples and was readily observed in immune-complex assays. The phosphorylation of angiotensin I by bovine thymus p56lck was weakly activated by polyionic compounds such as heparin and polylysine, and was strongly activated by high concentrations of NaCl. PMID:1953650

Wang, Q M; Srinivas, P R; Harrison, M L; Geahlen, R L

1991-10-15

59

Bovine seminal PDC-109 protein: An overview of biochemical and functional properties.  

PubMed

Although long-term storage of bovine semen is desirable for wider use, successful cryopreservation depends on several factors, including various proteins present in seminal plasma. One such group of proteins, viz. bovine seminal plasma (BSP) proteins represents the major protein fraction in bovine seminal plasma. They constitute three major heparin-binding (HB-) acidic proteins secreted by seminal vesicles, viz. BSP-A1/-A2 (PDC-109), BSP-A3 and BSP-30-kDa. By purification studies it was deduced that PDC-109 is a polypeptide of 109 amino acids and contains two tandem repeating fibronectin type-II (Fn-II) domains, preceded by a 23 residue N-terminal domain. Though BSP-A1 and BSP-A2 are biochemically similar they differ only in glycosylation and their mixture is called PDC-109 or gonadostatins. PDC-109 exists as a polydisperse, multimeric self-associated molecule and possesses multifunctional properties, viz. binding to the surface of plasma membrane of spermatozoa causing conformational change in the sperm surface proteins and enhances motility. Besides binding, PDC-109 protein provokes cholesterol efflux from sperm membrane and promotes sperm reservoir by interacting with oviductal membrane. Interaction of sperm with PDC-109 protein induces sperm capacitation and acrosome reaction. However, prolonged exposure of spermatozoa with free floating PDC-109 protein as during processing for preservation, increases cholesterol efflux from spermatozoa. The efflux of sperm membrane cholesterol and disturbance in cholesterol:phospholipids ratio causes destabilization of plasma membrane thereby inducing cryoinjury to the sperm. In this review, the biochemical, functional properties of PDC-109 protein and its role during semen cryopreservation is summarized. PMID:23489472

Srivastava, N; Jerome, A; Srivastava, S K; Ghosh, S K; Kumar, Amit

2013-02-22

60

Homology Modeling Study of Bovine ?-Calpain Inhibitor-Binding Domains  

PubMed Central

The activated mammalian CAPN-structures, the CAPN/CAST complex in particular, have become an invaluable target model using the structure-based virtual screening of drug candidates from the discovery phase to development for over-activated CAPN linked to several diseases, such as post-ischemic injury and cataract formation. The effect of Ca2+-binding to the enzyme is thought to include activation, as well as the dissociation, aggregation, and autolysis of small regular subunits. Unfortunately, the Ca2+-activated enzyme tends to aggregate when provided as a divalent ion at the high-concentration required for the protease crystallization. This is also makes it very difficult to crystallize the whole-length enzyme itself, as well as the enzyme-inhibitor complex. Several parameters that influence CAPN activity have been investigated to determine its roles in Ca2+-modulation, autoproteolysis, phosphorylation, and intracellular distribution and inhibition by its endogenous inhibitor CAST. CAST binds and inhibits CAPN via its CAPN-inhibitor domains (four repeating domains 1–4; CAST1–4) when CAPN is activated by Ca2+-binding. An important key to understanding CAPN1 inhibition by CAST is to determine how CAST interacts at the molecular level with CAPN1 to inhibit its protease activity. In this study, a 3D structure model of a CAPN1 bound bovine CAST4 complex was built by comparative modeling based on the only known template structure of a rat CAPN2/CAST4 complex. The complex model suggests certain residues of bovine CAST4, notably, the TIPPKYQ motif sequence, and the structural elements of these residues, which are important for CAPN1 inhibition. In particular, as CAST4 docks near the flexible active site of CAPN1, conformational changes at the interaction site after binding could be directly related to CAST4 inhibitory activity. These functional interfaces can serve as a guide to the site-mutagenesis in research on bovine CAPN1 structure-function relationships for the design of small molecules inhibitors to prevent uncontrolled and unspecific degradation in the proteolysis of key protease substrates.

Chai, Han-Ha; Lim, Dajeong; Lee, Seung-Hwan; Chai, Hee-Yeoul; Jung, Eunkyoung

2014-01-01

61

Cosecretion of secretory protein-I and parathormone by dispersed bovine parathyroid cells  

SciTech Connect

A RIA with a minimal sensitivity of 0.5 ng protein was developed for the measurement of bovine secretory protein-I (SP-I). With this assay and a previously established RIA for parathormone, the secretion and cell content of SP-I and parathormone were determined in dispersed bovine parathyroid cells. SP-I and parathormone were secreted in linear fashion over a 2-h period. The net secretion of both of these proteins diminished progressively as the concentration of calcium was raised from 0.25 mM to 3.0 mM. The molar ratio for the secreted proteins and those remaining in the cell varied from experiment to experiment but on average was 0.70 +/- 0.07 for the secreted proteins and 0.47 +/- 0.03 for the cellular proteins. Possible explanations for the difference in the ratio of SP-I to parathormone between cellular and secreted proteins include 1) a preferential secretion of SP-I; 2) a preferential intracellular degradation of SP-I; 3) a preferential postsecretory degradation of parathormone, or 4) differential affinities of potential fragments of either or both proteins for their antisera. These results suggest that SP-I and parathormone bear close but not identical metabolic and secretory fates.

Cohn, D.V. (Veterans Administration Medical Center, Kansas City, MO); Morrissey, J.J.; Shofstall, R.E.; Chu, L.L.H.

1982-02-01

62

Production and properties of health-promoting proteins and peptides from bovine colostrum and milk.  

PubMed

The high nutritive value and diverse functional properties of milk proteins are well known. Beyond these qualities, milk proteins have attracted growing scientific and commercial interest as a source of biologically active molecules. Such proteins are found in abundance in colostrum which is the initial milk secreted by mammalian species during late pregnancy and the first few days after birth of the offspring. The best characterized colostrum-based bioactive proteins include alpha-lactalbumin, beta-lactoglobulin, immunoglobulins, lactoferrin, lactoperoxidase and growth factors. All of them can nowadays be enriched and purified on an industrial scale from bovine colostral whey or cheese whey. These native proteins exhibit a wide range of biological activities that are known to affect the digestive function, metabolic responses to absorbed nutrients, growth and development of organs and disease resistance. Also, some of these proteins may prove beneficial in reduction of the risks of chronic human diseases reflected by the metabolic syndrome. It is speculated that such potentially beneficial effects are partially attributed to bioactive peptides derived from intact proteins. These peptides can be liberated during gastrointestinal digestion or fermentation of milk by starter cultures. The efficacy of a few peptides has been established in animal and human studies and the number of commercial products supplemented with specific milk peptides is envisaged to increase on global markets. Bovine colostrum appears as a highly potential source of biologically active native proteins and peptide fractions for inclusion as health-promoting ingredients in various food applications. PMID:24200017

Korhonen, H J

2013-01-01

63

A model of bovine tuberculosis control in domesticated cattle herds.  

PubMed Central

A typical strategy for disease control in domesticated animals involves regular field tests and quarantine of infected herds. This prevents disease spread beyond the herd, while slaughter of diseased animals removes the infection from within the herd. A model of bovine tuberculosis (Tb) control in cattle is examined, which includes 'test and slaughter' combined with herd isolation and vaccination. Herd status is represented by an integral equation expressing the duration of herd isolation. The current Tb situation in New Zealand is used as an example, and vaccination strategy discussed. Extrapolation of existing management strategies indicate that a vaccine of efficacy greater than 96% would be required, reaching 95% of target Tb levels within six years. These results suggest that a complementary strategy of vaccination and vector control may be more promising than vaccination alone.

Kao, R R; Roberts, M G; Ryan, T J

1997-01-01

64

Advanced oxidation protein products are generated by bovine neutrophils and inhibit free radical production in vitro.  

PubMed

Despite the recognised importance of oxidative stress in the health and immune function of dairy cows, protein oxidation markers have been poorly studied in this species. The current study aimed to characterise markers of protein oxidation generated by activated bovine neutrophils and investigate the biological effects of advanced oxidation protein products (AOPP) on bovine neutrophils. Markers of protein oxidation (AOPP, dityrosines and carbonyls) were measured in culture medium containing bovine serum albumin (BSA) exposed to neutrophils. The effect of AOPP-BSA on generation of reactive oxygen species (ROS) was assessed by chemiluminescence. Activation of caspases-3, -8 and -9 and the presence of DNA laddering were used as apoptosis markers. Greater amounts of AOPP were generated by phorbol myristate acetate (PMA)-activated than non-activated neutrophils (1.46 ± 0.13 vs. 0.75 ± 0.13 nmol/mg protein, respectively; P<0.05). Activated neutrophils and hypochlorous acid generated slightly different patterns of oxidized protein markers. Exposure to AOPP-BSA did not stimulate ROS production. Activated neutrophils generated a lesser amount of ROS when incubated with AOPP-BSA (P<0.001). Activation with PMA induced a loss of viable neutrophils after 3h, which was greater with AOPP-BSA incubation (P<0.05). Detectable amounts of active caspases-3, -8 and -9 were found in nearly all samples but differences in caspase activation or DNA laddering were not observed comparing treatment groups. Apoptosis was unlikely to be responsible for the greater loss of PMA-activated neutrophils cultured in AOPP-BSA and it is possible that primary necrosis occurred. The results suggest that accumulation of oxidized proteins at an inflammatory site might result in a progressive reduction of neutrophil viability. PMID:24291143

Bordignon, Milena; Da Dalt, Laura; Marinelli, Lieta; Gabai, Gianfranco

2014-01-01

65

Complex induction of bovine uterine proteins by interferon-tau.  

PubMed

Interferon-tau (IFN-tau) is released by the conceptus and induces two uterine proteins during early pregnancy: ubiquitin cross-reactive protein (UCRP) and granulocyte chemotactic protein-2 (GCP-2). The present experiments were designed to determine whether detection (Western blot) of cytosolic UCRP and release of GCP-2 could be used to examine IFN-tau signal transduction in cultured endometrial explants and primary epithelial cells. Recombinant (r) type 1 IFNs (rboIFN-tau and rboIFN-alpha; 5, 25, 100 nM) induced UCRP, but only rboIFN-tau induced GCP-2 in explant culture. Recombinant boIFN-tau and conceptus secretory proteins containing native IFN-tau induced UCRP and GCP-2 in cultured primary epithelial cells. All concentrations of rboIFN-alpha (25, 50, 100 nM) induced UCRP, but only the highest concentration induced GCP-2 in cultured primary epithelial cells. Interestingly, phorbol ester (100, 500, 1000 ng/ml) induced GCP-2, but it had no effect on UCRP. Because type 1 IFNs induce UCRP, IFN-tau probably interacts with the janus kinase (Jak)-associated IFN-alpha receptor to phosphorylate signal transducers and activators of transcription (STAT) and/or interferon regulatory factor-1 (IRF-1). However, IFN-tau-specific induction of GCP-2 may involve a variant type 1 receptor subunit or activators of transcription that are associated with protein kinase C and the Jak/STAT/IRF-1 pathway. PMID:9687298

Staggs, K L; Austin, K J; Johnson, G A; Teixeira, M G; Talbott, C T; Dooley, V A; Hansen, T R

1998-08-01

66

Insulin autoantibodies with high affinity to the bovine milk protein alpha casein  

PubMed Central

Insulin autoantibodies (IAA) can appear in children within months of introducing solid foods to the diet and before clinical type 1 diabetes. The aim of this study was to determine whether infant dietary antigens could be immunizing agents of IAA. To this end, IAA binding to [125I]insulin was competed with food preparations and extracts of foods encountered in the infant diet (milk formulas, bovine milk, wheat flour, fowl meal). Bovine milk powder extracts inhibited IAA-positive samples from six of 53 children (age 0·3–14·0 years) participating in German prospective cohorts. Inhibition in these sera ranged from 23 to 100%. Competition was abolished when hydrolyzed milk powder was used. Competition with protein components of bovine milk found that two of the six milk-reactive sera were inhibited strongly by alpha- and beta-casein; none were inhibited by the milk proteins bovine serum albumin or lactoglobulins. The two casein-reactive sera had high affinity to alpha-casein (1·7 × 109; 3·1 × 109 l/mol), and lesser affinity to beta-casein (4·0 × 108; 7·0 × 107 l/mol) and insulin (2·6 × 108; 1·6 × 108 l/mol). No children with milk-reactive IAA developed autoantibodies to other islet autoantigens or diabetes (median follow-up 9·8 years). These results suggest that autoimmunity to insulin can occur infrequently via cross-reactivity to food proteins, but this form of IAA immunization does not appear to be associated with progression to diabetes.

Adler, K; Mueller, D B; Achenbach, P; Krause, S; Heninger, A-K; Ziegler, A G; Bonifacio, E

2011-01-01

67

Antihypertensive Peptides Derived from Bovine Casein and Whey Proteins  

Microsoft Academic Search

Peptides play an important primary role as a supply of essential amino acids and a source of nitrogen. Recent studies have\\u000a reported on another role of peptides: having specific amino acid sequences that can express some biological functions in vivo. For an exhaustive study and supply of biologically active peptides, a large-scale screening of protein sources is necessary.\\u000a Various physiologically

Tadao Saito

68

Sequence variations of the bovine prion protein gene (PRNP) in native Korean Hanwoo cattle  

PubMed Central

Bovine spongiform encephalopathy (BSE) is one of the fatal neurodegenerative diseases known as transmissible spongiform encephalopathies (TSEs) caused by infectious prion proteins. Genetic variations correlated with susceptibility or resistance to TSE in humans and sheep have not been reported for bovine strains including those from Holstein, Jersey, and Japanese Black cattle. Here, we investigated bovine prion protein gene (PRNP) variations in Hanwoo cattle [Bos (B.) taurus coreanae], a native breed in Korea. We identified mutations and polymorphisms in the coding region of PRNP, determined their frequency, and evaluated their significance. We identified four synonymous polymorphisms and two non-synonymous mutations in PRNP, but found no novel polymorphisms. The sequence and number of octapeptide repeats were completely conserved, and the haplotype frequency of the coding region was similar to that of other B. taurus strains. When we examined the 23-bp and 12-bp insertion/deletion (indel) polymorphisms in the non-coding region of PRNP, Hanwoo cattle had a lower deletion allele and 23-bp del/12-bp del haplotype frequency than healthy and BSE-affected animals of other strains. Thus, Hanwoo are seemingly less susceptible to BSE than other strains due to the 23-bp and 12-bp indel polymorphisms.

Choi, Sangho

2012-01-01

69

Modelling N mineralization from bovine manure and sewage sludge composts.  

PubMed

Nitrogen mineralization kinetics were compared in three different soils (pH values: 5.2, 7.1 and 8.6) when treated with bovine manure (BM) and sewage sludge (SS) composts. The soil-compost mixtures were kept at a controlled moisture content of 60% of their water holding capacity (WHC) and were incubated in the dark at 25 °C for 2 years. Five mathematical models were compared (simple exponential, double exponential, special model, hyperbolic and parabolic), using as experimental data the mineralized N accumulated during 360 and 720 days of incubation. The results showed that the best fit for describing the mineralization of organic N from the compost after 1 year of experimentation was obtained with the simple exponential model. However, the special model showed the best fit for data from 2 years of incubation and thus better reflected organic N mineralization over a longer time-span. This suggested that the organic N in the two composts was made up of two organic pools of different degrees of stability. PMID:20951032

Gil, M V; Carballo, M T; Calvo, L F

2011-01-01

70

Identification of lipid synthesis and secretion proteins in bovine milk.  

PubMed

Lactation physiology is a process that is only partly understood. Proteomics techniques have shown to be useful to help advance the knowledge on lactation physiology in human and rodent species but have not been used as major tools for dairy cows, except for mastitis. In this paper, advanced non-targeted proteomics techniques (Filter aided sample preparation and NanoLC-Orbitrap-MS/MS) were applied to study the milk fat globule membrane and milk serum fraction, resulting in the identification of 246 proteins. Of these, 23 transporters and enzymes were related to lipid synthesis and secretion in mammary gland and their functions are discussed in detail. The identification of these intracellular transporters and enzymes in milk provides a possibility of using milk itself to study lipid synthesis and secretion pathways. This full-scale scan of milk proteins by using non-targeted proteomic analysis helps to reveal the important proteins involved in lipid synthesis and secretion for further examination in targeted studies. PMID:24433584

Lu, Jing; van Hooijdonk, Toon; Boeren, Sjef; Vervoort, Jacques; Hettinga, Kasper

2014-02-01

71

In vivo delivery of bovine viral diahorrea virus, E2 protein using hollow mesoporous silica nanoparticles  

NASA Astrophysics Data System (ADS)

Our work focuses on the application of mesoporous silica nanoparticles as a combined delivery vehicle and adjuvant for vaccine applications. Here we present results using the viral protein, E2, from bovine viral diarrhoea virus (BVDV). BVDV infection occurs in the target species of cattle and sheep herds worldwide and is therefore of economic importance. E2 is a major immunogenic determinant of BVDV and is an ideal candidate for the development of a subunit based nanovaccine using mesoporous silica nanoparticles. Hollow type mesoporous silica nanoparticles with surface amino functionalisation (termed HMSA) were characterised and assessed for adsorption and desorption of E2. A codon-optimised version of the E2 protein (termed Opti-E2) was produced in Escherichia coli. HMSA (120 nm) had an adsorption capacity of 80 ?g Opti-E2 per mg HMSA and once bound E2 did not dissociate from the HMSA. Immunisation studies in mice with a 20 ?g dose of E2 adsorbed to 250 ?g HMSA was compared to immunisation with Opti-E2 (50 ?g) together with the traditional adjuvant Quillaja saponaria Molina tree saponins (QuilA, 10 ?g). The humoral responses with the Opti-E2/HMSA nanovaccine although slightly lower than those obtained for the Opti-E2 + QuilA group demonstrated that HMSA particles are an effective adjuvant that stimulated E2-specific antibody responses. Importantly the cell-mediated immune responses were consistently high in all mice immunised with Opti-E2/HMSA nanovaccine formulation. Therefore we have shown the Opti-E2/HMSA nanoformulation acts as an excellent adjuvant that gives both T-helper 1 and T-helper 2 mediated responses in a small animal model. This study has provided proof-of-concept towards the development of an E2 subunit nanoparticle based vaccine.Our work focuses on the application of mesoporous silica nanoparticles as a combined delivery vehicle and adjuvant for vaccine applications. Here we present results using the viral protein, E2, from bovine viral diarrhoea virus (BVDV). BVDV infection occurs in the target species of cattle and sheep herds worldwide and is therefore of economic importance. E2 is a major immunogenic determinant of BVDV and is an ideal candidate for the development of a subunit based nanovaccine using mesoporous silica nanoparticles. Hollow type mesoporous silica nanoparticles with surface amino functionalisation (termed HMSA) were characterised and assessed for adsorption and desorption of E2. A codon-optimised version of the E2 protein (termed Opti-E2) was produced in Escherichia coli. HMSA (120 nm) had an adsorption capacity of 80 ?g Opti-E2 per mg HMSA and once bound E2 did not dissociate from the HMSA. Immunisation studies in mice with a 20 ?g dose of E2 adsorbed to 250 ?g HMSA was compared to immunisation with Opti-E2 (50 ?g) together with the traditional adjuvant Quillaja saponaria Molina tree saponins (QuilA, 10 ?g). The humoral responses with the Opti-E2/HMSA nanovaccine although slightly lower than those obtained for the Opti-E2 + QuilA group demonstrated that HMSA particles are an effective adjuvant that stimulated E2-specific antibody responses. Importantly the cell-mediated immune responses were consistently high in all mice immunised with Opti-E2/HMSA nanovaccine formulation. Therefore we have shown the Opti-E2/HMSA nanoformulation acts as an excellent adjuvant that gives both T-helper 1 and T-helper 2 mediated responses in a small animal model. This study has provided proof-of-concept towards the development of an E2 subunit nanoparticle based vaccine. Electronic supplementary information (ESI) available: Desorption studies of Opti-E2 bound HMSA. See DOI: 10.1039/c4nr01202j

Mahony, D.; Cavallaro, A. S.; Mody, K. T.; Xiong, L.; Mahony, T. J.; Qiao, S. Z.; Mitter, N.

2014-05-01

72

In vivo delivery of bovine viral diahorrea virus, E2 protein using hollow mesoporous silica nanoparticles.  

PubMed

Our work focuses on the application of mesoporous silica nanoparticles as a combined delivery vehicle and adjuvant for vaccine applications. Here we present results using the viral protein, E2, from bovine viral diarrhoea virus (BVDV). BVDV infection occurs in the target species of cattle and sheep herds worldwide and is therefore of economic importance. E2 is a major immunogenic determinant of BVDV and is an ideal candidate for the development of a subunit based nanovaccine using mesoporous silica nanoparticles. Hollow type mesoporous silica nanoparticles with surface amino functionalisation (termed HMSA) were characterised and assessed for adsorption and desorption of E2. A codon-optimised version of the E2 protein (termed Opti-E2) was produced in Escherichia coli. HMSA (120 nm) had an adsorption capacity of 80 ?g Opti-E2 per mg HMSA and once bound E2 did not dissociate from the HMSA. Immunisation studies in mice with a 20 ?g dose of E2 adsorbed to 250 ?g HMSA was compared to immunisation with Opti-E2 (50 ?g) together with the traditional adjuvant Quillaja saponaria Molina tree saponins (QuilA, 10 ?g). The humoral responses with the Opti-E2/HMSA nanovaccine although slightly lower than those obtained for the Opti-E2 + QuilA group demonstrated that HMSA particles are an effective adjuvant that stimulated E2-specific antibody responses. Importantly the cell-mediated immune responses were consistently high in all mice immunised with Opti-E2/HMSA nanovaccine formulation. Therefore we have shown the Opti-E2/HMSA nanoformulation acts as an excellent adjuvant that gives both T-helper 1 and T-helper 2 mediated responses in a small animal model. This study has provided proof-of-concept towards the development of an E2 subunit nanoparticle based vaccine. PMID:24811899

Mahony, D; Cavallaro, A S; Mody, K T; Xiong, L; Mahony, T J; Qiao, S Z; Mitter, N

2014-05-29

73

Isolation of genomic and cDNA clones encoding bovine poly(A) binding protein II.  

PubMed Central

cDNA clones for bovine poly(A) binding protein II (PAB II) were isolated. Their sequence predicts a protein of 32.8 kDa, revising earlier estimates of molecular mass. The protein contains one putative RNA-binding domain of the RNP type, an acidic N-terminal and a basic C-terminal domain. Analyses of authentic PAB II were in good agreement with all predictions from the cDNA sequence except that a number of arginine residues appeared to be post-translationally modified. Poly(A) binding protein II expressed in Escherichia coli was active in poly(A) binding and reconstitution of processive polyadenylation, including poly(A) tail length control. The cDNA clones showed a number of potential PAB II binding sites in the 3' untranslated sequence. Bovine poly(A)+RNA contained two mRNAs hybridizing to a PAB II-specific probe. Analysis of a genomic clone revealed six introns in the coding sequence. The revised molecular mass led to a demonstration of PAB II oligomer formation and a reinterpretation of earlier data concerning the protein's binding to poly(A). Images

Nemeth, A; Krause, S; Blank, D; Jenny, A; Jeno, P; Lustig, A; Wahle, E

1995-01-01

74

Complex proteinopathy with accumulations of prion protein, hyperphosphorylated tau, ?-synuclein and ubiquitin in experimental bovine spongiform encephalopathy of monkeys.  

PubMed

Proteins aggregate in several slowly progressive neurodegenerative diseases called 'proteinopathies'. Studies with cell cultures and transgenic mice overexpressing mutated proteins suggested that aggregates of one protein induced misfolding and aggregation of other proteins as well - a possible common mechanism for some neurodegenerative diseases. However, most proteinopathies are 'sporadic', without gene mutation or overexpression. Thus, proteinopathies in WT animals genetically close to humans might be informative. Squirrel monkeys infected with the classical bovine spongiform encephalopathy agent developed an encephalopathy resembling variant Creutzfeldt-Jakob disease with accumulations not only of abnormal prion protein (PrP(TSE)), but also three other proteins: hyperphosphorylated tau (p-tau), ?-synuclein and ubiquitin; ?-amyloid protein (A?) did not accumulate. Severity of brain lesions correlated with spongiform degeneration. No amyloid was detected. These results suggested that PrP(TSE) enhanced formation of p-tau and aggregation of ?-synuclein and ubiquitin, but not A?, providing a new experimental model for neurodegenerative diseases associated with complex proteinopathies. PMID:24769839

Piccardo, Pedro; Cervenak, Juraj; Bu, Ming; Miller, Lindsay; Asher, David M

2014-07-01

75

Epithelial and endothelial expression of the green fluorescent protein reporter gene under the control of bovine prion protein (PrP) gene regulatory sequences in transgenic mice  

NASA Astrophysics Data System (ADS)

The expression of the cellular form of the prion protein (PrPc) gene is required for prion replication and neuroinvasion in transmissible spongiform encephalopathies. The identification of the cell types expressing PrPc is necessary to understanding how the agent replicates and spreads from peripheral sites to the central nervous system. To determine the nature of the cell types expressing PrPc, a green fluorescent protein reporter gene was expressed in transgenic mice under the control of 6.9 kb of the bovine PrP gene regulatory sequences. It was shown that the bovine PrP gene is expressed as two populations of mRNA differing by alternative splicing of one 115-bp 5' untranslated exon in 17 different bovine tissues. The analysis of transgenic mice showed reporter gene expression in some cells that have been identified as expressing PrP, such as cerebellar Purkinje cells, lymphocytes, and keratinocytes. In addition, expression of green fluorescent protein was observed in the plexus of the enteric nervous system and in a restricted subset of cells not yet clearly identified as expressing PrP: the epithelial cells of the thymic medullary and the endothelial cells of both the mucosal capillaries of the intestine and the renal capillaries. These data provide valuable information on the distribution of PrPc at the cellular level and argue for roles of the epithelial and endothelial cells in the spread of infection from the periphery to the brain. Moreover, the transgenic mice described in this paper provide a model that will allow for the study of the transcriptional activity of the PrP gene promoter in response to scrapie infection.

Lemaire-Vieille, Catherine; Schulze, Tobias; Podevin-Dimster, Valérie; Follet, Jérome; Bailly, Yannick; Blanquet-Grossard, Françoise; Decavel, Jean-Pierre; Heinen, Ernst; Cesbron, Jean-Yves

2000-05-01

76

Bovine Papilloma Virus (BPV)Encoded E1 Protein Contains Multiple Activities Required for BPV DNA Replication  

Microsoft Academic Search

Replication of bovine papilloma virus (BPV) DNA requires two virus-encoded proteins, E1 and E2, while all other proteins are supplied by the host cell. Here, we describe the isolation of the E1 protein and show that it is a multifunctional protein. Purified E1 protein was required for the in vitro replication of BPV origin-containing DNA by extracts of mouse cells,

Yeon-Soo Seo; Freidemann Muller; Monika Lusky; Jerard Hurwitz

1993-01-01

77

Distribution of G/sub o. cap alpha. / mRNA and protein in bovine tissues  

SciTech Connect

G/sub o..cap alpha../ is a 39 kDa guanyl nucleotide-binding protein similar in structure and function to G/sub s..cap alpha../ and G/sub i..cap alpha../ in the adenylate cyclase complex and transducin (G/sub t..cap alpha../) in the retinal photon receptor system. A bovine retinal cDNA clone, lambdaG09, that encodes the complete amino acid sequence of G/sub o..cap alpha../ has been isolated. Nick-translated lambdaG09 cDNA and a 5' end-labeled oligonucleotide probe complementary to a 24 base sequence unique to G/sub o..cap alpha../ were used as probes for Northern analysis of poly(A)/sup +/ RNA from bovine tissues. A major 4.0 kb mRNA was detected in brain and retina and in lesser amounts in heart. Several smaller mRNAs also hybridized with both probes in these tissues and in liver and lung. G/sub o..cap alpha../ protein was identified using rabbit polyclonal antibodies directed against purified bovine G/sub o..cap alpha../ and pertussis toxin-catalyzed (/sup 32/P)ADP-ribosylation. Soluble and membrane proteins were incubated with toxin and (/sup 32/P)NAD and then separated by gel electrophoresis before transfer to nitrocellulose for immunoreaction and subsequent autoradiography. A radiolabeled and immunoreactive 39 kDa membrane protein was found principally in retina and brain, and to a lesser extent, in heart. Thus, in the tissues examined, distribution of the 4.0 kb mRNA parallels that of the immunoreactive G/sub o..cap alpha../ with relatively small amounts in heart and larger amounts in brain and retina.

Price, S.R.; Tsai, S.C.; Adamik, R.; Angus, C.W.; Van Meurs, K.P.; Czarnecki, S.; Bruckwick, E.C.; Moss, J.; Vaughan, M.

1987-05-01

78

Hemagglutination mediated by the spike protein of cell-adapted bovine torovirus.  

PubMed

Bovine torovirus (BToV)-Aichi, recently isolated in cultured cells, showed hemagglutination (HA) activity, although the virus has a truncated hemagglutinin-esterase (HE) protein, judging from its gene structure, indicating the existence of another viral protein with HA activity. We examined whether the spike (S) protein possesses HA activity. A BToV antiserum used in this study, reactive to S but not to HE, inhibited HA activity. Furthermore, cells infected with BToV and those expressing S showed hemadsorption (HAD) activity, which was inhibited by the anti-BToV serum; however, HAD activity by expressed HE was not blocked. These data indicate that the S protein of BToV-Aichi is responsible for its HA activity. PMID:23420207

Shimabukuro, Kozue; Ujike, Makoto; Ito, Toshihiro; Tsunemitsu, Hiroshi; Oshitani, Hitoshi; Taguchi, Fumihiro

2013-07-01

79

Serum-derived bovine immunoglobulin/protein isolate: postulated mechanism of action for management of enteropathy  

PubMed Central

The health and performance of the gastrointestinal tract is influenced by the interaction of a variety of factors, including diet, nutritional status, genetics, environment, stress, the intestinal microbiota, immune status, and gut barrier. Disruptions in one or more of these factors can lead to enteropathy or intestinal disorders that are known to occur in concert with certain disease states or conditions such as irritable bowel syndrome or human immunodeficiency virus (HIV) infection. Nutritional support in the form of a medical food along with current therapies could help manage the adverse effects of enteropathy, which include effects on nutrient digestion, absorption, and metabolism, as well as utilization of nutrients from foodstuffs. Numerous studies have demonstrated that oral administration of plasma- or serum-derived protein concentrates containing high levels of immunoglobulins can improve weight management, normalize gut barrier function, and reduce the severity of enteropathy in animals. Recent trials in humans provide preliminary evidence that a serum-derived bovine immunoglobulin/protein isolate is safe and improves symptoms, nutritional status, and various biomarkers associated with enteropathy in patients with HIV infection or diarrhea-predominant irritable bowel syndrome. This review summarizes data from preclinical and clinical studies with immunoglobulin-containing plasma/serum protein concentrates, with a focus on the postulated mode of action of serum-derived bovine immunoglobulin/protein isolate for patients with enteropathy.

Petschow, Bryon W; Burnett, Bruce; Shaw, Audrey L; Weaver, Eric M; Klein, Gerald L

2014-01-01

80

Protein solubility modeling  

NASA Technical Reports Server (NTRS)

A thermodynamic framework (UNIQUAC model with temperature dependent parameters) is applied to model the salt-induced protein crystallization equilibrium, i.e., protein solubility. The framework introduces a term for the solubility product describing protein transfer between the liquid and solid phase and a term for the solution behavior describing deviation from ideal solution. Protein solubility is modeled as a function of salt concentration and temperature for a four-component system consisting of a protein, pseudo solvent (water and buffer), cation, and anion (salt). Two different systems, lysozyme with sodium chloride and concanavalin A with ammonium sulfate, are investigated. Comparison of the modeled and experimental protein solubility data results in an average root mean square deviation of 5.8%, demonstrating that the model closely follows the experimental behavior. Model calculations and model parameters are reviewed to examine the model and protein crystallization process. Copyright 1999 John Wiley & Sons, Inc.

Agena, S. M.; Pusey, M. L.; Bogle, I. D.

1999-01-01

81

Cloning and characterization of the gene encoding the bovine BOULE protein.  

PubMed

The Deleted in Azoospermia (DAZ) genes encode potential RNA-binding proteins that are expressed exclusively in the germ-line. The bovine Deleted in Azoospermia-like gene is a strong candidate for male cattle-yak infertility. In this work, with the goe goal to further reveal the genetic cause of male cattle-yak sterility, another bovine DAZ family gene, b-boule, was isolated and characterized. The b-boule gene is predicted to encode a polypeptide of 295 amino acids with an RNP-type RNA recognition domain. Tertiary structure analysis shows that b-boule binds specifically to polypyrimidine RNAs and might act as a nuclear ribonucleoprotein particle auxiliary factor during germ cell formation and morphological changes of germ cells. RT-PCR assays revealed that b-boule was expressed specifically in the adult testis. However, an extremely low level of expression was detected in the testis of sterile male cattle-yaks. Microstructure of the testes from sterile males showed that type A spermatogonia were the only germ cells present and that few germ cells developed further than the stage of pachytene spermatocytes. These results suggest that b-boule may function in bovine spermatogenesis, and that low levels of b-boule expression might lead to male sterility in cattle-yaks. PMID:18987886

Zhang, Qingbo; Li, Jiahuang; Li, Qifa; Li, Xinfu; Liu, Zhenshan; Song, Dawei; Xie, Zhuang

2009-01-01

82

Transient ubiquitin cross-reactive protein gene expression in the bovine endometrium.  

PubMed

Bovine ubiquitin cross-reactive protein (boUCRP) is secreted by the endometrium from days 15 to 26 of pregnancy in response to conceptus-derived interferon-tau (IFN-tau). We hypothesized that the gene encoding boUCRP was under transcriptional control by the conceptus and IFN-tau. Northern blots using radiolabeled UCRP cDNA revealed a single UCRP transcript of approximately 700 b that was present (P < 0.05) in endometrial cells cultured with 25 nM rboIFN-tau. The UCRP mRNA was not detected in endometrium on days 15, 17, 18 or 19 of the estrous cycle (n = 4 cows on each day) or in spleen, kidney, liver, corpus luteum or muscle. Bovine UCRP mRNA was detectable (P < 0.05) in endometrium from pregnant cows by day 15, reached highest levels by day 17, remained elevated on days 18, 19 and 21, and then declined to amounts on day 26 that were not detectable. Northern blot using radiolabeled ubiquitin cDNA revealed presence of the two major ubiquitin transcripts UbB (1.2 Kb) and UbC (2.6 Kb) in all tissues examined. The bovine UCRP cDNA did not cross-hybridize with these ubiquitin transcripts. We conclude that transcription of the UCRP gene is transient during early pregnancy and regulated by IFN-tau. PMID:9348245

Hansen, T R; Austin, K J; Johnson, G A

1997-11-01

83

Characterization of two proteins of Staphylococcus aureus isolated from bovine clinical mastitis with homology to glyceraldehyde-3-phosphate dehydrogenase.  

PubMed

Staphylococcus aureus is the most common causative agent of bovine mastitis and vaccines developed to control this disease showed limited protection due in part to the lack of common antigens among the mastitis isolates. We isolated and identified two genes encoding proteins with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity from a S. aureus strain isolated from bovine clinical mastitis. The GapB and GapC proteins share considerable homology to the GapB and GapC products of human strains of S. aureus. These two proteins could be distinguished by their different GAPDH activities and binding to bovine transferrin properties. Both gapB and gapC genes were conserved in 11 strains tested, and the GapC protein was present on the surface of all S. aureus strains. PMID:15066729

Goji, Noriko; Potter, Andrew A; Perez-Casal, Jose

2004-04-19

84

Inhibition of protein kinase C results in decreased expression of bovine leukemia virus.  

PubMed Central

The in vitro expression of bovine leukemia virus (BLV) in short-term cultured bovine peripheral blood mononuclear cells (PBMC) is associated with increased spontaneous lymphocyte blastogenesis. The purpose of this study was to determine whether intracellular pathways responsible for antigen- or mitogen-induced lymphocyte blastogenesis were also responsible for induction of BLV expression. The protein kinase C (PKC) inhibitor 1-(5-isoquinolinylsulfonyl)-3-methylpiperazine dihydrochloride (3-methyl H7) decreased blastogenesis in a dose-dependent manner, as measured by [3H]thymidine incorporation, in unstimulated, lipopolysaccharide-stimulated and phorbol ester (PMA)-stimulated BLV-infected PBMC. Similarly, 3-methyl H7 decreased BLV expression, as measured by production of gp51 envelope antigen or p24gag antigen, in BLV-infected PBMC under the same conditions. Using an RNase protection assay, the inhibition of BLV expression by 3-methyl H7 was shown to be due to decreased transcriptional activity. The cyclic GMP-dependent protein kinase and cyclic AMP-dependent protein kinase inhibitor N-(2-guanidinoethyl)-5-isoquinolinesulfonamide (HA1004) did not inhibit either BLV expression or blastogenesis of BLV-infected bovine PBMC. Additional evidence for the PKC-dependent expression of BLV was obtained by using a persistently BLV-infected B-lymphocyte cell line, NBC-13. Activation of PKC by PMA in NBC-13 cells increased BLV expression. 3-methyl H7 decreased the PMA-induced expression of BLV in NBC-13 cells in a dose-dependent manner, whereas HA1004 did not inhibit this expression. These results identify a mechanism for the induction of BLV expression through PKC activation and therefore indicate that latency and replication of BLV is controlled by normal B-lymphocyte intracellular signaling pathways. Images

Jensen, W A; Wicks-Beard, B J; Cockerell, G L

1992-01-01

85

The structure of tubes of bovine rotavirus nucleocapsid protein (VP6) assembled in vitro.  

PubMed

The structure of tubes of reassembled nucleocapsid protein (VP6) from bovine rotavirus (BRV) was determined using optical diffraction of electron micrographs. The tubes consist of a five-start helix of hexagons, with 38 hexagons per helix in a true repeat of three turns. The morphological subunits comprising the hexagons are probably elongated trimers. The structure of naturally occurring tubes (D. Chasey and J. Labram, 1983, J. Gen. Virol. 64, 863-872) was also examined and shown to be similar but not identical to that of tubes assembled in vitro. Considerations of the assembly process are discussed. PMID:2847425

Ready, K F; Buko, K M; Whippey, P W; Alford, W P; Bancroft, J B

1988-11-01

86

Gene Networks Driving Bovine Mammary Protein Synthesis During the Lactation Cycle  

PubMed Central

A crucial role for both insulin and mTOR in the regulation of milk protein synthesis is emerging. Bovine mammary biopsies harvested during late-pregnancy through end of subsequent lactation were used to evaluate via quantitative PCR the expression of 44 genes involved in pathways of insulin, mTOR, AMPK, and Jak2-Stat5 signalling and also glucose and amino acid (AA) transporters. We observed an increased expression during lactation of ELF5, AA and glucose transporters, insulin signaling pathway components, MAPK14, FRAP1, EIF4EBP2, GSK3A and TSC1 among mTOR signaling-related genes. Among ribosomal components RPL22 was down-regulated. The overall data support a central role of AA and glucose transporters and insulin signaling through mTOR for the regulation of protein synthesis in bovine mammary gland. Furthermore, the existence of translational competition favoring the translation of milk protein transcripts was inferred from the combined dataset.

Bionaz, Massimo; Loor, Juan J.

2011-01-01

87

Differential extraction for the rapid purification of bovine surfactant protein B.  

PubMed

Surfactant protein B (SP-B), a peptide found in organic solvent extracts of mammalian surfactant, has been isolated from surfactant previously by column chromatography and/or preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS/PAGE). We have developed a method for isolation of SP-B from bovine surfactant utilizing differential organic extraction. Dried surfactant, isolated from lavage of excised cow lungs, was delipidated by extraction with diisopropyl ether-butanol (3:2). The aqueous layer, containing surfactant proteins, was dried and then was sequentially extracted with diethyl ether-ethanol (3:1) and CHCl3:MeOH:HCl (3:2:0.005 N). SP-B partitioned into chloroform-methanol, which was evaporated under N2. Purified SP-B, quantitated by Coomassie dye binding, represented 1% (wt/wt) of the original surfactant with a final phospholipid-to-protein ratio less than 1. Silver-stained SDS/PAGE of the SP-B extract revealed a single band at 9 kDa (reduced) and 18 kDa (nonreduced), which by immunoblotting reacted strongly with monospecific anti-SP-B antibody. Amino acid sequence analysis confirmed the presence of NH2 and N-1 terminal sequences of bovine SP-B. This procedure offers a rapid, reliable method for isolation of purified SP-B from whole surfactant. PMID:1616060

Beers, M F; Bates, S R; Fisher, A B

1992-06-01

88

Study of the protein-bound fraction of calcium, iron, magnesium and zinc in bovine milk  

NASA Astrophysics Data System (ADS)

Two approaches were used to study the interaction of Ca, Fe, Mg and Zn with bovine milk proteins by inductively coupled plasma optical emission spectrometry (ICPOES). Selective separations in bovine milk samples were accomplished employing an acid protein precipitation using 100 g l -1 trichloroacetic acid (TCA), and an enzymatic protein hydrolysis using 50 g l -1 pepsin (PEP) solution, respectively. The results were compared with total mineral contents determined after microwave-assisted acid digestion. The results obtained by enzymatic and acid precipitation evidenced the different interaction forms of Ca, Fe, Mg and Zn in the system formed by milk components. Iron was not solubilized by the TCA treatment, but was recovered completely after the enzymatic treatment. Quantitative recoveries of Ca, Mg and Zn were obtained using both approaches, showing that these analytes were bound to milk compounds affected by either treatment. Calcium, Mg and Zn are mainly associated with colloidal calcium phosphate and Fe is bound to the backbone of the casein polypeptide chain, cleaved by pepsin enzyme. The proposed approaches could be used to assess the complexity of these chemical interactions.

Silva, Fernando V.; Lopes, Gisele S.; Nóbrega, Joaquim A.; Souza, Gilberto B.; Nogueira, Ana Rita A.

2001-10-01

89

Osteogenic protein-1 promotes the synthesis and retention of extracellular matrix within bovine articular cartilage and chondrocyte cultures  

Microsoft Academic Search

Objective We have used recombinant osteogenic protein-1 to investigate our hypothesis that proper repair and maintenance of cartilage requires not only enhanced biosynthesis and replenishment of the extracellular matrix but also the enhancement of components necessary for matrix retention.Design The effects of osteogenic protein-1 were examined on bovine articular cartilage slices as well as isolated chondrocytes grown in alginate beads.

Y. Nishida; C. B. Knudson; K. E. Kuettner; W. Knudson

2000-01-01

90

Rapid Separation and Quantification of Major Caseins and Whey Proteins of Bovine Milk by Polyacrylamide Gel Electrophoresis  

Microsoft Academic Search

A rapid polyacrylamide gel electro- phoretic method was developed for separating and quantifying major pro- teins in casein and whey protein fractions of bovine milk. For casein separation, best results were achieved by an 8% poly- acrylamide gel containing 4 M urea and a top layer of large pore sample gel; for whey protein the most_ satisfactory separation was with

K. F. Ng-Kwai-Hang; E. M. Kroeker

1984-01-01

91

Relaxation kinetics and the glassiness of proteins: the case of bovine pancreatic trypsin inhibitor.  

PubMed Central

Folded proteins may be regarded as soft active matter under physiological conditions. The densely packed hydrophobic interior, the relatively molten hydrophilic exterior, and the spacer connecting these put together a large number of locally homogeneous regions. For the case of the bovine pancreatic trypsin inhibitor, with the aid of molecular dynamics simulations, we have demonstrated that the kinetics of the relaxation of the internal motions is highly concerted, manifesting the protein's heterogeneity, which may arise from variations in density, local packing, or the local energy landscape. This behavior is characterized in a stretched exponential decay described by an exponent of approximately 0.4 at physiological temperatures. Due to the trapped conformations, configurational entropy becomes smaller, and the associated stretch exponent drops to half of its value below the glass transition range. The temperature dependence of the inverse relaxation time closely follows the Vogel-Tamman-Fulcher expression when the protein is biologically active.

Baysal, Canan; Atilgan, Ali Rana

2002-01-01

92

Cell Arrest and Cell Death in Mammalian Preimplantation Development: Lessons from the Bovine Model  

Microsoft Academic Search

BackgroundThe causes, modes, biological role and prospective significance of cell death in preimplantation development in humans and other mammals are still poorly understood. Early bovine embryos represent a very attractive experimental model for the investigation of this fundamental and important issue.Methods and FindingsTo obtain reference data on the temporal and spatial occurrence of cell death in early bovine embryogenesis, three-dimensionally

Sandra Leidenfrost; Marc Boelhauve; Myriam Reichenbach; Tuna Güngör; Horst-Dieter Reichenbach; Fred Sinowatz; Eckhard Wolf; Felix A. Habermann

2011-01-01

93

Bovine Brain: An in vitro Translational Model in Developmental Neuroscience and Neurodegenerative Research  

PubMed Central

Animal models provide convenient and clinically relevant tools in the research on neurodegenerative diseases. Studies on developmental disorders extensively rely on the use of laboratory rodents. The present mini-review proposes an alternative translational model based on the use of fetal bovine brain tissue. The bovine (Bos taurus) possesses a large and highly gyrencephalic brain and the long gestation period (41?weeks) is comparable to human pregnancy (38–40?weeks). Primary cultures obtained from fetal bovine brain constitute a validated in vitro model that allows examinations of neurons and/or glial cells under controlled and reproducible conditions. Physiological processes can be also studied on cultured bovine neural cells incubated with specific substrates or by electrically coupled electrolyte-oxide-semiconductor capacitors that permit direct recording from neuronal cells. Bovine neural cells and specific in vitro cell culture could be an alternative in comparative neuroscience and in neurodegenerative research, useful for studying development of normal and altered circuitry in a long gestation mammalian species. Use of bovine tissues would promote a substantial reduction in the use of laboratory animals.

Peruffo, Antonella; Cozzi, Bruno

2014-01-01

94

Cloning and expression of a prion protein (PrP) gene from Korean bovine (Bos taurus coreanae) and production of rabbit anti-bovine PrP antibody.  

PubMed

A PrP gene, from a Korean bovine, exhibiting a nonsense and a missense polymorphism respectively at nucleotides 576 and 652 has been cloned. The latter resulted in Glu to Lys substitution at amino acid residue 218. After expression and purification of the recombinant bovine PrP (recBoPrP) with Glu218Lys substitution, a polyclonal antibody against this protein was raised. ELISA and Western blot analysis suggested that the recBoPrP obtained in this study had a unique conformation not presented in native PrP(C), and the polyclonal antibody recognized PrP in a conformation dependent manner. These reagents will be valuable tools for studying PrP conformation. PMID:18574558

Shin, Wooseok; Lee, Byungwoo; Hong, Sungyoul; Ryou, Chongsuk; Kwon, Moosik

2008-10-01

95

Protein changes in Trichinella spiralis muscle larvae in vitro induced by bovine bile.  

PubMed

The purpose of this study was to investigate bovine bile induced protein changes within Trichinella spiralis muscle larvae (ML) in vitro. The larvae were activated by 5% raw bovine bile diluted in saline and in serum-free RPMI-1640 medium at 37°C in 5% CO2 for 2h and, respectively. The crude and excretory-secretory (ES) antigens from ML were analyzed by SDS-PAGE and Western blot. Following activation and comparison to blots of non-activated ML, blots of activated T. spiralis crude worm extract gave rise to three new protein bands (133, 125, and 26 kDa) when screened with mouse infection sera, and four new bands (125, 116, 80, and 29 kDa) when screened with sera from mice immunized with ES antigen. In the same screenings, a loss of two bands migrating at 106 and 25 kDa, and three bands migrating at 76, 58, and 16 kDa, respectively, was observed. When ES antigens from activated ML were blotted and compared to non-activated ML, four new bands (136, 39, 38, and 36 kDa) and seven new bands (136, 120, 100, 39, 36, 34, and 31 kDa) appeared when screened with infection sera and ES immune sera, respectively. In the same comparison, two bands migrating at 67 and 20 kDa, and ten bands migrating at 132, 112, 33, 32, 26, 23, 21, 19, 16, and 15 kDa, were no longer recognized by the ML infection sera and immune sera, respectively. The results showed that after the ML were activated by bile, their protein profiles changed. It is not yet clear if this change is related to the induction or loss of synthesized proteins, or to changes in the migration profiles of existing proteins as a result of post-translational modifications. PMID:23433650

Wang, Li; Wang, Z Q; Cui, J

2013-05-20

96

Haptoglobin comprises about 10% of granule protein extracted from bovine granulocytes isolated from healthy cattle.  

PubMed

Haptoglobin (Hp) is a plasma protein with haemoglobin binding capacity important in maintaining the iron homeostasis and in disease processes influenced by iron metabolism. In cattle Hp is one of the major acute phase proteins, and increases rapidly during infectious disease. At acute clinical mastitis in dairy cows the Hp concentration increases markedly both in blood and milk. Hepatocytes are considered to be the main origin of Hp, but expression of Hp mRNA has also been found in the mammary gland and leukocytes in healthy cattle. In the present study we show that bovine granulocytes, isolated from peripheral blood of healthy cattle, contain abundant amounts of Hp within the granules. As shown by two-dimensional gel electrophoresis (2-DE) in combination with matrix assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-ToF-MS) bovine granulocyte Hp consists of two sets of peptides ca. 20 kDa (alpha-chains) and ca. 40 kDa (beta-chains) with multiple iso-forms. PMID:17681384

Cooray, Ruby; Waller, Karin Persson; Venge, Per

2007-10-15

97

Analysis of the bovine respiratory syncytial virus fusion protein (F) using monoclonal antibodies.  

PubMed

Seven monoclonal antibodies (MAbs) directed against bovine respiratory syncytial virus (BRSV) fusion (F) protein were produced and characterized by radioimmunoprecipitation and immunofluorescence assays. These seven MAbs together with the previously described MAbs (Beeler and Van Wyke Coelingh, 1989) to the F protein of human respiratory syncytial virus (HRSV) were used to study the antigenic variation of 12 strains of ungulate RSV. All except one MAbs specific for the HRSV-F protein reacted with ungulate RSV strains less efficiently, indicating that some epitopes are conserved, and others are not conserved on the F proteins of HRSV and BRSV strains. Three MAbs specific to the BRSV-F protein neutralized virus infectivity and reacted with all the ungulate RSV strains, suggesting that these epitopes are well conserved. Based on the reactivity of three other MAbs specific to the BRSV-F protein, ungulate RSVs could be grouped into two subgroups. The results indicated that there are antigenic variations in the F protein among ungulate RSV strains. PMID:9453129

Pastey, M K; Samal, S K

1997-11-01

98

A 32,000-molecular weight protein from bovine placenta with placental lactogen-like activity in radioreceptor assays  

SciTech Connect

Considerable discrepancies exist in the literature concerning the size and activity of bovine placental lactogen. Our bovine placental lactogen purification preparations were 20% as active as bovine PRL (bPRL) on a per weight basis when compared to bPRL in a lactogenic radioreceptor assay. To identify the active component in these preparations, the proteins were radioiodinated and bound to membrane receptors in the presence and absence of competing hormones, bPRL, and bovine GH (bGH). After centrifugation, membrane pellets were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the gels were autoradiographed. Only one radioiodinated protein band was present. This protein was displaced in the presence of competing hormone and comigrated with a major component of the purification preparations. In radioreceptor assays the active component was as active as bPRL and bGH. From the migration of protein standards included with the radioiodinated purification preparations in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, we estimate that the protein is 32,000 mol wt--about 10,000 mol wt larger than placental lactogens isolated in other species. The possibility that the active molecule was a precursor protein was investigated by examining proteins secreted by bovine placenta tissue cultures. The binding activity in these secretions, as well as in the purification preparations, eluted between ovalbumin (43,000 mol wt) and bPRL (22,000 mol wt) under nondenaturing conditions using high performance gel filtration chromatography. Analysis of this secreted protein, also by binding to membrane receptors, showed that the protein had the same molecular weight as that isolated from the purification preparations and was specifically displaced by the same hormones.

Eakle, K.A.; Arima, Y.; Swanson, P.; Grimek, H.; Bremel, R.D.

1982-05-01

99

87-kDa protein, a major specific substrate for protein kinase C: purification from bovine brain and characterization  

SciTech Connect

The 87-kDa protein, a major specific substrate for protein kinase C, has been purified 500-fold to apparent homogeneity from bovine forebrain supernatant. The purification procedure included batch adsorption to DE-52 (DEAE-cellulose), (NH/sub 4/)/sub 2/SO/sub 4/ precipitation, and chromatography on DEAE-Sephacel, Bio-Gel HTP (hydroxylapatite), Sephacryl S-400, and fast protein liquid chromatography ProRPC. The amino acid composition was notable for its high proportion of alanine (28.6 mol%) and its enrichment in glutamate/glutamine (18.1 mol%), glycine (12.6 mol%), and proline (11.3 mol%). The partial specific volume was 0.702 ml/g; the Stokes radius and sedimentation coefficient were 85 A and 2.11 S, respectively. Although the relative molecular mass of the protein on NaDodSO/sub 4//8% PAGE was 87-90 kDa, the molecular mass as determined from the above values was 68 kDa. The frictional ratio was 3.2, and the axial ratio was 60, indicating that the 87-kDa protein is an extremely elongated monomer. The purified 87-kDa protein was phosphorylated by purified protein kinase C to a stoichiometry of 2.2 mol of /sup 32/P per mol of 87-kDa protein (calculated using a value of 68 kDa for the molecular mass). Phosphorylation was exclusively on serine residues.

Albert, K.A.; Nairn, A.C.; Greengard, P.

1987-10-01

100

Relief of amplification inhibition in PCR with bovine serum albumin or T4 gene 32 protein  

SciTech Connect

The benefits of adding bovine serum albumin (BSA) or T4 gene 32 proteins (gp32) to PCR were evaluated with reaction mixtures containing substances that inhibit amplification. Whereas 10- to 1,000-fold more FeCl{sub 3}, hemin, fulvic acids, humic acids, tannic acids, or extracts from feces, freshwater, or marine water were accommodated in PCR when either 400 ng of BSA per {mu}l was included in the reactions, neither BSA nor gp32 relieved interference significantly when minimum inhibitory levels of bile salts, bilirubin, EDTA, NaCl, sodium dodecyl sulfate, or Triton X-100 were present. Use of BSA and gp32 together offered no more relief of inhibition than either alone at its optimal level, and neither protein had any noticeable effect on amplification in the absence of inhibitors. 21 refs., 3 figs.

Kreader, C.A. [Environmental Protection Agency, Cincinnati, OH (United States)

1996-03-01

101

Profile of bovine proteins in retained and normally expelled placenta in dairy cows.  

PubMed

Tissue-specific protein profile is determined by its function, structure, intensity of metabolism and usefulness. This profile remains under hormonal control. Any disturbance in the general metabolism may be reflected in changes in both protein quantity and quality. These changes can be of low or high specificity, and some can be used as clinical markers of pathological conditions. The aim of this study was to describe and to compare the protein profile of caruncle and foetal villi of bovine placenta that was either properly released or retained. Placental tissues were collected from healthy cows, divided into releasing and retaining foetal membranes, homogenized and subjected to 1D and 2D electrophoresis. Computer-aided analysis of gel images showed essential qualitative and quantitative alterations in protein profile between tissues that were properly released and retained. Alterations concerned both the number of fractions and spots as well as the intensity of staining. This preliminary study provides a general overview of the differences in the protein profile between released and retained foetal membranes. It may allow for selecting the group of proteins or single molecules, which should be further analysed in detail as possible markers differentiating the retention of foetal membranes in cows from placentas that were released spontaneously. The continuation of the study for the identification of particular spots detected in 2D gels is necessary. PMID:24325199

Kankofer, M; Wawrzykowski, J; Hoedemaker, M

2014-04-01

102

Application of an acid proteinase from Monascus purpureus to reduce antigenicity of bovine milk whey protein.  

PubMed

An acid proteinase from Monascus purpureus No. 3403, MpuAP, was previously purified and some characterized in our laboratory (Agric Biol Chem 48:1637-1639, 1984). However, further information about this enzyme is lacking. In this study, we investigated MpuAP's comprehensive substrate specificity, storage stability, and prospects for reducing antigenicity of whey proteins for application in the food industry. MpuAP hydrolyzed primarily five peptide bonds, Gln(4)-His(5), His(10)-Leu(11), Ala(14)-Leu(15), Gly(23)-Phe(24) and Phe(24)-Phe(25) in the oxidized insulin B-chain. The lyophilized form of the enzyme was well preserved at 30-40°C for 7 days without stabilizers. To investigate the possibility of reducing the antigenicity of the milk whey protein, enzymatic hydrolysates of the whey protein were evaluated by inhibition ELISA. Out of the three main components of whey protein, casein and ?-lactalbumin were efficiently degraded by MpuAP. The sequential reaction of MpuAP and trypsin against the whey protein successfully degraded casein, ?-lactalbumin and ?-lactoglobulin with the highest degree of hydrolysis. As a result, the hydrolysates obtained by using the MpuAP-trypsin combination showed the lowest antigenicity compared with the single application of pepsin, trypsin or pepsin-trypsin combination. Therefore, the overall result suggested that the storage-stable MpuAP and trypsin combination will be a productive approach for making hypoallergic bovine milk whey protein hydrolysates. PMID:21298320

Lakshman, P L Nilantha; Tachibana, Shinjiro; Toyama, Hirohide; Taira, Toki; Suganuma, Toshihiko; Suntornsuk, Worapot; Yasuda, Masaaki

2011-09-01

103

The Role and Content of Endogenous Insulin-Like Growth Factor-Binding Proteins in Bovine Articular Cartilage  

Microsoft Academic Search

Previous work identified insulin-like growth factor (IGF)-binding proteins (IGF-BPs) in chondrocyte culture fluids, but the relationship of these proteins to the composition of intact cartilage was not established. The aim of this work was to analyze the IGF-BP system resident in bovine articular cartilage and to examine its role in IGF-1-regulated proteoglycan (PG) metabolism. Protein extracts of freshly dissected or

Teresa I. Morales

1997-01-01

104

Bone Morphogenetic Protein 15 in the Pro-Mature Complex Form Enhances Bovine Oocyte Developmental Competence  

PubMed Central

Developmental competence of in vitro matured (IVM) oocytes needs to be improved and this can potentially be achieved by adding recombinant bone morphogenetic protein 15 (BMP15) or growth differentiation factor (GDF9) to IVM. The aim of this study was to determine the effect of a purified pro-mature complex form of recombinant human BMP15 versus the commercially available bioactive forms of BMP15 and GDF9 (both isolated mature regions) during IVM on bovine embryo development and metabolic activity. Bovine cumulus oocyte complexes (COCs) were matured in vitro in control medium or treated with 100 ng/ml pro-mature BMP15, mature BMP15 or mature GDF9 +/? FSH. Metabolic measures of glucose uptake and lactate production from COCs and autofluorescence of NAD(P)H, FAD and GSH were measured in oocytes after IVM. Following in vitro fertilisation and embryo culture, day 8 blastocysts were stained for cell numbers. COCs matured in medium +/? FSH containing pro-mature BMP15 displayed significantly improved blastocyst development (57.7±3.9%, 43.5±4.2%) compared to controls (43.3±2.4%, 28.9±3.7%) and to mature GDF9+FSH (36.1±3.0%). The mature form of BMP15 produced intermediate levels of blastocyst development; not significantly different to control or pro-mature BMP15 levels. Pro-mature BMP15 increased intra-oocyte NAD(P)H, and reduced glutathione (GSH) levels were increased by both forms of BMP15 in the absence of FSH. Exogenous BMP15 in its pro-mature form during IVM provides a functional source of oocyte-secreted factors to improve bovine blastocyst development. This form of BMP15 may prove useful for improving cattle and human artificial reproductive technologies.

Sudiman, Jaqueline; Sutton-McDowall, Melanie L.; Ritter, Lesley J.; White, Melissa A.; Mottershead, David G.; Thompson, Jeremy G.; Gilchrist, Robert B.

2014-01-01

105

Isolation of a calcium-binding protein of the acrosomal membrane of bovine spermatozoa  

PubMed Central

The mammalian sperm acrosome reaction is a calcium-dependent exocytotic event characterized by extensive fusion between the plasma and the outer acrosomal membrane. The mechanisms by which elevation of cytosolic calcium initiates the membrane fusion process are not understood and the present study was undertaken to identify calcium-binding proteins in the acrosomal membrane (AM) of bovine spermatozoa. Sperm heads, purified from sonicated spermatozoa, were used to isolate an acrosomal membrane-enriched fraction on Percoll density gradients. Using SDS-PAGE and a 45Ca2+-blot overlay assay, calcium-binding proteins of 64, 45, 43, and 39 kDa were identified in the AM enriched fraction. Phase separation analysis with Triton X-114 identified the 64 kDa polypeptide as an integral membrane protein. The 64 kDa polypeptide was purified and utilized to prepare a polyclonal antiserum. Both light and electron microscopic immunocytochemistry demonstrated that the protein was distributed throughout all domains of the acrosomal membrane. These results identify a 64 kDa calcium-binding integral membrane protein of the mammalian acrosome. Its potential function in calcium-dependent membrane fusion events of the acrosome reaction and in fertilization is discussed.

Nagdas, Subir K.; Buchanan, Teresa; McCaskill, Shaina; Mackey, Jared; Alvarez, George E.; Raychoudhury, Samir

2013-01-01

106

Association of the GTP-binding protein Rab3A with bovine adrenal chromaffin granules  

SciTech Connect

The Rab3A protein belongs to a large family of small GTP-binding proteins that are present in eukaryotic cells and that share amino acid identities with the Ras proteins (products of the ras protooncogenes). Rab3A, which is specifically located in nervous and endocrine tissues, is suspected to play a key role in secretion. Its localization was investigated in bovine adrenal gland by using a polyclonal antibody. Rab3A was detected in adrenal medulla but not in adrenal cortex. In cultured adrenal medulla cells, Rab3A was specifically expressed in the catecholamine-secreting chromaffin cells. Subcellular fractionation suggested that Rab3A is about 30% cytosolic and that particulate Rab3A is associated with the membrane of chromaffin granules (the catecholamine storage organelles) and with a second compartment likely to be the plasma membrane. The Rab3A localization on chromaffin granule membranes was confirmed by immunoadsorption with an antibody against dopamine {beta}-hydroxylase. Rab3A was not extracted from this membrane by NaCl or KBr but was partially extracted by urea and totally solubilized by Triton X-100, suggesting either an interaction with an intrinsic protein or a membrane association through fatty acid acylation. This study suggests that Rab3A, which may also be located on other secretory vesicles containing noncharacterized small GTP-binding proteins, is involved in their biogenesis or in the regulated secretion process.

Darchen, F.; Hammel, F.; Monteils, M.P.; Scherman, D. (Centre National de la Recherche Scientifique, Paris (France)); Zahraoui, A.; Tavitian, A. (Institut National de la Sante et de la Recherche Medicale, Paris (France))

1990-08-01

107

The quaternary structure of the recombinant bovine odorant-binding protein is modulated by chemical denaturants.  

PubMed

A large group of odorant-binding proteins (OBPs) has attracted great scientific interest as promising building blocks in constructing optical biosensors for dangerous substances, such as toxic and explosive molecules. Native tissue-extracted bovine OBP (bOBP) has a unique dimer folding pattern that involves crossing the ?-helical domain in each monomer over the other monomer's ?-barrel. In contrast, recombinant bOBP maintaining the high level of stability inherent to native tissue bOBP is produced in a stable native-like state with a decreased tendency for dimerization and is a mixture of monomers and dimers in a buffered solution. This work is focused on the study of the quaternary structure and the folding-unfolding processes of the recombinant bOBP in the absence and in the presence of guanidine hydrochloride (GdnHCl). Our results show that the recombinant bOBP native dimer is only formed at elevated GdnHCl concentrations (1.5 M). This process requires re-organizing the protein structure by progressing through the formation of an intermediate state. The bOBP dimerization process appears to be irreversible and it occurs before the protein unfolds. Though the observed structural changes for recombinant bOBP at pre-denaturing GdnHCl concentrations show a local character and the overall protein structure is maintained, such changes should be considered where the protein is used as a sensitive element in a biosensor system. PMID:24409322

Stepanenko, Olga V; Stepanenko, Olesya V; Staiano, Maria; Kuznetsova, Irina M; Turoverov, Konstantin K; D'Auria, Sabato

2014-01-01

108

Response of the rabbit metaphysis to implants of bovine bone morphogenetic protein (bBMP)  

SciTech Connect

The response of protodifferentiated and differentiated bone cells to bovine bone morphogenetic protein (bBMP) was observed in implants in the adult rabbit distal femoral metaphysis. Bovine serum albumin and denatured bBMP were implanted in the contralateral femur of controls. The changes of the bone marrow reflected the reaction of protodifferentiated cells. The changes in preexisting trabecular bone tissue reflected the reaction of differentiated cells to bBMP. /sup 45/Ca radioisotope quantitative methods demonstrated that the bone morphogenetic response was superimposed upon the reaction to the injury of surgical implantation. By the end of the fourth week, roentgenograms and histologic sections showed larger deposits of intrametaphyseal cartilage and bone in bBMP than in control implanted femurs. By the end of the eighth week, bone formation was associated with remodeling of the entire distal femur and expansion of the external diameter of the metaphysis. These observations indicate the need for investigation of perisinusoid and perivascular cells of periosteum, endosteum, and marrow stroma that are undifferentiated with respect to cartilage and bone but are principal target tissues for BMP.

Nilsson, O.; Urist, M.R.

1985-05-01

109

Molecular Dynamics Simulations Capture the Misfolding of the Bovine Prion Protein at Acidic pH.  

PubMed

Bovine spongiform encephalopathy (BSE), or mad cow disease, is a fatal neurodegenerative disease that is transmissible to humans and that is currently incurable. BSE is caused by the prion protein (PrP), which adopts two conformers; PrPC is the native innocuous form, which is ?-helix rich; and PrPSc is the ?-sheet rich misfolded form, which is infectious and forms neurotoxic species. Acidic pH induces the conversion of PrPC to PrPSc. We have performed molecular dynamics simulations of bovine PrP at various pH regimes. An acidic pH environment induced conformational changes that were not observed in neutral pH simulations. Putative misfolded structures, with nonnative ?-strands formed in the flexible N-terminal domain, were found in acidic pH simulations. Two distinct pathways were observed for the formation of nonnative ?-strands: at low pH, hydrophobic contacts with M129 nucleated the nonnative ?-strand; at mid-pH, polar contacts involving Q168 and D178 facilitated the formation of a hairpin at the flexible N-terminus. These mid- and low pH simulations capture the process of nonnative ?-strand formation, thereby improving our understanding of how PrPC misfolds into the ?-sheet rich PrPSc and how pH factors into the process. PMID:24970211

Cheng, Chin Jung; Daggett, Valerie

2014-01-01

110

Molecular Dynamics Simulations Capture the Misfolding of the Bovine Prion Protein at Acidic pH  

PubMed Central

Bovine spongiform encephalopathy (BSE), or mad cow disease, is a fatal neurodegenerative disease that is transmissible to humans and that is currently incurable. BSE is caused by the prion protein (PrP), which adopts two conformers; PrPC is the native innocuous form, which is ?-helix rich; and PrPSc is the ?-sheet rich misfolded form, which is infectious and forms neurotoxic species. Acidic pH induces the conversion of PrPC to PrPSc. We have performed molecular dynamics simulations of bovine PrP at various pH regimes. An acidic pH environment induced conformational changes that were not observed in neutral pH simulations. Putative misfolded structures, with nonnative ?-strands formed in the flexible N-terminal domain, were found in acidic pH simulations. Two distinct pathways were observed for the formation of nonnative ?-strands: at low pH, hydrophobic contacts with M129 nucleated the nonnative ?-strand; at mid-pH, polar contacts involving Q168 and D178 facilitated the formation of a hairpin at the flexible N-terminus. These mid- and low pH simulations capture the process of nonnative ?-strand formation, thereby improving our understanding of how PrPC misfolds into the ?-sheet rich PrPSc and how pH factors into the process.

Cheng, Chin Jung; Daggett, Valerie

2014-01-01

111

Association between Sscp Haplotypes at the Bovine Growth Hormone Gene and Milk Protein Percentage  

PubMed Central

The bovine Growth Hormone gene (bGH) is an attractive candidate gene for milk production in cattle. Single-strand conformation polymorphisms at bGH were identified and used to define haplotype configurations at this gene in the Israeli Holstein dairy cattle population (Bos taurus) and in the parent animals of the International Bovine Reference Family Panel (a collection of B. taurus and B. indicus crosses). B. taurus and B. indicus haplotypes at the bGH gene differed qualitatively, confirming the previously proposed long evolutionary separation of these cattle subraces. Only a small number of bGH haplotypes were present in the Israel Holstein population. One of the haplotypes, apparently of B. indicus origin, was found to have a highly significant positive effect on milk protein percentage. This illustrates the utility of the haplotype approach for uncovering candidate gene involvement in quantitative genetic variation in agricultural populations. The strong effect of an indicine haplotype in a taurine background raises the possibility that indicine alleles at other candidate genes may comprise a genetic resource for improvement of taurine populations. It is proposed that haplotype analysis may be a useful adjunct to measures of genetic distance for evaluating rare breeds with respect to gene conservation.

Lagziel, A.; Lipkin, E.; Soller, M.

1996-01-01

112

Modulated photophysics of a cationic DNA-staining dye inside protein bovine serum albumin: study of binding interaction and structural changes of protein.  

PubMed

The binding affinity of cationic DNA-staining dye, propidium iodide, with transport protein, bovine serum albumin, has been explored using UV-vis absorption, fluorescence, and circular dichroism spectroscopy. Steady state and time resolved fluorescence studies authenticate that fluorescence quenching of bovine serum albumin by propidium iodide is due to bovine serum albumin-propidium iodide complex formation. Thermodynamic parameters obtained from temperature dependent spectral studies cast light on binding interaction between the probe and protein. Site marker competitive binding has been encountered using phenylbutazone and flufenamic acid for site I and site II, respectively. Energy transfer efficiency and distance between bovine serum albumin and propidium iodide have been determined using Förster mechanism. Structural stabilization or destabilization of protein by propidium iodide has been investigated by urea denaturation study. The circular dichroism study as well as FT-IR measurement demonstrates some configurational changes of the protein in presence of the dye. Docking studies support the experimental data thereby reinforcing the binding site of the probe to the subdomain IIA of bovine serum albumin. PMID:24216153

Samanta, Anuva; Jana, Sankar; Ray, Debarati; Guchhait, Nikhil

2014-03-01

113

Modulation of immune responses to bovine herpesvirus-1 in cattle by immunization with a DNA vaccine encoding glycoprotein D as a fusion protein with bovine CD154  

PubMed Central

The objective of this study was to determine whether a DNA vaccine encoding bovine CD154 linked to a truncated version of bovine herpesvirus-1 (BHV-1) glycoprotein D (tgD-CD154) induces enhanced tgD-specific immune responses in cattle. In vitro characterization demonstrated that tgD and tgD-CD154 both bind to cultured bovine B cells, whereas only tgD-CD154 induces interleukin-4-dependent proliferation, suggesting that tgD-CD154 specifically binds the CD40 receptor and induces receptor signalling. Calves were immunized with plasmid encoding either tgD or tgD-CD154 by intradermal injection with a needle-free device. After two immunizations, tgD-specific immune responses were observed in both vaccinated groups and after challenge with BHV-1 these responses further increased. Animals immunized with plasmid encoding tgD-CD154 had significantly higher tgD-specific serum titres of immunoglobulins G and A but significantly lower numbers of tgD-specific interferon-?-secreting cells than animals immunized with plasmid encoding tgD after BHV-1 challenge. This suggests that the expression of an antigen as a chimeric protein with CD154 can qualitatively alter immune responses in cattle. Since we previously showed that plasmid encoding tgD-CD154 induces significantly enhanced secondary tgD-specific antibody responses in sheep, there appear to be interspecies differences in the immune responses induced by tgD-CD154, which suggests that both proteins in the chimeric molecule may influence protein targeting and the induction of an immune response.

Manoj, Sharmila; Griebel, Philip J; Babiuk, Lorne A; Van Drunen Littel-Van Den Hurk, Sylvia

2004-01-01

114

Effects of Dietary Spray-Dried Bovine Plasma Protein on Broiler Growth Performance and Breast-Meat Yield  

Microsoft Academic Search

SUMMARY Dietary spray-dried plasma protein (SDPP) is effective in improving growth performance of pigs raised in unsanitary conditions. However, little is known about the efficacy of SDPP in improving growth performance and carcass characteristics of broiler chickens. In the present study, graded levels of bovine SDPP (0 to 2% of the diet) were fed to male broiler chickens (Ross 308)

K. Bregendahl; D. U. Ahn; D. W. Trampel; J. M. Campbell

2005-01-01

115

Study on the interaction between bovine serum albumin and 4'-azido-2'-deoxyfluoroarabinocytidine or analogs by spectroscopy and molecular modeling.  

PubMed

The binding of 4'-azido-2'-deoxyfluoroarabinocytidine (FNC) or analogs (cytidine and 5'-cytidylate monophosphate) to bovine serum albumin (BSA) was investigated by fluorescence, UV-vis absorption spectroscopy and molecular modeling. The three compounds quenched the intrinsic fluorescence of BSA and the results revealed the presence of static quenching mechanism. The positive ?H and positive ?S for the systems suggested that the hydrophobic forces stabilized the interaction between the compounds and protein. Results also showed that FNC was the weakest quencher. PMID:24971719

Wang, Ruiyong; Wang, Xiaogai; Li, Zhigang; Xie, Yuanzhe; Yang, Lingling; Shi, Jie; Chang, Junbiao

2014-11-11

116

Calcium-binding protein of bovine intestine. The complete amino acid sequence.  

PubMed

The amino acid sequence for vitamin D-dependent bovine intestinal calcium binding protein has been established. It contains 85 amino acids in a single chain and lacks cysteine, tryptophan, methionine, histidine, and arginine. The NH2-terminal lysine is blocked by an N-acetyl group. Enzymatic digestion with trypsin, chymotrypsin, and pepsin yielded a number of peptides which were purified by two-dimensional high voltage paper electrophoresis. These peptides were examined by end group analysis and sequenced by the dansyl procedure. The absence of tryptophan permitted by a single cleavage of the molecule by N-bromosuccinimide at the tyrosine residue at position 8 and the larger fragment was subjected to automated Edman degradation. By these means, the following sequence was established: N-Ac-Lys-Gln-Ser-Pro-Leu-Glu-Tyr-Ala-Ala-Glu-Lys-Ser-Ile-Gln-Lys-Glu-Ile-Glu-Lys-Gly-Phe-Phe-Lys-Gln-Leu-Leu-Val-Ser-Val-Gln-Lys-Ala-Gly-Asp-Lys-Glu-Ser-Leu-Gln-Pro-Leu-Phe-Thr-Leu-Leu-Lys-Ser-Gly-Pro-Glu-Glu-Asn-Leu-Lys-Glu-Ser-Gln-Asn-Gly-Pro-Asp-Leu-Ls7-Ser-Gly-Pro-Gly-Asn-Asp-Leu-Glu-Glu-Lys-Gly-Thr-Asp-Val-Phe-Ser-Leu-Lys-Gln. Microheterogeneity may exist in the molecule at residue 76 in which position threonine may be replaced by serine. Comparison of the sequence of calcium-binding protein to the "test" sequence of Tufty and Kretsinger ((1975) Science 187, 167-169) proposed to identify E-F hands in muscle proteins suggests that intestinal calcium-binding protein may likewise contain one or possibly two E-F hands which could account for calcium-binding property. Dayhoff alignment scores, however, calculated for calcium-binding protein against nine E-F hands in muscle proteins parvalbumin, troponin and alkali light chains do not indicate that intestinal calcium-binding protein is homologous to these muscle protein chains. PMID:1176441

Huang, W Y; Cohn, D V; Hamilton, J W; Fullmer, C; Wasserman, R H

1975-10-10

117

Effect of trenbolone acetate on protein synthesis and degradation rates in fused bovine satellite cell cultures.  

PubMed

Although androgenic and estrogenic steroids are widely used to enhance muscle growth and increase feed efficiency in feedlot cattle, their mechanism of action is not well understood. Although in vivo studies have indicated that androgens affect protein synthesis and protein degradation rate in muscle, results from in vitro studies have been inconsistent. We have examined the effects of trenbolone acetate (TBA), a synthetic androgen, on protein synthesis and degradation rates in fused bovine satellite cell (BSC) cultures. Additionally, we have examined the effects of compounds that interfere with binding of TBA or insulin-like growth factor-1 (IGF-1) to their respective receptors on TBA-induced alterations in protein synthesis and degradation rates in BSC cultures. Treatment of fused BSC cultures with TBA results in a concentration-dependent increase (P < 0.05) in protein synthesis rate and a decrease (P < 0.05) in degradation rate, establishing that TBA directly affects these parameters. Flutamide, a compound that prevents androgen binding to the androgen receptor, suppresses (P < 0.05) TBA-induced alterations in protein synthesis and degradation in fused BSC cultures, indicating the androgen receptor is involved. JB1, a competitive inhibitor of IGF-1 binding to the type 1 IGF receptor (IGF1R), suppresses (P < 0.05) TBA-induced alterations in protein synthesis and degradation, indicating that this receptor also is involved in the actions of TBA on both synthesis and degradation. In summary, our data show that TBA acts directly to alter both protein synthesis and degradation rates in fused BSC cultures via mechanisms involving both the androgen receptor and IGF1R. PMID:20961723

Kamanga-Sollo, E; White, M E; Hathaway, M R; Weber, W J; Dayton, W R

2011-01-01

118

Probing protein structure by solvent perturbation of NMR spectra: the surface accessibility of bovine pancreatic trypsin inhibitor.  

PubMed Central

In the absence of specific interactions, the relative attenuation of protein NMR signals due to added stable free radicals such as TEMPOL should reflect the solvent accessibility of the molecular surface. The quantitative correlation between observed attenuation and surface accessibility was investigated with a model system, i.e., the small protein bovine pancreatic trypsin inhibitor. A detailed discussion is presented on the reliability and limits of the approach, and guidelines are provided for data acquisition, treatment, and interpretation. The NMR-derived accessibilities are compared with those obtained from x-ray diffraction and molecular dynamics data. Although the time-averaged accessibilities from molecular dynamics are ideally suited to fit the NMR data, better agreement was observed between the paramagnetic attenuations of the fingerprint cross-peaks of homonuclear proton spectra and the total NH and H alpha accessibilities calculated from x-ray coordinates, than from time-averaged molecular dynamics simulations. In addition, the solvent perturbation response appears to be a promising approach for detecting the thermal conformational evolution of secondary structure elements in proteins.

Molinari, H; Esposito, G; Ragona, L; Pegna, M; Niccolai, N; Brunne, R M; Lesk, A M; Zetta, L

1997-01-01

119

Statistical Modeling of protein spray drying at the lab scale  

Microsoft Academic Search

The objective of this study was to examine the effects of formulation and process variables on particle size and other characteristics\\u000a of a spray-dried model protein, bovine serum albumin (BSA), using a partial factorial design for experiments. Formulation\\u000a variables tested include concentration and zinc:protein complexation ratio. Process variables explored were inlet temperature,\\u000a liquid feed rate, drying air flow rate, and

Kristin B. Prinn; Henry R. Costantino; Mark Tracy

2002-01-01

120

Spontaneous and BSE-prion-seeded amyloid formation of full length recombinant bovine prion protein.  

PubMed

The conversion of the cellular isoform of the prion protein into the pathogenic isoform PrP(Sc) is the key event in prion diseases. The disease can occur spontaneously genetically or by infection. In earlier studies we presented an in vitro conversion system which simulates the structural transition in recPrP by varying low concentrations of SDS at constant NaCl. In the present study we adopted the conversion system from experimental Scrapie in hamster to bovine recPrP and generated amyloid fibrils. The intermediate state which is optimal for fibril formation is a soluble, beta-rich state. The system was extended using BSE-prions as seeds and led to an acceleration of fibril formation by orders of magnitude. This seeded amyloid formation assay avoids any PK-treatment, is therefore able to detect even PK-sensitive PrP(Sc) and does not require cellular components. PMID:18585368

Panza, Giannantonio; Stöhr, Jan; Dumpitak, Christian; Papathanassiou, Dimitrios; Weiss, Jürgen; Riesner, Detlev; Willbold, Dieter; Birkmann, Eva

2008-09-01

121

Evaluation of non-immunoaffinity methods for isolation of cellular prion protein from bovine brain.  

PubMed

Transmissible spongiform encephalopathies (TSEs) are progressive neurodegenerative diseases that affect the central nervous system of many animals, including humans. Research suggests that TSEs are caused by conversion of the cellular prion protein (PrP(C)), which is encoded in many tissues, especially brain, to the pathological form (PrP(Sc)). This conversion affects PrP(Sc) structure, conferring different biochemical properties, such as the increased resistance to proteinase K, that have been widely used for its purification. By contrast, PrP(C) is less resistant and its isolation is more challenging. Here, we propose a purification strategy to efficiently recover PrP(C) from healthy bovine brain using conventional non-immunoaffinity methods. The applicability of extraction using detergents, size exclusion chromatography, diafiltration with molecular weight cutoff (MWCO) filters, and immobilized metal affinity chromatography (IMAC) using Western blot (WB) analysis to detect the presence of PrP(C) is discussed in detail. PMID:24463017

Borges-Alvarez, M; Benavente, F; Márquez, M; Barbosa, J; Sanz-Nebot, V

2014-04-15

122

Purification and characterization of variants of acyl-CoA-binding protein in the bovine liver.  

PubMed Central

Four differently modified forms of acyl-CoA-binding protein (ACBP) were identified in ACBP purified from bovine liver. The majority of the purified ACBP was focused at pH 5.9 in isoelectric focusing and could be shown to be N-acetylated ACBP without any further modifications. Two minor peaks were focused at pH 5.25 and 4.85 respectively. Mass spectrometry and sequence determination showed that the pI 5.25 form was acetylated at Lys18 and that the pI 4.85 form was malonylated in the same position. Furthermore, it could be shown that non-enzymic glycosylation occurred during purification. The acetylated and malonylated variants of ACBP were only found in adult cattle. Images Fig. 5.

Jensen, M S; H?jrup, P; Rasmussen, J T; Knudsen, J

1992-01-01

123

Adhesion of Fusobacterium necrophorum to bovine endothelial cells is mediated by outer membrane proteins.  

PubMed

Fusobacterium necrophorum, a Gram-negative anaerobe, is frequently associated with suppurative and necrotic infections of animals and humans. The organism is a major bovine pathogen, and in cattle, the common fusobacterial infections are hepatic abscesses, foot rot, and necrotic laryngitis. The species comprises two subspecies: F. necrophorum subsp. necrophorum and F. necrophorum subsp. funduliforme. Bacterial adhesion to the host cell surface is a critical initial step in the pathogenesis, and outer membrane proteins (OMP) play an important role in adhesion and establishment of certain Gram-negative bacterial infections. The means by which F. necrophorum attaches to epithelial or endothelial cells has not been determined. We evaluated whether OMP of F. necrophorum, isolated from a liver abscess, mediated adhesion to bovine endothelial cells (adrenal gland capillary endothelial cell line). The extent of binding of subsp. necrophorum to the endothelial cells was higher than that of F. necrophorum subsp. funduliforme. Trypsin treatment of bacterial cells decreased their binding to endothelial cells indicating the protein nature of adhesins. Preincubation of endothelial cells with OMP extracted from F. necrophorum decreased the binding of bacterial cells. In addition, binding of each subspecies to endothelial cells was inhibited by polyclonal antibodies raised against respective OMP and the antibody-mediated inhibition was subspecies specific. The western blot analysis of OMP bound to endothelial cells with anti-OMP antibodies showed four OMP of 17, 24, 40 and 74 kDa. We conclude that OMP of F. necrophorum play a role in adhesion of bacterial cells to the endothelial cells. PMID:23153522

Kumar, Amit; Gart, Elena; Nagaraja, T G; Narayanan, Sanjeev

2013-03-23

124

Alterations in bovine platelet function and acute phase proteins induced by Pasteurella haemolytica A1.  

PubMed

Platelet function was assessed by aggregometry in 10 Holstein calves before and after exposure to Pasteurella haemolytica (biotype A, serotype 1) by intrabronchial challenge. At 24 h after exposure the platelets had become more reactive to stimulation with known platelet agonists such as adenosine diphosphate (ADP) and platelet-activating factor (PAF) and the platelet aggregates that formed were more resistant to disaggregation. The activation of platelets was an early response in the challenged calves as platelet function had returned to pretreatment levels 72 h after exposure to the bacteria while the acute phase reactant proteins, haptoglobin and fibrinogen, were approaching their peak values and alpha 2-macroglobulin levels had also risen significantly (P < 0.05) at this time. The plasma levels of these proteins were still elevated and albumin levels were depressed 6 d post-treatment. At post-mortem all calves exhibited pneumonic tissue damage. When P. haemolytica leukotoxin was added directly to bovine platelet suspensions both spontaneous aggregation and an increase in the aggregation response to ADP and PAF stimulation were observed. The morphological appearance of the platelet aggregates exhibited the typical pattern for bovine platelets with 2 distinct zones of cells being visible within each aggregate. One zone contained platelets in which the cytoplasmic granules were still evident and the other zone contained irregularly shaped platelets devoid of granular content. In the latter zone, discrete gaps, or pores, were evident in the plasma membrane of numerous platelets. This pore formation is characteristic of leukotoxin action and is not observed in ADP or PAF induced aggregates. PMID:9442932

Cheryk, L A; Hooper-McGrevy, K E; Gentry, P A

1998-01-01

125

Alterations in bovine platelet function and acute phase proteins induced by Pasteurella haemolytica A1.  

PubMed Central

Platelet function was assessed by aggregometry in 10 Holstein calves before and after exposure to Pasteurella haemolytica (biotype A, serotype 1) by intrabronchial challenge. At 24 h after exposure the platelets had become more reactive to stimulation with known platelet agonists such as adenosine diphosphate (ADP) and platelet-activating factor (PAF) and the platelet aggregates that formed were more resistant to disaggregation. The activation of platelets was an early response in the challenged calves as platelet function had returned to pretreatment levels 72 h after exposure to the bacteria while the acute phase reactant proteins, haptoglobin and fibrinogen, were approaching their peak values and alpha 2-macroglobulin levels had also risen significantly (P < 0.05) at this time. The plasma levels of these proteins were still elevated and albumin levels were depressed 6 d post-treatment. At post-mortem all calves exhibited pneumonic tissue damage. When P. haemolytica leukotoxin was added directly to bovine platelet suspensions both spontaneous aggregation and an increase in the aggregation response to ADP and PAF stimulation were observed. The morphological appearance of the platelet aggregates exhibited the typical pattern for bovine platelets with 2 distinct zones of cells being visible within each aggregate. One zone contained platelets in which the cytoplasmic granules were still evident and the other zone contained irregularly shaped platelets devoid of granular content. In the latter zone, discrete gaps, or pores, were evident in the plasma membrane of numerous platelets. This pore formation is characteristic of leukotoxin action and is not observed in ADP or PAF induced aggregates. Images Figure 2.

Cheryk, L A; Hooper-McGrevy, K E; Gentry, P A

1998-01-01

126

Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering.  

PubMed

In this work, an influence of bovine serum albumin proteins grafting on the surface properties of plasma-treated polyethylene and poly-l-lactic acid was studied. The interaction of the vascular smooth muscle cells with the modified polymer surface was determined. The surface properties were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, nano-LC-ESI-Q-TOF mass spectrometry, electrokinetic analysis, and goniometry. One of the motivations for this work is the idea that by the interaction of the cell with substrate surface, the proteins will form an interlayer between the cell and the substrate. It was proven that when interacting with the plasma-treated high-density polyethylene and poly-l-lactic acid, the bovine serum albumin protein is grafted on the polymer surface. Since the proteins are bonded to the substrate surface, they can stimulate cell adhesion and proliferation. PMID:24708858

Kasálková, Nikola Slepi?ková; Slepi?ka, Petr; Kolská, Zde?ka; Hoda?ová, Petra; Ku?ková, St?pánka; Svor?ík, Václav

2014-01-01

127

Grafting of bovine serum albumin proteins on plasma-modified polymers for potential application in tissue engineering  

NASA Astrophysics Data System (ADS)

In this work, an influence of bovine serum albumin proteins grafting on the surface properties of plasma-treated polyethylene and poly- l-lactic acid was studied. The interaction of the vascular smooth muscle cells with the modified polymer surface was determined. The surface properties were characterized by X-ray photoelectron spectroscopy, atomic force microscopy, nano-LC-ESI-Q-TOF mass spectrometry, electrokinetic analysis, and goniometry. One of the motivations for this work is the idea that by the interaction of the cell with substrate surface, the proteins will form an interlayer between the cell and the substrate. It was proven that when interacting with the plasma-treated high-density polyethylene and poly- l-lactic acid, the bovine serum albumin protein is grafted on the polymer surface. Since the proteins are bonded to the substrate surface, they can stimulate cell adhesion and proliferation.

Kasálková, Nikola Slepi?ková; Slepi?ka, Petr; Kolská, Zde?ka; Hoda?ová, Petra; Ku?ková, Št?pánka; Švor?ík, Václav

2014-04-01

128

Association of Bovine Papillomavirus Type 1 E6 oncoprotein with the focal adhesion protein paxillin through a conserved protein interaction motif  

Microsoft Academic Search

We have found that the E6 oncoprotein of Bovine Papillomavirus Type 1 (BE6) as well as the E6 protein of the cancer associated HPV-16 (16E6) interact with the focal adhesion protein paxillin. Mutational analysis of paxillin revealed that BE6 binds paxillin through small protein interaction motifs called LD motifs that have been previously identified as important in regulating association of

Scott B Vande Pol; Michael C Brown; Christopher E Turner; SB Vande Pol

1998-01-01

129

The influence of protein fractions from bovine colostrum digested in vivo and in vitro on human intestinal epithelial cell proliferation.  

PubMed

Colostrum consists of a number of biologically active proteins and peptides that influence physiological function and development of a neonate. The present study investigated the biological activity of peptides released from first day bovine colostrum through in vitro and in vivo enzymatic digestion. This was assessed for proliferative activity using a human intestinal epithelial cell line, T84. Digestion of the protein fraction of bovine colostrum in vitro was conducted with the enzymes pepsin, chymosin and trypsin. Pepsin and chymosin digests yielded protein fractions with proliferative activity similar to that observed with undigested colostrum and the positive control foetal calf serum (FCS). In contrast trypsin digestion significantly (P<0·05) decreased colostral proliferative activity when co-cultured with cells when compared with undigested colostrum. The proliferative activity of undigested colostrum protein and abomasal whey protein digesta significantly increased (P<0·05) epithelial cell proliferation in comparison to a synthetic peptide mix. Bovine colostrum protein digested in vivo was collected from different regions of the gastrointestinal tract (GIT) in newborn calves fed either once (n=3 calves) or three times at 12-h intervals (n=3 calves). Digesta collected from the distal duodenum, jejunum and colon of calves fed once, significantly (P<0·05) stimulated cell proliferation in comparison with comparable samples collected from calves fed multiple times. These peptide enriched fractions are likely to yield candidate peptides with potential application for gastrointestinal repair in mammalian species. PMID:24433585

Morgan, Alison J; Riley, Lisa G; Sheehy, Paul A; Wynn, Peter C

2014-02-01

130

Binding of (/sup 3/H)forskolin to platelet membranes and solubilized proteins from bovine brain  

SciTech Connect

(/sup 3/H)Forskolin ((/sup 3/H)FSK) bound to platelet membranes with a Kd of 20 nM and a Bmax of 125 fmol/mg protein. The Bmax was increased to 400 fmol/mg protein in the presence of GppNHp (or NaF) and MgCl/sub 2/ with no change in Kd. PGE/sub 1/ decreased the EC50 of GppNHp to increase the Bmax for (/sup 3/H)FSK binding from 600 nM to 35 nM. In contrast, PGE/sub 1/ had no effect on the EC50 of NaF to increase (/sup 3/H)FSK binding. (/sup 3/H)FSK binding increased slowly over 60 min when forskolin and GppNHp were added to membranes simultaneously at 20/sup 0/C. Preincubation of membranes with GppNHp at 20/sup 5/C also caused a linear increase in adenylate cyclase specific activity over 60 minutes. (/sup 3/H)FSK bound to solubilized protein from bovine brain membrane with a Kd of 22 nM. GppNHp increased the number of binding sites in solubilized proteins only if membranes were not preincubated with GppNHp prior to solubilization. In conclusion the number of binding sites for (/sup 3/H)FSK is increased by agents that activate adenylate cyclase through the Ns protein. These sites appear to be associated with an activated complex of the Ns protein and adenylate cyclase.

Nelson, C.A.; Seamon, K.B.

1986-05-01

131

Structure-function relationships of bovine pulmonary surfactant proteins: SP-B and SP-C.  

PubMed

Pulmonary surfactant contains at least three unique proteins: SP-A, SP-B and SP-C. SP-B and SP-C from bovine surfactant are markedly hydrophobic and have molecular masses between 3 and 26 kDa. We identify surfactant proteins under nonreducing conditions on polyacrylamide gels with approximate molecular mass of 5, 14, 26 kDa (SP-5, 14, 26) when organic solvent-soluble material is eluted from a Sephadex LH-20 size exclusion column followed by separation on a high-performance reverse-phase chromatography system. These bands correspond to monomeric SP-C, oligomeric SP-C and oligomeric SP-B, respectively. Computer analysis (Eisenberg-hydrophobic moment) of sequences for these proteins suggests that SP-B contains surface-seeking amphiphilic segments. In contrast, SP-C resembles a more hydrophobic transmembrane anchoring peptide. Dispersions containing dipalmitoylphosphatidylcholine, phosphatidylglycerol, palmitic acid and multimeric SP-B and SP-C duplicate the surface activity of natural surfactant when assayed in a pulsating bubble surfactometer. We speculate that oligomers of SP-B and monomers and oligomers of SP-C may act cooperatively in affecting surfactant function. An important function of SP-B and SP-C may be to affect the ordering of surfactant lipids so that rates of transport of surfactant lipids to the hypophase surface in the alveoli are enhanced. PMID:2160285

Takahashi, A; Waring, A J; Amirkhanian, J; Fan, B; Taeusch, H W

1990-05-01

132

Spectral [corrected] studies on the cadmium-ion-binding properties of bovine brain S-100b protein.  

PubMed

The effect of Cd2+ binding on bovine brain S-100b protein was studied using c.d. u.v. difference spectroscopy and fluorescence measurements. At pH 7.5, S-100b protein binds two Cd2+ ions per monomer with a Kd value of 3 x 10(-5) M. Addition of Cd2+ resulted in perturbing the single tyrosine residue (Tyr17) in the protein as indicated by u.v. difference spectroscopy and aromatic c.d. measurements. In the presence of Cd2+, the tyrosine residue moves to a more non-polar environment, since a red shift was observed in the u.v. difference spectrum. When the protein was excited at 278 nm, the tyrosine fluorescence emission maximum was centred at 306 nm. Cd2+ addition resulted in an increase in intrinsic fluorescence intensity. Fluorescence titration with Cd2+ indicated the protein binds Cd2+ with a Kd value of 3 x 10(-5) M. 2-p-Toluidinylnaphthalene-6-sulphonate-labelled protein, when excited at 345 nm, had a fluorescence emission maximum at 440 nm. Addition of Cd2+ to labelled protein resulted in a 5-fold increase in fluorescence intensity accompanied by a 5 nm blue shift in the emission maximum, suggesting that the probe, in the presence of Cd2+, moves to a hydrophobic domain. U.v. difference spectroscopic studies indicated a unique Cd2(+)-binding site on the protein, since Cd2+ addition yielded a large positive absorption band in the 240 nm region that is not found with either Ca2+ or Zn2- ions. Similar absorption bands have been observed in Cd-protein complexes such as Cd-metallothionein [Vasak, Kagi & Hill (1981) Biochemistry 20, 2852-2856] and also in model complexes of Cd2+ with 2-mercaptoethanol. This absorption band is believed to arise as a result of charge-transfer transitions between the thiolate and Cd2+. Of the two Cd2- -binding sites on the beta-chain, one must be located at the N-terminal end near the single tyrosine residue, since Cd2- and Zn2+ produced similar effects on the intrinsic protein fluorescence. The other Cd2+ site which is unique to Cd2+ must be Cys84, located at the C-terminal end. PMID:2039467

Donato, H; Mani, R S; Kay, C M

1991-05-15

133

Multiple protein biomarker assessment for recombinant bovine somatotropin (rbST) abuse in cattle.  

PubMed

Biomarker profiling, as a rapid screening approach for detection of hormone abuse, requires well selected candidate biomarkers and a thorough in vivo biomarker evaluation as previously done for detection of growth hormone doping in athletes. The bovine equivalent of growth hormone, called recombinant bovine somatotropin (rbST) is (il)legally administered to enhance milk production in dairy cows. In this study, first a generic sample pre-treatment and 4-plex flow cytometric immunoassay (FCIA) were developed for simultaneous measurement of four candidate biomarkers selected from literature: insulin-like growth factor 1 (IGF-1), its binding protein 2 (IGFBP2), osteocalcin and endogenously produced antibodies against rbST. Next, bovine serum samples from two extensive controlled rbST animal treatment studies were used for in vivo validation and biomarker evaluation. Finally, advanced statistic tools were tested for the assessment of biomarker combination quality aiming to correctly identify rbST-treated animals. The statistical prediction tool k-nearest neighbours using a combination of the biomarkers osteocalcin and endogenously produced antibodies against rbST proved to be very reliable and correctly predicted 95% of the treated samples starting from the second rbST injection until the end of the treatment period and even thereafter. With the same biomarker combination, only 12% of untreated animals appeared false-positive. This reliability meets the requirements of Commission Decision 2002/657/EC for screening methods in veterinary control. From the results of this multidisciplinary study, it is concluded that the osteocalcin - anti-rbST-antibodies combination represent fit-for-purpose biomarkers for screening of rbST abuse in dairy cattle and can be reliably measured in both the developed 4-plex FCIA as well as in a cost-effective 2-plex microsphere-based binding assay. This screening method can be incorporated in routine veterinary monitoring programmes: in the European Union for detection of rbST abuse and in the control of rbST-free dairy farms in the United States of America and other countries. PMID:23300820

Ludwig, Susann K J; Smits, Nathalie G E; van der Veer, Grishja; Bremer, Maria G E G; Nielen, Michel W F

2012-01-01

134

Multiple Protein Biomarker Assessment for Recombinant Bovine Somatotropin (rbST) Abuse in Cattle  

PubMed Central

Biomarker profiling, as a rapid screening approach for detection of hormone abuse, requires well selected candidate biomarkers and a thorough in vivo biomarker evaluation as previously done for detection of growth hormone doping in athletes. The bovine equivalent of growth hormone, called recombinant bovine somatotropin (rbST) is (il)legally administered to enhance milk production in dairy cows. In this study, first a generic sample pre-treatment and 4-plex flow cytometric immunoassay (FCIA) were developed for simultaneous measurement of four candidate biomarkers selected from literature: insulin-like growth factor 1 (IGF-1), its binding protein 2 (IGFBP2), osteocalcin and endogenously produced antibodies against rbST. Next, bovine serum samples from two extensive controlled rbST animal treatment studies were used for in vivo validation and biomarker evaluation. Finally, advanced statistic tools were tested for the assessment of biomarker combination quality aiming to correctly identify rbST-treated animals. The statistical prediction tool k-nearest neighbours using a combination of the biomarkers osteocalcin and endogenously produced antibodies against rbST proved to be very reliable and correctly predicted 95% of the treated samples starting from the second rbST injection until the end of the treatment period and even thereafter. With the same biomarker combination, only 12% of untreated animals appeared false-positive. This reliability meets the requirements of Commission Decision 2002/657/EC for screening methods in veterinary control. From the results of this multidisciplinary study, it is concluded that the osteocalcin – anti-rbST-antibodies combination represent fit-for-purpose biomarkers for screening of rbST abuse in dairy cattle and can be reliably measured in both the developed 4-plex FCIA as well as in a cost-effective 2-plex microsphere-based binding assay. This screening method can be incorporated in routine veterinary monitoring programmes: in the European Union for detection of rbST abuse and in the control of rbST-free dairy farms in the United States of America and other countries.

Ludwig, Susann K. J.; Smits, Nathalie G. E.; van der Veer, Grishja; Bremer, Maria G. E. G.; Nielen, Michel W. F.

2012-01-01

135

The interaction of bovine serum albumin with doxorubicin-loaded superparamagnetic iron oxide nanoparticles: spectroscope and molecular modelling identification.  

PubMed

To take a comprehensive evaluation of the bio-safety of doxorubicin-loaded superparamagnetic iron oxide nanoparticles (SPION), the interaction of bovine serum albumin (BSA) with the drug delivery was investigated by multi-spectroscopic techniques and molecular modelling calculation. Ultraviolet absorption and synchronous fluorescence results elucidate that DOX-SPION unfold the framework conformation of BSA, leading to changes in the microenvironment of amide moieties. Circular dichroism (CD) data show that the content of ?-helix decreases from 68.62% to 62.76%, which shows the changes of protein's secondary structure quantificationally. Through Stern-Volmer analysis, the quenching mode is determined to be static interaction, forming a stable bioconjugate. The molecular model illustrates that DOX prefers a highly polar binding site at the external region of domains ? of BSA, and the hydrogen bonds are marked. This work elucidates that the drug delivery has deleterious effects on the frame conformation of protein, affecting its physiological function. PMID:22087533

Liu, Yihong; Ji, Fanying; Liu, Rutao

2013-02-01

136

Identification of anti-adipogenic proteins in adult bovine serum suppressing 3T3-L1 preadipocyte differentiation.  

PubMed

Adipocyte differentiation is a complex developmental process forming adipocytes from various precursor cells. The murine 3T3-L1 preadipocyte cell line has been most frequently used in the studies of adipocyte differentiation. Differentiation of 3T3-L1 preadipocytes includes a medium containing fetal bovine serum (FBS) with hormonal induction. In this study, we observed that differentiation medium containing adult bovine serum (ABS) instead of FBS did not support differentiation of preadipocytes. Impaired adipocyte differentiation was due to the presence of a serum protein factor in ABS that suppresses differentiation of preadipocytes. Using a proteomic analysis, alpha-2-macroglobulin and paraoxonase/arylesterase 1, which were previously shown to suppress differentiation of preadipocytes, were identified as anti-adipogenic proteins. Although their functional mechanisms have not yet been elucidated, the anti-adipogenic effects of these proteins are discussed. PMID:24195790

Park, Jeongho; Park, Jihyun; Nahm, Sang-Soep; Choi, Inho; Kim, Jihoe

2013-12-01

137

Gz- and not Gi-proteins are coupled to pre-junctional ?-opioid receptors in bovine airways.  

PubMed

We investigated the signal transmission pathway by which activation of ?-opioid receptors attenuates acetylcholine (ACh) release in bovine trachealis. Electrical stimulation (ES)-induced [(3)H]-ACh release was determined in bovine tracheal smooth muscle strips pre-incubated with either the Gi-protein inhibitor pertussis toxin (PTX, 500 ng/ml and 1 ?g/ml) or the Gz-protein specific inhibitor arachidonic acid (AA, 10(-6)M and 10(-5)M) and then treated with DAMGO (D-Ala(2),N-MePhe(4),Gly-ol(5)-enkephalin) 10(-5)M. Indomethacin 10(-5)M was used to block AA cascade. The inhibitory effect of DAMGO on ES-induced [(3)H]-ACh release was PTX-insensitive, but, by contrast, ablated by AA in a concentration-dependent manner. AA 10(-5)M alone reduced [(3)H]-ACh release, an effect that was prevented by iberiotoxin 10(-7)M, suggesting an involvement of Ca(2+)-activated K(+)-channels. Western blot analysis consistently showed immunoreactive bands against a specific antibody anti-Gz-? subunit at ?40 kDa, consistent with the presence of Gz-protein. The present findings suggest that in isolated bovine trachealis, activation of ?-opioid receptors inhibits ACh-release through a signal transmission pathway involving Gz-protein rather than Gi-protein. PMID:23911590

Baroffio, Michele; Brichetto, Lorenzo; Franco, Luisa; Crimi, Emanuele; Rehder, Kai; Brusasco, Vito

2013-10-01

138

In vitro maturation, fertilization and culture to blastocysts of bovine oocytes in protein-free media.  

PubMed

This study examined the role of protein supplementation at the various steps of the in vitro production of bovine embryos derived from two different morphological categories of COC. The basic medium was TCM 199 and was supplemented with hormones during maturation in vitro and either estrous cow serum (ECS), bovine serum albumin (BSA) at various concentrations or polyvinyl-alcohol (PVA). Fertilization in vitro was carried out using frozen-thawed semen or one bull in Fert-talp containing heparin, hypotaurin and epinephrine and either 6 mg/ml BSA or 1 mg/ml PVA. In vitro culture up to the blastocyst stage was performed in TCM 199 supplemented with either ECS, BSA or PVA. The first experiment investigated the influence of different medium-supplements (ECS, BSA or PVA) on nuclear maturation and revealed no significant differences among treatment groups nor between categories of COC (63.9% to 74.9% and 48.9% to 77.0%, respectively). The time course of in vitro fertilization was elucidated in Experiment 2 in medium supplemented with either protein or PVA during maturation and fertilization. Penetration was not affected (70.9% to 79.3% penetration 12 h after onset of oocyte-sperm-co-incubation), but formation of pronuclei was decreased (P < 0.05) 12 and 19 h after onset of oocyte-sperm-co-incubation and was retarded in medium supplemented with PVA (12 h: 63.8 vs 21.4 %; 19 h: 57.5 vs 20.8 %, respectively) while cleavage was not affected. In Experiment 3, six treatment groups were formed in which the two different morphological categories of cumulus-oocyte-complexes (COC) were incubated in basic medium supplemented with 1) ECS during maturation and embryo culture and BSA during fertilization; 2) PVA during maturation and embryo culture, fertilization medium with PVA; 3) PVA during maturation and embryo culture, fertilization medium with BSA; 4) BSA (1 mg/ml) during maturation, fertilization and embryo culture; 5) BSA (6 mg/ml) during maturation, fertilization and embryo culture; and 6) BSA (10 mg/ml) during maturation, fertilization and embryo culture. The rates of cleavage and the development to morulae or blastocysts did not differ (P > 0.05) among treatment groups and between both categories of COC and were showing a high degree of variability (cleavage 54.0% to 65.1% and 41.3% to 55.7%, respectively; morulae 25.3% to 53.0% and 26.0% to 51.2%, respectively; blastocysts 5.4% to 24.7% and 0.6% to 20.3%, respectively). Parthenogenetic activation only rarely occurred in medium containing PVA throughout all steps of in vitro production of bovine embryos (Experiment 4) and led to early cleavage stages (8%), but no development to morula- or blastocyst-stages was observed. It is concluded that 1) formation of pronuclei was retarded in medium lacking protein-supplementation, indicating that BSA is required for regular fertilization in vitro and 2) under our experimental conditions, protein-supplementation is not necessary for maturation and development up to the blastocyst stage in vitro. PMID:16727707

Eckert, J; Niemann, H

1995-05-01

139

Structure and Dynamics of the Solvation of Bovine Pancreatic Trypsin Inhibitor in Explicit Water: A Comparative Study of the Effects of Solvent and Protein Polarizability  

SciTech Connect

The research described in this product was performed in part in the Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by the Department of Energy's Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory. To isolate the effects of the inclusion of polarizability in the force field model on the structure and dynamics of the solvating water in differing electrostatic environments of proteins, we present the results of molecular dynamics simulations of the bovine pancreatic trypsin inhibitor (BPTI) in water with force fields that explicitly include polarization for both the protein and the water. We use three model potentials for water and two model potentials for the protein. Two of the water models and one of the protein models are polarizable. A total of six systems were simulated representing all combinations of these polarizable and nonpolarizable protein and water force fields. We find that all six systems behave in a similar manner in regions of the protein that are weakly electrostatic (either hydrophobic or weakly hydrophilic). However, in the vicinity of regions of the protein with relatively strong electrostatic fields (near positively or negatively charged residues), we observe that the water structure and dynamics are dependent on both the model of the protein and the model of the water. We find that a large part of the dynamical dependence can be described by small changes in the local environments of each region that limit the local density of non-hydrogen-bonded waters, precisely the water molecules that facilitate the dynamical relaxation of the water-water hydrogen bonds. We introduce a simple method for rescaling for this effect. When this is done, we are able to effectively isolate the influence of polarizability on the dynamics. We find that the solvating water’s relaxation is most affected when both the protein and the water models are polarizable. However, when only one model (or neither) is polarizable, the relaxation is similar regardless of the models used.

Kim, Byoung Chan; Young, Tom; Harder, Edward; Friesner, Richard A.; Berne, Bruce J.

2005-09-01

140

Enantioselective quenching of room-temperature phosphorescence lifetimes of proteins: bovine and human serum albumins.  

PubMed

Enantioselective quenching of the room-temperature phosphorescence (RTP) lifetime of proteins was demonstrated due to the effects of various external chiral quenching agents. In the absence of quenchers, the RTP lifetimes for bovine serum albumin (BSA) and human serum albumin (HSA) were found to be 5.0 +/- 0.2 and 4.0 +/- 0.1 ms, respectively. The addition of various chiral quenchers (three pairs of binaphthols and two pairs of beta-blockers) into the deoxygenated sample solutions containing BSA and HSA reduced their RTP lifetimes significantly, i.e., from ca. 4-5 ms (in the absence) to an average lifetime of ca. 1-2 ms (in the presence) of the chiral quenchers. For the R and S enantiomers examined, marked differences in RTP lifetimes were observed, i.e., ranging from ca. 20-29% for the binaphthols to ca.14-16% for the beta-blockers. Such findings could lead to a better understanding of the relationship between chirality, dynamics/conformational changes, and biological functions of proteins. PMID:17274655

Wei, Yanli; Dong, Chuan; Liu, Diansheng; Shuang, Shaomin; Huie, Carmen W

2007-03-01

141

Human Milk Protein Production in Xenografts of Genetically Engineered Bovine Mammary Epithelial Stem Cells  

Microsoft Academic Search

BackgroundIn the bovine species milk production is well known to correlate with mammary tissue mass. However, most advances in optimizing milk production relied on improvements of breeding and husbandry practices. A better understanding of the cells that generate bovine mammary tissue could facilitate important advances in milk production and have global economic impact. With this possibility in mind, we show

Eugenio Martignani; Peter Eirew; Paolo Accornero; Connie J. Eaves; Mario Baratta; Joseph Najbauer

2010-01-01

142

Ultrarapid detection of bovine whey proteins in powdered soybean milk by perfusion reversed-phase high-performance liquid chromatography  

Microsoft Academic Search

A perfusion reversed-phase high-performance liquid chromatographic method has been developed to simultaneously separate soybean and bovine whey proteins (?-lactalbumin and ?-lactoglobulins (A+B)) in a very short analysis time (?5 min). The method consisted of a linear binary gradient water–acetonitrile–0.10% trifluoroacetic acid at a flow-rate of 3 ml\\/min, with the column thermostated at 60°C, and ultraviolet detection at 254 nm. This

M. C. Garc??a; M. L. Marina; M. Torre

1998-01-01

143

A rapid method for purifying osteopontin from bovine milk and interaction between osteopontin and other milk proteins  

Microsoft Academic Search

Osteopontin (OPN) was isolated from bovine milk whey using a two-step chromatographic procedure. Acid whey was batch-processed with DEAE-Sephacel at pH 5.0, and an OPN-rich fraction (approximately 85% purity) was obtained from the first chromatographic steps. Subsequent POROS® HQ anion exchange HPLC yielded approximately 11mg of purified OPN from 1L of whey. The identity of the protein was verified by

N. Azuma; A. Maeta; K. Fukuchi; C. Kanno

2006-01-01

144

Bone morphogenetic proteins in the bovine oviduct: differential expression of BMP-5 in the isthmus during the estrous cycle.  

PubMed

Bone morphogenetic proteins (BMPs) play a crucial role in mammalian reproduction, but little is known about their expression and function in the oviduct, where preimplantation events take place. In the present study, messenger RNA (mRNA) expression of BMPs was examined by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) in bovine oviduct epithelial cells obtained from ampulla and isthmus at different stages of the estrous cycle. Expression of BMP-2, -3, -4, -7, -10 and -15 mRNA was detected in epithelial cells of both anatomic regions, whereas BMP-5 mRNA was specifically expressed in isthmus epithelial cells throughout the estrous cycle. High expression levels for BMP-5 and for BMP-2, -4, and -7 mRNA were observed during the preovulatory stage. Considering the region-specific gene expression of BMP-5, its protein localization in the oviduct and its presence in the oviductal fluid were evaluated by immunohistochemistry and Western blot analysis. BMP-5 protein staining was observed in isthmus sections with a more intense signal in the luminal epithelial cell layer. In addition, a 21 kDa protein corresponding to the BMP-5 mature monomeric form was detected in bovine oviductal fluid throughout the estrous cycle. In conclusion, these results demonstrate that different members of the BMP family are expressed in the bovine oviduct during the estrous cycle, and reveal that BMP-5 is differentially expressed in the isthmus. The expression of this factor in the oviduct epithelium and its presence in the luminal fluid suggest a possible action of BMP-5 as a new autocrine and/or paracrine regulator of the reproductive events that occur in the bovine oviductal environment. PMID:24582268

García, Elina V; Valdecantos, Pablo A; Barrera, Daniel; Roldán-Olarte, Mariela; Miceli, Dora C

2014-05-01

145

Purification and characterization of a 190-kD microtubule-associated protein from bovine adrenal cortex  

Microsoft Academic Search

A heat-stable microtubule-associated protein (MAP) with molecular weight of 190,000, termed 190- kD MAP, was purified from bovine adrenal cortex. This MAP showed the same level of ability to promote tubulin polymerization as did MAP2 and tau from mammalian brains. Relatively high amounts of 190-kD MAP could bind to microtubules reconstituted in the presence of taxol. At maximum 1 mol

Hiromu Murofushi; Susumu Kotani; Hiroyuki Aizawa; Shin-ichi Hisanaga; Nobutaka Hirokawa; Hikoichi Sakai

1986-01-01

146

Role of a regulatory protein from the bovine cornea in activation of cell sources of corneal regeneration in vitro  

Microsoft Academic Search

A highly purified regulatory protein isolated from the bovine cornea (RPC) was tested for the effect on the rat and newt corneas\\u000a in vitro under different culture conditions. In the newt cornea, RPC stimulated limbus epithelial cells in roller cultures\\u000a and cells in the basal layer of corneal epithelium in both roller and stationary cultures. In roller cultures of the

D. V. Margasyuk; M. S. Krasnov; I. A. Yamskov; V. P. Yamskova

2008-01-01

147

Viscoelastic properties of bovine orbital connective tissue and fat: constitutive models  

Microsoft Academic Search

Reported mechanical properties of orbital connective tissue and fat have been too sparse to model strain–stress relationships\\u000a underlying biomechanical interactions in strabismus. We performed rheological tests to develop a multi-mode upper convected\\u000a Maxwell (UCM) model of these tissues under shear loading. From 20 fresh bovine orbits, 30 samples of connective tissue were\\u000a taken from rectus pulley regions and 30 samples

Lawrence Yoo; Vijay Gupta; Choongyeop Lee; Pirouz Kavehpore; Joseph L. Demer

148

The p66(Shc) adaptor protein controls oxidative stress response in early bovine embryos.  

PubMed

The in vitro production of mammalian embryos suffers from high frequencies of developmental failure due to excessive levels of permanent embryo arrest and apoptosis caused by oxidative stress. The p66Shc stress adaptor protein controls oxidative stress response of somatic cells by regulating intracellular ROS levels through multiple pathways, including mitochondrial ROS generation and the repression of antioxidant gene expression. We have previously demonstrated a strong relationship with elevated p66Shc levels, reduced antioxidant levels and greater intracellular ROS generation with the high incidence of permanent cell cycle arrest of 2-4 cell embryos cultured under high oxygen tensions or after oxidant treatment. The main objective of this study was to establish a functional role for p66Shc in regulating the oxidative stress response during early embryo development. Using RNA interference in bovine zygotes we show that p66Shc knockdown embryos exhibited increased MnSOD levels, reduced intracellular ROS and DNA damage that resulted in a greater propensity for development to the blastocyst stage. P66Shc knockdown embryos were stress resistant exhibiting significantly reduced intracellular ROS levels, DNA damage, permanent 2-4 cell embryo arrest and diminished apoptosis frequencies after oxidant treatment. The results of this study demonstrate that p66Shc controls the oxidative stress response in early mammalian embryos. Small molecule inhibition of p66Shc may be a viable clinical therapy to increase the developmental potential of in vitro produced mammalian embryos. PMID:24475205

Betts, Dean H; Bain, Nathan T; Madan, Pavneesh

2014-01-01

149

The p66Shc Adaptor Protein Controls Oxidative Stress Response in Early Bovine Embryos  

PubMed Central

The in vitro production of mammalian embryos suffers from high frequencies of developmental failure due to excessive levels of permanent embryo arrest and apoptosis caused by oxidative stress. The p66Shc stress adaptor protein controls oxidative stress response of somatic cells by regulating intracellular ROS levels through multiple pathways, including mitochondrial ROS generation and the repression of antioxidant gene expression. We have previously demonstrated a strong relationship with elevated p66Shc levels, reduced antioxidant levels and greater intracellular ROS generation with the high incidence of permanent cell cycle arrest of 2–4 cell embryos cultured under high oxygen tensions or after oxidant treatment. The main objective of this study was to establish a functional role for p66Shc in regulating the oxidative stress response during early embryo development. Using RNA interference in bovine zygotes we show that p66Shc knockdown embryos exhibited increased MnSOD levels, reduced intracellular ROS and DNA damage that resulted in a greater propensity for development to the blastocyst stage. P66Shc knockdown embryos were stress resistant exhibiting significantly reduced intracellular ROS levels, DNA damage, permanent 2–4 cell embryo arrest and diminished apoptosis frequencies after oxidant treatment. The results of this study demonstrate that p66Shc controls the oxidative stress response in early mammalian embryos. Small molecule inhibition of p66Shc may be a viable clinical therapy to increase the developmental potential of in vitro produced mammalian embryos.

Betts, Dean H.; Bain, Nathan T.; Madan, Pavneesh

2014-01-01

150

Inhibition of bovine kidney low molecular mass phosphotyrosine protein phosphatase by uric acid.  

PubMed

Uric acid inhibited 50% of the activity of bovine kidney low molecular mass phosphotyrosine protein phosphatase at concentrations of 1.0, 0.4, 1.3, and 0.2 mM, respectively for p-nitrophenyl phosphate (p-NPP), flavine mononucleotide, beta-naphthyl phosphate and tyrosine phosphate (Tyr-P) as substrates. The mixed type inhibition of p-NPP hydrolysis was fully reversible, with Kic and Kiu values of 0.4 and 1.1 mM, respectively; the inhibition by uric acid shifted the pH optimum from 5.0 to 6.5. When Tyr-P was the substrate, competitive inhibition was observed with a Ki value of 0.05 mM. Inhibition studies by uric acid in the presence of thiol compounds, and preincubation studies in the presence of inorganic phosphate suggest that the interaction of uric acid with the enzyme occurred at the active site, but did not involve SH residues, and that the mechanism of inhibition depended on the structure of the substrates. PMID:12683751

Granjeiro, José Mauro; Ferreira, Carmen Verissima; Granjeiro, Paulo Afonso; Da Silva, Cinthia Celestino; Taga, Eulázio Mikio; Volpe, Pedro Luiz Onofre; Aoyama, Hiroshi

2002-10-01

151

Ubiquitin cross-reactive protein is released by the bovine uterus in response to interferon during early pregnancy.  

PubMed

A 16-kDa protein has been identified that is secreted by the bovine endometrium in response to conceptus-derived interferon (IFN)-tau during early pregnancy. Because this uterine protein was similar in size to a human ubiquitin cross-reactive protein (hUCRP) that also was regulated by IFN, we suspected that they might be related. To test this hypothesis, uterine flushings, medium from cultured endometrium, and endometrial tissues were examined for the presence of ubiquitin-immunoreactive proteins. Immunoreacting proteins were detected through use of one-dimensional (1D)-PAGE and Western blotting with ubiquitin and hUCRP antiserum (1:500). A 16-kDa protein that cross-reacted with ubiquitin and hUCRP antisera was released by the endometrium and was present in uterine flushings from all Day 18 pregnant females examined (n = 12). The immunoreacting 16-kDa protein was absent in all nonpregnant females examined (n = 23). Regulation of this uterine protein by recombinant type I IFNs (rbIFN-tau, rbIFN-alpha, and roIFN-tau), using 0, 0.5, 5, and 25 nm of each IFN, was evaluated in nonpregnant (Day 12) heifers (n = 5) using 1D-PAGE and Western blotting. Release of the 16-kDa protein into medium was negligible in controls (0 nm IFN). For each IFN, a dose-dependent increase (p < 0.05) in release of the immunoreacting 16-kDa protein was noted. We conclude that the 16-kDa protein that is produced by the endometrium in response to IFN-tau during early pregnancy also shares epitopes with hUCRP and ubiquitin. The 16-kDa protein has been named bovine UCRP. PMID:8835381

Austin, K J; Ward, S K; Teixeira, M G; Dean, V C; Moore, D W; Hansen, T R

1996-03-01

152

Photochemistry and stereoselectivity of cellular retinaldehyde-binding protein from bovine retina  

SciTech Connect

11-cis-Retinaldehyde bound to cellular retinaldehyde-binding protein (CRALBP) is unaffected in bovine eyecup preparations by illumination that bleaches approximately 70% of the rhodopsin. Illumination of retinal homogenates to which CRALBP X (/sup 3/H)11-cis-retinaldehyde had been added did not result in a reduction of the specific activity of recovered 11-cis-retinaldehyde, ruling out a bleaching regeneration cycle. The quantum efficiency of photoisomerization for CRALBP X 11-cis-retinaldehyde was determined by comparing the rate of photoisomerization of 11-cis-retinaldehyde bound to purified CRALBP and opsin. The low value obtained (0.07), coupled with a low molar extinction coefficient (15,400 M-1 cm-1), results in a photosensitivity only about 4% that of rhodopsin. CRALBP binds 9-cis- and 11-cis-retinaldehyde, producing complexes with absorption maxima at 405 and 425 nm, respectively. No complexes were detected with 13-cis- and all-trans-retinaldehyde. Following incubation of CRALBP X 11-cis-retinol with an equimolar mixture of 9-, 11-, 13-cis-, and all-trans-retinaldehydes, only 11-cis-retinaldehyde and residual 11-cis-retinol are present on the protein following separation from excess retinoids. A similar result is obtained following incubation of CRALBP X 11-cis-retinol with mixtures of 9- and 11-cis-retinaldehyde ranging in composition from 9:1 to 1:9 (9-cis-:11-cis-,mol/mol). The results indicate that CRALBP X 11-cis-retinol is sufficiently stereoselective in its binding properties to warrant consideration as a component of the mechanism for the generation of 11-cis-retinaldehyde in the dark.

Saari, J.C.; Bredberg, D.L.

1987-06-05

153

Developmental expression of the cellular prion protein (PrPC) in bovine embryos  

PubMed Central

The mammalian cellular prion protein (PrPC) is a highly conserved glycoprotein that may undergo conversion into a conformationally altered isoform (scrapie prion protein or PrPSc), widely believed to be the pathogenic agent of transmissible spongiform encephalopathies (TSEs). Although much is known about PrPSc conversion and its role in TSEs, the normal function of PrPC has not been elucidated. In adult mammals, PrPC is most abundant in the central nervous tissue, with intermediate levels in the intestine and heart, and lower levels in the pancreas and liver. PrPC is expressed during neurogenesis throughout development, and it has recently been proposed that PrPC participates in neural cell differentiation during embryogenesis. In order to establish the developmental timing and to address the cell-specific expression of PrPC during mammalian development, we examined PrPC expression in bovine gametes and embryos through gestation Day 39. Our data revealed differential levels of Prnp mRNA at Days 4 and 18 in pre-attachment embryos. PrPC was detected in the developing central and peripheral nervous systems in Day 27, 32, and 39 embryos. PrPC was particularly expressed in differentiated neural cells located in the marginal regions of the central nervous system, but was absent from mitotically active, periventricular areas. Moreover, a PrPC cell-specific pattern of expression was detected in non-nervous tissues, including liver and mesonephros, during these stages. The potential participation of PrPC in neural cell differentiation is supported by its specific expression in differentiated states of neurogenesis.

Peralta, Oscar A.; Huckle, William R.; Eyestone, Willard H.

2012-01-01

154

Agouti Revisited: Transcript Quantification of the ASIP Gene in Bovine Tissues Related to Protein Expression and Localization  

PubMed Central

Beside its role in melanogenesis, the agouti signaling protein (ASIP) has been related to obesity. The potentially crucial role in adipocyte development makes it a tempting candidate for economic relevant, fat related traits in farm animals. The objective of our study was to characterize the mRNA expression of different ASIP transcripts and of putative targets in different bovine tissues, as well as to study consequences on protein abundance and localization. ASIP mRNA abundance was determined by RT-qPCR in adipose and further tissues of cattle representing different breeds and crosses. ASIP mRNA was up-regulated more than 9-fold in intramuscular fat of Japanese Black cattle compared to Holstein (p<0.001). Further analyses revealed that a transposon-derived transcript was solely responsible for the increased ASIP mRNA abundance. This transcript was observed in single individuals of different breeds indicating a wide spread occurrence of this insertion at the ASIP locus in cattle. The protein was detected in different adipose tissues, skin, lung and liver, but not in skeletal muscle by Western blot with a bovine-specific ASIP antibody. However, the protein abundance was not related to the observed ASIP mRNA over-expression. Immuno-histochemical analyses revealed a putative nuclear localization of ASIP additionally to the expected cytosolic signal in different cell types. The expression of melanocortin receptors (MCR) 1 to 5 as potential targets for ASIP was analyzed by RT-PCR in subcutaneous fat. Only MC1R and MC4R were detected indicating a similar receptor expression like in human adipose tissue. Our results provide evidence for a widespread expression of ASIP in bovine tissues at mRNA and, for the first time, at protein level. ASIP protein is detectable in adipocytes as well as in further cells of adipose tissue. We generated a basis for a more detailed investigation of ASIP function in peripheral tissues of various mammalian species.

Albrecht, Elke; Komolka, Katrin; Kuzinski, Judith; Maak, Steffen

2012-01-01

155

Cloning and cDNA sequence of a bovine submaxillary gland mucin-like protein containing two distinct domains.  

PubMed Central

A lambda gt11 cDNA library prepared from bovine submaxillary gland mRNA was screened with polyclonal anti-apo-bovine submaxillary mucin antibodies with the aim of obtaining the deduced amino acid sequence of the mucin core protein. One of the positive clones had a 1.8 kilobase (kb) cDNA insert and coded for an incomplete protein. A 2.0-kb cDNA clone was isolated by rescreening the library with the 1.8-kb cDNA. Nucleotide sequencing of the full-length 2.0-kb cDNA revealed an open reading frame that coded for a 563-amino acid protein. A striking feature of the cloned protein is the skewed distribution of the amino acids, most notably that of the hydroxy amino acids and cysteine. The amino-terminal domain of 339 residues is very rich in threonine, serine, and glycine and poor in cysteine, aspartic acid, tyrosine, phenylalanine, and tryptophan. In contrast, the carboxyl-terminal domain of 224 residues is rich in cysteine, aspartic acid, tyrosine, lysine, and asparagine and relatively poor in threonine, serine, and glycine. A search of the protein data bank for homologies to the deduced amino acid sequence revealed statistically significant matches to several proteins, including the porcine submaxillary apomucin fragment. The cysteine-rich domain by itself was not statistically homologous with any of the registered polypeptide sequences. RNA blot analysis using DNA probes corresponding to the mucin-like and cysteine-rich regions detected a nearly identical pattern of transcripts, demonstrating that the characterized clones are not artifacts of cDNA library construction. The blots also showed the presence of polydisperse transcripts in bovine submaxillary gland but no detectable hybridization signals in liver or brain RNA. Images

Bhargava, A K; Woitach, J T; Davidson, E A; Bhavanandan, V P

1990-01-01

156

Expression and distribution of cell adhesion-related proteins in bovine parthenogenetic embryos: The effects of oocyte vitrification.  

PubMed

The objective was to investigate expression of cell adhesion-related proteins (E-cadherin, ?-catenin, and the cytoskeletal protein F-actin) in bovine parthenogenetic embryos derived from vitrified-warmed oocytes. Bovine oocytes at metaphase II were randomly allocated into three groups: (1) untreated (control); (2) exposed to vitrification solution without freezing (toxicity); and (3) vitrified and warmed by the open-pulled straw method (vitrification). After parthenogenetic activation, in the vitrification group compared with the control, the timing of compaction was delayed in (108-120 vs. 96-108 hours, respectively), and the percentage of blastocysts that developed from eight-cell embryos was lower (32.08% vs. 61.03%; P < 0.05). To investigate whether vitrification delayed embryo compaction by affecting adhesion junction formation and function, immunostaining and quantitative reverse transcription polymerase chain reaction were done to characterize distribution patterns (E-cadherin, ?-catenin, and the cytoskeletal protein F-actin) and expression levels of cell adhesion-related proteins (?-catenin). Distribution of ?-catenin in eight-cell embryos from the vitrification group changed dramatically compared with the control and toxicity groups. Relative expression of ?-catenin at the mRNA and protein levels was lower (P < 0.05) than that of the fresh and toxicity groups. However, expression and distribution of E-cadherin were similar among groups. In conclusion, abnormal distribution and decreased expression of ?-catenin in bovine parthenogenetic eight-cell embryos derived from vitrified-warmed oocytes were associated with embryo compaction and reduced competence for subsequent embryo development. PMID:23602219

Zeng, Yan; Fu, Xiangwei; Zhou, Guangbin; Yue, Mingxing; Zhou, Yanhua; Zhu, Shien

2013-07-01

157

First Principles Predictions of the Structure and Function of G-Protein-Coupled Receptors: Validation for Bovine Rhodopsin  

PubMed Central

G-protein-coupled receptors (GPCRs) are involved in cell communication processes and with mediating such senses as vision, smell, taste, and pain. They constitute a prominent superfamily of drug targets, but an atomic-level structure is available for only one GPCR, bovine rhodopsin, making it difficult to use structure-based methods to design receptor-specific drugs. We have developed the MembStruk first principles computational method for predicting the three-dimensional structure of GPCRs. In this article we validate the MembStruk procedure by comparing its predictions with the high-resolution crystal structure of bovine rhodopsin. The crystal structure of bovine rhodopsin has the second extracellular (EC-II) loop closed over the transmembrane regions by making a disulfide linkage between Cys-110 and Cys-187, but we speculate that opening this loop may play a role in the activation process of the receptor through the cysteine linkage with helix 3. Consequently we predicted two structures for bovine rhodopsin from the primary sequence (with no input from the crystal structure)—one with the EC-II loop closed as in the crystal structure, and the other with the EC-II loop open. The MembStruk-predicted structure of bovine rhodopsin with the closed EC-II loop deviates from the crystal by 2.84 Ĺ coordinate root mean-square (CRMS) in the transmembrane region main-chain atoms. The predicted three-dimensional structures for other GPCRs can be validated only by predicting binding sites and energies for various ligands. For such predictions we developed the HierDock first principles computational method. We validate HierDock by predicting the binding site of 11-cis-retinal in the crystal structure of bovine rhodopsin. Scanning the whole protein without using any prior knowledge of the binding site, we find that the best scoring conformation in rhodopsin is 1.1 Ĺ CRMS from the crystal structure for the ligand atoms. This predicted conformation has the carbonyl O only 2.82 Ĺ from the N of Lys-296. Making this Schiff base bond and minimizing leads to a final conformation only 0.62 Ĺ CRMS from the crystal structure. We also used HierDock to predict the binding site of 11-cis-retinal in the MembStruk-predicted structure of bovine rhodopsin (closed loop). Scanning the whole protein structure leads to a structure in which the carbonyl O is only 2.85 Ĺ from the N of Lys-296. Making this Schiff base bond and minimizing leads to a final conformation only 2.92 Ĺ CRMS from the crystal structure. The good agreement of the ab initio-predicted protein structures and ligand binding site with experiment validates the use of the MembStruk and HierDock first principles' methods. Since these methods are generic and applicable to any GPCR, they should be useful in predicting the structures of other GPCRs and the binding site of ligands to these proteins.

Trabanino, Rene J.; Hall, Spencer E.; Vaidehi, Nagarajan; Floriano, Wely B.; Kam, Victor W. T.; Goddard, William A.

2004-01-01

158

Predicting bovine milk protein composition based on Fourier transform infrared spectra.  

PubMed

Phenotypic information on individual protein composition of cows is important for many aspects of dairy processing with cheese production as the center of gravity. However, measuring individual protein composition is expensive and time consuming. In this study, we investigated whether protein composition can be predicted based on inexpensive and routinely measured milk Fourier transform infrared (FTIR) spectra. Based on 900 calibration and 900 validation samples that had both capillary zone electrophoresis (CZE)-determined protein composition and FTIR spectra available, low to moderate validation R(2) were reached (from 0.18 for ?(S1)-casein to 0.56 for ?-lactoglobulin). The potential usefulness of this model on the phenotypic level was investigated by means of achieved selection differentials for 25% of the best animals. For ?-lactalbumin (R(2)=0.20), the selection differential amounted to 0.18 g/100g and for casein index (R(2)=0.50) to 1.24 g/100g. We concluded that predictions of protein composition were not accurate enough to enable selection of individual animals. However, for specific purposes when, for example, groups of animals that meet a certain threshold are to be selected, the presented model could be useful in practice on the phenotypic level. The potential usefulness of this model on the genetic level was investigated by means of genetic correlations between CZE-determined and FTIR-predicted protein composition traits. The genetic correlations ranged from 0.62 (?-casein) to 0.97 (whey). Thus, predictions of protein composition, when used as input to estimate breeding values, provide an excellent means for genetic improvement of protein composition. In addition, estimated repeatabilities based on 3 repeated observations of predicted protein composition showed that a considerable amount of prediction error can be removed using repeated observations. PMID:22032392

Rutten, M J M; Bovenhuis, H; Heck, J M L; van Arendonk, J A M

2011-11-01

159

Hydrogen peroxide-and fetal bovine serum-induced DNA synthesis in vascular smooth muscle cells: positive and negative regulation by protein kinase C isoforms  

Microsoft Academic Search

Hydrogen peroxide and fetal bovine serum stimulate DNA synthesis in growth-arrested smooth muscle cells with remarkably similar kinetics and cell density dependence. However, while stimulation with fetal bovine serum results in cell proliferation, that by H202 is followed by cell death. Depletion of conventional and novel protein kinase C isoforms, resulting from a long treatment with phorbol-l2-myristate- 13-acetate, further increases

Mara Fiorani; Orazio Cantoni; Andrea Tasinato; Daniel Boscoboinik; Angelo Azzi

1995-01-01

160

Recombinant Jembrana disease virus gag proteins identify several different antigenic domains but do not facilitate serological differentiation of JDV and nonpathogenic bovine lentiviruses  

Microsoft Academic Search

In Indonesia, it is suspected that there are two bovine lentiviruses circulating in the cattle population: a pathogenic Jembrana disease virus (JDV), and a nonpathogenic bovine immunodeficiency-like virus (BIV). Both viruses cross-react antigenically and cannot be differentiated by current serological tests using JDV antigens. To identify possible type-specific epitopes, a series of recombinant protein constructs including the matrix, capsid and

Moira Desport; Meredith E. Stewart; Carol A. Sheridan; William G. F. Ditcham; Surachmi Setiyaningsih; W. Masa Tenaya; Nining Hartaningsih; Graham E. Wilcox

2005-01-01

161

Ectopic induction of cartilage and bone by water-soluble proteins from bovine bone using a polyanhydride delivery vehicle.  

PubMed

Controlled release delivery vehicles for water-soluble osteogenic proteins from demineralized bovine bone matrix were constructed using polyanhydride polymers. The water-soluble proteins were isolated from a 4 M guanidine hydrochloride extract of bone matrix. The water-soluble proteins possessed Chondrogenic Stimulating Activity (CSA) when tested in stage 24 chick limb bud cell cultures, but were incapable of inducing cartilage or bone in vivo when implanted intramuscularly into mice by themselves. The polyanhydride polymers alone were also incapable of inducing ectopic cartilage or bone. However, when the water-soluble proteins were incorporated into the polymeric delivery vehicle, the combination was capable of inducing cartilage and bone up to 50% of the time. These results demonstrate that it is possible to use polyanhydride polymers as controlled-release delivery vehicles for soluble bioactive factors that interact with a local cell population. PMID:2398077

Lucas, P A; Laurencin, C; Syftestad, G T; Domb, A; Goldberg, V M; Caplan, A I; Langer, R

1990-07-01

162

Influence of metal ions on phosphatidylcholine bovine serum albumin model membrane, an FTIR study  

NASA Astrophysics Data System (ADS)

FTIR spectroscopy was used to study the interaction of K +, Ca 2+ and Eu 3+ ions and the Phosphatidylcholine (PC)-bovine serum albumin (BSA) complex. First, a PC-BSA interaction system was constructed. The analytical results of transmission electron microscope (TEM), quasi-elastic light scattering (QELS) techniques and FTIR-ATR spectroscopy indicated that PC molecules interacted with BSA in aqueous solutions. However, IR inspection was limited for aqueous solutions. Solid experimental condition was then employed, and FTIR spectra showed that the PC and BSA molecules incorporated with each other, which could represent their interactions in solutions. Then, the influence of metal ions on PC-BSA system was studied in solid experimental conditions, and FTIR spectroscopy was used in this study. The spectral results showed that: (1) K +, Ca 2+ and Eu 3+ ions all decreased the rigidities of acyl chains of PC in PC-BSA systems. (2) The interactions between Ca 2+, Eu 3+ ions and the hydrophilic phosphate ester and carbonyl ester groups of PC were stronger than that of K + ions, while the influent modes of Ca 2+ and Eu 3+ ions on these regions were different. (3) When the relative molar content of Eu 3+ ions to PC ( Ri/p) reached 2, the coordination effect between Eu 3+ ions and PO2- groups of PC was saturated. (4) The addition of these ions increased the content of ?-helix structures of BSA, and decreased the content of ?-turn structures. By comparing these results with the interactions of K +, Ca 2+, Eu 3+ ions with phospholipid system in the absence of protein, some special characters were discovered in the acyl regions of PC, while their interactions results with the hydrophilic regions of PC were alike. It might be interpreted that these metal ions influenced the acyl chains of PC mediated from BSA molecules, and coordinated directly with the hydrophilic regions of PC. As for biological membrane was a system included both phospholipid and proteins, these characters suggested that phospholipid-protein mixture system could be a better model in the studies of interaction between metal ions and biological membranes using FTIR spectroscopy.

Wang, Fan; Yang, Zhanlan; Zhou, Yong; Weng, Shifu; Zhang, Li; Wu, Jinguang

2006-08-01

163

Somatic cell mapping, polymorphism, and linkage analysis of bovine prolactin-related proteins and placental lactogen.  

PubMed

The bovine prolactin gene family includes novel members expressed in the fetal placenta that are distinct from placental lactogen. In this study, we investigated the genetic organization of four members of this gene family (PRP1, PRP3, PRP6, and PRP10) as well as placental lactogen (PL). Using a bovine-rodent hybrid somatic cell panel, all five genes were assigned to bovine chromosome 23, which contains prolactin and the major histocompatibility group (BOLA). Restriction fragment length polymorphisms were detected by all probes in breeding populations with the restriction enzyme MspI, whereas no polymorphisms were detected with BamHI. EcoRI, HindIII, TaqI, and PstI produced polymorphic fragments with some but not all of the probes tested. A PRP10 polymorphism, which is apparently the result of a insertion/deletion event, detected polymorphism frequency differences between Bos indicus and Bos taurus. No recombinational events were observed with these probes and prolactin using linkage analysis involving 91 American Holsteins. The bovine prolactin gene family was incorporated into a linkage group containing CYP21. Our studies demonstrate that members of the bovine prolactin gene family have a close physical association with each other, and all members demonstrate genetic variability in the breeding population. PMID:1358791

Dietz, A B; Georges, M; Threadgill, D W; Womack, J E; Schuler, L A

1992-09-01

164

Fibronectin type II-module proteins in the bovine genital tract and their putative role in cell volume control during sperm maturation.  

PubMed

The male reproductive tract of ungulates contains two protein families bearing tandemly arranged fibronectin II (Fn2) modules; one (small Fn2 proteins) bears two modules (e.g. BSP-A1/2), the other (long Fn2 proteins) bears four (e.g. epididymal sperm-binding protein 1 (ELSPBP1)). While it is well known that small Fn2 proteins are present in bull semen, nothing is known about long Fn2 proteins. In the present study, the presence of ELSPBP1 proteins in the bull epididymis and their association with maturing spermatozoa were investigated using a specific antibody against canine ELSPBP1. Analysis of western blots showed ELSPBP1 to be present in the caput, corpus and cauda regions of the epididymis. The protein, which bound phosphorylcholine (PC) strongly, appeared to associate with the spermatozoa during maturation because it was absent from caput spermatozoa but present on cauda spermatozoa. Immunocytochemistry of cauda spermatozoa showed the protein to be bound to the post-acrosomal and midpiece regions. ELSPBP1 could not be detected on freshly ejaculated spermatozoa but was revealed after a capacitating treatment. Our previous studies have shown differences between bovine caput and cauda spermatozoa in terms of their ability to control cell volume. Because of the close homology of BSP-A1/2 PC binding regions with Fn2 regions in ELSPBP1, BSP-A1/2 was used as a model to investigate the effect of a PC-binding Fn2 protein on cell volume control. While the protein had no effect on cauda spermatozoa, it caused caput spermatozoa to swell more in response to hypotonic stress, similarly to untreated cauda spermatozoa. PMID:19261225

Sahin, Evrim; Petrunkina, Anna M; Ekhlasi-Hundrieser, Mahnaz; Hettel, Christiane; Waberski, Dagmar; Harrison, Robin A P; Töpfer-Petersen, Edda

2009-01-01

165

Tight binding of inhibitors to bovine bc1 complex is independent of the Rieske protein redox state. Consequences for semiquinone stabilization in the quinol oxidation site.  

PubMed

To determine the effect of the redox state of the Rieske protein on ligand binding to the quinol oxidation site of the bc(1) complex, we measured the binding rate constants (k(1)) for stigmatellin and myxothiazol, at different concentrations of decylbenzoquinone or decylbenzoquinol, in the bovine bc(1) complex with the Rieske protein in the oxidized or reduced state. Stigmatellin and myxothiazol bound tightly and competitively with respect to quinone or quinol, independently of the redox state of the Rieske protein. In the oxidized bc(1) complex, the k(1) values for stigmatellin ( approximately 2.6 x 10(6) m(-1)s(-1)) and myxothiazol ( approximately 8 x 10(5) m(-1)s(-1)), and the dissociation constant (K(d)) for quinone, were similar between pH 6.5 and 9, indicating that ligand binding is independent of the protonation state of histidine 161 of the Rieske protein (pK(a) approximately 7.6). Reduction of the Rieske protein increased the k(1) value for stigmatellin and decreased the K(d) value for quinone by 50%, without modifying the k(1) for myxothiazol. These results indicate that reduction of the Rieske protein and protonation of histidine 161 do not induce a strong stabilization of ligand binding to the quinol oxidation site, as assumed in models that propose the existence of a highly stabilized semiquinone as a reaction intermediate during quinol oxidation. PMID:12364330

Covián, Raúl; Pardo, Juan Pablo; Moreno-Sánchez, Rafael

2002-12-13

166

Bovine binder-of-sperm protein BSP1 promotes protrusion and nanotube formation from liposomes  

SciTech Connect

Research highlights: {yields} Binder-of-sperm protein 1 (BSP1) modifies the morphology of lipidic vesicles inducing bead necklace-like and thread-like structures. {yields} In the presence of multilamellar liposomes, BSP1 leads to the formation of long nanotubes. {yields} The insertion of BSP1 in the external lipid leaflet of membranes induces local changes in bilayer curvature. -- Abstract: Binder-of-sperm (BSP) proteins interact with sperm membranes and are proposed to extract selectively phosphatidylcholine and cholesterol from these. This change in lipid composition is a key step in sperm capacitation. The present work demonstrates that the interactions between the protein BSP1 and model membranes composed with phosphatidylcholine lead to drastic changes in the morphology of the lipidic self-assemblies. Using cryo-electron microscopy and fluorescence microscopy, we show that, in the presence of the protein, the lipid vesicles elongate, and form bead necklace-like structures that evolve toward small vesicles or thread-like structures. In the presence of multilamellar vesicles, where a large reservoir of lipid is available, the presence of BSP proteins lead to the formation of long nanotubes. Long spiral-like threads, associated with lipid/protein complexes, are also observed. The local curvature of lipid membranes induced by the BSP proteins may be involved in lipid domain formation and the extraction of some lipids during the sperm maturation process.

Lafleur, Michel, E-mail: michel.lafleur@umontreal.ca [Department of Chemistry, Center for Self-Assembled Chemical Systems, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada)] [Department of Chemistry, Center for Self-Assembled Chemical Systems, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada); Courtemanche, Lesley [Department of Chemistry, Center for Self-Assembled Chemical Systems, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada)] [Department of Chemistry, Center for Self-Assembled Chemical Systems, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada); Karlsson, Goeran; Edwards, Katarina [Department of Physical and Analytical Chemistry, Uppsala University, Box 579, S-751 23 Uppsala (Sweden)] [Department of Physical and Analytical Chemistry, Uppsala University, Box 579, S-751 23 Uppsala (Sweden); Schwartz, Jean-Louis [Department of Physiology, Groupe d'etude des Proteines Membranaires, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada)] [Department of Physiology, Groupe d'etude des Proteines Membranaires, Universite de Montreal, C.P. 6128, Succ. Centre Ville, Montreal, Quebec, Canada H3C 3J7 (Canada); Manjunath, Puttaswamy [Maisonneuve-Rosemont Hospital Research Center and Faculty of Medecine, Universite de Montreal, 5415 L'Assomption Blvd, Montreal, Quebec, Canada H1T 2M4 (Canada)] [Maisonneuve-Rosemont Hospital Research Center and Faculty of Medecine, Universite de Montreal, 5415 L'Assomption Blvd, Montreal, Quebec, Canada H1T 2M4 (Canada)

2010-08-27

167

Cryopreservation for bovine embryos in serum-free freezing medium containing silk protein sericin.  

PubMed

Because the use of serum in the embryo cryopreservation increases the probability of animal health problems such as bovine spongiform encephalopathy (BSE) and viral infections, this study was conducted to examine the effects of sericin supplementation for serum-free freezing medium on the survival and development of bovine embryos after freezing-thawing and direct transfer to recipients. When in vitro-produced bovine embryos were frozen conventionally in the freezing medium supplemented with various concentrations (0.1%, 0.5%, and 1.0%) of sericin, the percentages of damaged zona pellucida, survival, and development of frozen-thawed embryos were similar to those of embryos frozen in freezing medium supplemented with 0.4% bovine serum albumin (BSA) and 20% fetal bovine serum (FBS) (0.4BSA/20F; control). When in vivo-derived embryos were frozen with 0.4BSA/20F (control), 0.5% sericin +20% FBS (0.5S/20F) or 0.5% sericin (0.5S) and were subsequently transferred directly to recipients, the percentages of recipients with pregnancy and normal calving in the 0.5S/20F group were higher than those in the control group (47.3% vs. 40.1% and 94.6% vs. 87.3%, respectively). Moreover, the percentages of recipients with pregnancy and normal calving (42.2% and 92.4%, respectively) in the 0.5S group were similar with those of other groups. In conclusion, these results indicate that serum-free freezing medium supplemented with sericin is available for the cryopreservation of bovine embryos and that it is beneficial for the elimination of a potential source of biological contamination by serum or BSA. PMID:23850826

Isobe, Tomohiro; Ikebata, Yoshihisa; Onitsuka, Takeshi; Do, Lanh Thi Kim; Sato, Yoko; Taniguchi, Masayasu; Otoi, Takeshige

2013-10-01

168

Sequences of the bovine and yeast ADP-ribosylation factor and comparison to other GTP-binding proteins.  

PubMed Central

The ADP-ribosylation factor (ARF) is a 21-kDa GTP-binding protein that serves as the cofactor in the cholera toxin-catalyzed activation of the stimulatory guanine nucleotide-binding protein of adenylate cyclase (Gs). An oligonucleotide probe based on the partial amino acid sequence was used to clone ARF from a bovine adrenal chromaffin cDNA library. The yeast (Saccharomyces cerevisiae) ARF gene was then cloned from a YCp50 genomic library by cross-species hybridization by using the coding region of the bovine gene. RNA gel blots of poly(A)+ RNA indicate that only one ARF message size (900 and 2000 base pairs) is present in yeast and cows, respectively. Comparison of the cDNA-derived amino acid sequences of ARF to other GTP-binding proteins reveals a structural relationship between ARF and the ras family of proteins. A slightly better structural relationship is detected when ARF is compared to the alpha subunits of the trimeric GTP-binding proteins, including Gs alpha. All of the biochemical characteristics of the purified ARF, including the lack of GTPase activity and the posttranslational myristoylation, are consistent with the derived sequences. Comparison of the ARF sequences to that of the chicken processed pseudogene (CPS-1), previously reported as a ras homologue, reveals that CPS-1 is actually an ARF-derived gene. These results demonstrate that ARF is a GTP-binding protein with structural features of both the ras and the trimeric GTP-binding protein families.

Sewell, J L; Kahn, R A

1988-01-01

169

Antithrombin III and its interaction with heparin. Comparison of the human, bovine, and porcine proteins by ÂąH NMR spectroscopy  

Microsoft Academic Search

ÂąH NMR has been used to characterize and compare the structures of antithrombin III from human, bovine, and porcine plasma as well as to investigate the interactions of each of these proteins with heparin fragments of defined length. The amino acid compositions of the three proteins are very similar, which is reflected in the gross features of their ÂąH NMR

Peter Gettins

1987-01-01

170

Polymer-Induced Heteronucleation for Protein Single Crystal Growth: Structural Elucidation of Bovine Liver Catalase and Concanavalin A Forms  

SciTech Connect

Obtaining single crystals for X-ray diffraction remains a major bottleneck in structural biology; when existing crystal growth methods fail to yield suitable crystals, often the target rather than the crystallization approach is reconsidered. Here we demonstrate that polymer-induced heteronucleation, a powerful technique that has been used for small molecule crystallization form discovery, can be applied to protein crystallization by optimizing the heteronucleant composition and crystallization formats for crystallizing a wide range of protein targets. Applying these advances to two benchmark proteins resulted in dramatically increased crystal size, enabling structure determination, for a half century old form of bovine liver catalase (BLC) that had previously only been characterized by electron microscopy, and the discovery of two new forms of concanavalin A (conA) from the Jack bean and accompanying structural elucidation of one of these forms.

Foroughi, Leila M.; Kang, You-Na; Matzger, Adam J. (Michigan)

2012-05-09

171

Structural equation models to estimate risk of infection and tolerance to bovine mastitis  

PubMed Central

Background One method to improve durably animal welfare is to select, as reproducers, animals with the highest ability to resist or tolerate infection. To do so, it is necessary to distinguish direct and indirect mechanisms of resistance and tolerance because selection on these traits is believed to have different epidemiological and evolutionary consequences. Methods We propose structural equation models with latent variables (1) to quantify the latent risk of infection and to identify, among the many potential mediators of infection, the few ones that influence it significantly and (2) to estimate direct and indirect levels of tolerance of animals infected naturally with pathogens. We applied the method to two surveys of bovine mastitis in the Walloon region of Belgium, in which we recorded herd management practices, mastitis frequency, and results of bacteriological analyses of milk samples. Results and discussion Structural equation models suggested that, among more than 35 surveyed herd characteristics, only nine (age, addition of urea in the rations, treatment of subclinical mastitis, presence of dirty liner, cows with hyperkeratotic teats, machine stripping, pre- and post-milking teat disinfection, and housing of milking cows in cubicles) were directly and significantly related to a latent measure of bovine mastitis, and that treatment of subclinical mastitis was involved in the pathway between post-milking teat disinfection and latent mastitis. These models also allowed the separation of direct and indirect effects of bacterial infection on milk productivity. Results suggested that infected cows were tolerant but not resistant to mastitis pathogens. Conclusions We revealed the advantages of structural equation models, compared to classical models, for dissecting measurements of resistance and tolerance to infectious diseases, here bovine mastitis. Using our method, we identified nine major risk factors that were directly associated with an increased risk of mastitis and suggested that cows were tolerant but not resistant to mastitis. Selection should aim at improved resistance to infection by mastitis pathogens, although further investigations are needed due to the limitations of the data used in this study.

2013-01-01

172

Modeling of protein interaction networks  

Microsoft Academic Search

We introduce a graph generating model aimed at representing the evolution of protein interaction networks. The model is based on the hypotesis of evolution by duplications and divergence of the genes which produce proteins. The obtained graphs shows multifractal properties recovering the absence of a characteristic connectivity as found in real data of protein interaction networks. The error tolerance of

Alexei Vazquez; A. Flammini; A. Maritan; A. Vespignani

2001-01-01

173

Modeling of Protein Interaction Networks  

Microsoft Academic Search

We introduce a graph-generating model aimed at representing the evolution of protein interaction networks. The model is based on the hypothesis of evolution by duplication and divergence of the genes which produce proteins. The obtained graphs have multifractal properties recovering the absence of a characteristic connectivity as found in real data of protein interaction networks. The error tolerance of the

Alexei Vázquez; Alessandro Flammini; Amos Maritan; Alessandro Vespignani

2003-01-01

174

Capillary permeability-increasing enzyme from the venom of Agkistrodon caliginosus (kankoku-mamushi): activity due to the release of peptide material from a protein in bovine plasma.  

PubMed

When a mixture of the purified capillary permeability-increasing enzyme from A. caliginosus venom and bovine plasma or heated bovine plasma was injected into the depilated skin of the back of a rabbit, the capillary permeability-increasing activity was much greater than that induced by injection of the enzyme alone. The substance which increases capillary permeability was extracted from the incubated mixture of bovine plasma and enzyme with 50-70% ethanol. Its activity was lost when treated with carboxypeptidase A. Thus, it is supposed that the increase in capillary permeability induced by the enzyme is due to a low molecular weight peptide released from a protein in bovine plasma by the action of the enzyme. No liberation by the enzyme of histamine or anaphylatoxins of the complement system was found. PMID:4039478

Ohtani, Y; Suda, T; Takahashi, H

1985-01-01

175

A phenomenological model for predicting fatigue life in bovine trabecular bone.  

PubMed

Cyclic loading of bone during daily activities can lead to fatigue degradation and increased risk of fracture in both the young and elderly population. Damage processes under cyclic loading in trabecular bone result in the reduction of the elastic modulus and accumulation of residual strain. These effects increase with increasing stress levels, leading to a progressive reduction in fatigue life. The present work analyzes the effect of stress and strain variation on the above damage processes in bovine trabecular bone, and develops a phenomenological model relating fatigue life to the imposed stress level. The elastic modulus reduction of the bone specimens was observed to depend on the maximum compressive strain, while the rate of residual strain accumulation was a function of the stress level. A model was developed for the upper and lower bounds of bone elastic modulus reduction with increasing number of cycles, at each stress range. The experimental observations were described well by the model. The model predicted the bounds of the fatigue life with change in fatigue stress. The decrease in the fatigue life with increasing stress was related to corresponding increases in the residual strain accumulation rates at the elevated stress levels. The model shows the validity of fatigue predictions from relatively few cyclic experiments, by combining trends observed in the monotonic and the cyclic tests. The model also presents a relatively simple procedure for predicting the endurance limit for bovine trabecular bone specimens. PMID:15341169

Ganguly, P; Moore, T L A; Gibson, L J

2004-06-01

176

Harmonic Dynamics of Proteins: Normal Modes and Fluctuations in Bovine Pancreatic Trypsin Inhibitor  

Microsoft Academic Search

A normal mode analysis making use of an empirical potential function including local and nonlocal (nonbonded) interactions is performed for the bovine pancreatic trypsin inhibitor in the full conformational space of the molecule (1,740 degrees of freedom); that is, all bond lengths and angles, as well as dihedral angles, are included for the 580-atom system consisting of all heavy atoms

Bernard Brooks; Martin Karplus

1983-01-01

177

Effect of endocrine and paracrine factors on protein synthesis and cell proliferation in bovine hoof tissue culture.  

PubMed

Laminitis is a major cause of lameness in dairy cattle, and is widely attributed to a defect in the horny tissue that gives the hoof its mechanical strength. Defective horn is associated with, and may be preceded by, impaired keratin deposition in the hoof epidermis. The cause of abnormal keratin deposition is not easily identified but, like epidermal keratinization in other tissues, is likely to be controlled by hormones and the paracrine action of locally produced growth factors. The hormonal regulation of keratin synthesis and cell proliferation in the bovine hoof was studied using tissue explants in organ culture. As the highest incidence of laminitis is in early lactation, the study focused on insulin, cortisol and prolactin, three hormones implicated in lactogenesis and galactopoiesis. Incubation of tissue explants for 24 h in medium containing insulin (10-5000 ng/ml) stimulated protein synthesis measured by incorporation of 35S-labelled amino acids. Histochemical examination showed that insulin binding co-localized with the site of protein synthesis. Insulin also stimulated DNA synthesis, an index of cell proliferation, which was measured by incorporation of [3H]methyl thymidine. Cortisol (10-5000 ng/ml) decreased protein synthesis, whereas prolactin (10-5000 ng/ml) had no significant effect on protein or DNA synthesis. Epidermal growth factor (10-200 ng/ml), a potent inhibitor of keratinization in other tissues, stimulated protein synthesis compared with untreated controls. Epidermal growth factor binding was located microscopically to the germinal and differentiating epidermal layers. SDS-PAGE and fluorography showed that the population of proteins synthesized in the presence of any hormone or growth factor combination did not differ from that in untreated controls and included the keratins involved in horn deposition. The results show that bovine hoof keratinization is under endocrine and growth factor control, and suggest that systemic changes in lactogenic hormones may act to inhibit keratin deposition. PMID:10191470

Hendry, K A; MacCallum, A J; Knight, C H; Wilde, C J

1999-02-01

178

The Mouse Homolog of the Bovine Leukemia Virus Receptor Is Closely Related to the d Subunit of Adaptor-Related Protein Complex AP3, Not Associated with the Cell Surface  

Microsoft Academic Search

A mouse cDNA (mBLVR1) which was highly homologous to the bovine cDNA of the bovine leukemia virus receptor (BLVR) gene was cloned. The mBLVR1 cDNA, of 4,730 bp, covered nearly the full length of the mRNA (about 5 kb) and included an open reading frame (ORF) encoding a protein of 1,199 amino acids. While the bovine BLVR protein was thought

TAKAKO SUZUKI; HIDETOSHI IKEDA

1998-01-01

179

A novel in vitro bovine cartilage punch model for assessing the regeneration of focal cartilage defects with biocompatible bacterial nanocellulose  

PubMed Central

Introduction Current therapies for articular cartilage defects fail to achieve qualitatively sufficient tissue regeneration, possibly because of a mismatch between the speed of cartilage rebuilding and the resorption of degradable implant polymers. The present study focused on the self-healing capacity of resident cartilage cells in conjunction with cell-free and biocompatible (but non-resorbable) bacterial nanocellulose (BNC). This was tested in a novel in vitro bovine cartilage punch model. Methods Standardized bovine cartilage discs with a central defect filled with BNC were cultured for up to eight weeks with/without stimulation with transforming growth factor-?1 (TGF-?1. Cartilage formation and integrity were analyzed by histology, immunohistochemistry and electron microscopy. Content, release and neosynthesis of the matrix molecules proteoglycan/aggrecan, collagen II and collagen I were also quantified. Finally, gene expression of these molecules was profiled in resident chondrocytes and chondrocytes migrated onto the cartilage surface or the implant material. Results Non-stimulated and especially TGF-?1-stimulated cartilage discs displayed a preserved structural and functional integrity of the chondrocytes and surrounding matrix, remained vital in long-term culture (eight weeks) without signs of degeneration and showed substantial synthesis of cartilage-specific molecules at the protein and mRNA level. Whereas mobilization of chondrocytes from the matrix onto the surface of cartilage and implant was pivotal for successful seeding of cell-free BNC, chondrocytes did not immigrate into the central BNC area, possibly due to the relatively small diameter of its pores (2 to 5 ?m). Chondrocytes on the BNC surface showed signs of successful redifferentiation over time, including increase of aggrecan/collagen type II mRNA, decrease of collagen type I mRNA and initial deposition of proteoglycan and collagen type II in long-term high-density pellet cultures. Although TGF-?1 stimulation showed protective effects on matrix integrity, effects on other parameters were limited. Conclusions The present bovine cartilage punch model represents a robust, reproducible and highly suitable tool for the long-term culture of cartilage, maintaining matrix integrity and homoeostasis. As an alternative to animal studies, this model may closely reflect early stages of cartilage regeneration, allowing the evaluation of promising biomaterials with/without chondrogenic factors.

2013-01-01

180

Acoustic wave propagation in bovine cancellous bone: Application of the Modified Biot-Attenborough model  

NASA Astrophysics Data System (ADS)

Acoustic wave propagation in bovine cancellous bone is experimentally and theoretically investigated in the frequency range of 0.5-1 MHz. The phase velocity, attenuation coefficient, and broadband ultrasonic attenuation (BUA) of bovine cancellous bone are measured as functions of frequency and porosity. For theoretical estimation, the Modified Biot-Attenborough (MBA) model is employed with three new phenomenological parameters: the boundary condition, phase velocity, and impedance parameters. The MBA model is based on the idealization of cancellous bone as a nonrigid porous medium with circular cylindrical pores oriented normal to the surface. It is experimentally observed that the phase velocity is approximately nondispersive and the attenuation coefficient linearly increases with frequency. The MBA model predicts a slightly negative dispersion of phase velocity linearly with frequency and the nonlinear relationships of attenuation and BUA with porosity. The experimental results are in good agreement with the theoretical results estimated with the MBA model. It is expected that the MBA model can be usefully employed in the field of clinical bone assessment for the diagnosis of osteoporosis.

Lee, Kang Il; Roh, Heui-Seol; Yoon, Suk Wang

2003-10-01

181

Acoustic wave propagation in bovine cancellous bone: application of the Modified Biot-Attenborough model.  

PubMed

Acoustic wave propagation in bovine cancellous bone is experimentally and theoretically investigated in the frequency range of 0.5-1 MHz. The phase velocity, attenuation coefficient, and broadband ultrasonic attenuation (BUA) of bovine cancellous bone are measured as functions of frequency and porosity. For theoretical estimation, the Modified Biot-Attenborough (MBA) model is employed with three new phenomenological parameters: the boundary condition, phase velocity, and impedance parameters. The MBA model is based on the idealization of cancellous bone as a nonrigid porous medium with circular cylindrical pores oriented normal to the surface. It is experimentally observed that the phase velocity is approximately nondispersive and the attenuation coefficient linearly increases with frequency. The MBA model predicts a slightly negative dispersion of phase velocity linearly with frequency and the nonlinear relationships of attenuation and BUA with porosity. The experimental results are in good agreement with the theoretical results estimated with the MBA model. It is expected that the MBA model can be usefully employed in the field of clinical bone assessment for the diagnosis of osteoporosis. PMID:14587625

Lee, Kang Il; Roh, Heui-Seol; Yoon, Suk Wang

2003-10-01

182

Agouti revisited: transcript quantification of the ASIP gene in bovine tissues related to protein expression and localization.  

PubMed

Beside its role in melanogenesis, the agouti signaling protein (ASIP) has been related to obesity. The potentially crucial role in adipocyte development makes it a tempting candidate for economic relevant, fat related traits in farm animals. The objective of our study was to characterize the mRNA expression of different ASIP transcripts and of putative targets in different bovine tissues, as well as to study consequences on protein abundance and localization. ASIP mRNA abundance was determined by RT-qPCR in adipose and further tissues of cattle representing different breeds and crosses. ASIP mRNA was up-regulated more than 9-fold in intramuscular fat of Japanese Black cattle compared to Holstein (p<0.001). Further analyses revealed that a transposon-derived transcript was solely responsible for the increased ASIP mRNA abundance. This transcript was observed in single individuals of different breeds indicating a wide spread occurrence of this insertion at the ASIP locus in cattle. The protein was detected in different adipose tissues, skin, lung and liver, but not in skeletal muscle by Western blot with a bovine-specific ASIP antibody. However, the protein abundance was not related to the observed ASIP mRNA over-expression. Immuno-histochemical analyses revealed a putative nuclear localization of ASIP additionally to the expected cytosolic signal in different cell types. The expression of melanocortin receptors (MCR) 1 to 5 as potential targets for ASIP was analyzed by RT-PCR in subcutaneous fat. Only MC1R and MC4R were detected indicating a similar receptor expression like in human adipose tissue. Our results provide evidence for a widespread expression of ASIP in bovine tissues at mRNA and, for the first time, at protein level. ASIP protein is detectable in adipocytes as well as in further cells of adipose tissue. We generated a basis for a more detailed investigation of ASIP function in peripheral tissues of various mammalian species. PMID:22530003

Albrecht, Elke; Komolka, Katrin; Kuzinski, Judith; Maak, Steffen

2012-01-01

183

Purified bovine plasma blocking factor decreases Bovine leukemia virus p24 expression while increasing protein synthesis and transcriptional activity of peripheral blood mononuclear cells in short-term culture  

PubMed Central

Abstract Bovine leukemia virus (BLV) induces a persistent infection in the B-cells causing polyclonal expansion of B-cells in one-third of infected cattle and lymphosarcoma in less than 5% of infected cattle. While BLV is difficult to detect in vivo, it is readily produced by cultured lymphocytes and is diminished when supplemented by bovine plasma. This phenomenon is attributed to a poorly characterized plasma blocking factor (PBF). We assessed the effects of bovine plasma on cell viability and BLV p24 expression, and the effects of purified PBF on protein synthesis and gene expression of short-term cultures of bovine lymphocytes. The addition of 25% plasma or semi-purified PBF to cultures had no significant effect on cell viability but caused significant decreases in BLV p24 production and significantly increased de novo protein synthesis. Utilizing a human microarray, the RNA messages of 83 genes involved in cell division, cell metabolism, and gene regulation were up-regulated.

2005-01-01

184

Sclera-Choroid-RPE Transport of Eight ?-Blockers in Human, Bovine, Porcine, Rabbit, and Rat Models  

PubMed Central

Purpose. To determine the influence of drug lipophilicity, ocular pigmentation, and species differences on transscleral solute transport. Methods. The transport of eight ?-blockers across excised sclera/sclera-choroid-RPE (SCRPE) of albino rabbit, pigmented rabbit, human, porcine, and bovine eyes was determined over 6 hours. The ex vivo transscleral ?-blocker transport to the vitreous at the end of 6 hours was determined in euthanatized, pigmented Brown Norway rats. The thicknesses of the sclera and SCRPE and the melanin content in choroid-RPE (CRPE) were measured to determine whether species differences in drug transport can be explained on this basis. Results. Solute lipophilicity inversely correlated with the SCRPE cumulative percentage of transport in all species (R2 ? 0.80). The CRPE impeded the SCRPE transport of all ?-blockers (51%–64% resistance in the rabbits; 84%–99.8% in the bovine and porcine eyes) more than the sclera, with the impedance increasing with lipophilicity. SCRPE transport followed the trend albino rabbit > pigmented rabbit > human > porcine > bovine, and a cross-species comparison showed good Spearman's rho correlation (R2 ? 0.85). Bovine (R2 = 0.84), porcine (R2 = 0.84), and human (R2 = 0.71) SCRPE transport was more predictive than that in the rabbit models (R2 = 0.60–0.61) of transscleral solute transport to the vitreous in rats. The CRPE concentrations were higher in pigmented rabbits than in albino rabbits. The melanin content of the CRPE exhibited the trend albino rabbit ? pigmented rabbit < porcine ? bovine < rat. Normalization to scleral thickness abolished the species differences in scleral transport. Normalization to SCRPE thickness and melanin content significantly reduced species differences in SCRPE transport. Conclusions. Owing to the presence of pigment and drug binding, choroid-RPE is the principal barrier to transscleral ?-blocker transport, with the barrier being more significant for lipophilic ?-blockers. Although different in magnitude between species, sclera/SCRPE transport can be correlated between species. Tissue thickness accounts for the species differences in scleral transport. Differences in tissue thickness and melanin content largely account for the species differences in SCRPE transport.

Kadam, Rajendra S.; Cheruvu, Narayan P. S.; Edelhauser, Henry F.

2011-01-01

185

Bovine ?1-acid glycoprotein, a thermostable version of its human counterpart: Insights from Fourier transform infrared spectroscopy and in silico modelling.  

PubMed

?1-Acid glycoprotein (AGP) is a plasma protein and a member of the acute phase response. AGP is known to bind and carry several biologically active compounds, as well as to down-modulate the immune system activities. In this work, the structure of bovine AGP has been investigated by Fourier-Transform infrared spectroscopy. A model structure has been obtained on the basis of human AGP and refined by molecular dynamics. In spite of the similar structure, bovine AGP shows an unexpectedly higher (?20 °C) thermostability than its human counterpart. Inspection of the model structure has pointed out the presence of 12 ionic bridges and 2 sulphur-aromatic interactions, whereas only 6 ionic bridges were detected in human AGP. The high number (9) of glutamic acid residues involved in the ionic interactions might explain the significantly decreased thermostability measured at pH 5.5 (Tm ? 71 °C) with respect to pH 7.4 (Tm ? 81 °C), whereas thermostability of human AGP was only slightly affected by lowering the pH. As in human AGP and several other lipocalins, a temperature-induced molten globule state has been observed in the denaturation pathway of bovine AGP. PMID:24530968

Baldassarre, Maurizio; Galeazzi, Roberta; Maggiore, Beatrice; Tanfani, Fabio; Scirč, Andrea

2014-07-01

186

Baculovirus Expression of the Fusion Protein Gene of Bovine Respiratory Syncytial Virus and Utility of the Recombinant Protein in a Diagnostic Enzyme Immunoassay  

PubMed Central

The fusion (F) protein of bovine respiratory syncytial virus (BRSV) was expressed by using a baculovirus vector. Antigenicity was tested by immunofluorescence analysis with F-specific monoclonal and polyclonal antibodies. Antibodies to recombinant F protein raised in a rabbit neutralized BRSV and human respiratory syncytial virus infectivity when tested in a plaque reduction assay. The recombinant F protein was evaluated as a source of antigen in an enzyme-linked immunosorbent assay (ELISA), and this ELISA was compared with the virus neutralization (VN) test for detecting BRSV antibodies in 10 consecutive serum samples from four calves vaccinated with a live modified BRSV vaccine and from two nonvaccinated control calves. The ELISA with the baculovirus-expressed F protein as an antigen compared favorably with the VN test and is a rapid, sensitive, and specific method for detecting serum antibodies to BRSV.

Pastey, Manoj K.; Samal, Siba K.

1998-01-01

187

Modeling Protein Self Assembly  

ERIC Educational Resources Information Center

Understanding the structure and function of proteins is an important part of the standards-based science curriculum. Proteins serve vital roles within the cell and malfunctions in protein self assembly are implicated in degenerative diseases. Experience indicates that this topic is a difficult one for many students. We have found that the concept…

Baker, William P.; Jones, Carleton Buck; Hull, Elizabeth

2004-01-01

188

Modeling Protein Domain Function  

ERIC Educational Resources Information Center

This simple but effective laboratory exercise helps students understand the concept of protein domain function. They use foam beads, Styrofoam craft balls, and pipe cleaners to explore how domains within protein active sites interact to form a functional protein. The activity allows students to gain content mastery and an understanding of the…

Baker, William P.; Jones, Carleton "Buck"; Hull, Elizabeth

2007-01-01

189

MATER protein expression and intracellular localization throughout folliculogenesis and preimplantation embryo development in the bovine  

Microsoft Academic Search

BACKGROUND: Mater (Maternal Antigen that Embryos Require), also known as Nalp5 (NACHT, leucine rich repeat and PYD containing 5), is an oocyte-specific maternal effect gene required for early embryonic development beyond the two-cell stage in mouse. We previously characterized the bovine orthologue MATER as an oocyte marker gene in cattle, and this gene was recently assigned to a QTL region

Sophie Pennetier; Christine Perreau; Svetlana Uzbekova; Aurore Thélie; Bernadette Delaleu; Pascal Mermillod; Rozenn Dalbičs-Tran

2006-01-01

190

Bovine and rabbit models for the study of a Staphylococcus aureus avirulent mutant strain, RC122  

PubMed Central

Staphylococcus aureus is the main etiological agent of bovine mastitis. Intramammary infections are difficult to cure and vaccination appears to be an alternative to prevent the disease. Research has focused on the development of mutants affected in the synthesis of pathogenicity determinants. We constructed a mutant strain (RC122) after chemical mutagenesis. In a mouse model, the strain was shown to be 1500 times less virulent, showed similar kinetics of disappearance in the kidney as its parental strain, and a good degree of protection against a challenge from homologous and heterologous strains. The objective of the present report was to study the avirulent RC122 S. aureus mutant strain in rabbit and bovine infection models. The results clearly show that RC122 was less virulent than its parental strain in a rabbit skin model, and was also correlated with its avirulence as an udder pathogen. These traits make the RC122 mutant strain interesting as a potential strain for an experimental vaccine trial in dairy herds.

Reinoso, Elina; Magnano, Gabriel; Giraudo, Jose; Calzolari, Aldo; Bogni, Cristina

2002-01-01

191

Protein-coated polymer as a matrix for enzyme immobilization: immobilization of trypsin on bovine serum albumin-coated allyl glycidyl ether-ethylene glycol dimethacrylate copolymer.  

PubMed

Allyl glycidyl ether (AGE)-ethylene glycol dimethacrylate (EGDM) copolymer with 25% crosslink density (AGE-25) shows excellent bovine serum albumin (BSA) adsorption (up to 16% (w/w)) at pH 8.0 and the adsorbed BSA is strongly bound. This protein-coated polymer provides a novel matrix with naturally existing functional groups such as thiol, amino, and carboxylic acid that are available for covalent immobilization of functional enzymes. Employing appropriate strategies, trypsin as a model protein was covalently bound to BSA-coated matrix both independently, and in a stepwise manner on the same matrix, with less than 5% loss of enzyme activity during immobilization. Glutaraldehyde crosslinking after immobilization provide stable enzyme preparation with activity of 510 units/g recycled up to six times without loss of enzyme activity. AFM studies reveal that the polymer surface has protein peaks and valleys rather than a uniform monolayer distribution of the protein and the immobilized enzyme preparation can best be described as polymer supported cross-linked enzyme aggregates (CLEAs). PMID:24449609

Jasti, Lakshmi Swarnalatha; Dola, Sandhya Rani; Kumaraguru, Thenkrishnan; Bajja, Sreedhar; Fadnavis, Nitin W; Addepally, Uma; Rajdeo, Kishor; Ponrathnam, Surendra; Deokar, Sarika

2014-01-01

192

An Immunoglobulin Binding Protein (Antigen 5) of the Stable Fly (Diptera: Muscidae) Salivary Gland Stimulates Bovine Immune Responses  

PubMed Central

The stable fly, Stomoxys calcitrans, is an economically important pest of livestock. Prior studies demonstrated lymphocyte suppression by crude salivary gland extract (SGE) of the stable fly. A dominant 27 kDa protein identified in the SGE was reported to stimulate immunodominant antibody responses in exposed cattle. The purpose of this study was to determine if this protein, now identified as a homolog of insect proteins named antigen 5 (Ag5), was responsible for the lymphocyte suppression and if naďve calves can mount an immune response to it. Calves raised in the winter months were immunized with recombinant Ag5 (rAg5) expressed in Drosophila S2 cells or with “natural” Ag5 protein isolated by preparative gel electrophoresis of SGE. Control calves were immunized with adjuvant alone. Rising antibody concentrations to rAg5 were detected in two of three calves immunized with rAg5 and one of three calves immunized with natural Ag5. Recall lymphocyte responses to rAg5 were detected at 21 and 28 DPI in calves immunized with rAg5 but not in calves immunized with the natural Ag5 or those exposed to adjuvant alone. Mitogen-stimulated bovine lymphocyte responses were not suppressed by rAg5. Further investigation using immunoblotting revealed that rAg5 binds to the Fc and F(ab’)2 portions of bovine IgG, but not to an Fab fragment. These findings suggest that Ag5 of the stable fly salivary gland is not immunosuppressive, but has immunoglobulin binding properties and can invoke specific antibody and memory lymphocyte responses in immunized calves.

AMERI, M.; WANG, X.; WILKERSON, M. J.; KANOST, M. R.; BROCE, A. B.

2008-01-01

193

Commercial Bovine Proteoglycan Is Highly Arthritogenic and Can Be Used as an Alternative Antigen Source for PGIA Model  

PubMed Central

Rheumatoid arthritis (RA) is the most common systemic autoimmune disease. It affects mainly the joints, causing synovitis, cartilage destruction, and bone erosion. Many experimental models are used to study the mechanisms involved in immunopathogenesis and new therapies for this disease. Proteoglycan-induced arthritis (PGIA) is a widely used model based on the cross-reactivity of injected foreign (usually human) PG and mice self-PG. Considering the complexity of the extraction and purification of human PG, in this study we evaluated the arthritogenicity of bovine PG that is commercially available. Bovine PG was highly arthritogenic, triggering 100% incidence of arthritis in female BALB/c retired breeder mice. Animals immunized with bovine PG presented clinical symptoms and histopathological features similar to human RA and other experimental models. Moreover, bovine PG immunization determined higher levels of proinflammatory and anti-inflammatory cytokines in arthritic mice compared to healthy ones. As expected, only the arthritic group produced IgG1 and IgG2a antibodies against PG. Thus, commercial bovine PG can be used as an alternative antigenic source to PGIA for the study of many RA aspects, including the immunopathogenesis of the disease and also the development of new therapies.

Ishikawa, Larissa Lumi Watanabe; Colavite, Priscila Maria; da Rosa, Larissa Camargo; Franca, Thais Graziela Donega; Zorzella-Pezavento, Sofia Fernanda Goncalves; Chiuso-Minicucci, Fernanda; Sartori, Alexandrina

2014-01-01

194

Cell Arrest and Cell Death in Mammalian Preimplantation Development: Lessons from the Bovine Model  

PubMed Central

Background The causes, modes, biological role and prospective significance of cell death in preimplantation development in humans and other mammals are still poorly understood. Early bovine embryos represent a very attractive experimental model for the investigation of this fundamental and important issue. Methods and Findings To obtain reference data on the temporal and spatial occurrence of cell death in early bovine embryogenesis, three-dimensionally preserved embryos of different ages and stages of development up to hatched blastocysts were examined in toto by confocal laser scanning microscopy. In parallel, transcript abundance profiles for selected apoptosis-related genes were analyzed by real-time reverse transcriptase-polymerase chain reaction. Our study documents that in vitro as well as in vivo, the first four cleavage cycles are prone to a high failure rate including different types of permanent cell cycle arrest and subsequent non-apoptotic blastomere death. In vitro produced and in vivo derived blastocysts showed a significant incidence of cell death in the inner cell mass (ICM), but only in part with morphological features of apoptosis. Importantly, transcripts for CASP3, CASP9, CASP8 and FAS/FASLG were not detectable or found at very low abundances. Conclusions In vitro and in vivo, errors and failures of the first and the next three cleavage divisions frequently cause immediate embryo death or lead to aberrant subsequent development, and are the main source of developmental heterogeneity. A substantial occurrence of cell death in the ICM even in fast developing blastocysts strongly suggests a regular developmentally controlled elimination of cells, while the nature and mechanisms of ICM cell death are unclear. Morphological findings as well as transcript levels measured for important apoptosis-related genes are in conflict with the view that classical caspase-mediated apoptosis is the major cause of cell death in early bovine development.

Leidenfrost, Sandra; Boelhauve, Marc; Reichenbach, Myriam; Gungor, Tuna; Reichenbach, Horst-Dieter; Sinowatz, Fred; Wolf, Eckhard; Habermann, Felix A.

2011-01-01

195

A heterogeneous population model for contagious bovine pleuropneumonia transmission and control in pastoral communities of East Africa  

Microsoft Academic Search

Pastoral cattle live in highly structured communities characterized by complex contact patterns. The present paper describes a spatially heterogeneous model for the transmission of contagious bovine pleuropneumonia (CBPP) developed specifically for pastoral communities of East Africa. The model is validated against serological data on the prevalence of CBPP infection in several communities of southern Sudan and against livestock owner information

J. C. Mariner; J. McDermott; J. A. P. Heesterbeek; G. Thomson; P. L. Roeder; S. W. Martin

2006-01-01

196

Proteins other than the locus of enterocyte effacement-encoded proteins contribute to Escherichia coli O157:H7 adherence to bovine rectoanal junction stratified squamous epithelial cells  

PubMed Central

Background In this study, we present evidence that proteins encoded by the Locus of Enterocyte Effacement (LEE), considered critical for Escherichia coli O157 (O157) adherence to follicle-associated epithelial (FAE) cells at the bovine recto-anal junction (RAJ), do not appear to contribute to O157 adherence to squamous epithelial (RSE) cells also constituting this primary site of O157 colonization in cattle. Results Antisera targeting intimin-?, the primary O157 adhesin, and other essential LEE proteins failed to block O157 adherence to RSE cells, when this pathogen was grown in DMEM, a culture medium that enhances expression of LEE proteins. In addition, RSE adherence of a DMEM-grown-O157 mutant lacking the intimin protein was comparable to that seen with its wild-type parent O157 strain grown in the same media. These adherence patterns were in complete contrast to that observed with HEp-2 cells (the adherence to which is mediated by intimin-?), assayed under same conditions. This suggested that proteins other than intimin-? that contribute to adherence to RSE cells are expressed by this pathogen during growth in DMEM. To identify such proteins, we defined the proteome of DMEM-grown-O157 (DMEM-proteome). GeLC-MS/MS revealed that the O157 DMEM-proteome comprised 684 proteins including several components of the cattle and human O157 immunome, orthologs of adhesins, hypothetical secreted and outer membrane proteins, in addition to the known virulence and LEE proteins. Bioinformatics-based analysis of the components of the O157 DMEM proteome revealed several new O157-specific proteins with adhesin potential. Conclusion Proteins other than LEE and intimin-? proteins are involved in O157 adherence to RSE cells at the bovine RAJ. Such proteins, with adhesin potential, are expressed by this human pathogen during growth in DMEM. Ongoing experiments to evaluate their role in RSE adherence should provide both valuable insights into the O157-RSE interactions and new targets for more efficacious anti-adhesion O157 vaccines.

2012-01-01

197

The molecular cloning of the complementary deoxyribonucleic acid for bovine vitamin D-dependent calcium-binding protein: structure of the full-length protein and evidence for homologies with other calcium-binding proteins of the troponin-C superfamily of proteins.  

PubMed

We have cloned the cDNA for bovine intestinal vitamin D-dependent calcium-binding protein and, based on the sequence of the DNA, have deduced the structure of the full-length protein. The sequence of the cDNA clone predicts a protein comprised of 78 amino acids with a mol wt of 8788. The mRNA for the protein in bovine duodenum is about 500-600 bases in length. The protein sequence of bovine intestinal calcium-binding protein is 87% homologous with the sequence of porcine intestinal vitamin D-dependent calcium-binding protein and 81% homologous with the sequence of rat intestinal vitamin D-dependent calcium-binding protein. Hydrophilicity plots of the proteins noted above show that despite differences in amino acid sequence the proteins have similar patterns. In addition, the predicted secondary structure of the proteins is similar. Bovine intestinal calcium-binding protein shows 48.6% homology with the alpha-chain and 38.2% homology with the beta-chain of bovine S-100 protein and a similar high degree of homology with the beta-chain of human S-100 protein. The protein also demonstrates 36-43% homology with parvalbumin alpha and beta from various species and with troponin-C. There is some homology with the 28K vitamin D-dependent calcium-binding proteins. Vitamin D-dependent bovine intestinal calcium-binding protein is closely related to other mammalian intestinal calcium-binding proteins and to the S-100 proteins, parvalbumins, and troponin-C. PMID:2710141

Kumar, R; Wieben, E; Beecher, S J

1989-02-01

198

Conserved Arginines of Bovine Adenovirus-3 33K Protein Are Important for Transportin-3 Mediated Transport and Virus Replication  

PubMed Central

The L6 region of bovine adenovirus (BAdV)-3 encodes a spliced protein designated 33K. The 33K specific sera detected five major proteins and three minor proteins in transfected or virus infected cells, which could arise by internal initiation of translation and alternative splicing. The 33K protein is predominantly localized to the nucleus of BAdV-3 infected cells. The 33K nuclear transport utilizes both classical importin-?/-? and importin-? dependent nuclear import pathways and preferentially binds to importin-?5 and transportin-3 receptors, respectively. Analysis of mutant 33K proteins demonstrated that amino acids 201–240 of the conserved C-terminus of 33K containing RS repeat are required for nuclear localization and, binding to both importin-?5 and transportin-3 receptors. Interestingly, the arginine residues of conserved RS repeat are required for binding to transportin-3 receptor but not to importin-?5 receptor. Moreover, mutation of arginines residues of RS repeat proved lethal for production of progeny virus. Our results suggest that arginines of RS repeat are required for efficient nuclear transport of 33K mediated by transportin-3, which appears to be essential for replication and production of infectious virion.

Islam, Azharul; Tikoo, Suresh K.

2014-01-01

199

Identification and characterization of an intracellular protein complex that binds fibroblast growth factor-2 in bovine brain.  

PubMed Central

The fibroblast growth factor (FGF) family is composed of polypeptides with sequence identity which signal through transmembrane tyrosine kinase receptors. We report here the purification from bovine brain microsomes of an FGF-2-binding complex composed of three proteins of apparent molecular masses 150 kDa, 79 kDa and 46 kDa. Only the 150 kDa and 79 kDa proteins bound FGF-2 in cross-linking and ligand-blotting experiments. Binding of FGF-2 to p79 is enhanced in the presence of calcium. Peptide sequences allowed the identification of p150 and the cloning of the cDNAs encoding p79 and p46. The deduced amino acid sequence of p79 reveals high similarity to those of gastrin-binding protein and mitochondrial enoyl-CoA hydratase/hydroxyacyl-CoA dehydrogenase. p46 is similar to mitochondrial ketoacyl-CoA thiolase. Stable transfection of FR3T3 rat fibroblast cells with p79 cDNA analysed by electron microscopy following immunolabelling of ultra-thin cryosections revealed a localization of p79 in the secretory pathway, mainly in the endoplasmic reticulum and the Golgi region, where it is specifically associated with the molecular chaperone calnexin. In vivo a protein similar to the Golgi protein MG-160 forms a complex with FGF-2 and p79.

Chevet, E; Lemaitre, G; Cailleret, K; Dahan, S; Bergeron, J J; Katinka, M D

1999-01-01

200

Protein-Specific Analysis of Humoral Immune Responses in a Clinical Trial for Vaccines against Contagious Bovine Pleuropneumonia? †  

PubMed Central

Specific humoral immune responses in a clinical trial on cattle for vaccines against contagious bovine pleuropneumonia (CBPP) were investigated. The trial included a subunit vaccine consisting of five recombinant putative variable surface proteins of the infectious agent Mycoplasma mycoides subsp. mycoides small colony type (M. mycoides SC) compared to the currently approved attenuated vaccine strain T1/44 and untreated controls. Humoral immune responses to 65 individual recombinant surface proteins of M. mycoides SC were monitored by a recently developed bead-based array assay. Responses to the subunit vaccine components were found to be weak. Animals vaccinated with this vaccine were not protected and had CBPP lesions similar to those of the untreated controls. In correlating protein-specific humoral responses to T1/44-induced immunity, five proteins associated with a protective immune response were identified by statistical evaluation, namely, MSC_1046 (LppQ), MSC_0271, MSC_0136, MSC_0079, and MSC_0431. These five proteins may be important candidates in the development of a novel subunit vaccine against CBPP.

Hamsten, Carl; Tjipura-Zaire, Georgina; McAuliffe, Laura; Huebschle, Otto J. B.; Scacchia, Massimo; Ayling, Roger D.; Persson, Anja

2010-01-01

201

Structural and functional similarities of bovine alpha-crystallin and mouse small heat-shock protein. A family of chaperones.  

PubMed

alpha-Crystallin, composed of the subunits alpha A and alpha B, is a major vertebrate eye lens protein, accomplishing a structural role in maintaining lens stability and transparency. Both subunits also occur in low amounts outside the lens, where their precise function is unknown. They are structurally related to the small heat-shock proteins (HSPs), and increasing evidence indicates that they have also functional similarities with the small HSPs. To extend our insight into these structural and functional relationships, the mouse small HSP (HSP25) was compared with bovine alpha-crystallin, with respect to several known properties of the latter. We show that alpha-crystallin and HSP25 resemble each other in secondary structure and have similar stability toward urea dissociation at pH 7.0. Mixed polymers can be formed from any combination of alpha A-crystallin, alpha B-crystallin, and HSP25 subunits. Furthermore, we demonstrate that HSP25, like alpha-crystallin, can function as a molecular chaperone, by suppressing heat-induced aggregation of other proteins, and is an efficient inhibitor of elastase. Finally, HSP25 is found to be a substrate for protein cross-linking by tissue-type transglutaminase, like alpha B-crystallin. Our results thus corroborate that alpha-crystallin and the small HSPs have comparable functions, probably being involved in the protection of other proteins under conditions of stress. PMID:8093449

Merck, K B; Groenen, P J; Voorter, C E; de Haard-Hoekman, W A; Horwitz, J; Bloemendal, H; de Jong, W W

1993-01-15

202

Acrosin activity regulation by protein kinase C and tyrosine kinase in bovine sperm acrosome exocytosis induced by lysophosphatidylcholine.  

PubMed

Acrosin is an important proteolytic enzyme that is capable of hydrolysing the zona pellucida in bovine oocyte. Lysophosphatydic acid (LPA) derivated from lysophosphatidylcholine (LPC) is known to trigger the acrosome exocytosis. The present study was aimed at examining the acrosin activity variations in LPC-induced acrosome exocytosis and its regulation by tyrosine kinase, protein kinase C (PKC) and voltage-dependent calcium channels (VDCC) in spermatozoa previously capacitated with heparin or quercetin. The enzyme activities were spectrophotometrically measured using N-?-benzoyl-DL-arginine p-nitroanilide as an acrosin-specific substrate. The capacitation and acrosomal reaction were evaluated by chlorotetracycline assay, and the viability and acrosome integrity were evaluated by the trypan blue stain/differential interference contrast. It was observed that LPC induced acrosome exocytosis and increased the activity of acrosin in spermatozoa previously capacitated with heparin. In heparin/LPC-treated samples, it was observed that the inhibition of tyrosine kinase and PKC blocked the acrosome exocytosis and the acrosin activity (p < 0.05). Under these conditions, in heparin-capacitated spermatozoa, the LPC provokes an acrosin activity increase that is independent of calcium influx through VDCC Type L. In cryopreserved bovine spermatozoa, LPC might require modulation, mainly tyrosine kinase participation with respect to PKC activity to induce acrosome exocytosis and increase acrosin activity. PMID:22335484

Pérez Aguirreburualde, M S; Fernández, S; Córdoba, M

2012-12-01

203

Identification of immune genes and proteins involved in the response of bovine mammary tissue to Staphylococcus aureus infection  

PubMed Central

Background Mastitis in dairy cattle results from infection of mammary tissue by a range of micro-organisms but principally coliform bacteria and Gram positive bacteria such as Staphylococcus aureus. The former species are often acquired by environmental contamination while S. aureus is particularly problematic due to its resistance to antibiotic treatments and ability to reside within mammary tissue in a chronic, subclinical state. The transcriptional responses within bovine mammary epithelial tissue subjected to intramammary challenge with S. aureus are poorly characterised, particularly at the earliest stages of infection. Moreover, the effect of infection on the presence of bioactive innate immune proteins in milk is also unclear. The nature of these responses may determine the susceptibility of the tissue and its ability to resolve the infection. Results Transcriptional profiling was employed to measure changes in gene expression occurring in bovine mammary tissues sampled from three dairy cows after brief and graded intramammary challenges with S. aureus. These limited challenges had no significant effect on the expression pattern of the gene encoding ?-casein but caused coordinated up-regulation of a number of cytokines and chemokines involved in pro-inflammatory responses. In addition, the enhanced expression of two genes, S100 calcium-binding protein A12 (S100A12) and Pentraxin-3 (PTX3) corresponded with significantly increased levels of their proteins in milk from infected udders. Both genes were shown to be expressed by mammary epithelial cells grown in culture after stimulation with lipopolysaccharide. There was also a strong correlation between somatic cell count, a widely used measure of mastitis, and the level of S100A12 in milk from a herd of dairy cows. Recombinant S100A12 inhibited growth of Escherichia coli in vitro and recombinant PTX3 bound to E. coli as well as C1q, a subunit of the first component of the complement cascade. Conclusion The transcriptional responses in infected bovine mammary tissue, even at low doses of bacteria and short periods of infection, probably reflect the combined contributions of gene expression changes resulting from the activation of mammary epithelial cells and infiltrating immune cells. The secretion of a number of proinflammatory cytokines and chemokines from mammary epithelial cells stimulated by the bacteria serves to trigger the recruitment and activation of neutrophils in mammary tissue. The presence of S100A12 and PTX3 in milk from infected udder quarters may increase the anti-bacterial properties of milk thereby helping to resolve the mammary tissue infection as well as potentially contributing to the maturation of the newborn calf epithelium and establishment of the newborn gut microbial population.

Lutzow, Ylva C Strandberg; Donaldson, Laurelea; Gray, Christian P; Vuocolo, Tony; Pearson, Roger D; Reverter, Antonio; Byrne, Keren A; Sheehy, Paul A; Windon, Ross; Tellam, Ross L

2008-01-01

204

Bovine Somatotropin and Rumen-Undegradable Protein Effects in Prepubertal Dairy Heifers: Effects on Body Composition and Organ and Tissue Weights  

Microsoft Academic Search

The objectives of this study were to determine the effect of recombinant bovine somatotropin (bST) and added dietary rumen undegradable protein (RUP) on organand tissueweights andbody compositionin grow- ing dairy heifers. Thirty-two Holstein heifers were in the experiment, 8 killed initially at 3 mo of age, with the remaining 24 Holstein heifers randomly assigned to treatments (n = 6) consisting

U. Moallem; G. E. Dahl; E. K. Duffey; A. V. Capuco; D. L. Wood; K. R. McLeod; R. L. Baldwin VI; R. A. Erdman

2004-01-01

205

Overexpression of binding protein and disruption of the PMR1 gene synergistically stimulate secretion of bovine prochymosin but not plant Thaumatin in yeast  

Microsoft Academic Search

When the heterologous proteins thaumatin and bovine prochymosin are produced in yeast cells as a fusion with the yeast invertase secretory signal peptide, less than 2% of the product is secreted in a biologically active form into the medium. The remainder accumulates intracellularly in a misfolded conformation. We investigated whether this poor secretion can be improved by overexpression of binding

M. M. Harmsen; M. I. Bruyne; H. A. Raué; J. Maat

1996-01-01

206

Detection of multiple retroviral infections in cattle and cross-reactivity of bovine immunodeficiency-like virus and human immunodeficiency virus type 1 proteins using bovine and human sera in a western blot assay.  

PubMed Central

Bovine antibovine immunodeficiency-like virus (BIV) antibodies were detected by Western blot analysis (WBA) using a chemiluminescence protocol. Bovine sera with anti-BIV activity, obtained from cows in two dairy herds, had antibodies directed against a variety of BIV-specific antigens indicating chronic infections. These sera were also tested for serological reactivity against bovine leukemia virus (BLV) and bovine syncytial virus (BSV). Cows most commonly had anti-BSV antibodies (12 of 39). Evidence for infection with BSV and BIV or BSV and BLV occurred with almost equal frequency (5 of 39 and 4 of 39, respectively) while only one instance of BIV and BLV coseropositivity was detected. The high prevalence of BSV seropositivity is consistent with a relatively infectious virus, which, as is known, may be transferred congenitally. Similar rates of coseropositivity of BIV or BLV with BSV in this population suggest that BIV is no more infectious than BLV and probably requires prolonged close contact for transmission. Seven of nine cows with anti-BIV antibodies detected primarily human immunodeficiency virus type 1 (HIV-1) p51 and p63 antigens by WBA using an alkaline phosphatase detection system, suggesting that HIV-1 proteins have potential usefulness in screening cattle for BIV seropositivity. Six human sera that showed strong reactivity against multiple HIV-1 proteins and the serum from one of three patients considered to be an "indeterminate" HIV-1 reactor, cross-reacted primarily with BIV p26. This is the first report of human sera with antibody to BIV-specific proteins.(ABSTRACT TRUNCATED AT 250 WORDS) Images Fig. 1. Fig. 2. Fig. 3.

Jacobs, R M; Smith, H E; Gregory, B; Valli, V E; Whetstone, C A

1992-01-01

207

Interaction of bisphenol A with bovine hemoglobin using spectroscopic and molecular modeling methods.  

PubMed

The interaction of bisphenol A with bovine hemoglobin (BHb) under physiological conditions was investigated by using fluorescence, ultraviolet-visible (UV-Vis) absorption, circular dichroism (CD), and molecular modeling. The experimental results showed that BPA can bind with BHb to form a complex. The binding constant Ka and the number of binding sites n were calculated to be 1.49 × 10(5) L mol(-1) and 1, respectively. Molecular modeling study revealed that BPA bound into BHb central cavity, and the binding mode of BPA-BHb complex could be hydrogen bonding. The UV-Vis absorption and CD spectra indicated that the secondary structure of BHb was altered, which may affect physiological functions of hemoglobin. This work is helpful for clarifying the molecular toxic mechanism of BPA in vivo. PMID:22054083

Fang, Xiaoyan; Cao, Shutao; Liu, Rutao

2011-11-01

208

Characterization of the Interaction between Eupatorin and Bovine Serum Albumin by Spectroscopic and Molecular Modeling Methods  

PubMed Central

This study investigated the interaction between eupatorin and bovine serum albumin (BSA) using ultraviolet-visible (UV-vis) absorption, fluorescence, synchronous fluorescence, circular dichroism (CD) spectroscopies, and molecular modeling at pH 7.4. Results of UV-vis and fluorescence spectroscopies illustrated that BSA fluorescence was quenched by eupatorin via a static quenching mechanism. Thermodynamic parameters revealed that hydrophobic and electrostatic interactions played major roles in the interaction. Moreover, the efficiency of energy transfer, and the distance between BSA and acceptor eupatorin, were calculated. The effects of eupatorin on the BSA conformation were analyzed using UV-vis, CD, and synchronous fluorescence. Finally, the binding of eupatorin to BSA was modeled using the molecular docking method.

Xu, Hongliang; Yao, Nannan; Xu, Haoran; Wang, Tianshi; Li, Guiying; Li, Zhengqiang

2013-01-01

209

Protein crystal growth in microgravity review of large scale temperature induction method: Bovine insulin, human insulin and human ?-interferon  

NASA Astrophysics Data System (ADS)

The Protein Crystal Growth Facility (PCF) is space-flight hardware that accommodates large scale protein crystal growth experiments using temperature change as the inductive step. Recent modifications include specialized instrumentation for monitoring crystal nucleation with laser light scattering. This paper reviews results from its first seven flights on the Space Shuttle, the last with laser light scattering instrumentation in place. The PCF's objective is twofold: (1) the production of high quality protein crystals for x-ray analysis and subsequent structure-based drug design and (2) preparation of a large quantity of relatively contaminant free crystals for use as time-release protein pharmaceuticals. The first three Shuttle flights with bovine insulin constituted the PCF's proof of concept, demonstrating that the space-grown crystals were larger and diffracted to higher resolution than their earth-grown counterparts. The later four PCF missions were used to grow recombinant human insulin crystals for x-ray analysis and continue productions trials aimed at the development of a processing facility for crystalline recombinant a-interferon.

Long, Marianna M.; Bishop, John Bradford; Delucas, Lawrence J.; Nagabhushan, Tattanhalli L.; Reichert, Paul; Smith, G. David

1997-01-01

210

Protein crystal growth in microgravity review of large scale temperature induction method: bovine insulin, human insulin and human alpha interferon  

NASA Astrophysics Data System (ADS)

The protein crystal growth facility (PCF) is space-flight hardware that accommodates large scale protein crystal growth experiments using temperature change as the inductive step. Recent modifications include specialized instrumentation for monitoring crystal nucleation with laser light scattering. This paper reviews results from the PCF's first seven flights on the Space Shuttle, the last with laser light scattering instrumentation. The PCF's objective is twofold: (1) production of high quality protein crystals for X-ray analysis and subsequent structure based drug design and (2) preparation of a large quantity of relatively contaminant free crystals for use as time-release protein pharmaceuticals. The first three Shuttle flights with bovine insulin constituted the PCF's proof of concept, demonstrating that the space-grown crystals were larger and diffracted to higher resolution than their earth-grown counterparts. The later four PCF missions were used to grow recombinant human insulin crystals for X-ray analysis and to continue productions trials aimed at the development of a processing facility for crystalline recombinant alpha interferon.

Long, Marianna M.; Bishop, John Bradford; Nagabhushan, Tattanahalli L.; Reichert, Paul; Smith, G. David; DeLucas, Lawrence J.

1996-10-01

211

Thermodynamic and structural analysis of homodimeric proteins: model of ?-lactoglobulin.  

PubMed

The energetics of protein homo-oligomerization was analyzed in detail with the application of a general thermodynamic model. We have studied the thermodynamic aspects of protein-protein interaction employing ?-lactoglobulin A from bovine milk at pH=6.7 where the protein is mainly in its dimeric form. We performed differential calorimetric scans at different total protein concentration and the resulting thermograms were analyzed with the thermodynamic model for oligomeric proteins previously developed. The thermodynamic model employed, allowed the prediction of the sign of the enthalpy of dimerization, the analysis of complex calorimetric profiles without transitions baselines subtraction and the obtainment of the thermodynamic parameters from the unfolding and the association processes and the compared with association parameters obtained with Isothermal Titration Calorimetry performed at different temperatures. The dissociation and unfolding reactions were also monitored by Fourier-transform infrared spectroscopy and the results indicated that the dimer of ?-lactoglobulin (N(2)) reversibly dissociates into monomeric units (N) which are structurally distinguishable by changes in their infrared absorbance spectra upon heating. Hence, it is proposed that ?-lactoglobulin follows the conformational path induced by temperature:N(2)?2N?2D. The general model was validated with these results indicating that it can be employed in the study of the thermodynamics of other homo-oligomeric protein systems. PMID:22172914

Burgos, Inés; Dassie, Sergio A; Villarreal, Marcos A; Fidelio, Gerardo D

2012-02-01

212

The Presence of Disease-Associated Prion Protein in Skeletal Muscle of Cattle Infected with Classical Bovine Spongiform Encephalopathy  

PubMed Central

ABSTRACT The aim of this study was to investigate the presence of disease-associated prion protein (PrPSc) in the skeletal muscle of cattle infected with classical bovine spongiform encephalopathy (C-BSE). The study was carried out systematically in 12 different muscle samples from 43 (3 field and 40 experimental) cases of C-BSE; however, muscle spindles were not available in many of these cases. Therefore, analysis became restricted to a total of 31 muscles in 23 cattle. Even after this restriction, low levels of PrPSc were detected in the muscle spindles of the masseter, intercostal, triceps brachii, psoas major, quadriceps femoris and semitendinosus muscles from 3 field and 6 experimental clinical-stage cases. The present data indicate that small amounts of PrPSc are detectable by immunohistochemistry in the skeletal muscles of animals terminally affected with C-BSE.

OKADA, Hiroyuki; MIYAZAWA, Kohtaro; FUKUDA, Shigeo; IWAMARU, Yoshifumi; IMAMURA, Morikazu; MASUJIN, Kentaro; MATSUURA, Yuichi; FUJII, Takashi; FUJII, Kei; KAGEYAMA, Soichi; YOSHIOKA, Miyako; MURAYAMA, Yuichi; YOKOYAMA, Takashi

2013-01-01

213

Endothelin stimulates a sustained 1,2-diacylglycerol increase and protein kinase C activation in bovine aortic smooth muscle cells  

SciTech Connect

Endothelin is a long-lasting potent vasoconstrictor peptide. We report here that in bovine aortic smooth muscle cells, endothelin biphasically increased total cellular diacylglycerol (DAG) content. When cellular DAG was labeled with (/sup 14/C) glycerol for 48h, endothelin stimulated (/sup 14/C)DAG formation in a biphasic pattern. Only one prolonged phase of DAG accumulation was observed when cells were labeled with (/sup 3/H)glycerol for 2 h. Endothelin induced an increase in the membranous protein kinase C (PKC) activities, which lasted for more than 20 min. These data suggest that (i) endothelin stimulates a sustained generation of DAG, (ii) this accumulation of DAG results in a sustained translocation of cytosolic PKC activities to the membrane.

Lee, T.S.; Chao, T.; Hu, K.Q.; King, G.L.

1989-07-14

214

A dynamic model of bovine tuberculosis spread and control in Great Britain.  

PubMed

Bovine tuberculosis (TB) is one of the most complex, persistent and controversial problems facing the British cattle industry, costing the country an estimated Ł100 million per year. The low sensitivity of the standard diagnostic test leads to considerable ambiguity in determining the main transmission routes of infection, which exacerbates the continuing scientific debate. In turn this uncertainty fuels the fierce public and political disputes on the necessity of controlling badgers to limit the spread of infection. Here we present a dynamic stochastic spatial model for bovine TB in Great Britain that combines within-farm and between-farm transmission. At the farm scale the model incorporates stochastic transmission of infection, maintenance of infection in the environment and a testing protocol that mimics historical government policy. Between-farm transmission has a short-range environmental component and is explicitly driven by movements of individual cattle between farms, as recorded in the Cattle Tracing System. The resultant model replicates the observed annual increase of infection over time as well as the spread of infection into new areas. Given that our model is mechanistic, it can ascribe transmission pathways to each new case; the majority of newly detected cases involve several transmission routes with moving infected cattle, reinfection from an environmental reservoir and poor sensitivity of the diagnostic test all having substantive roles. This underpins our findings on the implications of control measures. Very few of the control options tested have the potential to reverse the observed annual increase, with only intensive strategies such as whole-herd culling or additional national testing proving highly effective, whereas controls focused on a single transmission route are unlikely to be highly effective. PMID:25008532

Brooks-Pollock, Ellen; Roberts, Gareth O; Keeling, Matt J

2014-07-10

215

Exploring causal networks of bovine milk fatty acids in a multivariate mixed model context  

PubMed Central

Background Knowledge regarding causal relationships among traits is important to understand complex biological systems. Structural equation models (SEM) can be used to quantify the causal relations between traits, which allow prediction of outcomes to interventions applied to such a network. Such models are fitted conditionally on a causal structure among traits, represented by a directed acyclic graph and an Inductive Causation (IC) algorithm can be used to search for causal structures. The aim of this study was to explore the space of causal structures involving bovine milk fatty acids and to select a network supported by data as the structure of a SEM. Results The IC algorithm adapted to mixed models settings was applied to study 14 correlated bovine milk fatty acids, resulting in an undirected network. The undirected pathway from C4:0 to C12:0 resembled the de novo synthesis pathway of short and medium chain saturated fatty acids. By using prior knowledge, directions were assigned to that part of the network and the resulting structure was used to fit a SEM that led to structural coefficients ranging from 0.85 to 1.05. The deviance information criterion indicated that the SEM was more plausible than the multi-trait model. Conclusions The IC algorithm output pointed towards causal relations between the studied traits. This changed the focus from marginal associations between traits to direct relationships, thus towards relationships that may result in changes when external interventions are applied. The causal structure can give more insight into underlying mechanisms and the SEM can predict conditional changes due to such interventions.

2014-01-01

216

Characterization of insulin-like growth factor binding proteins secreted by cultured bovine theca and granulosa cells.  

PubMed

Insulin-like growth factor binding proteins (IGFBPs) secreted by bovine granulosa and theca interna cells cultured in the presence of different luteinizing factors--insulin (2 micrograms/ml), forskolin (10 microM), or a combination of the two were examined and characterized. Direct binding of [125I]IGF to the conditioned media was compared to progesterone production under these different treatments. In theca cells, maximal secretion of IGFBPs was achieved using forskolin alone, whereas maximal progesterone production was induced by the insulin+forskolin treatment. In contrast, maximal secretion of both IGFBPs and progesterone in granulosa cells was achieved using forskolin alone. IGFBP species secreted by the two cell types under the different treatments were detected by ligand blotting. Conditioned media from theca cells in serum-free medium collected on the seventh day of culture exhibited three bands of 34, 40 and 44 kDa when treated with insulin or forskolin. The intensity of the 40-44 kDa complex was enhanced and a 21 kDa band appeared when cells were treated with a combination of insulin plus forskolin. Conditioned media of granulosa cells stimulated with insulin or forskolin exhibited 21, 27, 29, 34 and 40-44 kDa bands. Treatment with insulin+forskolin greatly increased the intensity of a 40-44 kDa complex. A similar shift towards high molecular weight binding proteins was observed when these media were analyzed by high-performance liquid chromatography gel filtration. These findings substantiate the secretion of IGFBPs by bovine theca and granulosa cells and show it to be dependent on culture treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1284495

Sakal, E; Gertler, A; Aflalo, L; Meidan, R

1992-12-01

217

Molecular modelling of protein-protein/protein-solvent interactions  

NASA Astrophysics Data System (ADS)

The inner workings of individual cells are based on intricate networks of protein-protein interactions. However, each of these individual protein interactions requires a complex physical interaction between proteins and their aqueous environment at the atomic scale. In this thesis, molecular dynamics simulations are used in three theoretical studies to gain insight at the atomic scale about protein hydration, protein structure and tubulin-tubulin (protein-protein) interactions, as found in microtubules. Also presented, in a fourth project, is a molecular model of solvation coupled with the Amber molecular modelling package, to facilitate further studies without the need of explicitly modelled water. Basic properties of a minimally solvated protein were calculated through an extended study of myoglobin hydration with explicit solvent, directly investigating water and protein polarization. Results indicate a close correlation between polarization of both water and protein and the onset of protein function. The methodology of explicit solvent molecular dynamics was further used to study tubulin and microtubules. Extensive conformational sampling of the carboxy-terminal tails of 8-tubulin was performed via replica exchange molecular dynamics, allowing the characterisation of the flexibility, secondary structure and binding domains of the C-terminal tails through statistical analysis methods. Mechanical properties of tubulin and microtubules were calculated with adaptive biasing force molecular dynamics. The function of the M-loop in microtubule stability was demonstrated in these simulations. The flexibility of this loop allowed constant contacts between the protofilaments to be maintained during simulations while the smooth deformation provided a spring-like restoring force. Additionally, calculating the free energy profile between the straight and bent tubulin configurations was used to test the proposed conformational change in tubulin, thought to cause microtubule destabilization. No conformational change was observed but a nucleotide dependent 'softening' of the interaction was found instead, suggesting that an entropic force in a microtubule configuration could be the mechanism of microtubule collapse. Finally, to overcome much of the computational costs associated with explicit soIvent calculations, a new combination of molecular dynamics with the 3D-reference interaction site model (3D-RISM) of solvation was integrated into the Amber molecular dynamics package. Our implementation of 3D-RISM shows excellent agreement with explicit solvent free energy calculations. Several optimisation techniques, including a new multiple time step method, provide a nearly 100 fold performance increase, giving similar computational performance to explicit solvent.

Luchko, Tyler

218

Sperm viability is influenced in vitro by the bovine seminal protein aSFP: Effects on motility, mitochondrial activity and lipid peroxidation  

Microsoft Academic Search

The 13 kDa acidic seminal fluid protein (aSFP) is a major component of bovine semen exerting growth factor-like activity. The influence of the pure protein on sperm viability was observed by evaluating sperm motility using computer-assisted semen analysis. Furthermore, mitochondrial dehydrogenase activity as a parameter of sperm metabolism and the integrity of sperm membranes using a metal catalyzed lipid peroxidation

C. Schöneck; J. Braun; R. Einspanier

1996-01-01

219

A peptide derived from human bactericidal\\/permeability-increasing protein (BPI) exerts bactericidal activity against Gram-negative bacterial isolates obtained from clinical cases of bovine mastitis  

Microsoft Academic Search

Gram-negative bacteria are responsible for approximately one-third of the clinical cases of bovine mastitis and can elicit a life-threatening, systemic inflammatory response. Lipopolysaccharide (LPS) is a membrane component of Gram-negative bacteria and is largely responsible for evoking the inflammatory response. Antibiotic and anti-inflammatory therapy for treating Gram-negative infections remains suboptimal. Bactericidal\\/permeability-increasing protein (BPI) is a neutrophil-derived protein with antimicrobial and

Annapoorani Chockalingam; Cindy E. McKinney; Manuela Rinaldi; Dante S. Zarlenga; Douglas D. Bannerman

2007-01-01

220

A VERSATILE MODEL OF DISEASE TRANSMISSION APPLIED TO FORECASTING BOVINE TUBERCULOSIS DYNAMICS IN WHITE-TAILED DEER POPULATIONS  

Microsoft Academic Search

A model was derived for disease transmission in dynamic host populations and its application was demonstrated in forecasting possible outcomes of a bovine tuberculosis (Myco- bacterium boris) epidemic in a white-tailed deer (Odocoileus virginianus) population. The ap- proach was mechanistic, based disease transmission on the probability of each susceptible indi- vidual becoming infected per unit time, and afforded the flexibility

C. W. McCarty; M. W. Miller

1998-01-01

221

Unraveling the binding mechanism of polyoxyethylene sorbitan esters with bovine serum albumin: A novel theoretical model based on molecular dynamic simulations.  

PubMed

To gain a better understanding of the interactions governing the binding mechanism of proteins with non-ionic surfactants, the association processes of Tween 20 and Tween 80 with the bovine serum albumin (BSA) protein were investigated using molecular dynamics (MD) simulations. Protein:surfactant molar ratios were chosen according to the critical micelle concentration (CMC) of each surfactant in the presence of BSA. It was found that both the hydrophilic and the hydrophobic groups of the BSA equally contribute to the surface area of interaction with the non-ionic surfactants. A novel theoretical model for the interactions between BSA and these surfactants at the atomic level is proposed, where both surfactants bind to non-specific domains of the BSA three-dimensional structure mainly through their polyoxyethylene groups, by hydrogen bonds and van der Waals interactions. This is well supported by the strong electrostatic and van der Waals interaction energies obtained in the calculations involving surfactant polyoxyethylene groups and different protein regions. The results obtained from the MD simulations suggest that the formation of surfactant clusters over the BSA structure, due to further cooperative self-assembly of Tween molecules, could increase the protein conformational stability. These results extend the current knowledge on molecular interactions between globular proteins and non-ionic surfactants, and contribute to the fine-tuning design of protein formulations using polysorbates as excipients for minimizing the undesirable effects of protein adsorption and aggregation. PMID:24309134

Delgado-Magnero, Karelia H; Valiente, Pedro A; Ruiz-Peńa, Miriam; Pérez-Gramatges, Aurora; Pons, Tirso

2014-04-01

222

Validation of bovine glycomacropeptide as an intestinal anti-inflammatory nutraceutical in the lymphocyte-transfer model of colitis.  

PubMed

Milk ?-casein-derived bovine glycomacropeptide (GMP) exerts immunomodulatory effects. It exhibits intestinal anti-inflammatory activity in chemically induced models of colitis. However, to validate its clinical usefulness as a nutraceutical, it is important to assess its effects in a model with a closer pathophysiological connection with human inflammatory bowel disease. Therefore, in the present study, we used the lymphocyte-transfer model of colitis in mice and compared the effects of GMP in this model with those obtained in the dextran sulphate sodium (DSS) model. GMP (15 mg/d) resulted in higher body-weight gain and a reduction of the colonic damage score and myeloperoxidase (MPO) activity in Rag1(-/-) mice with colitis induced by the transfer of naďve T cells. The colonic and ileal weight:length ratio was decreased by approximately 25%, albeit non-significantly. GMP treatment reduced the percentage of CD4? interferon (IFN)-?? cells in mesenteric lymph nodes (MLN). The basal production of IL-6 by MLN obtained from the GMP-treated mice ex vivo was augmented. However, concanavalin A-evoked production was similar. The colonic expression of regenerating islet-derived protein 3?, S100A8, chemokine (C-X-C motif) ligand 1 and IL-1? was unaffected by GMP, while that of TNF-? and especially IFN-? was paradoxically increased. In the DSS model, GMP also reduced the activity of colonic MPO, but it failed to alter weight gain or intestinal weight:length ratio. GMP augmented the production of IL-10 by MLN cells and was neutral towards other cytokines, except exhibiting a trend towards increasing the production of IL-6. The lower effect was attributed to the lack of the effect of GMP on epithelial cells. In conclusion, GMP exerts intestinal anti-inflammatory effects in lymphocyte-driven colitis. PMID:24229852

Ortega-González, Mercedes; Capitán-Cańadas, Fermín; Requena, Pilar; Ocón, Borja; Romero-Calvo, Isabel; Aranda, Carlos; Suárez, María Dolores; Zarzuelo, Antonio; Sánchez de Medina, Fermín; Martínez-Augustin, Olga

2014-04-14

223

Adsorption of bovine serum albumin on CoCrMo surface: effect of temperature and protein concentration.  

PubMed

The adsorption of bovine serum albumin (BSA) onto CoCrMo surface has been studied as a function of concentration of BSA and temperature by electrochemical techniques. The electrochemical impedance spectroscopy (EIS) technique was used to investigate the interfacial behaviour of BSA at open circuit potential (OCP). The charge transfer resistance was very sensitive to the amount of adsorbed protein, indicating that the adsorption process was accompanied by the transfer of charge and influenced the mechanism and kinetics of the corrosion reaction. At all the temperatures studied, adsorption of BSA onto the CoCrMo surface was successfully described with a Langmuir adsorption isotherm. EIS study was also carried out for determine the surface charge density, resulting from protein adsorption, and it was shown to be directly proportional to the amount of adsorbed protein (surface concentration). Thermodynamic data of adsorption was obtained for analyzing the adsorption of BSA onto CoCrMo surface. Gibbs free energy of adsorption, DeltaG(ADS) values, for BSA in the investigated temperature range (-51kJmol(-1)) showed that the molecules have a strong affinity for the CoCrMo surface. Enthalpy (DeltaH(ADS)) and entropy (DeltaS(ADS)) of adsorption suggested that the adsorption process of BSA onto the CoCrMo surface is an endothermic process and the molecule suffers structural changes when adsorbing on the metallic surface. PMID:20554436

Valero Vidal, C; Olmo Juan, A; Igual Muńoz, A

2010-10-01

224

Primary structure of bovine pituitary secretory protein I (chromogranin A) deduced from the cDNA sequence  

SciTech Connect

Secretory protein I (SP-I), also referred to as chromogranin A, is an acidic glycoprotein that has been found in every tissue of endocrine and neuroendocrine origin examined but never in exocrine or epithelial cells. Its co-storage and co-secretion with peptide hormones and neurotransmitters suggest that it has an important endocrine or secretory function. The authors have isolated cDNA clones from a bovine pituitary lambdagt11 expression library using an antiserum to parathyroid SP-I. The largest clone (SP4B) hybridized to a transcript of 2.1 kilobases in RNA from parathyroid, pituitary, and adrenal medulla. Immunoblots of bacterial lysates derived from SP4B lysognes demonstrated specific antibody binding to an SP4B/..beta..-galactosidase fusion protein (160 kDa) with a cDNA-derived component of 46 kDa. Radioimmunoassay of the bacterial lystates with SP-I antiserum yielded parallel displacement curves of /sup 125/I-labeled SP-I by the SP4B lysate and authentic SP-I. SP4B contains a cDNA of 1614 nucleotides that encodes a 449-amino acid protein (calculated mass, 50 kDa). The nucleotide sequences of the pituitary SP-I cDNA and adrenal medullary SP-I cDNAs are nearly identical. Analysis of genomic DNA suggests that pituitary, adrenal, and parathyroid SP-I are products of the same gene.

Ahn, T.G.; Cohn, D.V.; Gorr, S.U.; Ornstein, D.L.; Kashdan, M.A.; Levine, M.A.

1987-07-01

225

Primary structure of bovine pituitary secretory protein I (chromogranin A) deduced from the cDNA sequence.  

PubMed Central

Secretory protein I (SP-I), also referred to as chromogranin A, is an acidic glycoprotein that has been found in every tissue of endocrine and neuroendocrine origin examined but never in exocrine or epithelial cells. Its co-storage and co-secretion with peptide hormones and neurotransmitters suggest that it has an important endocrine or secretory function. We have isolated cDNA clones from a bovine pituitary lambda gt11 expression library using an antiserum to parathyroid SP-I. The largest clone (SP4B) (approximately equal to 1.6 kilobases) hybridized to a transcript of 2.1 kilobases in RNA from parathyroid, pituitary, and adrenal medulla. Immunoblots of bacterial lysates derived from SP4B lysogens demonstrated specific antibody binding to an SP4B/beta-galactosidase fusion protein (160 kDa) with a cDNA-derived component of 46 kDa. Radioimmunoassay of the bacterial lysates with SP-I antiserum yielded parallel displacement curves of 125I-labeled SP-I by the SP4B lysate and authentic SP-I. SP4B contains a cDNA of 1614 nucleotides that encodes a 449-amino acid protein (calculated mass, 50 kDa). The nucleotide sequences of the pituitary SP-I cDNA and adrenal medullary SP-I cDNAs are nearly identical. Analysis of genomic DNA suggests that pituitary, adrenal, and parathyroid SP-I are products of the same gene. Images

Ahn, T G; Cohn, D V; Gorr, S U; Ornstein, D L; Kashdan, M A; Levine, M A

1987-01-01

226

Mapping the calcification of bovine pericardium in rat model by enhanced micro-computed tomography.  

PubMed

The calcification initiation and progression of bioprosthetic heart valve were investigated in a rat model by enhanced micro-computed tomography, together with histologic study and scanning electron microscope analysis. The implantation data at early stage showed apparent dendritic patterns in the radiographic images for the glutaraldehyde-treated bovine pericardium and this dendritic pattern was verified to be associated with the vessel distribution in the tissue. Histologic study and scanning electron microscope analysis both indicated that the calcium deposits in the pericardium vessels regions were more grievous than those scattered in the collagen fibers in the first two weeks after implantation. Subsequently, calcification spreaded and the entire sample was severely calcified in 60 days. PMID:24973299

Liu, Jing; Zhong, Shengping; Lan, Hualin; Meng, Xu; Zhang, Haibo; Fan, Yubo; Wang, Yuxing; Wang, Chunren; Wang, Zhaoxu

2014-09-01

227

Proteolytic cleavage and shedding of the bovine prion protein in two cell culture systems.  

PubMed

We have compared the processing, turnover and release of bovine PrP (boPrP) in transfected baby hamster kidney (BHK) and mouse neuroblastoma (N2a) cells. In BHK cells, boPrP was subjected to two distinct proteolytic cleavage events, the first was mapped between K(121) and H(122) generating an N-terminal and a C-terminal PrP fragment. Transport block experiments, cell surface biotinylation and PIPLC analyses showed that the bulk of boPrP on the cell surface was the C-terminal fragment and indicated that the first cleavage of boPrP took place prior to or very soon after it appears at the cell surface. The second cleavage was situated at the extreme C-terminus of the boPrP GPI-anchored C-terminal fragment and as a result of this was shed into the medium rapidly. The kinetics, the migration in SDS-PAGE of the released fragment and protease inhibition studies indicate that a proteolytic activity was responsible for the release of the boPrP fragment from its GPI-anchor. Both N- and C-terminal fragments of boPrP could be detected in the medium. Moreover, in normal bovine brain, a C-terminal fragment was identified, suggesting that similar proteolytic processing events occur in vivo. In N2a cells, the majority of boPrP was subjected to a more complete degradation process, and only trace amounts of full length boPrP was shed into cell culture medium in a process which also indicated a release by proteolytic cleavage. PMID:16140411

Zhao, Hongxing; Klingeborn, Mikael; Simonsson, Magnus; Linné, Tommy

2006-01-01

228

Modeling of protein interaction networks  

Microsoft Academic Search

We introduce a graph generating model aimed at representing the evolution of\\u000aprotein interaction networks. The model is based on the hypotesis of evolution\\u000aby duplications and divergence of the genes which produce proteins. The\\u000aobtained graphs shows multifractal properties recovering the absence of a\\u000acharacteristic connectivity as found in real data of protein interaction\\u000anetworks. The error tolerance of

Alexei Vazquez; Alessandro Flammini; Amos Maritan; Alessandro Vespignani

2001-01-01

229

Pattern-matching models for the differential diagnosis of bovine spongiform encephalopathy.  

PubMed

This study assessed the performance of a system for making decisions about the diagnosis of bovine spongiform encephalopathy (BSE). The system consisted of four pattern-matching models. The sensitivity, specificity, likelihood ratios and accuracy of each model were determined by using clinical descriptions of 100 suspect BSE cases which had been submitted for brain histopathology by veterinary officers of the Ministry of Agriculture, Fisheries and Food, 50 of which were true positive cases (confirmed by histopathology) and 50 false positive cases (not confirmed by histopathology). The clinical description of each case consisted of 14 clinical signs, each of which was defined as either present or absent. The system compared the case descriptions with the profiles of possible differential diagnoses, each profile consisting of the frequency of occurrence of the same 14 clinical signs. The pattern-matching models used the sums of the sign frequencies to rank the differential diagnoses. Models 1 and 2 derived information only from the presence of signs; models 3 and 4 derived information from the presence and absence of signs. Models 2 and 4 excluded diagnoses which did not have in their profile a sign which was observed, and diagnoses which had a sign in their profile which should always be present according to the profile description but which was not observed. The best performances by the models were: sensitivity 96 per cent (model 1 and model 2), specificity 72 per cent (model 4), accuracy 72 per cent (model 4), likelihood ratio of a positive test 2.00 (model 4), likelihood ratio of a negative test 0.21 (model 4). PMID:10390800

Cockcroft, P D

1999-05-29

230

Application of an exponential model to early postmortem bovine muscle pH decline.  

PubMed

An exponential decay equation was used to describe the decline of pH in bovine M. longissimus thoracis et lumborum (LTL) muscle postmortem. Goodness of fit of the equation was tested by convergence of model criterion and contrasting predicted against expected pH values of LTL muscles from 24 Bos taurus steer carcasses measured 40, 60, 80, 120, 185 and 1440 min after stunning. The model was fitted to pH values measured, as well as to the measured pH values adjusted to arbitrary constant temperatures of 10, 20 or 30°C. The model failed to converge for 46% of the muscles sampled in the data sets with measured pH. The model converged for 100% of the muscles with pH values adjusted to 10, 20 or 30°C. Limits of agreement, heteroscedasticity and intraclass correlation analyses showed little difference in the fit of the model to pH values adjusted to either 10, 20 or 30°C. Fitting this model to pH data adjusted to 20°C will enable comparison of rates of pH decline between muscles of different temperatures postmortem or laboratory homogenates and facilitate relation of pH decline rate to aspects of beef quality. PMID:22061917

Bruce, H L; Scott, J R; Thompson, J M

2001-05-01

231

Bovine herpesvirus 1 glycoprotein M forms a disulfide-linked heterodimer with the U(L)49.5 protein.  

PubMed

Nine glycoproteins (gB, gC, gD, gE, gG, gH, gI, gK, and gL) have been identified in bovine herpesvirus 1 (BHV-1). gM has been identified in many other alpha-, beta-, and gammaherpesviruses, in which it appears to play a role in membrane penetration and cell-to-cell fusion. We sought to express BHV-1 open reading frame U(L)10, which encodes gM, and specifically identify the glycoprotein. We corrected a frameshift error in the published sequence and used the corrected sequence to design coterminal peptides from the C terminus. These were expressed as glutathione S-transferase fusion proteins in Escherichia coli. The fusion protein containing the 63 C-terminal amino acids from the corrected gM sequence engendered antibodies that immunoprecipitated a 30-kDa protein from in vitro translation reactions programmed with the U(L)10 gene. Proteins immunoprecipitated by this antibody from virus-infected cells ran at 36 and 43 kDa in reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and 43 and 48 kDa in nonreducing SDS-PAGE. Only the larger of the pair was present in virions. A 7-kDa protein was released from gM by reducing agents. The 7-kDa protein was not recognized in Western blots probed with the anti-gM antibody but reacted specifically with antibodies prepared against BHV-1 U(L)49.5, previously reported to be a 9-kDa protein associated with an unidentified 39-kDa protein (X. Liang, B. Chow, C. Raggo, and L. A. Babiuk, J. Virol. 70:1448-1454, 1996). This is the first report of a small protein covalently bound to any herpesvirus gM. Similar patterns of hydrophobic domains and cysteines in all known gM and U(L)49.5 homologs suggest that these two proteins may be linked by disulfide bonds in all herpesviruses. PMID:9525625

Wu, S X; Zhu, X P; Letchworth, G J

1998-04-01

232

Predicting bovine milk protein composition based on Fourier transform infrared spectra  

Microsoft Academic Search

Phenotypic information on individual protein composition of cows is important for many aspects of dairy processing with cheese production as the center of gravity. However, measuring individual protein composition is expensive and time consuming. In this study, we investigated whether protein composition can be predicted based on inexpensive and routinely measured milk Fourier transform infrared (FTIR) spectra. Based on 900

M. J. M. Rutten; H. Bovenhuis; J. M. L. Heck; J. A. M. van Arendonk

2011-01-01

233

Protein designs in HP models  

NASA Astrophysics Data System (ADS)

The inverse protein folding problem is that of designing an amino acid sequence which folds into a prescribed shape. This problem arises in drug design where a particular structure is necessary to ensure proper protein-protein interactions and could have applications in nanotechnology. A major challenge in designing proteins with native folds that attain a specific shape is to avoid proteins that have multiple native folds (unstable proteins). In this technical note we present our results on protein designs in the variant of Hydrophobic-Polar (HP) model introduced by Dill [6] on 2D square lattice. The HP model distinguishes only polar and hydrophobic monomers and only counts the number of hydrophobic contacts in the energy function. To achieve better stability of our designs we use the Hydrophobic-Polar-Cysteine (HPC) model which distinguishes the third type of monomers called ``cysteines'' and incorporates also the disulfid bridges (SS-bridges) into the energy function. We present stable designs in 2D square lattice and 3D hexagonal prism lattice in the HPC model.

Gupta, Arvind; Khodabakhshi, Alireza Hadj; Ma?uch, Ján; Rafiey, Arash; Stacho, Ladislav

2009-07-01

234

Effects of dietary cottonseed oil and tannin supplements on protein and fatty acid composition of bovine milk.  

PubMed

This experiment was conducted to determine the effects of diets supplemented with cottonseed oil, Acacia mearnsii-condensed tannin extract, and a combination of both on composition of bovine milk. Treatment diets included addition of cottonseed oil (800 g/d; CSO), condensed tannin from Acacia mearnsii (400 g/d; TAN) or a combination of cottonseed oil (800 g/d) and condensed tannin (400 g/d; CPT) with a diet consisting of 6·0 kg dry matter (DM) of concentrates and alfalfa hay ad libitum, which also served as the control diet (CON). Relative to the CON diet, feeding CSO and CPT diets had a minor impact on feed intake and yield of lactose in milk. These diets increased yields of milk and protein in milk. In contrast to the TAN diet, the CSO and CPT diets significantly decreased milk fat concentration and altered milk fatty acid composition by decreasing the proportion of saturated fatty acids but increasing proportions of monounsaturated and polyunsaturated fatty acids. The CPT diet had a similar effect to the CSO diet in modifying fatty acid profile. Overall, reduction in milk fat concentration and changes in milk fatty acid profile were probably due to supplementation of linoleic acid-rich cottonseed oil. The TAN diet had no effect on feed intake, milk yield and milk protein concentration. However, a reduction in the yields of protein and lactose occurred when cows were fed this diet. Supplemented tannin had no significant effect on fat concentration and changes in fatty acid profile in milk. All supplemented diets did not affect protein concentration or composition, nitrogen concentration, or casein to total protein ratio of the resulting milk. PMID:24594257

Aprianita, Aprianita; Donkor, Osaana N; Moate, Peter J; Williams, S Richard O; Auldist, Martin J; Greenwood, Jae S; Hannah, Murray C; Wales, William J; Vasiljevic, Todor

2014-05-01

235

Short communication: Bovine ?-casein is a ferritin-binding protein and inhibitory factor of milk ferritin immunoassay  

Microsoft Academic Search

Commercial bovine milk ?-casein, but not ?- and ?-caseins, bound to bovine spleen ferritin, as determined by an immunoassay for ferritin. In contrast, ?-casein did not bind to apoferritin. The binding of ?-casein to bovine spleen ferritin was strongly inhibited by increas- ing ionic strength by the addition of 0.5 M (NH4)2SO4. The addition of ?-casein to a known amount

G. Sugawara; R. Inoue; K. Watanabe; H. Ohtsuka; K. Orino

2009-01-01

236

Device induced thromboembolism in a bovine in vitro coronary stent model.  

PubMed

The potential of a new bovine in vitro model to evaluate various aspects of device induced thromboembolism was studied using two test modes. First, the effect of an antithrombotic drug on stent induced thromboembolism was assessed. The antithrombotic potential of an antiplatelet agent was compared with that of the other conventional antithrombotic agents (aspirin, dipyridamole) used in the past with this in vitro model. Stent associated thrombus was assessed gravimetrically at the end of the experiment. Emboli were assessed continuously using a light scattering microemboli detection system. Second, the sensitivity of the model to flow induced thromboembolism was studied using a combination of surface roughness and stenosis. Thrombus was assessed visually, and emboli were assessed as described earlier. The results show that 1) this in vitro model is sensitive to the action of antithrombotic drugs, and to the effect of hemodynamics on thromboembolism; 2) the antiplatelet drug used in this study was effective in attenuating thromboembolism; 3) a stenosis in combination with roughness produced more emboli than roughness alone; and 4) the model was useful for the study of physical and biochemical aspects of thromboembolism. PMID:9804458

Sukavaneshvar, S; Solen, K A; Mohammad, S F

1998-01-01

237

Generation of the bovine viral diarrhea virus e0 protein in transgenic astragalus and its immunogenicity in sika deer.  

PubMed

The bovine viral diarrhea virus (BVDV), a single-stranded RNA virus, can cause fatal diarrhea syndrome, respiratory problems, and reproductive disorders in herds. Over the past few years, it has become clear that the BVDV infection rates are increasing and it is likely that an effective vaccine for BVDV will be needed. In this study, transgenic Astragalus was used as an alternative productive platform for the expression of glycoprotein E0. The immunogenicity of glycoprotein E0 expressed in transgenic Astragalus was detected in deer. The presence of pBI121-E0 was confirmed by polymerase chain reaction (PCR), transcription was verified by reverse transcription- (RT-) PCR, and recombinant protein expression was confirmed by ELISA and Western blot analyses. Deer that were immunized subcutaneously with the transgenic plant vaccine developed specific humoral and cell-mediated immune responses against BVDV. This study provides a new method for a protein with weak immunogenicity to be used as part of a transgenic plant vaccine. PMID:24963321

Gao, Yugang; Zhao, Xueliang; Zang, Pu; Liu, Qun; Wei, Gongqing; Zhang, Lianxue

2014-01-01

238

Generation of the Bovine Viral Diarrhea Virus E0 Protein in Transgenic Astragalus and Its Immunogenicity in Sika Deer  

PubMed Central

The bovine viral diarrhea virus (BVDV), a single-stranded RNA virus, can cause fatal diarrhea syndrome, respiratory problems, and reproductive disorders in herds. Over the past few years, it has become clear that the BVDV infection rates are increasing and it is likely that an effective vaccine for BVDV will be needed. In this study, transgenic Astragalus was used as an alternative productive platform for the expression of glycoprotein E0. The immunogenicity of glycoprotein E0 expressed in transgenic Astragalus was detected in deer. The presence of pBI121-E0 was confirmed by polymerase chain reaction (PCR), transcription was verified by reverse transcription- (RT-) PCR, and recombinant protein expression was confirmed by ELISA and Western blot analyses. Deer that were immunized subcutaneously with the transgenic plant vaccine developed specific humoral and cell-mediated immune responses against BVDV. This study provides a new method for a protein with weak immunogenicity to be used as part of a transgenic plant vaccine.

Gao, Yugang; Zang, Pu; Liu, Qun; Wei, Gongqing

2014-01-01

239

Characteristics of Several Minor-Protein Fractions Isolated from Bovine Milk1  

Microsoft Academic Search

SUMMARY Four minor-protein fractions were isolated from the same source of skimmilk by methods similar to those reported by Rowland for proteose-peptone, by Aschaffenberg for a-proteose, by Jenness for milk component 5, and by Weinstein for the minor-protein fraction. A fifth fraction, the soluble membrane-protein, a constituent of the fat\\/plasma interfacial layer, was isolated from washed cream. These fractions were

J. R. Brunner; M. P. Thompson

1961-01-01

240

Modelling transmission of bovine tuberculosis in red deer and wild boar in Normandy, France.  

PubMed

In early 2001, Mycobacterium bovis infection was confirmed in red deer (RD) (Cervus elaphus) shot in Normandy region, France. An epidemiological survey conducted during the following hunting season in two connected forests confirmed the occurrence of the disease in both free-ranging RD and wild boar (WB) (Sus scrofa). This was the first detected bovine tuberculosis outbreak in wildlife in France. We present a simple deterministic age-structured model of the within- and between-species M. bovis transmission in RD and WB populations that distinguishes direct transmission (horizontal and pseudo-vertical) and indirect transmission through contaminated offal left behind by hunters. Results issued from the epidemiological surveys conducted in Normandy forests were used to estimate transmission parameters. Because data for RD and WB populations were not available, population sizes at demographic equilibrium were estimated and used to run the model. We qualitatively tested different control measure scenarios with our model, considering different mortality rates and offal harvesting, to determine which ones affect the success of infection control. The most realistic control scenario would combine the total depopulation of RD and good compliance with offal harvesting, because the model suggests that infected offal left by hunters represents the main transmission source of M. bovis in the field. PMID:22958262

Zanella, G; Bar-Hen, A; Boschiroli, M-L; Hars, J; Moutou, F; Garin-Bastuji, B; Durand, B

2012-09-01

241

Influences of different thermal processings in milk, bovine meat and frog protein structure.  

PubMed

Several studies have associated the digestibility of proteins to its imunogenic potential. Though, it was objectified to evaluate the impact of the thermal processing with high and low temperatures on the proteins structure of three types of foods, by means of the digestibility in vitro and electroforesis en gel de poliacrilamida. The pasteurize was observed in such a way, firing 95 şC during 15 minutes, how much freeze dried causes qualitative and quantitative modifications of constituent proteins of the food. The most sensible proteins to the increasing thermal processing order were beef, frog meat, and the last, cow milk. PMID:23848117

Coura Oliveira, Tatiana; Lopes Lima, Samuel; Bressan, Josefina

2013-01-01

242

Regional variations in the distribution and colocalization of extracellular matrix proteins in the juvenile bovine meniscus  

PubMed Central

A deeper understanding of the composition and organization of extracellular matrix molecules in native, healthy meniscus tissue is required to fully appreciate the degeneration that occurs in joint disease and the intricate environment in which an engineered meniscal graft would need to function. In this study, regional variations in the tissue-level and pericellular distributions of collagen types I, II and VI and the proteoglycans aggrecan, biglycan and decorin were examined in the juvenile bovine meniscus. The collagen networks were extensively, but not completely, colocalized, with tissue-level organization that varied with radial position across the meniscus. Type VI collagen exhibited close association with large bundles composed of type I and II collagen and, in contrast to type I and II collagen, was further concentrated in the pericellular matrix. Aggrecan was detected throughout the inner region of the meniscus but was restricted to the pericellular matrix and sheaths of collagen bundles in the middle and outer regions. The small proteoglycans biglycan and decorin exhibited regional variations in staining intensity but were consistently localized in the intra- and/or peri-cellular compartments. These results provide insight into the complex hierarchy of extracellular matrix organization in the meniscus and provide a framework for better understanding meniscal degeneration and disease progression and evaluating potential repair and regeneration strategies.

Vanderploeg, Eric J; Wilson, Christopher G; Imler, Stacy M; Ling, Carrie Hang-Yin; Levenston, Marc E

2012-01-01

243

Protein Self-Association in Solution: The Bovine Pancreatic Trypsin Inhibitor Decamer  

PubMed Central

We have used magnetic relaxation dispersion to study bovine pancreatic trypsin inhibitor (BPTI) self-association as a function of pH, salt type and concentration, and temperature. The magnetic relaxation dispersion method sensitively detects stable oligomers without being affected by other interactions. We find that BPTI decamers form cooperatively under a wide range of solution conditions with no sign of dimers or other small oligomers. Decamer formation is opposed by electrostatic repulsion among numerous cationic residues confined within a narrow channel. Accordingly, the decamer population increases with increasing pH, as cationic residues are deprotonated, and with increasing salt concentration. The salt effect cannot be described in terms of Debye screening, but involves the ion-specific sequestering of anions within the narrow channel. The lifetime of the BPTI decamer is 101 ± 4 min at 27°C. We propose that the BPTI decamer, with a heparin chain threading the decamer channel, plays a functional role in the mast cell. We also detect a higher oligomer that appears to be a subcritical nucleation cluster of 3–5 decamers. We argue that monomeric crystals form at high pH despite a high decamer population in solution, because the ion pairs that provide the critical decamer-decamer contacts are disrupted at high pH.

Gottschalk, Michael; Venu, Kandadai; Halle, Bertil

2003-01-01

244

Effect of trenbolone acetate on protein synthesis and degradation rates in fused bovine satellite cell cultures  

Microsoft Academic Search

Although androgenic and estrogenic steroids are widely used to enhance muscle growth and increase feed efficiency in feedlot cattle, their mechanism of action is not well understood. Although in vivo studies have indicated that androgens affect protein synthesis and protein degradation rate in muscle, results from in vitro studies have been inconsistent. We have examined the effects of trenbolone acetate

E. Kamanga-Sollo; M. E. White; M. R. Hathaway; W. J. Weber; W. R. Dayton

2011-01-01

245

Comparison of PTFE, pericardium bovine and fascia lata for repair of incisional hernia in rat model, experimental study.  

PubMed

Incisional hernia is a frequent complication of abdominal surgery developing in 11-20 % of patients undergoing an abdominal operation. Regarding morbidity and loss of manpower, incisional hernias continue to be a fundamental problem for surgeons. In this experimental study, three commonly used mesh materials (Goretex PTFE; Tutoplast Fascia lata; Tutopatch Pericardium bovine) were compared according to effectiveness, strength, adhesion formation, histological changes, and early complications. Three groups, each consisting of 14 rats, have been formed as group A: polytetrafluoroethylene (PTFE), group B: pericardium bovine and group C: fascia lata. Evaluations were achieved at the end of the first and second postoperative week, respectively. Adhesion formation, wound maturation, bursting pressure, and tensile strength were evaluated. No statistically significant difference regarding adhesion formation was observed between groups although adhesion formation was less significant in PTFE and pericardium bovine groups than in the fascia lata group. Bursting pressure and tensile strength values were significantly higher in PTFE group than in the fascia lata group ( P<0.05). No statistically significant difference was observed between groups regarding wound maturation. In this experimental model, PTFE and pericardium bovine were found to be superior to fascia lata in abdominal wall repair. PMID:12612797

Kapan, S; Kapan, M; Goksoy, E; Karabicak, I; Oktar, H

2003-03-01

246

Fatty acid-binding protein activities in bovine muscle, liver and adipose tissue  

SciTech Connect

Subcutaneous adipose tissue, sternomandibularis muscle and liver were obtained from steers immediately postmortem. Muscle strips and adipose tissue snips were incubated with 0.75 mM (1- UC)palmitate and 5 mM glucose. Muscle strips esterified palmitate at the rate of 2.5 nmol/min per gram tissue, which was 30% of the rate observed for adipose tissue. Fatty acid-binding protein activity was measured in 104,000 x g supernatant fractions of liver, muscle and adipose tissue homogenates. Muscle and adipose tissue fractions bound 840 and 140 pmol (1- UC)palmitoyl-CoA per gram tissue, respectively. Fatty acid-binding protein activity was greater in adipose tissue than in muscle when data were expressed per milligram protein. Fatty acid binding-protein activity was correlated with the rate of palmitate esterification within each tissue. Liver contained the highest fatty acid-binding protein activity.

Smith, S.B.; Ekeren, P.A.; Sanders, J.O.

1985-11-01

247

Two cellular single-strand-specific DNA-binding proteins interact with two regions of the bovine papillomavirus type 1 genome, including the origin of DNA replication.  

PubMed Central

We have identified and purified to near homogeneity two specific single-stranded DNA-binding factors (SPSF I and II) with molecular masses of 42 and 39 kDa, respectively, from calf thymus. Gel retention analysis and competition experiments demonstrate that the ubiquitous proteins SPSF I and II specifically interact with single-stranded DNA derived from the minimal in vitro origin of replication of bovine papillomavirus type 1 and a region of the viral genome proposed to be involved in plasmid maintenance. Bovine papillomavirus type 1 proteins do not interfere with DNA binding of SPSF I and II. The exact location of the binding domains of SPSF I and II on the DNA has been determined by methylation interference and T4 DNA polymerase footprinting. A potential cellular binding site for SPSF I and II is the major promoter (P2) of the human c-myc gene. Images

Habiger, C; Stelzer, G; Schwarz, U; Winnacker, E L

1992-01-01

248

Blocking of CD4 cell receptors for the human immunodeficiency virus type 1 (HIV-1) by chemically modified bovine milk proteins: potential for AIDS prophylaxis.  

PubMed

The chemical transformation of synthetic combinatorial libraries to increase the diversity of compounds of medicinal interest was reported recently. Chemical modification of natural products represents a complementary approach to accomplish this aim. Modification of lysines by aromatic acid anhydrides, preferentially by 3-hydroxyphthalic and trimellitic anhydrides and trimellitic anhydride chloride, converted commonly available proteins (human and bovine serum albumin and casein) into potent inhibitors of (i) binding between the HIV-1 gp 120 envelope glycoprotein and the CD4 cell receptor, probably owing to their binding to CD4, and (ii) infection by HIV-1. Modified bovine milk proteins are also potent HIV-1 inhibitors and may have potential for anti-HIV-1 prophylaxis. PMID:8619951

Neurath, A R; Debnath, A K; Strick, N; Li, Y Y; Lin, K; Jiang, S

1995-01-01

249

Ca2+ and Zn2+-binding properties of nitrated S-100b protein from bovine brain.  

PubMed Central

The single tyrosine residue in S-100b protein was nitrated by treatment with tetranitromethane in 0.1 M-Tris/HCl buffer, pH 8.0, containing 2 mM-EDTA. The nitrated protein did not differ significantly in secondary structure from its native unmodified counterpart, as revealed by far-u.v. c.d. measurements. The effect of Ca2+ on the modified protein was different from that on the native protein, e.g. addition of Ca2+ resulted in a loss of helical content from 55 to 47% with the native protein whereas Ca2+ had no significant effect on the gross conformation of the nitrated derivative. Near-u.v. c.d. studies also indicated a very minimal effect on the tyrosine residue and this was also reflected in the u.v.-absorption difference spectrum. Polyacrylamide-gel electrophoresis in the absence of SDS showed the nitrated S-100b to move faster in the presence of EDTA compared with the calcium-bound state, suggesting that the modified protein does bind Ca2+ although it does not undergo a major conformational change in response to Ca2+ addition. In contradistinction, Zn2+ binding was not influenced by nitration, as demonstrated by aromatic c.d. and u.v.-difference spectroscopy. It is clear from this study that the single tyrosine residue in S-100b is critical to sense the Ca2+-induced conformational changes in the protein. Images Fig. 1.

Mani, R S; Kay, C M

1986-01-01

250

From explanation to prediction: a model for recurrent bovine tuberculosis in Irish cattle herds.  

PubMed

There is a good understanding of factors associated with bovine tuberculosis (BTB) risk in Irish herds. As yet, however, this knowledge has not been incorporated into predictive models with the potential for improved, risk-based surveillance. The goal of the study was to enhance the national herd scoring system for BTB risk, thus leading to improved identification of cattle herds at high risk of recurrent BTB episodes. A retrospective cohort study was conducted to develop a statistical model predictive of recurrent bovine tuberculosis episodes in cattle herds in the Republic of Ireland. Herd-level disease history data for the previous 12 years, the previous 3 years, the previous episode, and the current-episode were used in survival analyses to determine the aspects of disease history that were predictive of a recurrent breakdown within 3 years of a cleared BTB episode. Relative to herds with 0-1 standard reactors in the current BTB episode, hazard ratios increased to 1.3 and 1.6 for herds with 2-5 and >5 standard reactors, respectively. Compared to herds with <30 animals, hazard ratios increased from 1.8 to 2.5 and then to 3.1 for herds with 30-79, 80-173, and >174 animals respectively. Relative to herds with <4 herd-level tests in the previous 3 years, herds with 4-5 and >5 tests had 1.1 and 1.4 times greater hazard of a BTB breakdown. Herds that did not have a BTB episode in the 5 years prior to their 2001 episode were 0.8 times less likely to breakdown in the next 3 years than herds that did. Herds breaking down in the spring or summer were 0.8 times less likely to suffer a recurrent breakdown than herds breaking down in autumn or winter (this was likely due to seasonality in testing regimes). The presence of a confirmed BTB lesion was not predictive of increased risk of recurrent BTB. Despite the availability of detailed disease history, the predictive ability of the model was poor. One explanation for this was that herds suffering a recurrence of BTB on their first test after clearing a BTB episode were different from herds that broke down later in the period at risk. Future research might need to include additional variables to identify which subsets of herd BTB episodes, if any, have identifiable features that are predictive of recurrent breakdowns. PMID:20236717

Wolfe, Dianna M; Berke, Olaf; Kelton, David F; White, Paul W; More, Simon J; O'Keeffe, James; Martin, S Wayne

2010-05-01

251

Immunohistochemical study of seminiferous epithelium in adult bovine testis using monoclonal antibodies against Ki67 protein and proliferating cell nuclear antigen (PCNA)  

Microsoft Academic Search

The distribution pattern of proliferating cell nuclear antigen (PCNA) and Ki-67 protein was studied in adult bovine seminiferous epithelium by means of immunohistochemistry using monoclonal antibodies. Tailoring the methodological protocol for each of the two proliferation markers was a necessary prerequisite for obtaining optimal results in tubular sections and whole-mounts. A-, I- and B-spermatogonia displayed PCNA-positive nuclei, except during meta-,

Karl-Heinz Wrobel; Daniela Bickel; Richard Kujat

1996-01-01

252

cDNA Cloning of S100 Calcium-binding Proteins from Bovine Periodontal Ligament and Their Expression in Oral Tissues  

Microsoft Academic Search

The periodontal ligament (PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. To characterize PDL cells at the molecular level, we constructed a cDNA library from bovine PDL tissue. We then focused on the isolation of S100 calcium-binding proteins (CaBPs), because they mediate Ca2+ signaling and control

W. R. Duarte; S. Kasugai; T. Iimura; S. Oida; K. Takenaga; K. Ohya; I. Ishikawa

1998-01-01

253

Process standardization for optimal virus recovery and removal of substrate DNA and bovine serum proteins in Vero cell-derived rabies vaccine.  

PubMed

Purification of a rabies vaccine by a single zonal centrifugation run was replaced by two runs with optimal standardization of the sucrose density gradient. As a result, significant reductions in the levels of substrate DNA and bovine serum protein in the Vero cell-derived human rabies vaccine were achieved. Following many trials, for the first run, loading of the 3.2-l capacity K-3 rotor with 1800 ml of 60% sucrose solution and 1400 ml of vaccine PBS buffer solution gave a satisfactory linear gradient. However, after the first run, the substrate DNA and bovine serum contents exceeded the required levels. After protamine sulphate and Tween-80 treatment of the concentrated inactivated material, a second run using the same procedure as in the first run was tried. However, these purification procedures resulted in low virus recovery. To achieve optimal virus recovery, and removal of substrate DNA and bovine serum protein, the peak fractions from the first run as indicated by the haemagglutination, sucrose concentration, and optical density values were pooled and the sucrose concentration of the pooled fractions was increased to 60%. A second (flotation) run was then carried out. Using this method, the virus recovery rate was more than 95% that of the first run, and the levels of cellular DNA and bovine serum protein were well within the acceptable limits of less than 100 pg/dose and one part per million, respectively. The substrate DNA was quantified by both radioactive labeling and non-radioactive biotin labeling methods. For the quantification of calf serum protein, a counter-immunoelectrophoresis method was developed and effectively applied. A potency assay was performed using the National Institutes of Health (NIH) and well-standardized in vitro single radial immuno diffusion (SRD) methods. Finally, an immunogenicity study was conducted with human volunteers and the results were confirmed by a rapid fluorescent focus inhibition test (RFFIT). PMID:16233321

Kumar, Ananda Arone Prem; Rao, Yarlagadda Udaya Bhaskara; Joseph, Arokiaswami Leo William; Mani, Kavaratty Raju; Swaminathan, Krishnaswami

2002-01-01

254

Induction of humoral and cellular immune responses in mice by a recombinant fowlpox virus expressing the E2 protein of bovine viral diarrhea virus  

Microsoft Academic Search

A recombinant fowlpox virus (rFPV\\/E2) expressing the E2 protein of bovine viral diarrhea virus (BVDV) was constructed and characterized. Mice were immunized with recombinant virus and both humoral and cellular immune responses were studied. rFPV\\/E2 induced BVDV-specific antibodies which were detected by ELISA. In addition, mouse sera were shown to neutralize BVDV. A cytokine ELISA assay revealed that mice vaccinated

Seyyed Mehdy Elahi; Jean Bergeron; Éva Nagy; Brian Geoffrey Talbot; Serge Harpin; Shi-Hsiang Shen; Youssef Elazhary

1999-01-01

255

ARPP-2 1, a Cyclic AMP-Regulated Phosphoprotein Enriched in Dopamine-Innervated Brain Regions. I. Purification and Characterization of the Protein from Bovine Caudate Nucleus  

Microsoft Academic Search

ARPP-21 (CAMP-regulated phosphoprotein, M, = 27,000 as determined by SDS\\/PAGE) is a major cytosolic substrate for CAMP-stimulated protein phosphorylation in dopamine-in- nervated regions of rat CNS (Walaas et al., 1983~). This acidic phosphoprotein has now been identified in bovine caudate nucleus cytosol and purified to homogeneity from this source. The purification procedure involved diethyl- aminoethyl-cellulose chromatography, ammonium sulfate fractionation, phenyl-Sepharose

Hugh C. Hemmings; Paul Greengard

256

Molecular characterization of RNA and protein synthesis during a one-step growth curve of bovine viral diarrhoea virus in ovine (SFT-R) cells  

Microsoft Academic Search

The aim of this study was to determine the kinetics of noncytopathic bovine viral diarrhoea virus (BVDV) multiplication and synthesis of BVDV specific RNA and proteins in ovine cells (SFT-R) during a one-step growth curve. The virus titre and RNA level were determined by focus-forming assay and real time RT-PCR. The RNA synthesis was detected by Northern blot while synthesis

N. Mishra; B. S. Mathapati; K. Rajukumar; R. K. Nema; S. P. Behera; S. C. Dubey

2010-01-01

257

Effects of Environmental Factors and Milk Protein Polymorphism on Composition of Casein Fraction in Bovine Milk  

Microsoft Academic Search

Test-day samples were collected from individual Holstein cows in 62 herds enrolled in the Quebec Dairy Herd Analysis Service. Samples were analyzed for protein, fat, casein, and serum protein content, somatic cell count, and relative percentages of as- , \\/3-, and K-casein and a-lactalbumin. Cows included in the study were phenotyped for the genetic variants of as1- , t3-, and

E. M. Kroeker; K. F. Ng-Kwai-Hang; J. F. Hayes; J. E. Moxley

1985-01-01

258

Bovine Serum Albumin Glutaraldehyde for Completely Sutureless Laparoscopic Heminephrectomy in a Survival Porcine Model  

PubMed Central

Abstract Introduction Laparoscopic partial nephrectomy (LPN) has not received widespread clinical application because of its technical challenge. Bovine serum albumin glutaraldehyde (BSAG) is a hemostatic agent that is independent of the clotting cascade. We evaluated the use of BSAG as the sole agent for parenchymal and collecting system closure during LPN in a survival porcine model. Methods Eighteen pigs underwent hilar clamping and LPN by longitudinal excision of the lateral one-third of the right kidney. The opened collecting system was covered with oxidized cellulose to prevent BSAG seepage into the urinary tract. BSAG was allowed to set for 10 or 5 minutes. Twelve animals underwent survival LPN BSAG only closure; six control pigs were acutely studied using saline. Urinary extravasation was evaluated by injection of furosemide and indigo carmine, and then evaluating the renal surface and bladder catheter drainage for dye. A subjective bleeding score was assigned after hilum unclamping. At 6 weeks, BSAG kidneys were harvested for burst pressure testing and histopathological analysis. Results All 12 pigs survived for 6 weeks. No pigs had urinary extravasation. Mean percentage of kidney removed by weight was 19%. Mean warm ischemia time was 29 minutes. Five pigs required a second BSAG application to achieve a bleeding score of 0. Mean arterial and collecting system burst pressures were 301.8 and 322.4?mm Hg, respectively. Mean postoperative creatinine increase was 0.07?mg/dL. Conclusion BSAG for completely sutureless LPN in a survival porcine model was feasible.

Gamboa, Aldrin Joseph R.; Kaplan, Adam G.; Khosravi, Amanda; Truong, Hung; Andrade, Lorena; Lin, Rachelle; Alipanah, Reza; Ortiz, Cervando; McCormick, David; Box, Geoffrey N.; Lee, Hak J.; Deane, Leslie A.; Edwards, Robert A.; McDougall, Elspeth M.; Clayman, Ralph V.

2010-01-01

259

Bovine serum albumin oligomers in the E- and B-forms at low protein concentration and ionic strength.  

PubMed

The manuscript describes the study of the oligomerization process of bovine serum albumin (BSA) in two different structural monomeric forms: the extended-form (E) at pH 2.0 and the basic-form (B) at pH 9.0. The study was conducted at low protein concentration (1mg/ml) and relatively short incubation time (maximum 56 days) in order to investigate early oligomerization events rather than the formation of mature fibrils. The comparison between the two isoforms show that oligomers form much faster (?6 days) in the E-form than in the B-form where formation of oligomers requires ?4 weeks. The oligomers appear to be limited to a maximum of tetramers with size <30 nm. Hydrophobic interactions from exposed neutral amino acid residues in the elongated E-form are the likely cause for the quick formation of aggregates at acidic pH. We used an array of biophysical techniques for the study and determined that oligomerization occurs without further large changes in the secondary structure of the monomers. Under the conditions adopted in this study, aggregation does not seem to exceed the formation of tetramers, even though a very small amount of much larger aggregates seem to form. PMID:23148944

Babcock, Jeremiah J; Brancaleon, Lorenzo

2013-02-01

260

Some nutritional effects of folate-binding protein in bovine milk on the bioavailability of folate to rats  

SciTech Connect

The excretions of folate compounds into both the urine and bile were investigated in rats after the administration of pteroylglutamic acid (PteGlu) with or without the folate-binding protein (FBP) prepared from bovine milk. When the sample solution, containing either free or bound (/sup 3/H)PteGlu (i.e., bound to the FBP from milk), was delivered to rats intragastrically via oral intubation, the amounts of (/sup 3/H)PteGlu excreted into the feces did not change. On the other hand, the urinary excretion of /sup 3/H-labeled folate compounds, especially (/sup 3/H)5-methyltetrahydrofolic acid (5-CH/sub 3/-H/sub 4/PteGlu), after the administration of bound (/sup 3/H)PteGlu was significantly lower (P less than 0.01) than that after the administration of free (/sup 3/H)PteGlu. The urinary excretion of (/sup 3/H)5-CH/sub 3/-H/sub 4/PteGlu was directly proportional to the initial amount of free (/sup 3/H)PteGlu administered. The similar effect of FBP was also observed when the biliary excretion of /sup 3/H-labeled folate compounds was investigated in situ. Furthermore, the incorporation of (/sup 3/H)PteGlu into folate-requiring intestinal microorganisms was considerably reduced when it was bound to FBP. These results suggest that milk FBP has some nutritional effects on the bioavailability of folate in vivo.

Tani, M.; Iwai, K.

1984-04-01

261

Synthesis of nano-bioactive glass-ceramic powders and its in vitro bioactivity study in bovine serum albumin protein  

NASA Astrophysics Data System (ADS)

Bioactive glasses and ceramics have proved to be able to chemically bond to living bone due to the formation of an apatite-like layer on its surface. The aim of this work was preparation and characterization of bioactive glass-ceramic by sol-gel method. Nano-bioglass-ceramic material was crushed into powder and its bioactivity was examined in vitro with respect to the ability of hydroxyapatite layer to form on the surface as a result of contact with bovine serum albumin (BSA) protein. The obtained nano-bioactive glass-ceramic was analyzed before and after contact with BSA solution. This study used scanning electron microscope (SEM), transmission electron microscope (TEM), X-ray powder diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) analysis to examine its morphology, crystallinity and composition. The TEM images showed that the NBG particles size were 10-40 nm. Bioactivity of nanopowder was confirmed by SEM and XRD due to the presence of a rich bone-like apatite layer. Therefore, this nano-BSA-bioglass-ceramic composite material is promising for medical applications such as bone substitutes and drug carriers.

Nabian, Nima; Jahanshahi, Mohsen; Rabiee, Sayed Mahmood

2011-07-01

262

Recombinant Jembrana disease virus gag proteins identify several different antigenic domains but do not facilitate serological differentiation of JDV and nonpathogenic bovine lentiviruses.  

PubMed

In Indonesia, it is suspected that there are two bovine lentiviruses circulating in the cattle population: a pathogenic Jembrana disease virus (JDV), and a nonpathogenic bovine immunodeficiency-like virus (BIV). Both viruses cross-react antigenically and cannot be differentiated by current serological tests using JDV antigens. To identify possible type-specific epitopes, a series of recombinant protein constructs including the matrix, capsid and nucleocapsid proteins were produced from JDV gag and the expressed proteins were tested by Western blot using JDV and BIV hyperimmune sera. JDV matrix and truncated capsid proteins were recognised by both JDV and BIV hyperimmune sera indicating that there were multiple cross-reactive epitopes present in JDV gag. At least three epitopic regions were identified in these constructs, including the major homology region, by monoclonal antibody binding studies. JDV nucleocapsid recombinant protein was not recognised by either JDV or BIV hyperimmune sera and none of the recombinant gag proteins were able to differentiate between JDV positive sera from Jembrana disease endemic and Jembrana disease-free areas. Additionally, a 40 amino acid recombinant subunit protein encompassing the region recently found to contain an epitope unique to BIV [Zheng, L., Zhang, S., Wood, C., Kapil, S., Wilcox, G.E., Loughin, T.A., Minocha, H.C., 2001. Differentiation of two bovine lentiviruses by a monoclonal antibody on the basis of epitope specificity. Clin. Diagn. Lab. Immunol. 8, 283-287] was tested but was not recognised by either JDV positive sera from Jembrana disease-endemic or Jembrana disease-free areas. PMID:15664061

Desport, Moira; Stewart, Meredith E; Sheridan, Carol A; Ditcham, William G F; Setiyaningsih, Surachmi; Tenaya, W Masa; Hartaningsih, Nining; Wilcox, Graham E

2005-03-01

263

Solution structures and model membrane interactions of lactoferrampin, an antimicrobial peptide derived from bovine lactoferrin  

Microsoft Academic Search

Bovine lactoferrampin (LFampinB) has been identified as a novel antimicrobial peptide, which is derived from the N-terminal lobe of bovine lactoferrin. In this study, the solution structure of LFampinB bound to negatively charged sodium dodecyl sulphate micelles and zwitterionic dodecyl phosphocholine micelles was determined using 2-dimensional nuclear magnetic resonance (NMR) spectroscopy. The interaction between LFampinB and multilamellar phospholipid vesicles, containing

Evan F. Haney; Fanny Lau; Hans J. Vogel

2007-01-01

264

Bovine colostrum increases pore-forming claudin-2 protein expression but paradoxically not ion permeability possibly by a change of the intestinal cytokine milieu.  

PubMed

An impaired intestinal barrier function is involved in the pathogenesis of inflammatory bowel disease (IBD). Several nutritional factors are supposed to be effective in IBD treatment but scientific data about the effects on the intestinal integrity remain scarce. Bovine colostrum was shown to exert beneficial effects in DSS-induced murine colitis, and the present study was undertaken to explore the underlying molecular mechanisms. Western blot revealed increased claudin-2 expression in the distal ileum of healthy mice after feeding with colostrum for 14 days, whereas other tight junction proteins (claudin-3, 4, 10, 15) remained unchanged. The colostrum-induced claudin-2 induction was confirmed in differentiated Caco-2 cells after culture with colostrum for 48 h. Paradoxically, the elevation of claudin-2, which forms a cation-selective pore, was neither accompanied by increased ion permeability nor impaired barrier function. In an in situ perfusion model, 1 h exposure of the colonic mucosa to colostrum induced significantly increased mRNA levels of barrier-strengthening cytokine transforming growth factor-?, while interleukine-2, interleukine-6, interleukine-10, interleukine-13, and tumor-necrosis factor-? remained unchanged. Thus, modulation of the intestinal transforming growth factor-? expression might have compensated the claudin-2 increase and contributed to the observed barrier strengthening effects of colostrum in vivo and in vitro. PMID:23717570

Bodammer, Peggy; Kerkhoff, Claus; Maletzki, Claudia; Lamprecht, Georg

2013-01-01

265

Investigating the Use of Protein Saver Cards for Storage and Subsequent Detection of Bovine Anti-Brucella abortus Smooth Lipopolysaccharide Antibodies and Gamma Interferon  

PubMed Central

Brucella abortus, a smooth strain of the genus Brucella, is the causative agent of bovine brucellosis. To support the ongoing development of diagnostic tests for bovine brucellosis, the use of Protein Saver cards (Whatman) for bovine blood serum and plasma sample collection has been evaluated. These cards offer significant logistical and safety alternatives to transporting and storing liquid samples and may aid in diagnostic programs and validation studies. To evaluate the utility of these cards, 204 bovine blood serum samples from Brucella-infected and noninfected animals were stored on and eluted from the Protein Saver cards. Anti-Brucella smooth lipopolysaccharide (sLPS) antibody titers for the serum eluates were compared to those of the unprocessed original serum samples by indirect enzyme-linked immunosorbent assay (ELISA). The results showed a highly significant correlation between titers from the serum eluates and the unprocessed sera. Therefore, under these circumstances, serum eluates and unprocessed serum samples may be used interchangeably. Blood plasma from 113 mitogen-stimulated whole-blood samples was added to and eluted from the Protein Saver cards. The gamma interferon (IFN-?) titers in the plasma eluates were compared to those of the unprocessed plasma samples obtained by IFN-? ELISA. The results showed a significant correlation between the plasma eluates and the unprocessed plasma samples. To derive a signal in the plasma eluate, it was necessary to develop a novel and highly sensitive ELISA for the detection of IFN-?. The serum samples stored on cards at room temperature over a 10-day period showed little variation in antibody titers. However, the plasma eluates showed a progressive loss of IFN-? recovery over 10 days when stored at room temperature.

Commander, Nicola J.; Erdenlig, Sevil; McGiven, John A.; Stack, Judy

2013-01-01

266

Protein Production in the Bovine. Daily Production of the Specific Milk Proteins during the Lactation Period1  

Microsoft Academic Search

The synthesis of a-casein, B-casein, a-lactalbumin, and B-lactoglobulin was mutually related and reached a maximum level about five days after parturition. The daily levels of these proteins were unrelated to those of Tcasein, the immune globulins, and milk serum albumin. Maximum protein synthesis occurred about five days after parturition; that of fat, lactose, and total milk volume required about one

B. L. Larson; K. A. Kendall

1957-01-01

267

Stage-specific expression and effect of bone morphogenetic protein 2 on bovine granulosa cell estradiol production: regulation by cocaine and amphetamine regulated transcript.  

PubMed

Members of the bone morphogenetic protein (BMP) family regulate follicular development and granulosa cell function. However, changes in expression of BMP2 and its receptors during follicular waves in cattle and ability of BMP2 to modulate bovine granulosa cell estradiol production are not well understood. The objectives of this study were to determine temporal regulation of mRNA for BMP2 and its type I and II receptors (BMPR1A and BMPR2) in bovine follicles collected at specific stages of a follicular wave (predeviation, early dominance, mid dominance, preovulatory), ability of BMP2 to modulate bovine granulosa cell steroidogenesis, and whether effects of BMP2 on granulosa cell estradiol production are influenced by cotreatment with cocaine- and amphetamine-regulated transcript (CART), an intrafollicular regulatory peptide shown to inhibit estradiol production in response to other trophic hormones (FSH and IGF1). Relative abundance of mRNAs for Bmp2 and Bmpr2 was elevated at the mid dominance stage relative to earlier stages of the follicular wave and further increased at the preovulatory stage. Abundance of mRNA for Bmpr1a was lowest at early dominance stage and highest at preovulatory stage relative to other stages of the follicular wave examined. Treatment of bovine granulosa cells in vitro with BMP2 increased estradiol but decreased progesterone concentrations. Co-incubation with CART reduced the BMP2-stimulated increase in granulosa cell estradiol production. Results suggest that BMP2 may play a regulatory role in development of bovine follicles to the preovulatory stage and that CART can inhibit granulosa cell estradiol production in response to multiple hormones/growth factors, including BMP2. PMID:23313114

Selvaraju, S; Folger, J K; Gupta, P S P; Ireland, J J; Smith, G W

2013-04-01

268

Recapture after exocytosis causes differential retention of protein in granules of bovine chromaffin cells  

PubMed Central

After exocytosis, chromaffin granules release essentially all their catecholamines in small fractions of a second, but it is unknown how fast they release stored peptides and proteins. Here we compare the exocytic release of fluorescently labelled neuropeptide Y (NPY) and tissue plasminogen activator from single granules. Exocytosis was tracked by measuring the membrane capacitance, and single granules in live cells were imaged by evanescent field microscopy. Neuropeptide Y left most granules in small fractions of a second, while tissue plasminogen activator remained in open granules for minutes. Taking advantage of the dependence on pH of the fluorescence of green fluorescent protein, we used rhythmic external acidification to determine whether and when granules re-sealed. One-third of them re-sealed within 100 s and retained significant levels of tissue plasminogen activator. Re-sealing accounts for only a fraction of the endocytosis monitored in capacitance measurements. When external [Ca2+] was raised, even neuropeptide Y remained in open granules until they re-sealed. It is concluded that a significant fraction of chromaffin granules re-seal after exocytosis, and retain those proteins that leave granules slowly. We suggest that granules vary the stoichiometry of release by varying both granule re-sealing and the association of proteins with the granule matrix.

Perrais, David; Kleppe, Ingo C; Taraska, Justin W; Almers, Wolfhard

2004-01-01

269

Impact of antifreeze proteins and antifreeze glycoproteins on bovine sperm during freeze-thaw  

Microsoft Academic Search

There are no reports on the use of antifreeze proteins (AFP) and antifreeze glycoproteins (AFGP) for the use of bull sperm cryopreservation despite studies in the ram, mouse and chimpanzee. The effect of freezing and thawing on bull sperm viability, osmotic resistance and acrosome integrity were observed following the addition of AFP1, AFPIII and AFGP at four concentrations (0.1, 1,

N. S. Prathalingam; W. V. Holt; S. G. Revell; S. Mirczuk; R. A. Fleck; P. F. Watson

2006-01-01

270

Nonstructural Proteins NS1 and NS2 of Bovine Respiratory Syncytial Virus Block Activation of Interferon Regulatory Factor 3  

PubMed Central

We have previously shown that the nonstructural (NS) proteins NS1 and NS2 of bovine respiratory syncytial virus (BRSV) mediate resistance to the alpha/beta interferon (IFN)-mediated antiviral response. Here, we show that they, in addition, are able to prevent the induction of beta IFN (IFN-?) after virus infection or double-stranded RNA stimulation. In BRSV-infected MDBK cells upregulation of IFN-stimulated genes (ISGs) such as MxA did not occur, although IFN signaling via JAK/STAT was found intact. In contrast, infection with recombinant BRSVs lacking either or both NS genes resulted in efficient upregulation of ISGs. Biological IFN activity and IFN-? were detected only in supernatants of cells infected with the NS deletion mutants but not with wild-type (wt) BRSV. Subsequent analyses of IFN-? promoter activity showed that infection of cells with the double deletion mutant BRSV ?NS1/2, but not with BRSV wt, resulted in a significant increase in IFN-? gene promoter activity. Induction of the IFN-? promoter depends on the activation of three distinct transcription factors, NF-?B, ATF-2/c-Jun, and IFN regulatory factor 3 (IRF-3). Whereas NF-?B and ATF-2/c-Jun activities were readily detectable and comparable in both wt BRSV- and BRSV ?NS1/2-infected cells, phosphorylation and transcriptional activity of IRF-3, however, were observed only after BRSV ?NS1/2 infection. NS protein-mediated inhibition of IRF-3 activation and IFN induction should have considerable impact on the pathogenesis and immunogenicity of BRSV.

Bossert, Birgit; Marozin, Sabrina; Conzelmann, Karl-Klaus

2003-01-01

271

Functional Characterization of Bovine Viral Diarrhea Virus Nonstructural Protein 5A by Reverse Genetic Analysis and Live Cell Imaging  

PubMed Central

Nonstructural protein 5A (NS5A) of bovine viral diarrhea virus (BVDV) is a hydrophilic phosphoprotein with RNA binding activity and a critical component of the viral replicase. In silico analysis suggests that NS5A encompasses three domains interconnected by two low-complexity sequences (LCSs). While domain I harbors two functional determinants, an N-terminal amphipathic helix important for membrane association, and a Zn-binding site essential for RNA replication, the structure and function of the C-terminal half of NS5A are still ill defined. In this study, we introduced a panel of 10 amino acid deletions covering the C-terminal half of NS5A. In the context of a highly efficient monocistronic replicon, deletions in LCS I and the N-terminal part of domain II, as well as in domain III, were tolerated with regard to RNA replication. When introduced into a bicistronic replicon, only deletions in LCS I and the N-terminal part of domain II were tolerated. In the context of the viral full-length genome, these mutations allowed residual virion morphogenesis. Based on these data, a functional monocistronic BVDV replicon coding for an NS5A variant with an insertion of the fluorescent protein mCherry was constructed. Live cell imaging demonstrated that a fraction of NS5A-mCherry localizes to the surface of lipid droplets. Taken together, this study provides novel insights into the functions of BVDV NS5A. Moreover, we established the first pestiviral replicon expressing fluorescent NS5A-mCherry to directly visualize functional viral replication complexes by live cell imaging.

Isken, Olaf; Langerwisch, Ulrike; Schonherr, Robert; Lamp, Benjamin; Schroder, Kristin; Duden, Rainer; Rumenapf, Tillmann H.

2014-01-01

272

Nonstructural proteins NS1 and NS2 of bovine respiratory syncytial virus block activation of interferon regulatory factor 3.  

PubMed

We have previously shown that the nonstructural (NS) proteins NS1 and NS2 of bovine respiratory syncytial virus (BRSV) mediate resistance to the alpha/beta interferon (IFN)-mediated antiviral response. Here, we show that they, in addition, are able to prevent the induction of beta IFN (IFN-beta) after virus infection or double-stranded RNA stimulation. In BRSV-infected MDBK cells upregulation of IFN-stimulated genes (ISGs) such as MxA did not occur, although IFN signaling via JAK/STAT was found intact. In contrast, infection with recombinant BRSVs lacking either or both NS genes resulted in efficient upregulation of ISGs. Biological IFN activity and IFN-beta were detected only in supernatants of cells infected with the NS deletion mutants but not with wild-type (wt) BRSV. Subsequent analyses of IFN-beta promoter activity showed that infection of cells with the double deletion mutant BRSV DeltaNS1/2, but not with BRSV wt, resulted in a significant increase in IFN-beta gene promoter activity. Induction of the IFN-beta promoter depends on the activation of three distinct transcription factors, NF-kappaB, ATF-2/c-Jun, and IFN regulatory factor 3 (IRF-3). Whereas NF-kappaB and ATF-2/c-Jun activities were readily detectable and comparable in both wt BRSV- and BRSV DeltaNS1/2-infected cells, phosphorylation and transcriptional activity of IRF-3, however, were observed only after BRSV DeltaNS1/2 infection. NS protein-mediated inhibition of IRF-3 activation and IFN induction should have considerable impact on the pathogenesis and immunogenicity of BRSV. PMID:12885884

Bossert, Birgit; Marozin, Sabrina; Conzelmann, Karl-Klaus

2003-08-01

273

Modeling of bovine type-I collagen fibrils: interaction with pickling and retanning agents.  

PubMed

Bovine Type I collagen was investigated, building on a large scale computer model of a collagen fibril in water, and focusing on two stages of the leather manufacturing process. The effects of different salts (NaCl, CaCl(2), and Na(2)SO(4)) on the swelling behavior of collagen at low pH (the pickling process) were studied. The salts suppress the swelling of the fibrils at low pH and we find specific stabilizing influences for CaCl(2) and Na(2)SO(4), due to weak Ca(2+)/Cl(-) and strong SO(4) (2-)/lysine/arginine interactions, respectively. Using state-of-the-art sampling techniques, such as the metadynamics algorithm, to allow an efficient exploration of configuration space, we were able to investigate the effect of polyacrylate and poly(methyl acrylate) - two polymeric retanning agents - on the fibril. Both polymers interact with the ammonium groups on the surface, but polyacrylate shows significantly stronger interactions. We suggest that it is this stronger interaction that contributes to the reduced suitability of PAA as a tanning agent. PMID:17295396

Bulo, Rosa E; Siggel, Lorenz; Molnar, Ferenc; Weiss, Horst

2007-02-12

274

Gene expression analysis of protein synthesis pathways in bovine mammary epithelial cells purified from milk during lactation and short-term restricted feeding.  

PubMed

The objective of the study was to investigate selected key regulatory pathways of milk protein biosynthesis in primary bovine mammary epithelial cells (MECs) of dairy cows during the first 155 days of lactation. In addition, cows were exposed to feed restriction for a short period (FR) during different stages of lactation (week 4 and 21 pp) to study adjustment processes of molecular protein biosynthesis to metabolic challenge. Morning milk samples from twenty-four Holstein-Friesian cows were collected throughout the experimental period (n = 10 per animal). MEC from raw milk were purified using an immunomagnetic separation technique and used for real-time quantitative PCR analyses. As was seen in transcript abundances of all major milk proteins, mRNA levels of E74-like factor 5 (ELF5), an enhancer of signal transducer and activator of transcription (STAT) action, concomitantly decreased towards mid-lactation. Expression of ELF5 as well as of all milk protein genes showed a similar increase during FR in early lactation. Occasional changes in expression could be seen in other Janus kinase (JAK)/STAT factors and in mammalian target of rapamycin (mTOR) pathway elements. Amino acid transfer and glucose transporter and the ?-casein expression were also partially affected. In conclusion, our findings suggest a pivotal role of the transcription factor ELF5 in milk protein mRNA expression with complementary JAK/STAT and mTOR signalling for the regulation of protein biosynthesis in the bovine mammary gland. PMID:23402545

Sigl, T; Meyer, H H D; Wiedemann, S

2014-02-01

275

A theoretical model for the collective motion of proteins by means of principal component analysis  

Microsoft Academic Search

A coarse grained model in the frame work of principal component analysis is presented. We used a bath of harmonic oscillators\\u000a approach, based on classical mechanics, to derive the generalized Langevin equations of motion for the collective coordinates.\\u000a The dynamics of the protein collective coordinates derived from molecular dynamics simulations have been studied for the Bovine\\u000a Pancreatic Trypsin Inhibitor. We

Hiqmet Kamberaj

2011-01-01

276

Mesoporous calcium silicate for controlled release of bovine serum albumin protein  

Microsoft Academic Search

The purpose of this study is to synthesize mesoporous calcium silicate (CS or wollastonite, CaSiO3) and evaluate its possible application in protein\\/drug delivery. First, calcium silicate was synthesized by wet chemical method and then mesoporosity was created by acid modification of the synthesized CS particle using hydrochloric acid at pH 7, 4.5, and 0.5. The results showed that a hydrated

Weichang Xue; Amit Bandyopadhyay; Susmita Bose

2009-01-01

277

Influence of the pore fluid on the phase velocity in bovine trabecular bone In Vitro: Prediction of the biot model  

NASA Astrophysics Data System (ADS)

The present study aims to investigate the influence of the pore fluid on the phase velocity in bovine trabecular bone in vitro. The frequency-dependent phase velocity was measured in 20 marrow-filled and water-filled bovine femoral trabecular bone samples. The mean phase velocities at frequencies between 0.6 and 1.2 MHz exhibited significant negative dispersions for both the marrow-filled and the water-filled samples. The magnitudes of the dispersions showed no significant differences between the marrow-filled and the water-filled samples. In contrast, replacement of marrow by water led to a mean increase in the phase velocity of 27 m/s at frequencies from 0.6 to 1.2 MHz. The theoretical phase velocities of the fast wave predicted by using the Biot model for elastic wave propagation in fluid-saturated porous media showed good agreements with the measurements.

Lee, Kang Il

2013-01-01

278

Limited proteolysis of bovine alpha-lactalbumin: isolation and characterization of protein domains.  

PubMed Central

The partly folded states of alpha-lactalbumin (alpha-LA) exposed to acid solution at pH 2.0 (A-state) or at neutral pH upon EDTA-mediated removal of the single protein-bound calcium ion (apo form) have been probed by limited proteolysis experiments. These states are nowadays commonly considered to be molten globules and thus protein-folding intermediates. Pepsin was used for proteolysis at acid pH, while proteinase K and chymotrypsin at neutral pH. The expectations were that these proteolytic probes would detect sites and/or chain regions in the partly folded states of alpha-LA sufficiently dynamic, or even unfolded, capable of binding and adaptation to the specific stereochemistry of the protease's active site. A time-course analysis of the proteolytic events revealed that the fast, initial proteolytic cuts of the 123-residue chain of alpha-LA in its A-state or apo form by the three proteases occur at the same chain region 39-54, the actual site(s) of cleavage depending upon the protease employed. This region in native alpha-LA encompasses the beta-sheets of the protein. Subsequent cleavages occur mostly at chain regions 31-35 and 95-105. Four fragment species of alpha-LA have been isolated by reverse-phase high-performance liquid chromatography, and their conformational properties examined by circular dichroism and fluorescence emission spectroscopy. The single chain fragment 53-103, containing all the binding sites for calcium in native alpha-LA and cross-linked by two disulfide bridges, maintains in aqueous buffer and in the presence of calcium ions a folded structure characterized by the same content of alpha-helix of the corresponding chain segment in native alpha-LA. Evidence for some structure was also obtained for the two-chain species 1-40 and 104-123, as well as 1-31 and 105-123, both systems being covalently linked by two disulfide bonds. In contrast, the protein species given by fragment 1-34 connected to fragment 54-123 or 57-123 via four disulfide bridges adopts in solution a folded structure with the helical content expected for a native-like conformation. Of interest, the proteolytic fragment species herewith isolated correspond to the structural domains and subdomains of alpha-LA that can be identified by computational analysis of the three-dimensional structure of native alpha-LA (Siddiqui AS, Barton GI, 1995, Protein Sci 4:872-884). The fast, initial cleavages at the level of the beta-sheet region of native alpha-LA indicate that this region is highly mobile or even unfolded in the alpha-LA molten globule(s), while the rest of the protein chain maintains sufficient structure and rigidity to prevent extensive proteolysis. The subsequent cleavages at chain segment 95-105 indicate that also this region is somewhat mobile in the A-state or apo form of the protein. It is concluded that the overall domain topology of native alpha-LA is maintained in acid or at neutral pH upon calcium depletion. Moreover, the molecular properties of the partly folded states of alpha-LA deduced here from proteolysis experiments do correlate with those derived from previous NMR and other physicochemical measurements.

Polverino de Laureto, P.; Scaramella, E.; Frigo, M.; Wondrich, F. G.; De Filippis, V.; Zambonin, M.; Fontana, A.

1999-01-01

279

Expression of minute virus of mice structural proteins in murine cell lines transformed by bovine papillomavirus-minute virus of mice plasmid chimera.  

PubMed Central

Recombinant plasmids containing the genomes of both bovine papillomavirus type I and minute virus of mice (MVM) were constructed and used to transform mouse C127 cells. Transformed lines that express MVM gene products with high efficiency were isolated and characterized. These transformants synthesize large amounts of MVM structural polypeptides and spontaneously assemble them into empty virion particles that are released into the culture medium. These lines were, however, genetically unstable; they slowly generated subpopulations that failed to express MVM-specific proteins, and they possessed episomal DNA in which both MVM and bovine papillomavirus sequences were deleted or rearranged, or both. Clonal isolates of these transformants were also superinfectible by infectious MVM virus. Therefore, in spite of their instability, they should be useful host cell lines for transcomplementing mutations introduced into the MVM genome and for growing defective viruses as virions. Images

Pintel, D; Merchlinsky, M J; Ward, D C

1984-01-01

280

Overexpression of a monomeric form of the bovine odorant-binding protein protects Escherichia coli from chemical-induced oxidative stress.  

PubMed

Abstract Mammalian odorant-binding proteins (OBPs) are soluble lipocalins produced in the nasal mucosa and in other epithelial tissues of several animal species, where they are supposed to serve as scavengers for small structurally unrelated hydrophobic molecules. These would include odorants and toxic aldehydes like 4-hydroxy-2-nonenal (HNE), which are end products of lipid peroxidation; therefore OBP might physiologically contribute to preserve the integrity of epithelial tissues under oxidative stress conditions by removing toxic compounds from the environment and, eventually, driving them to the appropriate degradative pathways. With the aim of developing a biological model based on a living organism for the investigation of the antioxidant properties of OBP, here we asked whether the overexpression of the protein could confer protection from chemical-induced oxidative stress in Escherichia coli. To this aim, bacteria were made to overexpress either GCC-bOBP, a redesigned monomeric mutant of bovine OBP, or its amino-terminal 6-histidine-tagged version 6H-GCC-bOBP. After inducing overexpression for 4 h, bacterial cells were diluted in fresh culture media, and their growth curves were followed in the presence of hydrogen peroxide (H2O2) and tert-Butyl hydroperoxide (tBuOOH), two reactive oxygen species whose toxicity is mainly due to lipid peroxidation, and menadione, a redox-cycling drug producing the superoxide ion. GCC-bOBP and 6H-GCC-bOBP were found to protect bacterial cells from the insulting agents H2O2 and tBuOOH but not from menadione. The obtained data led us to hypothesize that the presence of overexpressed OBP may contribute to protect bacterial cells against oxidative stress probably by sequestering toxic compounds locally produced during the first replication cycles by lipid peroxidation, before bacteria activate their appropriate enzyme-based antioxidative mechanisms. PMID:24697800

Macedo-Márquez, A; Vázquez-Acevedo, M; Ongay-Larios, L; Miranda-Astudillo, H; Hernández-Muńoz, R; González-Halphen, D; Grolli, S; Ramoni, R

2014-07-01

281

Complementary deoxyribonucleic acid sequence encoding bovine ubiquitin cross-reactive protein : A comparison with ubiquitin and a 15-kDa ubiquitin homolog.  

PubMed

Pregnancy in the cow depends on secretion of interferon-tau (IFN-?) by the conceptus (trophoblast and embryo) and the actions of this cytokine on the uterine endometrium. A novel 17-kDa uterine protein that is regulated by IFN-? during early pregnancy and crossreacts with ubiquitin antiserum on Western blots, has been named bovine ubiquitin cross-reactive protein (bUCRP). We suspected that bUCRP might be structurally related to ubiquitin, and to a human UCRP (ISG15 product) that has been described in several cell lines to be regulated by Type I IFNs. In this study, immunoscreening of a bovine endometrial cDNA library with ubiquitin antiserum resulted in the isolation of cDNAs encoding bUCRP. Nucleotide sequence of the bUCRP cDNA shared 70% identity with hUCRP and 30% identity with a tandem ubiquitin repeat. Computer translation revealed that bUCRP shared the Leu-Arg-Gly-Gly (LRGG) C-terminal sequence with ubiquitin and hUCRP that has been implicated in the modulation of intracellular proteins. However, some ubiquitin residues known to function in the ligation (Arg-54) to targeted proteins and in the processing of conjugates through the proteasome (His-68), have been lost through mutation in bUCRP. Lys-48, known to function in formation of ubiquitin polymers, was present in hUCRP, but mutated to Arg in bUCRP. Because bUCRP is secreted and retains the LRGG sequence, it may have both intracellular and secreted endocrine roles in maintaining pregnancy. Bovine UCRP also may have very different intracellular roles when compared with ubiquitin and hUCRP because of mutations in residues known to form polymers and to target proteins to degradation. PMID:21153111

Austin, K J; Pru, J K; Hansen, T R

1996-10-01

282

The comparative specificity of 3 oestradiol-binding proteins. Rat alpha-foetoprotein, rat liver 17beta-hydroxy steroid dehydrogenase and anti-(oestradiol-6-carboxymethyloxime-bovine serum albumin) antiserum.  

PubMed Central

1. The specificity of 3 oestradiol-binding proteins was studied. Two of these proteins are naturally occurring (rat alpha-foetoprotein and rat liver microsomal 17beta-hydroxy steroid dehydrogenase) and the third is an artificially induced model, anti-(oestradiol-6-carboxymethyloxime-bovine serum albumin) gamma-globulins. 2. A specific binding procedure for each protein model permitted a determination of its affinity for oestradiol and for 30 other steroids. 3. The results obtained have brought to light the different areas of the steroid molecule that are important for its recognition by each of the three proteins. The two naturally occurring proteins (alpha-foetoprotein and 17beta-hydroxy steroid dehydrogenase) recognize the edge of the steroid defined by C-4, C-6, C-8 and C-15. On the other hand, the gamma-globulins recognize the opposite edge, i.e. that defined by C-2, C-10, C-11 and C-17. 4. Diethylstilboestrol, whose structure is analogous to that of a steroid, is only recognized by the two naturally occurring proteins.

Laurant, C; de Lauzon, S D; Cittanova, N; Nunez, E; Jayle, M F

1975-01-01

283

The protection of bovine skeletal myofibrils from proteolytic damage post mortem by small heat shock proteins.  

PubMed

This study aimed to determine how small heat shock proteins (sHSPs) protect myofibrillar proteins from ?-calpain degradation during ageing. Immunoprecipitation experiments with M. longissimus dorsi (LD) from Angus heifers (n=14) examined the interaction between ??-crystallin, desmin, titin, HSP20, HSP27 and ?-calpain. Results showed that ??-crystallin associated with desmin, titin, HSP20, HSP27 and ?-calpain. Exogenous ??-crystallin reduced desmin and titin degradations in myofibrillar extracts and attenuated ?-calpain activity. In a second experiment, bull LD (n=94) were aged at -1.5°C for up to 28days post mortem. ?-Calpain autolysed faster in high ultimate pH (pHu) meat (pHu?6.2) and this was concomitant with the more rapid degradation of titin and filamin in this pHu group. Desmin stability in intermediate pHu meat (pHu 5.8 to 6.19) may be due to the protection of myofibril-bound sHSPs combined with the competitive inhibition of ?-calpain by sHSPs. PMID:24769876

Lomiwes, D; Hurst, S M; Dobbie, P; Frost, D A; Hurst, R D; Young, O A; Farouk, M M

2014-08-01

284

Jumonji domain-containing protein 3 regulates histone 3 lysine 27 methylation during bovine preimplantation development.  

PubMed

Understanding the mechanisms of epigenetic remodeling that follow fertilization is a fundamental step toward understanding the bases of early embryonic development and pluripotency. Extensive and dynamic chromatin remodeling is observed after fertilization, including DNA methylation and histone modifications. These changes underlie the transition from gametic to embryonic chromatin and are thought to facilitate embryonic genome activation. In particular, trimethylation of histone 3 lysine 27 (H3K27me3) is associated with gene-specific transcription repression. Global levels of this epigenetic mark are high in oocyte chromatin and decrease to minimal levels at the time of embryonic genome activation. We provide evidence that the decrease in H3K27me3 observed during early development is cell-cycle independent, suggesting an active mechanism for removal of this epigenetic mark. Among H3K27me3-specific demethylases, Jumonji domain-containing protein 3 (JMJD3), but not ubiquitously transcribed tetratricopeptide repeat X (UTX), present high transcript levels in oocytes. Soon after fertilization JMJD3 protein levels increase, concurrent with a decrease in mRNA levels. This pattern of expression suggests maternal inheritance of JMJD3. Knockdown of JMJD3 by siRNA injection in parthenogenetically activated metaphase II oocytes resulted in inhibition of the H3K27me3 decrease normally observed in preimplantation embryos. Moreover, knockdown of JMJD3 in oocytes reduced the rate of blastocyst development. Overall, these results indicate that JMJD3 is involved in active demethylation of H3K27me3 during early embryo development and that this mark plays an important role during the progression of embryos to blastocysts. PMID:22308433

Canovas, Sebastian; Cibelli, Jose B; Ross, Pablo J

2012-02-14

285

Prediction of Protein-protein Interactions Using Alpha Shape Modeling  

NASA Astrophysics Data System (ADS)

Protein-protein interactions play important roles in a lot of biological progress. Previous studies about protein-protein interactions were mainly based on sequence analysis. As more 3D structural information can be obtained from protein-protein complexes, structural analysis becomes feasible and useful. In this study, we used structural alignment to predict the protein-binding site and apply 3D alpha shape modeling to analyze the interface characteristics. We have developed a method for protein-protein interaction prediction. The result indicates good performance of our method in discriminating protein-binding structures from non-protein binding structures. Our method outperforms the previous methods based on the Matthews correlation coefficient.

Zhou, Weiqiang; Yan, Hong; Fan, Xiaodan; Hao, Quan

2011-06-01

286

The refined structure of vitamin D-dependent calcium-binding protein from bovine intestine. Molecular details, ion binding, and implications for the structure of other calcium-binding proteins.  

PubMed

The structure of bovine intestinal calcium-binding protein (ICaBP) has been determined crystallographically at a resolution of 2.3 A and refined by a least squares technique to an R factor of 17.8%. The refined structure includes all 600 non-hydrogen protein atoms, two bound calcium ions, and solvent consisting of one sulfate ion and 36 water molecules. The molecule consists of two helix-loop-helix calcium-binding domains known as EF hands, connected by a linker containing a single turn of helix. Helix-helix interactions are primarily hydrophobic, but also include a few strategic hydrogen bonds. Most of the hydrogen bonds, however, are found in the calcium-binding loops, where they occur both within a single loop and between the two. Examination of the hydrogen bonding patterns in the calcium-binding loops of ICaBP and the related protein, parvalbumin, reveals several conserved hydrogen bonds which are evidently important for loop stabilization. The primary and tertiary structural features which promote the formation of an EF hand were originally identified from the structure of parvalbumin. They are modified in light of the ICaBP structure and considered as they apply to other calcium-binding proteins. The C-terminal domain of ICaBP is a normal EF hand, with ion binding properties similar to those of the calmodulin hands, but the N-terminal domain is a variant hand whose calcium ligands are mostly peptide carbonyls. Relative to a normal EF hand, this domain exhibits a similar KD for calcium binding but a greatly reduced affinity for calcium analogs such as cadmium and the lanthanide series. Lanthanides in particular may be inappropriate models for calcium in this system. PMID:3722173

Szebenyi, D M; Moffat, K

1986-07-01

287

Vitamin D Signaling in the Bovine Immune System: A Model for Understanding Human Vitamin D Requirements  

PubMed Central

The endocrine physiology of vitamin D in cattle has been rigorously investigated and has yielded information on vitamin D requirements, endocrine function in health and disease, general metabolism, and maintenance of calcium homeostasis in cattle. These results are relevant to human vitamin D endocrinology. The current debate regarding vitamin D requirements is centered on the requirements for proper intracrine and paracrine vitamin D signaling. Studies in adult and young cattle can provide valuable insight for understanding vitamin D requirements as they relate to innate and adaptive immune responses during infectious disease. In cattle, toll-like receptor recognition activates intracrine and paracrine vitamin D signaling mechanism in the immune system that regulates innate and adaptive immune responses in the presence of adequate 25-hydroxyvitamin D. Furthermore, experiments with mastitis in dairy cattle have provided in vivo evidence for the intracrine vitamin D signaling mechanism in macrophages as well as vitamin D mediated suppression of infection. Epidemiological evidence indicates that circulating concentrations above 32 ng/mL of 25-hydroxyvitamin D are necessary for optimal vitamin D signaling in the immune system, but experimental evidence is lacking for that value. Experiments in cattle can provide that evidence as circulating 25-hydroxyvitamin D concentrations can be experimentally manipulated within ranges that are normal for humans and cattle. Additionally, young and adult cattle can be experimentally infected with bacteria and viruses associated with significant diseases in both cattle and humans. Utilizing the bovine model to further delineate the immunomodulatory role of vitamin D will provide potentially valuable insights into the vitamin D requirements of both humans and cattle, especially as they relate to immune response capacity and infectious disease resistance.

Nelson, Corwin D.; Reinhardt, Timothy A.; Lippolis, John D.; Sacco, Randy E.; Nonnecke, Brian J.

2012-01-01

288

Influence of non ionizing radiation of base stations on the activity of redox proteins in bovines  

PubMed Central

Background The influence of electromagnetic fields on the health of humans and animals is still an intensively discussed and scientifically investigated issue (Prakt Tierarzt 11:15-20, 2003; Umwelt Medizin Gesellschaft 17:326-332, 2004; J Toxicol Environment Health, Part B 12:572–597, 2009). We are surrounded by numerous electromagnetic fields of variable strength, coming from electronic equipment and its power cords, from high-voltage power lines and from antennas for radio, television and mobile communication. Particularly the latter cause’s controversy, as everyone likes to have good mobile reception at anytime and anywhere, whereas nobody wants to have such a basestation antenna in their proximity. Results In this experiment, the NIR has resulted in changes in the enzyme activities. Certain enzymes were disabled, others enabled by NIR. Furthermore, individual behavior patterns were observed. While certain cows reacted to NIR, others did not react at all, or even inversely. Conclusion The present results coincide with the information from the literature, according to which NIR leads to changes in redox proteins, and that there are individuals who are sensitive to radiation and others that are not. However, the latter could not be distinctly attributed – there are cows that react clearly with one enzyme while they do not react with another enzyme at all, or even the inverse. The study approach of testing ten cows each ten times during three phases has proven to be appropriate. Future studies should however set the post-exposure phase later on.

2014-01-01

289

The osteoinductive activity of bone morphogenetic protein (BMP) purified by repeated extracts of bovine bone.  

PubMed

Native bone morphogenetic proteins (BMPs) extracted from bone have been used clinically to stimulate bone regeneration and repair. However, preparation of purified BMP is a laborious process. This study investigated the yield, activity and cost effectiveness of repeatedly extracting the same bone matrix to produce purified BMP. While repeated extraction was able to increase the yield 62% the activity of the partially purified BMP in later extracts decreased both in vitro and in vivo. This decline in activity appears to be due to an increase in non-BMP contaminants, such as collagen, in the extracts. When the first three extracts were combined and processed together activity was equivalent to that of the first extract. A simple analysis based on the cost of reagents used and the time required for purification indicates that separate processing of the extracts is inefficient while combining the first and second extracts and processing them together would result in a small cost saving. Based on this study we would recommend that the demineralized bone matrix be extracted no more than twice and that the extracts be combined for further processing. PMID:15176456

Hu, Zhen Ming; Peel, Sean A F; Sandor, George K B; Clokie, Cameron M L

2004-03-01

290

Bovine factor XIIa inhibitor.  

PubMed

Bovine factor XIIa inhibitor was purified by an improved method employing affinity for heparin. N-terminal amino acid sequencing revealed a unique sequence without homology to any other known protein sequences. Peptide sequencing, however, showed that a part of the bovine factor XIIa inhibitor was homologous to human C1-inhibitor with a fraction of identical amino acid residues around 70%. Deglycosylation studies and carbohydrate analysis showed the presence of N- and O-linked carbohydrate. Bovine factor XIIa inhibitor did not inhibit plasma kallikrein and trypsin. The reactive site comprised an Arg-Asn bond, and represents the first example of asparagine as a P1' residue in Serpins with well documented inhibitory activity. PMID:8457651

Muldbjerg, M; Markussen, S; Magnusson, S; Halkier, T

1993-02-01

291

Sensitivity-dependent model of protein protein interaction networks  

NASA Astrophysics Data System (ADS)

The scale free structure p(k) ~ k-? of protein-protein interaction networks can be reproduced by a static physical model in simulation. We inspect the model theoretically, and find the key reason for the model generating apparent scale free degree distributions. This explanation provides a generic mechanism of 'scale free' networks. Moreover, we predict the dependence of ? on experimental protein concentrations or other sensitivity factors in detecting interactions, and find experimental evidence to support the prediction.

Zhang, Jingshan; Shakhnovich, Eugene I.

2008-09-01

292

Conserved epitopes on the hemagglutinin-neuraminidase proteins of human and bovine parainfluenza type 3 viruses: nucleotide sequence analysis of variants selected with monoclonal antibodies.  

PubMed

We have previously identified 11 epitopes located in two topologically nonoverlapping antigenic sites (A and B) and a third bridging site (C) on the human type 3 parainfluenza virus (PIV3) hemagglutinin-neuraminidase (HN) glycoprotein by using monoclonal antibodies (MAbs) which inhibit hemagglutination and virus infectivity (K. L. Coelingh, C. C. Winter, and B. R. Murphy, Virology 143:569-582, 1985). We have identified three additional antigenic sites (D, E, and F) on the HN molecule by competitive-binding assays of anti-HN MAbs which have no known biological activity. Epitopes in sites A, D, and F are conserved on the bovine PIV3 HN glycoprotein and also among a wide range of human isolates. The dideoxy method was used to identify nucleotide substitutions in the HN genes of antigenic variants selected with neutralizing MAbs representing epitopes in site A which are shared by human and bovine PIV3. The deduced amino acid substitutions in the variants were located in separate hydrophilic stretches of HN residues which are conserved in the primary structures of the HN proteins of both human and bovine PIV3 strains. PMID:2427750

Coelingh, K J; Winter, C C; Murphy, B R; Rice, J M; Kimball, P C; Olmsted, R A; Collins, P L

1986-10-01

293

The Importance of Protein-Protein Interactions on the pH-Induced Conformational Changes of Bovine Serum Albumin: A Small-Angle X-Ray Scattering Study  

PubMed Central

Abstract The combined effects of concentration and pH on the conformational states of bovine serum albumin (BSA) are investigated by small-angle x-ray scattering. Serum albumins, at physiological conditions, are found at concentrations of ?35–45 mg/mL (42 mg/mL in the case of humans). In this work, BSA at three different concentrations (10, 25, and 50 mg/mL) and pH values (2.0–9.0) have been studied. Data were analyzed by means of the Global Fitting procedure, with the protein form factor calculated from human serum albumin (HSA) crystallographic structure and the interference function described, considering repulsive and attractive interaction potentials within a random phase approximation. Small-angle x-ray scattering data show that BSA maintains its native state from pH 4.0 up to 9.0 at all investigated concentrations. A pH-dependence of the absolute net protein charge is shown and the charge number per BSA is quantified to 10(2), 8(1), 13(2), 20(2), and 26(2) for pH values 4.0, 5.4, 7.0, 8.0, and 9.0, respectively. The attractive potential diminishes as BSA concentration increases. The coexistence of monomers and dimers is observed at 50 mg/mL and pH 5.4, near the BSA isoelectric point. Samples at pH 2.0 show a different behavior, because BSA overall shape changes as a function of concentration. At 10 mg/mL, BSA is partially unfolded and a strong repulsive protein-protein interaction occurs due to the high amount of exposed charge. At 25 and 50 mg/mL, BSA undergoes some re-folding, which likely results in a molten-globule state. This work concludes by confirming that the protein concentration plays an important role on the pH-unfolded BSA state, due to a delicate compromise between interaction forces and crowding effects.

Barbosa, Leandro R.S.; Ortore, Maria Grazia; Spinozzi, Francesco; Mariani, Paolo; Bernstorff, Sigrid; Itri, Rosangela

2010-01-01

294

Role of bovine pulmonary surfactant-associated proteins in the surface-active property of phospholipid mixtures.  

PubMed

The surfactant-associated proteins, SP-A, SP-B and SP-C have been isolated from bovine pulmonary surfactant. The biophysical roles of SP-B and SP-C in reconstituted surfactants, with various phospholipid mixtures subjected to different thermal treatments, have been examined using a pulsating bubble surfactometer. The phospholipid mixtures were: (A) dipalmitoylphosphatidylcholine (DPPC)/egg phosphatidylcholine (PC)/egg phosphatidylglycerol (PG) (6:2:2, w/w); (B) DPPC/PG (9:1); and (C) DPPC/PG (7:3). Thermal treatments involved mixing SP-B or SP-C, at room temperature, with lipids in chloroform/methanol (9:1, v/v) and removing the solvent under N2 by (1) evaporation at room temperature; (2) evaporation at 45 degrees C; or (3) incubation at 45 degrees C overnight prior to evaporation at 45 degrees C. In all cases, 45 degrees C solvent evaporation was the most effective treatment. DPPC/egg PG (7:3) was the most favourable lipid composition. With either a static or a pulsating bubble, SP-C promoted a rapid decrease in surface tension with little change thereafter. This implies that SP-C is effective in enhancing phospholipid adsorption but does not play an important role in the removal of non-DPPC lipid from the monolayer. While SP-B was not as effective in facilitating phospholipid absorption, samples containing this protein could achieve near zero surface tension upon pulsation. A very low surface tension could also be attained during the initial pulsation of DPPC/PG plus SP-B mixtures which had been allowed to adsorb until equilibrium. This observation indicates that SP-B promotes the removal of PG from the monolayer. SP-A alone had only a slight effect on the surface activity of the DPPC/PG (7:3) mixture, and did not accelerate adsorption of samples containing SP-C. However, SP-A facilitated phospholipid adsorption and may also enhance the removal of PG from monolayers in the presence of SP-B. PMID:2223863

Yu, S H; Possmayer, F

1990-10-01

295

Modeling Enzymatic Reactions in Proteins.  

NASA Astrophysics Data System (ADS)

We will discuss application of our density functional (DFT)-based QM/MM methodology to modeling a variety of protein active sites, including methane monooxygenase, myoglobin, and cytochrome P450. In addition to the calculation of intermediates, transition states, and rate constants, we will discuss modeling of reactions requiring protein conformational changes. Our methodology reliably achieves small errors as a result of imposition of the QM/MM boundary. However, the accuracy of DFT methods can vary significantly with the type of system under study. We will discuss a novel approach to the reduction of errors in gradient corrected and hybrid DFT functionals, using empirical localized orbital corrections (DFT-LOC), which addresses this problem effectively. For example, the mean unsigned error in atomization energies for the G3 data set using the B3LYP-LOC model is 0.8 kcal/mole, as compared with 4.8 kcal/mole for B3LYP and 1.0 kcal/mole for G3 theory.

Friesner, Richard

2007-03-01

296

The efficiency of trypsin digestion for mass-spectrometry-based identification and quantification of oxidized proteins: evaluation of the digestion of oxidized bovine serum albumin.  

PubMed

In bottom-up proteomics approaches, the enzymatic proteolysis step before mass spectrometry (MS) analysis is of crucial importance, as only the efficient digestion of the protein will ensure its accurate quantification. The structural and chemical alterations occurring upon protein oxidation may decrease the efficiency of trypsin digestion, compromising the ensuing MS analysis. Herein, the efficiency of the trypsin digestion of oxidized bovine serum albumin (BSA) was assessed by protein-sequence coverage and the exponentially modified protein abundance index (emPAI) algorithm, allowing a comparison of protein abundance in samples with different levels of oxidation. Despite the extensive oxidation induced to BSA, verified by analysis of protein carbonyls, no significant difference in the yield of tryptic peptides from oxidized samples could be observed by nano-high-performance liquid chromatography (HPLC) and nano-HPLC7-electrospray ionization-MS analysis. After a database search, similar protein-sequence coverage rates were obtained for both treated and control samples. Thus, exponentially modified protein abundance index scores confirmed that, regardless of being oxidized, the same amount of BSA was present in the sodium dodecyl sulfate/polyacrylamide gel electrophoresis bands excised for digestion. The obtained results show that the digestion of the control and oxidized samples were similar, leading to the conclusion that in-gel proteolysis is not a main hindrance for the identification and quantification of oxidized proteins by MS. PMID:24892298

Gouveia, Duarte D; Silva, André M N; Vitorino, Rui; Domingues, M Rosário M; Domingues, Pedro

2014-01-01

297

Herbal adaptogens combined with protein fractions from bovine colostrum and hen egg yolk reduce liver TNF-? expression and protein carbonylation in Western diet feeding in rats  

PubMed Central

Background We examined if a purported anti-inflammatory supplement (AF) abrogated Western-diet (WD)-induced liver pathology in rats. AF contained: 1) protein concentrates from bovine colostrum and avian egg yolk; 2) herbal adaptogens and antioxidants; and 3) acetyl-L-carnitine. Methods Nine month-old male Brown Norway rats were allowed ad libitum access to WD for 41–43 days and randomly assigned to WD?+?AF feeding twice daily for the last 31–33 days (n?=?8), or WD and water-placebo feeding twice daily for the last 31–33 days (n?=?8). Rats fed a low-fat/low-sucrose diet (CTL, n?=?6) for 41–43 days and administered a water-placebo twice daily for the last 31–33 days were also studied. Twenty-four hours following the last gavage-feed, liver samples were analyzed for: a) select mRNAs (via RT-PCR) as well as genome-wide mRNA expression patterns (via RNA-seq); b) lipid deposition; and, c) protein carbonyl and total antioxidant capacity (TAC). Serum was also examined for TAC, 8-isoprostane and clinical chemistry markers. Results WD?+?AF rats experienced a reduction in liver Tnf-? mRNA (-2.8-fold, p?protein carbonyl content differed between WD?+?AF versus WD rats, although liver protein carbonyls tended to be lower in WD?+?AF versus CTL rats (p?=?0.08). RNA-seq revealed that 19 liver mRNAs differed between WD?+?AF versus WD when both groups were compared with CTL rats (+/- 1.5-fold, p?

2014-01-01

298

Evaluation of Serum-Derived Bovine Immunoglobulin Protein Isolate in Subjects with Diarrhea-Predominant Irritable Bowel Syndrome  

PubMed Central

BACKGROUND There is increased interest in combining nutritional modalities with pharmacological therapies for managing patients with diarrhea-predominant IBS (IBS-D). AIM A randomized, double-blind, placebo-controlled study to evaluate the impact of oral serum-derived bovine immunoglobulin/protein isolate (SBI) on gastrointestinal symptom scores and quality of life (QoL) in subjects with IBS-D. METHODS Study subjects previously diagnosed with IBS-D according to ROME II criteria were recruited from London, Ontario, Canada and assigned to receive 5 g/day SBI, 10 g/day SBI, or placebo for 6 weeks. Daily symptom frequency and severity scores and a modified IBS-36 questionnaire assessed the impact of nutritional intervention. Laboratory assessments were performed at screening and end of treatment (EOT) to evaluate safety. Within-group comparisons of changes in number of days per week with symptoms and symptom severity were conducted on the per-protocol population of subjects using a t-test. RESULTS Subjects who received SBI at 10 g/day (N = 15) had statistically significant within-group reductions in abdominal pain (p < 0.01), loose stools (p < 0.01), bloating (p < 0.05), flatulence (p < 0.01), urgency (p < 0.05) and any symptom (p < 0.01) at EOT vs. baseline. Subjects receiving 5 g/day of SBI (N = 15) realized statistically significant within-group reductions in days with flatulence (p < 0.035), incomplete evacuation (p < 0.05), and any symptom (p < 0.01). There were no significant changes in QoL scores or in hematology or clinical chemistry among treatment groups. CONCLUSIONS This pilot study showed that nutritional therapy with either 10 g/day or 5 g/day of SBI in 30 patients was well tolerated and resulted in statistically significant within group improvements in both symptom days and in daily symptom scores in subjects with IBS-D. Additional studies are underway with larger numbers of subjects to validate these findings.

Wilson, Dale; Evans, Malkanthi; Weaver, Eric; Shaw, Audrey L.; Klein, Gerald L.

2013-01-01

299

Insulin-like activity and immunoreactive insulin content of gel-filtered protein fractions of bovine serum  

Microsoft Academic Search

Summary  Serum globulin fractions isolated by gel filtration from fasting bovine sera with simultaneous exclusion of low molecular endogenous immunoreactive insulin (IRI), were found to contain immunoreactive insulin-like material in quantities which equal or surpass the directly assayable IRI content of native fasting sera. In one additional and clearly aberrant serum, independently isolated globulin fractions were found to contain 306 U

W. Konijnendijk; P. R. Bouman

1970-01-01

300

Bovine Pericardium Patch Wrapping Intestinal Anastomosis Improves Healing Process and Prevents Leakage in a Pig Model  

PubMed Central

Failure of intestinal anastomosis is a major complication following abdominal surgery. Biological materials have been introduced as reinforcement of abdominal wall hernia in contaminated setting. An innovative application of biological patch is its use as reinforcement of gastrointestinal anastomosis. The aim of study was to verify whether the bovine pericardium patch improves the healing of anastomosis, when in vivo wrapping the suture line of pig intestinal anastomosis, avoiding leakage in the event of deliberately incomplete suture. Forty-three pigs were randomly divided: Group 1 (control, n?=?14): hand-sewn ileo-ileal and colo-colic anastomosis; Group 2 (n?=?14): standard anastomosis wrapped by pericardium bovine patch; Group 3 (n?=?1) and 4 (n?=?14): one suture was deliberately incomplete and also wrapped by patch in the last one. Intraoperative evaluation, histological, biochemical, tensiometric and electrophysiological studies of intestinal specimens were performed at 48 h, 7 and 90 days after. In groups 2 and 4, no leak, stenosis, abscess, peritonitis, mesh displacement or shrinkage were found and adhesion rate decreased compared to control. Biochemical studies showed mitochondrial function improvement in colic wrapped anastomosis. Tensiometric evaluations suggested that the patch preserves the colic contractility similar to the controls. Electrophysiological results demonstrated that the patch also improves the mucosal function restoring almost normal transport properties. Use of pericardium bovine patch as reinforcement of intestinal anastomosis is safe and effective, significantly improving the healing process. Data of prevention of acute peritonitis and leakage in cases of iatrogenic perforation of anastomoses, covered with patch, is unpublished.

Testini, Mario; Gurrado, Angela; Portincasa, Piero; Scacco, Salvatore; Marzullo, Andrea; Piccinni, Giuseppe; Lissidini, Germana; Greco, Luigi; De Salvia, Maria Antonietta; Bonfrate, Leonilde; Debellis, Lucantonio; Sardaro, Nicola; Staffieri, Francesco; Carratu, Maria Rosaria; Crovace, Antonio

2014-01-01

301

Modeling the topology of protein interaction networks  

NASA Astrophysics Data System (ADS)

A major issue in biology is the understanding of the interactions between proteins. These interactions can be described by a network, where the proteins are modeled by nodes and the interactions by edges. The origin of these protein networks is not well understood yet. Here we present a two-step model, which generates clusters with the same topological properties as networks for protein-protein interactions, namely, the same degree distribution, cluster size distribution, clustering coefficient, and shortest path length. The biological and model networks are not scale-free but exhibit small-world features. The model allows the fitting of different biological systems by tuning a single parameter.

Schneider, Christian M.; de Arcangelis, Lucilla; Herrmann, Hans J.

2011-07-01

302

Assessment of a Protein Cocktail-Based Skin Test for Bovine Tuberculosis in a Double-Blind Field Test in Cattle  

PubMed Central

Bovine tuberculosis (bTB) is a worldwide zoonosis caused mainly by Mycobacterium bovis. The traditional diagnostic method used often is the tuberculin skin test, which uses bovine purified protein derivatives (PPD-B). However, it is difficult to maintain uniformity of PPD-B from batch to batch, and it shares common antigens with nonpathogenic environmental mycobacteria. To overcome these problems, M. bovis-specific antigens that showed good T cell stimulation, such as CFP-10, ESAT-6, Rv3615c, etc., have been used in the skin test, but there have been no large-scale clinical studies on these antigens. In this study, two combinations (CFP-10/ESAT-6/TB10.4 protein cocktail and CFP-10/ESAT-6/Rv3872/MPT63 protein cocktail) were developed and used as stimuli in the skin test. Cattle were double-blind tested to assess the efficiency of the protein cocktail-based skin tests. The results showed that the CFP-10/ESAT-6/TB10.4 protein cocktail-based skin test can differentiate TB-infected cattle from Mycobacterium avium-infected ones and that it shows a high degree of agreement with the traditional tuberculin skin test (? = 0.8536) and gamma interferon (IFN-?) release assay (? = 0.8154). Compared to the tuberculin skin test, the relative sensitivity and relative specificity of the CFP-10/ESAT-6/TB10.4-based skin test were 87% and 97%, respectively., The relative sensitivity and relative specificity of the CFP-10/ESAT-6/TB10.4-based skin test were 93% and 92%, respectively, on comparison with the IFN-? release assay. The correlation between the increases in skin thickness observed after the inoculation of stimuli was high (PPD-B versus CFP-10/ESAT-6/TB10.4, Spearman r of 0.8435). The correlation between the optical density at 450 nm (OD450) obtained after blood stimulation with PPD-B and the increase in skin thickness observed after inoculation of the CFP-10/ESAT-6/TB10.4 protein cocktail was high (Spearman r = 0.7335). Therefore, the CFP-10/ESAT-6/TB10.4-based skin test responses correlate to traditional measures of bovine TB evaluation, including skin test and gamma interferon release assay.

Xin, Ting; Jia, Hong; Ding, Jiabo; Li, Pingjun; Yang, Hongjun; Hou, Shaohua; Yuan, Weifeng; Guo, Xiaoyu; Wang, Haichun; Liang, Qianqian; Li, Ming

2013-01-01

303

Relationships of the phase velocity with the microarchitectural parameters in bovine trabecular bone in vitro: Application of a stratified model  

NASA Astrophysics Data System (ADS)

The present study aims to provide insight into the relationships of the phase velocity with the microarchitectural parameters in bovine trabecular bone in vitro. The frequency-dependent phase velocity was measured in 22 bovine femoral trabecular bone samples by using a pair of transducers with a diameter of 25.4 mm and a center frequency of 0.5 MHz. The phase velocity exhibited positive correlation coefficients of 0.48 and 0.32 with the ratio of bone volume to total volume and the trabecular thickness, respectively, but a negative correlation coefficient of -0.62 with the trabecular separation. The best univariate predictor of the phase velocity was the trabecular separation, yielding an adjusted squared correlation coefficient of 0.36. The multivariate regression models yielded adjusted squared correlation coefficients of 0.21-0.36. The theoretical phase velocity predicted by using a stratified model for wave propagation in periodically stratified media consisting of alternating parallel solid-fluid layers showed reasonable agreements with the experimental measurements.

Lee, Kang Il

2012-08-01

304

A stochastic risk-analysis model for the spread of bovine viral diarrhea virus after introduction to naďve cow–calf herds  

Microsoft Academic Search

A stochastic SIR model was developed to simulate the spread of bovine viral diarrhea virus (BVDV) through a cow–calf herd and estimate the effect of the virus on the herd, including abortions, calf morbidity, and calf mortality. The model was applied with three herd sizes (400, 100, and 50 head) and four control strategies (no intervention, vaccination of breeding stock,

Rebecca L. Smith; Michael W. Sanderson; David G. Renter; Robert Larson; Bradley White

2010-01-01

305

Differences in Levels of Secreted Locus of Enterocyte Effacement Proteins between Human Disease-Associated and Bovine Escherichia coli O157  

PubMed Central

Ongoing extensive epidemiological studies of verotoxin-carrying Escherichia coli O157 (stx+ eae+) have shown this bacterial pathogen to be common in cattle herds in the United States and the United Kingdom. However, the incidence of disease in humans due to this pathogen is still very low. This study set out to investigate if there is a difference between strains isolated from human disease cases and those isolated from asymptomatic cattle which would account for the low disease incidence of such a ubiquitous organism. The work presented here has compared human disease strains from both sporadic and outbreak cases with a cross-section, as defined by pulsed-field gel electrophoresis, of E. coli O157 strains from cattle. Human (n = 22) and bovine (n = 31) strains were genotyped for carriage of the genes for Shiga-like toxin types 1, 2, and 2c; E. coli secreted protein genes espA, espB, and espP; the enterohemolysin gene; eae (intimin); ast (enteroaggregative E. coli stable toxin [EAST]); and genes for common E. coli adhesins. Strains were also phenotyped for hemolysin, EspP, Tir, and EspD expression as well as production of actin and cytoskeletal rearrangement associated with attaching and effacing (A/E) lesions on HeLa cells. The genotyping confirmed that there was little difference between the two groups, including carriage of stx2 and stx2c, which was similar in both sets. ast alleles were confirmed to all contain mutations that would prevent EAST expression. espP mutations were found only in cattle strains (5 of 30). Clear differences were observed in the expression of locus of enterocyte effacement (LEE)-encoded factors between strains and in different media. EspD, as an indicator of LEE4 (espA, -B, and -D) expression, and Tir levels in supernatants were measured. Virtually all strains from both sources could produce EspD in Luria-Bertani broth, although at very different levels. Standard trichloroacetic acid precipitation of secreted proteins from tissue culture medium produced detectable levels of EspD from the majority of strains of human origin (15 of 20) compared with only a few (4 of 20) bovine strains (P < 0.001), which is indicative of much higher levels of protein secretion from the human strains. Addition of bovine serum albumin carrier protein before precipitation and enhanced detection techniques confirmed that EspD could be detected after growth in tissue culture medium for all strains, but levels from strains of human origin were on average 90-fold higher than those from strains of bovine origin. In general, levels of secretion also correlated with ability to form A/E lesions on HeLa cells, with only the high-level protein secretors in tissue culture medium exhibiting a localized adherence phenotype. This research shows significant differences between human- and bovine-derived E. coli O157 (stx+ eae+) strains and their production of certain LEE-encoded virulence factors. These data support the recent finding of Kim et al. (J. Kim, J. Nietfeldt, and A. K. Benson, Proc. Natl. Acad. Sci. USA 96:13288–13293, 1999) proposing different E. coli O157 lineages in cattle and humans and extend the differential to the regulation of virulence factors. Potentially only a subset of E. coli O157 isolates (stx+ eae+) in cattle may be capable of causing severe disease in humans.

McNally, Alan; Roe, Andrew J.; Simpson, Sally; Thomson-Carter, Fiona M.; Hoey, D. E. Elaine; Currie, Carol; Chakraborty, Trinad; Smith, David G. E.; Gally, David L.

2001-01-01

306

Monoclonal antibodies to surface-exposes proteins of Mycoplasma mycoides subsp. mycoides (small-colony strain), which causes contagious bovine pleuropneumonia.  

PubMed Central

Outbreaks of bovine pleuropneumonia caused by small-colony strains of Mycoplasma mycoides subsp. mycoides occur in Africa, and vaccination is used for control. Since protein subunits are needed to improve multivalent vaccines, monoclonal antibodies (MAbs) were made to facilitate protein identification and isolation. Eleven immunoglobulin M MAbs derived from mouse spleen donors immunized with disrupted whole organisms bound periodate-sensitive epitopes on externally exposed polysaccharide. Seven of these MAbs caused in vitro growth inhibition of M. mycoides subsp. mycoides; however, reaction with carbohydrate epitopes prevented their use in identifying proteins. Ten additional MAbs from mouse spleen donors immunized with Triton X-114-phase integral membrane proteins reacted with periodate-insensitive, proteinase K-sensitive epitopes. These MAbs were classified into three groups based on immunoblots of Triton X-114-phase proteins. One group reacted with 96-, 16-, and 15-kDa proteins. Another group reacted with 26-, 21-, and 16-kDa proteins, while a third group reacted only with 26- and 21-kDa proteins. One MAb from each group reacted with trypsinsensitive epitopes on live organisms, yet none caused in vitro growth inhibition. Representative MAbs reacted with all small-colony strains in immunoblots and did not react with large colony strains. However, these MAbs were not specific for small-colony strains, as proteins from two other M. mycoides cluster organisms were identified. Nevertheless, MAbs to surface-exposed epitopes on integral membrane proteins will be useful for isolation of these proteins for immunization, since one or more might induce growth-inhibiting antibodies or other protective responses.

Kiarie, M N; Rurangirwa, F R; Perryman, L E; Jasmer, D P; McGuire, T C

1996-01-01

307

The effect of dietary intake of the acidic protein fraction of bovine colostrum on influenza A (H1N1) virus infection.  

PubMed

Acidic protein levels in the milk decrease markedly as lactation progresses, suggesting that it is an important part of the colostrum. However, little attention has been paid to their biological function. In this study, we isolated the acidic protein fraction of bovine colostrum (AFC, isoelectric point <5) by anion-exchange chromatography, and investigated the effect of its dietary intake on influenza A (H1N1) virus infection. 100% of mice infected with 1 LD50 of the virus survived when administered AFC for 14 days prior to infection, compared with 33% survival when administered phosphate buffered saline (PBS). Moreover, consumption of AFC reduced the weight loss associated with infection. We propose that dietary intake of AFC has a prophylactic effect on influenza A virus infection. PMID:23620352

Xu, Mei Ling; Kim, Hyoung Jin; Chang, Don Yong; Kim, Hong-Jin

2013-06-01

308

Bovine pericardium patch wrapping intestinal anastomosis improves healing process and prevents leakage in a pig model.  

PubMed

Failure of intestinal anastomosis is a major complication following abdominal surgery. Biological materials have been introduced as reinforcement of abdominal wall hernia in contaminated setting. An innovative application of biological patch is its use as reinforcement of gastrointestinal anastomosis. The aim of study was to verify whether the bovine pericardium patch improves the healing of anastomosis, when in vivo wrapping the suture line of pig intestinal anastomosis, avoiding leakage in the event of deliberately incomplete suture. Forty-three pigs were randomly divided: Group 1 (control, n = 14): hand-sewn ileo-ileal and colo-colic anastomosis; Group 2 (n = 14): standard anastomosis wrapped by pericardium bovine patch; Group 3 (n = 1) and 4 (n = 14): one suture was deliberately incomplete and also wrapped by patch in the last one. Intraoperative evaluation, histological, biochemical, tensiometric and electrophysiological studies of intestinal specimens were performed at 48 h, 7 and 90 days after. In groups 2 and 4, no leak, stenosis, abscess, peritonitis, mesh displacement or shrinkage were found and adhesion rate decreased compared to control. Biochemical studies showed mitochondrial function improvement in colic wrapped anastomosis. Tensiometric evaluations suggested that the patch preserves the colic contractility similar to the controls. Electrophysiological results demonstrated that the patch also improves the mucosal function restoring almost normal transport properties. Use of pericardium bovine patch as reinforcement of intestinal anastomosis is safe and effective, significantly improving the healing process. Data of prevention of acute peritonitis and leakage in cases of iatrogenic perforation of anastomoses, covered with patch, is unpublished. PMID:24489752

Testini, Mario; Gurrado, Angela; Portincasa, Piero; Scacco, Salvatore; Marzullo, Andrea; Piccinni, Giuseppe; Lissidini, Germana; Greco, Luigi; De Salvia, Maria Antonietta; Bonfrate, Leonilde; Debellis, Lucantonio; Sardaro, Nicola; Staffieri, Francesco; Carratů, Maria Rosaria; Crovace, Antonio

2014-01-01

309

PreImplantation Factor (PIF) correlates with early mammalian embryo development-bovine and murine models  

PubMed Central

Background PreImplantation Factor (PIF), a novel peptide secreted by viable embryos is essential for pregnancy: PIF modulates local immunity, promotes decidual pro-adhesion molecules and enhances trophoblast invasion. To determine the role of PIF in post-fertilization embryo development, we measured the peptide's concentration in the culture medium and tested endogenous PIF's potential trophic effects and direct interaction with the embryo. Methods Determine PIF levels in culture medium of multiple mouse and single bovine embryos cultured up to the blastocyst stage using PIF-ELISA. Examine the inhibitory effects of anti-PIF-monoclonal antibody (mAb) added to medium on cultured mouse embryos development. Test FITC-PIF uptake by cultured bovine blastocysts using fluorescent microscopy. Results PIF levels in mouse embryo culture medium significantly increased from the morula to the blastocyst stage (ANOVA, P = 0.01). In contrast, atretic embryos medium was similar to the medium only control. Detectable - though low - PIF levels were secreted already by 2-cell stage mouse embryos. In single bovine IVF-derived embryos, PIF levels in medium at day 3 of culture were higher than non-cleaving embryos (control) (P = 0.01) and at day 7 were higher than day 3 (P = 0.03). In non-cleaving embryos culture medium was similar to medium alone (control). Anti-PIF-mAb added to mouse embryo cultures lowered blastocyst formation rate 3-fold in a dose-dependent manner (2-way contingency table, multiple groups, X2; P = 0.01) as compared with non-specific mouse mAb, and medium alone, control. FITC-PIF was taken-up by cultured bovine blastocysts, but not by scrambled FITC-PIF (control). Conclusions PIF is an early embryo viability marker that has a direct supportive role on embryo development in culture. PIF-ELISA use to assess IVF embryo quality prior to transfer is warranted. Overall, our data supports PIF's endogenous self sustaining role in embryo development and the utility of PIF- ELISA to detect viable embryos in a non-invasive manner.

2011-01-01

310

Microscopic model of protein crystal growth  

Microsoft Academic Search

A microscopic, reversible model to study protein crystal nucleation and growth is presented. The probability of monomer attachment to the growing crystal was assumed to be proportional to the protein volume fraction and the orientational factor representing the anisotropy of protein molecules. The rate of detachment depended on the free energy of association of the given monomer in the lattice,

Andrzej M. Kierzek; Piotr Pokarowski; Piotr Zielenkiewicz

2000-01-01

311

Bovine endometrial epithelial cells as a model system to study oxytocin receptor regulation.  

PubMed

Endometrial epithelial cell cultures were established from bovine uterine tissue collected during the oestrous cycle from commercially slaughtered animals. These cells were shown to express moderately high levels of oxytocin receptors (OTR) (up to 30000 per cell) after about one week in culture. These receptors have been characterized at the molecular, pharmacological and functional level and shown to be identical to those expressed in the bovine endometrium in vivo. Preliminary experiments to investigate the regulation of the OTR and its gene using this system, have shown that expression is to a large degree constitutive, the receptors being spontaneously upregulated during culture. Sex steroids at concentrations close to or above the serum limits observed in vivo appeared to have no effect, although the cells were shown to express mRNA for the specific steroid receptors throughout culture. Only the blastocyst product, interferon-tau, showed a significant effect, downregulating both OTR and their gene transcripts in the cultured endometrial epithelial cells. Although more extensive studies are necessary, these results support the view that the OTR gene is controlled in part at least by a combination of constitutive and inhibitory elements. PMID:10027614

Horn, S; Bathgate, R; Lioutas, C; Bracken, K; Ivell, R

1998-01-01

312

Stochastic model for protein flexibility analysis  

NASA Astrophysics Data System (ADS)

Protein flexibility is an intrinsic property and plays a fundamental role in protein functions. Computational analysis of protein flexibility is crucial to protein function prediction, macromolecular flexible docking, and rational drug design. Most current approaches for protein flexibility analysis are based on Hamiltonian mechanics. We introduce a stochastic model to study protein flexibility. The essential idea is to analyze the free induction decay of a perturbed protein structural probability, which satisfies the master equation. The transition probability matrix is constructed by using probability density estimators including monotonically decreasing radial basis functions. We show that the proposed stochastic model gives rise to some of the best predictions of Debye-Waller factors or B factors for three sets of protein data introduced in the literature.

Xia, Kelin; Wei, Guo-Wei

2013-12-01

313

Foot-and-Mouth Disease Virus, but Not Bovine Enterovirus, Targets the Host Cell Cytoskeleton via the Nonstructural Protein 3Cpro?  

PubMed Central

Foot-and-mouth disease virus (FMDV), a member of the Picornaviridae, is a pathogen of cloven-hoofed animals and causes a disease of major economic importance. Picornavirus-infected cells show changes in cell morphology and rearrangement of cytoplasmic membranes, which are a consequence of virus replication. We show here, by confocal immunofluorescence and electron microscopy, that the changes in morphology of FMDV-infected cells involve changes in the distribution of microtubule and intermediate filament components during infection. Despite the continued presence of centrosomes in infected cells, there is a loss of tethering of microtubules to the microtubule organizing center (MTOC) region. Loss of labeling for ?-tubulin, but not pericentrin, from the MTOC suggests a targeting of ?-tubulin (or associated proteins) rather than a total breakdown in MTOC structure. The identity of the FMDV protein(s) responsible was determined by the expression of individual viral nonstructural proteins and their precursors in uninfected cells. We report that the only viral nonstructural protein able to reproduce the loss of ?-tubulin from the MTOC and the loss of integrity of the microtubule system is FMDV 3Cpro. In contrast, infection of cells with another picornavirus, bovine enterovirus, did not affect ?-tubulin distribution, and the microtubule network remained relatively unaffected.

Armer, Hannah; Moffat, Katy; Wileman, Thomas; Belsham, Graham J.; Jackson, Terry; Duprex, W. Paul; Ryan, Martin; Monaghan, Paul

2008-01-01

314

Isolation of a cDNA clone for the type I regulatory subunit of bovine cAMP-dependent protein kinase.  

PubMed Central

A cDNA clone for the type I regulatory subunit (RI) of cAMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) was isolated from bovine testis by a differential screening method. mRNA coding for RI was enriched 50- to 100-fold by polysome immunoadsorption chromatography with affinity-purified rabbit anti-RI and protein A-Sepharose. Poly(A)+ RNA from these polysomes was utilized to construct a cDNA library in pBR322, and this library was screened for hybridization to 32P-labeled cDNAs synthesized from either total or RI-enriched poly(A)+ RNA. Plasmids isolated from colonies showing preferential hybridization to the latter probe were further characterized by hybrid selection and DNA sequence analysis. One of these plasmids (designated 62C12) contains a 1,350-nucleotide insert that hybridized to RI mRNA; partial nucleotide sequence analysis confirmed its identity and indicated that it may contain the entire RI coding region. We also have identified a recombinant plasmid with a 1,550-nucleotide insert that selected through hybridization a mRNA coding for a 55,000-dalton protein that crossreacts with anti-RI antibodies. The function of this latter protein is unknown. Images

Lee, D C; Carmichael, D F; Krebs, E G; McKnight, G S

1983-01-01

315

Functional assay, expression of growth factors and proteins modulating bone-arrangement in human osteoblasts seeded on an anorganic bovine bone biomaterial.  

PubMed

The basic aspects of bone tissue engineering include chemical composition and geometry of the scaffold design, because it is very important to improve not only cell attachment and growth but especially osteodifferentiation, bone tissue formation, and vascularization. Geistlich Bio-Oss (GBO) is a xenograft consisting of deproteinized, sterilized bovine bone, chemically and physically identical to the mineral phase of human bone. In this study, we investigated the growth behaviour and the ability to form focal adhesions on the substrate, using vinculin, a cytoskeletal protein, as a marker. Moreover, the expression of bone specific proteins and growth factors such as type I collagen, osteopontin, bone sialoprotein, bone morphogenetic protein-2 (BMP-2), BMP-7 and de novo synthesis of osteocalcin in normal human osteoblasts (NHOst) seeded on xenogenic GBO were evaluated. Our observations suggest that after four weeks of culture in differentiation medium, the NHOst showed a high affinity for the three dimensional biomaterial; in fact, cellular proliferation, migration and colonization were clearly evident. The osteogenic differentiation process, as demonstrated by morphological, histochemical, energy dispersive X-ray microanalysis and biochemical analysis was mostly obvious in the NHOst grown on three-dimensional inorganic bovine bone biomaterial. Functional studies displayed a clear and significant response to calcitonin when the cells were differentiated. In addition, the presence of the biomaterial improved the response, suggesting that it could drive the differentiation of these cells towards a more differentiated osteogenic phenotype. These results encourage us to consider GBO an adequate biocompatible three-dimensional biomaterial, indicating its potential use for the development of tissue-engineering techniques. PMID:20648427

Trubiani, O; Fulle, S; Traini, T; Paludi, M; la Rovere, R; Orciani, M; Caputi, S; Piattelli, A

2010-01-01

316

Effects of Saturated Long-chain Fatty Acid on mRNA Expression of Genes Associated with Milk Fat and Protein Biosynthesis in Bovine Mammary Epithelial Cells  

PubMed Central

This study was conducted to determine the effects of saturated long-chain fatty acids (LCFA) on cell proliferation and triacylglycerol (TAG) content, as well as mRNA expression of ?s1-casein (CSN1S1) and genes associated with lipid and protein synthesis in bovine mammary epithelial cells (BMECs). Primary cells were isolated from the mammary glands of Holstein dairy cows, and were passaged twice. Then cells were cultured with different levels of palmitate or stearate (0, 200, 300, 400, 500, and 600 ?M) for 48 h and fetal bovine serum in the culture solution was replaced with fatty acid-free BSA (1 g/L). The results showed that cell proliferation tended to be increased quadratically with increasing addition of stearate. Treatments with palmitate or stearate induced an increase in TAG contents at 0 to 600 ?M in a concentration-dependent manner, and the addition of 600 ?M was less effective in improving TAG accumulation. The expression of acetyl-coenzyme A carboxylase alpha, fatty acid synthase and fatty acid-binding protein 3 was inhibited when palmitate or stearate were added in culture medium, whereas cluster of differentiation 36 and CSN1S1 mRNA abundance was increased in a concentration-dependent manner. The mRNA expressions of peroxisome proliferator-activated receptor gamma, mammalian target of rapamycin and signal transducer and activator of transcription 5 with palmitate or stearate had no significant differences relative to the control. These results implied that certain concentrations of saturated LCFA could stimulate cell proliferation and the accumulation of TAG, whereas a reduction may occur with the addition of an overdose of saturated LCFA. Saturated LCFA could up-regulate CSN1S1 mRNA abundance, but further studies are necessary to elucidate the mechanism for regulating milk fat and protein synthesis.

Qi, Lizhi; Yan, Sumei; Sheng, Ran; Zhao, Yanli; Guo, Xiaoyu

2014-01-01

317

Hidden Markov Models in Computational Biology Applications to Protein Modeling  

Microsoft Academic Search

Ridden .Markov Models (HMMs) are applied t.0 the problems of statistical modeling, database searching and multiple sequence alignment of protein families and protein domains. These methods are demonstrated on the globin family, the protein kinase catalytic domain, and the EF-hand calcium binding motif. In each case the parameters of an HMM are estimated from a training set of unaligned sequences.

Anders Krogh; Michael Brown; I. Saira

1994-01-01

318

Energy depletion of bovine mammary epithelial cells activates AMPK and suppresses protein synthesis through inhibition of mTORC1 signaling.  

PubMed

The molecular mechanisms by which cellular energy status regulates global protein synthesis in mammary epithelial cells have not been characterized. The objective of this study was to examine the effect of AMP-activated protein kinase (AMPK) activation by 2-deoxyglucose on protein synthesis and the mammalian target of rapamycin complex 1 (mTORC1) signaling pathway in bovine mammary epithelial cells. Phosphorylation of AMPK at Thr172 increased by 1.4-fold within 5 min, and remained elevated throughout a 30-min time course, in response to 2-deoxyglucose. Global rates of protein synthesis declined by 78% of control values. The decline in protein synthesis was associated with repression of mTORC1 signaling, as indicated by reduced phosphorylation of ribosomal protein S6 kinase 1 and eIF4E binding protein-1 (4E-BP1). Phosphorylation of ER-stress marker eIF2? was also increased but only at 30 min of 2-deoxyglucose exposure. 2-Deoxyglucose increased phosphorylation of tuberous sclerosis complex 2 (TSC2) on AMPK consensus sites but did not change the amount of TSC1 bound to TSC2. Activation of AMPK did not result in changes in the amount of raptor bound to mTOR. The inhibitory effects of AMPK activation on mTORC1 signaling were associated with a marked increase in Ser792 phosphorylation on raptor. Collectively, the results suggest that activation of AMPK represses global protein synthesis in mammary epithelial cells through inhibition of mTORC1 signaling. PMID:22972179

Burgos, S A; Kim, J J M; Dai, M; Cant, J P

2013-03-01

319

3D structural models of transmembrane proteins  

PubMed Central

Summary Transmembrane proteins are macromolecules implicated in major biological process and diseases. Due to their specific neighborhood, few transmembrane protein structures are nowadays available. The building of structural models of transmembrane proteins is a major research area. Due to the lack of available 3D structures, automatic homology modeling is not an efficient way to propose pertinent structural models. Hence, most of the structural models of transmembrane proteins are done through a more complex protocol. This latter comprises the use of secondary structure prediction to complete the comparative modeling process. Then, refinement and assessment steps are performed go often to a novel comparative modeling process. Nowadays, it is also possible to take attention to the helix – helix and helix – lipid interactions, and, to build even quaternary structures. In all cases, the taking into account of experimental data is the most important factor to proceed to correct structural models.

De Brevern, Alexandre G.

2010-01-01

320

Pathogenicity of yeasts and algae isolated from bovine mastitis secretions in a murine model.  

PubMed

The pathogenicity of strains of yeasts and algae isolated from bovine mastitis secretions was evaluated in immunosuppressed mice. Strains of Candida tropicalis (n = 3) were the most pathogenic, but all strains of Geotrichum capitatum (n = 5) and the colourless alga Prototheca zopfii (n = 5) were also lethal to mice at the highest dose of 1 x 10(7) CFU per mouse. Reisolation of the inoculated micro-organisms and the occurrence of histopathological lesions within organs of mice challenged with 1 x 10(3) to 1 x 10(7) CFU per animal revealed strains of C. krusei (n = 17), C. kefyr (n = 6) and C. rugosa (n = 6) to have a moderate pathogenicity, whereas strains of C. valida (n = 1) and C. catenulata (n = 2) were weakly pathogenic and non-pathogenic respectively. PMID:7845414

Jensen, H E; Aalbaek, B

1994-01-01

321

Protein crystal growth in microgravity review of large scale temperature induction method: Bovine insulin, human insulin and human {alpha}-interferon  

SciTech Connect

The Protein Crystal Growth Facility (PCF) is space-flight hardware that accommodates large scale protein crystal growth experiments using temperature change as the inductive step. Recent modifications include specialized instrumentation for monitoring crystal nucleation with laser light scattering. This paper reviews results from its first seven flights on the Space Shuttle, the last with laser light scattering instrumentation in place. The PCF's objective is twofold: (1) the production of high quality protein crystals for x-ray analysis and subsequent structure-based drug design and (2) preparation of a large quantity of relatively contaminant free crystals for use as time-release protein pharmaceuticals. The first three Shuttle flights with bovine insulin constituted the PCF's proof of concept, demonstrating that the space-grown crystals were larger and diffracted to higher resolution than their earth-grown counterparts. The later four PCF missions were used to grow recombinant human insulin crystals for x-ray analysis and continue productions trials aimed at the development of a processing facility for crystalline recombinant a-interferon.

Long, Marianna M.; Bishop, John Bradford; DeLucas, Lawrence J.; Nagabhushan, Tattanhalli L.; Reichert, Paul; Smith, G. David [Center for Macromolecular Crystallography University of Alabama at Birmingham, Birmingham, Alabama (United States); Schering Plough Research Institute Kenilworth, New Jersey (United States); Hauptman-Woodward Medical Research Institute Buffalo, New York and Roswell Park Cancer Institute Buffalo, New York (United States)

1997-01-10

322

Fourier transform infrared spectroscopic analysis of the intact zona pellucida of the mammalian egg: changes in the secondary structure of bovine zona pellucida proteins during fertilization.  

PubMed

The zona pellucida is the acellular transparent envelope surrounding the mammalian oocyte. An analysis of the changes in the structures of zona pellucida proteins is essential for understanding the molecular mechanisms underlying the important physiological roles of the zona during fertilization and preimplantation. The hardening of the zona caused by the structural changes during fertilization is generally accepted to be responsible for blocking polyspermy. In this study, we analyzed changes in the secondary structure of the zona during fertilization by Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy. The predominance of beta-sheet structure in porcine ovarian egg zona proteins in water was ascertained using FTIR spectra. Alpha-helix structure was also present. The attenuated total reflection (ATR)-FTIR spectrum of intact, unsolubilized porcine zonae pellucidae from ovarian eggs indicated that the zona proteins in the native zona pellucida also have beta-structure as the main constituent. Attenuated total reflection-FTIR spectroscopy of intact bovine zona pellucida obtained from ovarian and fertilized eggs at the blastocyst stage revealed that the beta-structure content increased during fertilization. Furthermore, a reduction of the thickness of the zona during fertilization was observed using transmission electron microscopy. Therefore, the change in the zona architecture that causes hardening of the zona during fertilization is accompanied by changes in the secondary structure of the zona proteins. PMID:16446492

Nara, Masayuki; Yonezawa, Naoto; Shimada, Takeshi; Takahashi, Kazuya; Tanokura, Masaru; Yumoto, Fumiaki; Nakagawa, Hiroyuki; Ohashi, Kazuyo; Hamano, Seizo; Nakano, Minoru

2006-02-01

323

The carboxy-terminal domain shared by the bovine papillomavirus E2 transactivator and repressor proteins contains a specific DNA binding activity.  

PubMed Central

The E2 open reading frame of bovine papilloma virus 1 (BPV-1) has been shown to encode both positive and negative acting transcriptional regulatory factors. The DNA binding properties of these factors were analysed to investigate the mechanism by which they might regulate viral gene expression. Polypeptides corresponding to the full-length E2 product and a shorter protein thought to represent the repressor function were synthesized in vitro by translation of T7 polymerase generated transcripts. Using rabbit antisera generated against synthetic peptides from the E2 open reading frame, it was possible to immunoprecipitate each of these products and show that each was capable of binding the same specific sequence located at several sites in the BPV-1 genome. This DNA binding property was mapped to a conserved carboxy-terminal domain of 101 amino acids by analysis of truncated polypeptides synthesized from the E2 open reading frame. Images

McBride, A A; Schlegel, R; Howley, P M

1988-01-01

324

Hyperoxia-induced ciliary loss and oxidative damage in an in vitro bovine model: The protective role of antioxidant vitamins E and C  

SciTech Connect

Highlights: Black-Right-Pointing-Pointer A new bovine bronchial model for studying hyperoxia-induced cilia loss is presented. Black-Right-Pointing-Pointer Hyperoxia-induced cilia loss was associated with increased sloughing of cells. Black-Right-Pointing-Pointer Hyperoxia led to higher epithelial glutathione levels, evidence of oxidative stress. Black-Right-Pointing-Pointer Hyperoxia led to increased DNA damage (Comet), and lipid peroxidation (TBARS). Black-Right-Pointing-Pointer Vitamins C and E partially protected against hyperoxia-induced cilia loss. -- Abstract: Although elevated oxygen fraction is used in intensive care units around the world, pathological changes in pulmonary tissue have been shown to occur with prolonged exposure to hyperoxia. In this work a bovine bronchus culture model has been successfully used to evaluate the effects of hyperoxia on ciliated epithelium in vitro. Samples were cultured using an air interface method and exposed to normoxia, 21% O{sub 2} or hyperoxia, 95% O{sub 2}. Cilial coverage was assessed using scanning electron microscopy (SEM). Tissue damage (lactate dehydrogenase, LDH, in the medium), lipid peroxidation (thiobarbituric acid reactive substances, TBARS), DNA damage (comet assay), protein oxidation (OxyBlot kit) and antioxidant status (total glutathione) were used to assess whether the hyperoxia caused significant oxidative stress. Hyperoxia caused a time-dependent decline (t{sub Vulgar-Fraction-One-Half} = 3.4 d compared to 37.1 d under normoxia) in cilial coverage (P < 0.0001). This was associated with a significant increase in the number of cells (2.80 {+-} 0.27 Multiplication-Sign 10{sup 6} compared to 1.97 {+-} 0.23 Multiplication-Sign 10{sup 6} ml{sup -1} after 6 d), many apparently intact, in the medium (P < 0.05); LDH release (1.06 {+-} 0.29 compared to 0.83 {+-} 0.36 {mu}mol min{sup -1} g{sup -1} after 6 d; P < 0.001); lipid peroxidation (352 {+-} 16 versus 247 {+-} 11 {mu}mol MDA g{sup -1} for hyperoxia and normoxia, respectively); % tail DNA (18.7 {+-} 2.2 versus 11.1 {+-} 1.5); protein carbonyls (P < 0.05); and total glutathione (229 {+-} 20 {mu}mol g{sup -1} versus 189 {+-} 15 {mu}mol g{sup -1}). Vitamins E (10{sup -7} M) and C (10{sup -6} or 10{sup -7} M) alone or in combination (10{sup -7} M and 10{sup -6} M, respectively) had a significant protective effect on the hyperoxia-induced reduction in percentage cilial coverage (P < 0.05). In conclusion, hyperoxia caused damage to cultured bovine bronchial epithelium and denudation of cilia. The antioxidant vitamins E and C significantly protected against hyperoxia-induced cilia loss.

Al-Shmgani, Hanady S.; Moate, Roy M. [School of Biomedical and Biological Sciences, University of Plymouth (United Kingdom)] [School of Biomedical and Biological Sciences, University of Plymouth (United Kingdom); Sneyd, J. Robert [Plymouth University Peninsula Schools of Medicine and Dentistry, Plymouth (United Kingdom)] [Plymouth University Peninsula Schools of Medicine and Dentistry, Plymouth (United Kingdom); Macnaughton, Peter D. [Derriford Critical Care Unit, Plymouth (United Kingdom)] [Derriford Critical Care Unit, Plymouth (United Kingdom); Moody, A. John, E-mail: jmoody@plymouth.ac.uk [School of Biomedical and Biological Sciences, University of Plymouth (United Kingdom)

2012-12-14

325

Effect of additives on the release of a model protein from PLGA microspheres  

Microsoft Academic Search

The purpose of this study was to investigate the effect of 2 additives, poly(ethylene glycol (PEG) 1000 and 1,2,3-tridecanoyl\\u000a glycerol (tricaprin), on the physico-chemical characteristics and in vitro release of a model protein, bovine serum albumin\\u000a (BSA), form poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres. BSA-loaded microspheres were prepared by the double emulsion\\u000a solvent evaporation method. Additives were incorporated into microspheres to modify

Feirong Kang; Jagdish Singh

2001-01-01

326

Physical interaction between bovine viral diarrhea virus nonstructural protein 4A and adenosine deaminase acting on RNA (ADAR).  

PubMed

Bovine viral diarrhea virus (BVDV) is a positive-sense RNA virus known to produce double-stranded RNA (dsRNA) during its replication in the cytoplasm. Extended dsRNA duplexes can be hyperedited by adenosine deaminase acting on RNA (ADAR), which catalyzes adenosine (A)-to-inosine (I) editing. A-to-I editing has been reported for various viruses. A number of cellular antiviral defense strategies are stimulated by dsRNA, and this may involve hyperediting of dsRNA by ADARs, followed by targeted cleavage by cytoplasmic endonucleases. Here, we identify ADAR as a binding partner of BVDV NS4A in vitro and in vivo and show that the N-terminal domain of NS4A is the ADAR-binding domain. We also show that ADAR has an inhibitory effect on BVDV replication when overexpressed in BVDV-infected bovine cells. Our findings suggest a role of NS4A in the interaction of BVDV with ADAR that favors virus replication. PMID:24500065

Mohamed, Yassir Mahgoub; Bangphoomi, Norasuthi; Yamane, Daisuke; Suda, Yuto; Kato, Kentaro; Horimoto, Taisuke; Akashi, Hiroomi

2014-07-01

327

Enhanced virulence of sheep-passaged bovine spongiform encephalopathy agent is revealed by decreased polymorphism barriers in prion protein conversion studies.  

PubMed

Bovine spongiform encephalopathy (BSE) can be efficiently transmitted to small ruminants (sheep and goats) with certain prion protein (PrP) genotypes. Polymorphisms in PrP of both the host and donor influence the transmission efficiency of transmissible spongiform encephalopathies (TSEs) in general. These polymorphisms in PrP also modulate the PrP conversion underlying TSE agent replication. Here we demonstrate that single-round protein misfolding cyclic amplification (PMCA) can be used to assess species and polymorphism barriers at the molecular level. We assessed those within and between the ovine and bovine species in vitro using a variety of natural scrapie and experimentally generated cross-species BSE agents. These BSE agents include ovBSE-ARQ isolates (BSE derived from sheep having the ARQ/ARQ PrP genotype), and two unique BSE-derived variants: BSE passaged in VRQ/VRQ sheep and a cow BSE agent isolate generated by back-transmission of ovBSE-ARQ into its original host. PMCA allowed us to quantitatively determine PrP conversion profiles that correlated with known in vivo transmissibility and susceptibility in the two ruminant species in which strain-specific molecular signatures, like its molecular weight after protease digestion, were maintained. Furthermore, both BSE agent isolates from ARQ and VRQ sheep demonstrated a surprising transmission profile in which efficient transmissions to both sheep and bovine variants was combined. Finally, all data support the notion that ARQ-derived sheep BSE points to a significant increase in virulence compared to all other tested scrapie- and BSE-derived variants reflected by the increased conversion efficiencies of previously inefficient convertible PrP variants (including the so-called "resistant" sheep ARR variant). Importance: Prion diseases such as scrapie in sheep and goats, BSE in cattle, and Creutzfeldt-Jakob disease (CJD) in humans are fatal neurodegenerative diseases caused by prions. BSE is known to be transmissible to a variety of hosts, including sheep and humans. Based on the typical BSE agent strain signatures and epidemiological data, the occurrence of a novel variant of CJD in humans was linked to BSE occurrence in the United Kingdom. Measures, including genetic selection of sheep toward less susceptible PrP genotypes, have been implemented to lower the risk of BSE transmission into sheep, since the disease could potentially spread into a natural reservoir. In this study, we demonstrated using molecular PrP conversion studies that when BSE is first transmitted through sheep, the host range is modified significantly and the PrP converting potency increased, allowing the ovine BSE to transmit more efficiently than cow BSE into supposedly less susceptible hosts. PMID:24371051

Priem, Jan; Langeveld, Jan P M; van Keulen, Lucien J M; van Zijderveld, Fred G; Andreoletti, Olivier; Bossers, Alex

2014-03-01

328

A Recombinant Human Parainfluenza Virus Type 3 (PIV3) in Which the Nucleocapsid N Protein Has Been Replaced by That of Bovine PIV3 Is Attenuated in Primates  

PubMed Central

The shipping fever (SF) and Kansas (Ka) strains of bovine parainfluenza virus type 3 (BPIV3) are restricted in their replication in rhesus monkeys 100- to 1,000-fold compared to human parainfluenza virus type 3 (HPIV3), and the Ka strain also was shown to be attenuated in humans. To initiate an investigation of the genetic basis of the attenuation of BPIV3 in primates, we produced viable chimeric HPIV3 recombinants containing the nucleoprotein (N) open reading frame (ORF) from either BPIV3 Ka or SF in place of the HPIV3 N ORF. These chimeric recombinants were designated cKa-N and cSF-N, respectively. Remarkably, cKa-N and cSF-N grew to titers comparable to those of their HPIV3 and BPIV3 parents in LLC-MK2 monkey kidney and Madin-Darby bovine kidney cells. Thus, the heterologous nature of the N protein did not impede replication in vitro. However, cKa-N and cSF-N were each restricted in replication in rhesus monkeys to a similar extent as Ka and SF, respectively. This identified the BPIV3 N protein as a determinant of the host range restriction of BPIV3 in primates. These chimeras thus combine the antigenic determinants of HPIV3 with the host range restriction and attenuation phenotype of BPIV3. Despite their restricted replication in rhesus monkeys, the chimeric viruses induced a level of resistance to HPIV3 challenge in these animals which was indistinguishable from that conferred by immunization with HPIV3. The infectivity, attenuation, and immunogenicity of these BPIV3/HPIV3 chimeras suggest that the modified Jennerian approach described in the present report represents a novel method to design vaccines to protect against HPIV3-induced disease in humans.

Bailly, Jane E.; McAuliffe, Josephine M.; Durbin, Anna P.; Elkins, William R.; Collins, Peter L.; Murphy, Brian R.

2000-01-01

329

Determination by photoaffinity labelling of the hydrophobic part of the binding site for acyl-CoA esters on acyl-CoA-binding protein from bovine liver.  

PubMed Central

Acyl-CoA esters containing the photoreactive acids 12-(4'-azido-2'-nitrophenoxy)[1-14C]dodecanoic acid ([14C]AND-acid) or N-(4'-azido-2'-nitro-[3'-5'-3H]phenyl)-12-aminododecanoic acid ([3H]NANPA-acid) were synthesized. The photoreactive acyl-CoA esters could be bound to bovine acyl-CoA-binding protein (ACBP) and photocrosslinked to the protein. The photocrosslinked acyl-CoA-ACBP complex was separated from unlabelled ACBP on reverse-phase h.p.l.c. and the purified complex was digested with trypsin, Staphylococcus aureus V8 proteinase or endoproteinase Asp-N. By four independent peptide maps it was shown that the amino acids taking part in forming the hydrophobic binding site for acyl-CoA esters in bovine ACBP are located on the peptide segment from Asp21 to Asp38. Both photoreactive acyl-CoA esters used in this study labelled strongly in the segment from Tyr28 to Ala34. 12-(4'-Azido-2'-nitrophenoxy)[1-14C]-dodecanoyl-CoA ([14C]AND-CoA) also introduced a label at position Asp38, but o labelling was found before Ser29. In contrast, N-(4'-azido-2'-nitro[3',5'-3H]phenyl)-12-aminododecanoyl-CoA [3H]NANPA-CoA) also labelled the segment from Asp21 to Tyr28. The difference in labelling by the two photoreactive ligands is most likely caused by different mobility of the arylazido group when linked to the fatty acid either through a phenolic O- or an anilinic N- bond.

Hach, M; Pedersen, S N; Borchers, T; H?jrup, P; Knudsen, J

1990-01-01

330

Peroxynitrite-mediated oxidative protein modifications  

Microsoft Academic Search

Proteins are targets of reactive species and detection of oxidatively modified proteins is often used as an index of oxidative stress. Peroxynitrite is a strong oxidant formed by reaction of nitric oxide with superoxide. Using fatty acid-free bovine serum albumin as a model we examined peroxynitrite-mediated protein modifications. The reaction of protein with peroxynitrite resulted in the oxidation of tryptophan

Harry Ischiropoulos; Abu B. Al-Mehdi

1995-01-01

331

Molecular analysis of non structural rotavirus group A enterotoxin gene of bovine origin from India.  

PubMed

The rotavirus enterotoxin NSP4 (nonstructural protein 4), plays a pivotal role in viral morphogenesis as well as pathogenesis. In this study, the NSP4 gene of rotavirus group A (RVA) isolates of bovine origin isolated in several states of India from 2008 to 2011 were characterized. The complete open reading frame of 23 RVA strains were sequenced and analyzed phylogenetically. Genotype E1 was detected for the first time in bovines from India, in addition to the more common bovine genotype E2. Sequence similarity analysis of the E1 sequences showed a close genetic relatedness to human strains. Six of the bovine E2 genotypes strains clustered near bovine and unusual human strains (possible human animal reassortant) from Thailand, while the remaining E2 sequences clustered with Indian bovine strains. Analysis pointed out one positively selected site (154aa), believe to be part of an antigenic region and 123 negatively selected sites. Unexpectedly, a pentameric NSP4 structure of the coiled coil domain in the E1 carrying strains and a monomeric NSP4 in RVA strain P14 (E2) was predicted based on homology modeling, potentially affecting the biological properties of NSP4. The close relationship between bovine and human rotavirus strains further highlights the complex interaction among rotaviruses of different species. PMID:24747605

Malik, Yashpal Singh; Kumar, Naveen; Sharma, Kuldeep; Ghosh, Souvik; Bányai, Krisztián; Balasubramanian, Ganesh; Kobayashi, Nobumichi; Matthijnssens, Jelle

2014-07-01

332

Screening of Mycobacterium avium subsp. paratuberculosis mutants for attenuation in a bovine monocyte-derived macrophage model  

PubMed Central

Vaccination remains a major tool for prevention and progression of Johne's disease, a chronic enteritis of ruminants worldwide. Currently there is only one licensed vaccine within the United States and two vaccines licensed internationally against Johne's disease. All licensed vaccines reduce fecal shedding of Mycobacterium avium subsp. paratuberculosis (MAP) and delay disease progression. However, there are no available vaccines that prevent disease onset. A joint effort by the Johne's Disease Integrated Program (JDIP), a USDA-funded consortium, and USDA—APHIS/VS sought to identify transposon insertion mutant strains as vaccine candidates in part of a three phase study. The focus of the Phase I study was to evaluate MAP mutant attenuation in a well-defined in vitro bovine monocyte-derived macrophage (MDM) model. Attenuation was determined by colony forming unit (CFUs) counts and slope estimates. Based on CFU counts alone, the MDM model did not identify any mutant that significantly differed from the wild-type control, MAP K-10. Slope estimates using mixed models approach identified six mutants as being attenuated. These were enrolled in protection studies involving murine and baby goat vaccination-challenge models. MDM based approach identified trends in attenuation but this did not correlate with protection in a natural host model. These results suggest the need for alternative strategies for Johne's disease vaccine candidate screening and evaluation.

Lamont, Elise A.; Talaat, Adel M.; Coussens, Paul M.; Bannantine, John P.; Grohn, Yrjo T.; Katani, Robab; Li, Ling-ling; Kapur, Vivek; Sreevatsan, Srinand

2014-01-01

333

Recombinant protein and DNA vaccines derived from hsp60 Trichophyton mentagrophytes control the clinical course of trichophytosis in bovine species and guinea-pigs.  

PubMed

Heat shock proteins (hsp) were identified in many infectious agents as immunodominant antigens with a protective effect. Immunization of laboratory animals by selected representants of hsp60, hsp70 and hsp90 isolated from several pathogens induced protective host immunity and significantly reduced clinical manifestation of infection. The present study involves preparation of a recombinant protein vaccine and a DNA vaccine. Both vaccines were derived from the hsp60 of the dermatophyte, Trichophyton mentagrophytes. Challenge trials with evaluation of the protective effect of vaccination were performed on calves and guinea-pigs. Both vaccination procedures reduced, in statistically significant fashion, the clinical course of skin mycosis in calves experimentally inoculated with the dermatophyte, T. mentagrophytes. In experiments with guinea-pigs, increased protection was only seen with DNA vaccination. After DNA vaccine application, no paravaccination side-effects (granulomas at the injection site, changed total state of the animal) were observed. Only vaccination with the recombinant protein in calf's experiment induced specific serum antibodies. This observation indicates that antibodies are not associated with protection. In summary, DNA vaccine hsp60 is the most promising for prevention of bovine trichophytosis. PMID:15504125

Milan, Raska; Alois, Rybnikar; Josef, Chumela; Jana, Belakova; Evzen, Weigl

2004-10-01

334

Choline and acetylcholine detection based on peroxidase-like activity and protein antifouling property of platinum nanoparticles in bovine serum albumin scaffold.  

PubMed

Platinum nanoparticles (PtNPs) in the scaffold of bovine serum albumin (BSA) through biomineralization are found to possess excellent peroxidase-like activity that can catalyze N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium salt (TOPS) coupled with 4-amino-antipyrine (4-AAP) by the action of hydrogen peroxide to give an obvious purple product. Based on this phenomenon, acetylcholinesterase (AChE) and choline oxidase (ChOx) are used to catalyze ACh and choline to form the active product H2O2 and the as-produced H2O2 is detected optically. Owning to the protection effect of the protein shell, BSA-PtNPs turn out to be very stable and preserve the catalytic activity in the presence of protein and even in the real plasma samples. This protein antifouling property makes the BSA-PtNPs suitable for a wide range of applications in sensors for biological samples. Choline in infant formula and ACh in plasma have been successfully detected. PMID:25038538

He, Shao-Bin; Wu, Gang-Wei; Deng, Hao-Hua; Liu, Ai-Lin; Lin, Xin-Hua; Xia, Xing-Hua; Chen, Wei

2014-12-15

335

Antigenic variation among bovine enteric coronaviruses (BECV) and bovine respiratory coronaviruses (BRCV) detected using monoclonal antibodies  

Microsoft Academic Search

Summary.  ?Bovine coronavirus (BCV) causes neonatal calf diarrhea (CD) and is associated with winter dysentery (WD) in adult dairy cattle.\\u000a It can also be isolated from the respiratory tracts of cattle entering feedlots. Monoclonal antibodies (MAbs) specific for\\u000a the hemagglutinin esterase (HE) and spike (S) surface proteins of 2 bovine enteric coronavirus (BECV) strains and two bovine\\u000a respiratory coronavirus (BRCV) strains

M. Hasoksuz; S. Lathrop; M. A. Al-dubaib; P. Lewis; L. J. Saif

1999-01-01

336

A Mesoscopic Model for Protein-Protein Interactions in Solution  

PubMed Central

Protein self-association may be detrimental in biological systems, but can be utilized in a controlled fashion for protein crystallization. It is hence of considerable interest to understand how factors like solution conditions prevent or promote aggregation. Here we present a computational model describing interactions between protein molecules in solution. The calculations are based on a molecular description capturing the detailed structure of the protein molecule using x-ray or nuclear magnetic resonance structural data. Both electrostatic and van der Waals interactions are included and the salt particles are explicitly treated allowing investigations of systems containing mono-, di-, and trivalent ions. For three different proteins—lysozyme, ?-chymotrypsinogen, and calbindin D9k—we have investigated under which conditions (salt concentration, ion valency, pH, and/or solvent) the proteins are expected to aggregate via evaluation of the second virial coefficient. Good agreement is found with experimental data where available. Calbindin is investigated in more detail, and it is demonstrated how changes in solvent and/or counterion valency lead to attractive ion-ion correlation effects. For high valency counterions we have found abnormal trends in the second virial coefficient. With trivalent counterions, attraction of two negatively charged protein molecules can be favored because the repulsive term is decreased for entropic reasons due to the low number of particles present.

Lund, Mikael; Jonsson, Bo

2003-01-01

337

Computational model for protein unfolding simulation  

NASA Astrophysics Data System (ADS)

The protein folding problem is one of the fundamental and important questions in molecular biology. However, the all-atom molecular dynamics studies of protein folding and unfolding are still computationally expensive and severely limited by the time scale of simulation. In this paper, a simple and fast protein unfolding method is proposed based on the conformational stability analyses and structure modeling. In this method, two structure-based conditions are considered to identify the unstable regions of proteins during the unfolding processes. The protein unfolding trajectories are mimicked through iterative structure modeling according to conformational stability analyses. Two proteins, chymotrypsin inhibitor 2 (CI2) and ? -spectrin SH3 domain (SH3) were simulated by this method. Their unfolding pathways are consistent with the previous molecular dynamics simulations. Furthermore, the transition states of the two proteins were identified in unfolding processes and the theoretical ? values of these transition states showed significant correlations with the experimental data (the correlation coefficients are >0.8). The results indicate that this method is effective in studying protein unfolding. Moreover, we analyzed and discussed the influence of parameters on the unfolding simulation. This simple coarse-grained model may provide a general and fast approach for the mechanism studies of protein folding.

Tian, Xu-Hong; Zheng, Ye-Han; Jiao, Xiong; Liu, Cai-Xing; Chang, Shan

2011-06-01

338

Strain activation of bovine aortic smooth muscle cell proliferation and alignment: study of strain dependency and the role of protein kinase A and C signaling pathways  

NASA Technical Reports Server (NTRS)

Smooth muscle cell (SMC) phenotype can be altered by physical forces as demonstrated by cyclic strain-induced changes in proliferation, orientation, and secretion of macromolecules. However, the magnitude of strain required and the intracellular coupling pathways remain ill defined. To examine the strain requirements for SMC proliferation, we selectively seeded bovine aortic SMC either on the center or periphery of silastic membranes which were deformed with 150 mm Hg vacuum (0-7% center; 7-24% periphery). SMC located in either the center or peripheral regions showed enhanced proliferation compared to cells grown under the absence of cyclic strain. Moreover, SMC located in the center region demonstrated significantly (P < 0.005) greater proliferation as compared to those in the periphery. In contrast, SMC exposed to high strain (7-24%) demonstrated alignment perpendicular to the strain gradient, whereas SMC in the center (0-7%) remained aligned randomly. To determine the mechanisms of these phenomena, we examined the effect of cyclic strain on bovine aortic SMC signaling pathways. We observed strain-induced stimulation of the cyclic AMP pathway including adenylate cyclase activity and cyclic AMP accumulation. In addition, exposure of SMC to cyclic strain caused a significant increase in protein kinase C (PKC) activity and enzyme translocation from the cytosol to a particulate fraction. Further study was conducted to examine the effect of strain magnitude on signaling, particularly protein kinase A (PKA) activity as well as cAMP response element (CRE) binding protein levels. We observed significantly (P < 0.05) greater PKA activity and CRE binding protein levels in SMC located in the center as compared to the peripheral region. However, inhibition of PKA (with 10 microM Rp-cAMP) or PKC (with 5-20 ng/ml staurosporine) failed to alter either the strain-induced increase in SMC proliferation or alignment. These data characterize the strain determinants for activation of SMC proliferation and alignment. Although strain activated both the AC/cAMP/PKA and the PKC pathways in SMC, singular inhibition of PKA and PKC failed to prevent strain-induced alignment and proliferation, suggesting either their lack of involvement or the multifactorial nature of these responses.

Mills, I.; Cohen, C. R.; Kamal, K.; Li, G.; Shin, T.; Du, W.; Sumpio, B. E.

1997-01-01

339

Enthalpy convergence temperatures: proteins and model compounds  

Microsoft Academic Search

It is suggested that the classical model of non-polar hydration in proteins does not take into account a large negative enthalpy due to the solvation of polar surface, mostly represented by the peptide group. Analysis of the dissolution thermodynamics of several organic compounds, containing functional groups typical of proteins, clarifies that the penalty associated with the burial of polar surface

Raffaele Ragone; Paola Stiuso; Giovanni Colonna

1995-01-01

340

Optimal local propensities for model proteins  

Microsoft Academic Search

Lattice models of proteins were used to examine the role of local propen- sities in stabilizing the native state of a protein, using techniques drawn from spin-glass theory to characterize the free-energy landscapes. In the strong evolutionary limit, optimal condi- tions for folding are achieved when the contri- butions from local interactions to the stability of the native state is

Sridhar Govindarajan; Richard A. Goldstein

1995-01-01

341

Removal of model proteins by means of low-pressure inductively coupled plasma discharge  

NASA Astrophysics Data System (ADS)

Surgical instruments are intended to come into direct contact with the patients' tissues and thus interact with their first immune defence system. Therefore they have to be cleaned, sterilized and decontaminated, in order to prevent any kind of infections and inflammations or to exclude the possibility of transmission of diseases. From this perspective, the removal of protein residues from their surfaces constitutes new challenges, since certain proteins exhibit high resistance to commonly used sterilization and decontamination techniques and hence are difficult to remove without inducing major damages to the object treated. Therefore new approaches must be developed for that purpose and the application of non-equilibrium plasma discharges represents an interesting option. The possibility to effectively remove model proteins (bovine serum albumin, lysozyme and ubiquitin) from surfaces of different materials (Si wafer, glass, polystyrene and gold) by means of inductively coupled plasma discharges sustained in different argon containing mixtures is demonstrated and discussed in this paper.

Kylián, O.; Rauscher, H.; Gilliland, D.; Brétagnol, F.; Rossi, F.

2008-05-01

342

Non-classical mechanisms of steroid sensing in the ovary: lessons from the bovine oxytocin model.  

PubMed

Steroidogenic tissues such as the ovary, testes or adrenal glands are paradoxical in that they often indicate actions of steroid hormones within a dynamic range of ligand concentration in a high nanomolar or even micromolar level, i.e. at the natural concentrations existing within those organs. Yet ligand-activated nuclear steroid receptors act classically by direct interaction with DNA in the picomolar or low nanomolar range. Moreover, global genomic studies suggest that less than 40% of steroid-regulated genes involve classical responsive elements in gene promoter regions. The bovine oxytocin gene is a key element in the maternal recognition of pregnancy in ruminants and is regulated via an SF1 site in its proximal promoter. This gene is also regulated by steroids acting in a non-classical manner, involving nuclear receptors which do not interact directly with DNA. Dose-response relationships for these actions are in the high nanomolar range. Similar 'steroid sensing' mechanisms may prevail for other SF1-regulated genes and predict alternative pathways by which environmental endocrine disruptors might influence the functioning of steroid-producing organs and hence indirectly the steroid-dependent control of physiology and development. PMID:23632104

Ivell, Richard; Dai, Yanzhenzi; Mann, Navdeep; Anand-Ivell, Ravinder

2014-01-25

343

Template-Based Protein Structure Modeling  

PubMed Central

Functional characterization of a protein is often facilitated by its 3D structure. However, the fraction of experimentally known 3D models is currently less than 1% due to the inherently time-consuming and complicated nature of structure determination techniques. Computational approaches are employed to bridge the gap between the number of known sequences and that of 3D models. Template-based protein structure modeling techniques rely on the study of principles that dictate the 3D structure of natural proteins from the theory of evolution viewpoint. Strategies for template-based structure modeling will be discussed with a focus on comparative modeling, by reviewing techniques available for all the major steps involved in the comparative modeling pipeline.

Fiser, Andras

2014-01-01

344

Temporal regulation of mRNAs for select bone morphogenetic proteins (BMP), BMP receptors and their associated SMAD proteins during bovine early embryonic development: effects of exogenous BMP2 on embryo developmental progression  

PubMed Central

Background We previously demonstrated embryotrophic actions of maternal (oocyte-derived) follistatin during bovine early embryogenesis. Classical actions of follistatin are attributed to inhibition of activity of growth factors including activins and bone morphogenetic proteins (BMP). However, temporal changes in BMP mRNA in early bovine embryos and the effects of exogenous BMP on embryo developmental progression are not understood. The objectives of present studies were to characterize mRNA abundance for select BMP, BMP receptors and BMP receptor associated SMADs during bovine oocyte maturation and early embryogenesis and determine effects of addition of exogenous BMP protein on early development. Methods Relative abundance of mRNA for BMP2, BMP3, BMP7, BMP10, SMAD1, SMAD5, ALK3, ALK6, ALK2, BMPR2, ACVR2A and ACVR2B was determined by RT-qPCR analysis of germinal vesicle (GV) and in vitro matured metaphase II (MII) oocytes and in vitro produced embryos collected at pronuclear, 2-cell (C), 4C, 8C, 16C, morula and blastocyst stages. Effects of addition of recombinant human BMP2 (0, 1, 10 and 100 ng/ml) during initial 72 h of embryo culture on early cleavage (within 30 h post insemination), total cleavage, development to 8C-16C and blastocyst stages and blastocyst mRNA abundance for markers of inner cell mass (NANOG) and trophectoderm (CDX2) were also determined. Results Abundance of mRNA for BMP2, BMP10, SMAD1, SMAD5, ALK3, ALK2, BMPR2 and ACVR2B was elevated in MII oocytes and/or pronuclear stage embryos (relative to GV) and remained elevated through the 8C -16C stages, whereas BMP3, BMP7 and ALK2 mRNAs were transiently elevated. Culture of embryos to the 8C stage in the presence of ?-amanitin resulted in increased abundance for all of above transcripts examined relative to untreated 8C embryos. Effects of addition of exogenous BMP2 on early cleavage rates and rates of development to 8C-16C and blastocyst stages were not observed, but BMP2 treatment increased blastocyst mRNA for CDX2 and NANOG. Conclusions Abundance of maternally derived mRNAs for above BMP system components are dynamically regulated during oocyte maturation and early embryogenesis. Exogenous BMP2 treatment does not influence progression to various developmental endpoints, but impacts characteristics of resulting blastocysts. Results support a potential role for BMPs in bovine early embryogenesis.

2014-01-01

345

A model of consumers' risk perceptions toward recombinant bovine growth hormone (rbGH): the impact of risk characteristics.  

PubMed

This study estimates the effect risk characteristics, described as outrage factors by Hadden, have on consumers' risk perceptions toward the food-related biotechnology, recombinant bovine growth hormone (rbGH). The outrage factors applicable to milk from rbGH treated herds are involuntary risk exposure, unfamiliarity with the product's production process, unnatural product characteristics, lack of trust in regulator's ability to protect consumers in the marketplace, and consumers' inability to distinguish milk from rbGH treated herds compared to milk from untreated herds. An empirical analysis of data from a national survey of household food shoppers reveals that outrage factors mediate risk perceptions. The results support the inclusion of outrage factors into the risk perception model for the rbGH product, as they add significantly to the explanatory power of the model and therefore reduce bias compared to a simpler model of attitudinal and demographic factors. The study indicates that outrage factors which have a significant impact on risk perceptions are the lack of trust in the FDA as a food-related information source, and perceiving no consumer benefits from farmers' use of rbGH. Communication strategies to reduce consumer risk perceptions therefore could utilize agencies perceived as more trustworthy and emphasize the benefits of rbGH use to consumers. PMID:10765429

Grobe, D; Douthitt, R; Zepeda, L

1999-08-01

346

Evaluation of protein-protein docking model structures using all-atom molecular dynamics simulations combined with the solution theory in the energy representation  

NASA Astrophysics Data System (ADS)

We propose a method to evaluate binding free energy differences among distinct protein-protein complex model structures through all-atom molecular dynamics simulations in explicit water using the solution theory in the energy representation. Complex model structures are generated from a pair of monomeric structures using the rigid-body docking program ZDOCK. After structure refinement by side chain optimization and all-atom molecular dynamics simulations in explicit water, complex models are evaluated based on the sum of their conformational and solvation free energies, the latter calculated from the energy distribution functions obtained from relatively short molecular dynamics simulations of the complex in water and of pure water based on the solution theory in the energy representation. We examined protein-protein complex model structures of two protein-protein complex systems, bovine trypsin/CMTI-1 squash inhibitor (PDB ID: 1PPE) and RNase SA/barstar (PDB ID: 1AY7), for which both complex and monomer structures were determined experimentally. For each system, we calculated the energies for the crystal complex structure and twelve generated model structures including the model most similar to the crystal structure and very different from it. In both systems, the sum of the conformational and solvation free energies tended to be lower for the structure similar to the crystal. We concluded that our energy calculation method is useful for selecting low energy complex models similar to the crystal structure from among a set of generated models.

Takemura, Kazuhiro; Guo, Hao; Sakuraba, Shun; Matubayasi, Nobuyuki; Kitao, Akio

2012-12-01

347

A flavin-dependent sulfhydryl oxidase in bovine milk.  

PubMed

Both metal and flavin-dependent sulfhydryl oxidases catalyze the net generation of disulfide bonds with the reduction of oxygen to hydrogen peroxide. The first mammalian sulfhydryl oxidase to be described was an iron-dependent enzyme isolated from bovine milk whey (Janolino, V.G., and Swaisgood, H.E. (1975) J. Biol. Chem. 250, 2532-2537). This protein was reported to contain 0.5 atoms of iron per 89 kDa subunit and to be completely inhibited by ethylenediaminetetraacetate (EDTA). However the present work shows that a soluble 62 kDa FAD-linked and EDTA-insensitive sulfhydryl oxidase apparently constitutes the dominant disulfide bond-generating activity in skim milk. Unlike the metalloenzyme, the flavoprotein is not associated tightly with skim milk membranes. Sequencing of the purified bovine enzyme (>70% coverage) showed it to be a member of the Quiescin-sulfhydryl oxidase (QSOX) family. Consistent with its solubility, this bovine QSOX1 paralogue lacks the C-terminal transmembrane span of the long form of these proteins. Bovine milk QSOX1 is highly active toward reduced RNase and with the model substrate dithiothreitol. The significance of these new findings is discussed in relation to the earlier reports of metal-dependent sulfhydryl oxidases. PMID:17944490

Jaje, Jennifer; Wolcott, Holly N; Fadugba, Olajumoke; Cripps, Diane; Yang, Austin J; Mather, Ian H; Thorpe, Colin

2007-11-13

348

A Flavin-dependent Sulfhydryl Oxidase in Bovine Milk†  

PubMed Central

Both metal and flavin-dependent sulfhydryl oxidases catalyze the net generation of disulfide bonds with the reduction of oxygen to hydrogen peroxide. The first mammalian sulfhydryl oxidase to be described was an iron-dependent enzyme isolated from bovine milk whey. This protein was reported to contain 0.5 atoms of iron per 89 kDa subunit and to be completely inhibited by ethylenediaminetetraacetate (EDTA). However the present work shows that a soluble 62 kDa FAD-linked and EDTA-insensitive sulfhydryl oxidase apparently constitutes the dominant disulfide bond-generating activity in skim milk. Unlike the metalloenzyme, the flavoprotein is not associated tightly with skim milk membranes. Sequencing of the purified bovine enzyme (< 70% coverage) showed it to be a member of the Quiescin-sulfhydryl oxidase (QSOX) family. Consistent with its solubility, this bovine QSOX1 paralog lacks the C-terminal transmembrane span of the long form of these proteins. Bovine milk QSOX1 is highly active towards reduced RNase and with the model substrate dithiothreitol. The significance of these new findings is discussed in relation to the earlier reports of metal-dependent sulfhydryl oxidases.

Jaje, Jennifer; Wolcott, Holly N.; Fadugba, Olajumoke; Cripps, Diane; Yang, Austin J.; Mather, Ian H.; Thorpe, Colin

2008-01-01

349

Bovine Mammary Progenitor Cells: Current Concepts and Future Directions  

Microsoft Academic Search

Although cell number is positively correlated with milk production, much remains to be learned about the bovine mammary stem cell and progenitor cells. Bovine mammary development is driven by many of the same classic mammogenic hormones studied in murine models, yet histologic features of bovine mammary development differ from that of rodent models. Most notably, terminal end buds, as they

A. V. Capuco; S. Ellis

2005-01-01

350

Bovine Herpesvirus 1 Tegument Protein VP22 Interacts with Histones, and the Carboxyl Terminus of VP22 Is Required for Nuclear Localization  

PubMed Central

The bovine herpesvirus 1 (BHV-1) UL49 gene encodes a viral tegument protein termed VP22. UL49 homologs are conserved among alphaherpesviruses. Interestingly, the BHV-1 VP22 deletion mutant virus is asymptomatic and avirulent in infected cattle but produces only a slight reduction in titer in vitro. Attenuation of the BHV-1 VP22 deletion mutant virus in vivo suggests that VP22 plays a functional role in BHV-1 replication. In herpes simplex virus type 1, the VP22 homolog was previously shown to interact with another tegument protein,VP16, the ?-transinducing factor in vitro. In this report, we show that (i) the nuclear targeting of VP22 is independent of other viral factors, (ii) the carboxyl terminus of VP22 is required for its nuclear localization, (iii) VP22 associates with histones and nucleosomes, (iv) an antihistone monoclonal antibody cross-reacts with VP22, and (v) acetylation of histone H4 is decreased in VP22-expressing cells as well as virus-infected cells. Our data suggest that VP22 may have a modulatory function during BHV-1 infection.

Ren, Xiaodi; Harms, Jerome S.; Splitter, Gary A.

2001-01-01

351

Analysis of bovine selenoprotein P-like protein gene and availability of metal responsive element (MRE) located in its promoter  

Microsoft Academic Search

Selenoprotein P-like protein, similar to selenoprotein P, uses multiple TGAs for incorporation of selenocysteines but not as stop codons. It is also characterized by having a His–Pro-rich domain and a regionally differential expression pattern. Hence, in addition to selenium metabolism, this protein is considered to have a developmental function. In the present study, the structure of the selenoprotein P-like protein

Motoko Fujii; Kiyofumi Saijoh; Tatsuya Kobayashi; Shigeki Fujii; Myeong Jin Lee; Kimiaki Sumino

1997-01-01

352

The distribution of heat in bone during radiofrequency ablation of an ex vivo bovine model of osteoid osteoma.  

PubMed

Osteoid osteoma is treated primarily by radiofrequency (RF) ablation. However, there is little information about the distribution of heat in bone during the procedure and its safety. We constructed a model of osteoid osteoma to assess the distribution of heat in bone and to define the margins of safety for ablation. Cavities were drilled in cadaver bovine bones and filled with a liver homogenate to simulate the tumour matrix. Temperature-sensing probes were placed in the bone in a radial fashion away from the cavities. RF ablation was performed 107 times in tumours < 10 mm in diameter (72 of which were in cortical bone, 35 in cancellous bone), and 41 times in cortical bone with models > 10 mm in diameter. Significantly higher temperatures were found in cancellous bone than in cortical bone (p < 0.05). For lesions up to 10 mm in diameter, in both bone types, the temperature varied directly with the size of the tumour (p < 0.05), and inversely with the distance from it. Tumours of > 10 mm in diameter showed a trend similar to those of smaller lesions. No temperature rise was seen beyond 12 mm from the edge of a cortical tumour of any size. Formulae were developed to predict the expected temperature in the bone during ablation. PMID:24788505

Greenberg, A; Berenstein Weyel, T; Sosna, J; Applbaum, J; Peyser, A

2014-05-01

353

Effect of cyclin-dependent kinase (CDK) inhibition on expression, localization and activity of maturation promoting factor (MPF) and mitogen activated protein kinase (MAPK) in bovine oocytes.  

PubMed

This study aimed to assess the effects of cyclin-dependent kinase (CDK) inhibition on factors involved in the control of meiosis in bovine oocytes: maturation promoting factor (MPF) (p34(cdc2) and cyclin B1) and mitogen activated protein kinase (MAPK). Oocytes were maintained at germinal vesicle (GV) stage in vitro with 10??M of the CDK inhibitor butyrolactone I (BLI) for 24?h (inhibited). After this period, some of the oocytes were transferred to in vitro maturation (IVM) culture for 24?h (inhibited and matured). Control oocytes were assessed immediately after follicle aspiration (immature) or after in vitro maturation for 24?h (matured). Real-time PCR analyses showed that transcripts for p34(cdc2) and MAPK were detected in immature and inhibited oocytes and decreased after maturation, irrespective of CDK inhibition with BLI. Cyclin B1 was detected at similar levels in all oocyte groups. The p34(cdc2) and MAPK proteins were detected by Western blotting at similar levels in all oocyte groups, and cyclin B1 protein was detected only after maturation. Immunofluorescence detection showed that p34(cdc2) was localized in the cytoplasm and GV of immature oocytes, and then throughout the cytoplasm after maturation. Cyclin B1 and MAPK were detected in the cytoplasm in all oocyte groups. Maturation promoting factor and MAPK activities were similar throughout most of maturation for oocytes treated with or without BLI. In conclusion, CDK inhibition did not affect the expression (mRNA and protein levels) and localization of MPF and MAPK, and had nearly no effect on kinase activities during maturation. PMID:19602178

Quetglas, M D; Adona, P R; de Bem, T H C; Pires, P R L; Leal, C L V

2010-12-01

354

Simple Hydrophobic/Hydrophilic Protein Folding Model  

NSDL National Science Digital Library

the Simple Hydrophobic/Hydrophilic Protein Folding Model implements a 2-Dimensional hydrophobic/hydrophilic (HP) folding protein to test the energetics of protein folding, the change in size of a protein relative to energy, and the computation time for complex protein problems. A Metropolis Monte-Carlo algorithm is used to determine if randomly guided movements are carried out by monomers (non-specific amino acids) within the polymer (protein tertiary structure). The Monte-Carlo steps are run in a worker task (separate to EJS) while size and energy are periodically updated according to the Frames Per Second (FPS). A worker task isolates a single thread from the operating computer and uses that to cycle through a block of code as quickly as possible. In this case, one cycle constitutes one Monte-Carlo time step, the time unit. To accurately update data from a thread that runs outside of the EJS module, atomic variables are used. Every time the program updates the evolution page, the atomic variables for size and energy are recorded and added to the view (i.e. plots) for the polymer structure that occurred in the nearest worker task cycle to that evolution update. The Simple Hydrophobic/Hydrophilic Protein Folding Model was created using the Easy Java/JavaScript Simulations (EjsS) version 5 modeling tool. It is distributed as a ready-to-run (compiled) Java archive.

Kozlowski, Ryan

2014-06-01

355

Regeneration of bovine and octopus opsins in situ with natural and artificial retinals  

SciTech Connect

The authors consider the problem of color regulation in visual pigments for both bovine rhodopsin and octopus rhodopsin. Both pigments have 11-cis-retinal as their chromophore. These rhodopsins were bleached in their native membranes, and the opsins were regenerated with natural and artificial chromophores. Both bovine and octopus opsins were regenerated with the 9-cis- and 11-cis-retinal isomers, but the octopus opsin was additionally regenerated with the 13-cis and all-trans isomers. Titration of the octopus opsin with 11-cis-retinal gave an extinction coefficient for octopus rhodopsin of 27,000 {plus minus} 3,000 M{sup {minus}1} cm{sup {minus}1} at 475 nm. The absorption maxima of bovine artificial pigments formed by regenerating opsin with the 11-cis dihydro series of chromophores support a color regulation model for bovine rhodopsin in which the chromophore-binding site of the protein has two negative charges: one directly hydrogen bonded to the Schiff base nitrogen and another near carbon-13. Formation of octopus artificial pigments with both all-trans and 11-cis dihydro chromophores leads to a similar model for octopus rhodopsin and metarhodopsin: there are two negative charges in the chromophore-binding site, one directly hydrogen bonded to the Schiff base nitrogen and a second near carbon-13. The interaction of this second charge with the chromophore in octopus rhodopsin is weaker than in bovine, while in metarhodopsin it is as strong as in bovine.

Koutalos, Y.; Ebrey, T.G.; Tsuda, M.; Odashima, K.; Lien, T.; Park, M.H.; Shimizu, N.; Derguini, F.; Nakanishi, K.; Gilson, H.R.; Honig, B. (Univ. of Illinois, Urbana-Champaign (USA))

1989-03-21

356

Separation of bovine spermatozoa proteins using 2D-PAGE revealed the relationship between tektin-4 expression patterns and spermatozoa motility.  

PubMed

Poor semen quality has long been associated with bull infertility. However, the molecular basis in spermatozoa cells underlying the mechanisms of bull infertility remain unknown. The purpose of this study was to determine whether there is any protein in bovine spermatozoa related to semen quality. Semen samples from 18 Brahman bulls, 3 to 10 yrs of age, were assessed for semen quality in terms of spermatozoa motility and spermatozoa morphology. Spermatozoa extracts were separated using 2D-PAGE followed by staining with Coomassie blue. At least one duplicate gel was performed for each sample. Each gel was scanned with an ImageScanner System and analyzed for spots by ImageMaster 2D platinum software. The related protein spot(s) with semen quality was cut from the gel and identified by LC MS/MS. The results showed that at least 600 protein spots were detected in the spermatozoa extracts of the Brahman bulls. Of all these spots, there were 3 of 56 kDa at pI 6.4, 6.6 and 6.8 (Z(1), Z(2) and Z(3), respectively) that clearly showed different expression pattern among 18 Brahman bulls. Of 18 bulls (a) five showed the presence of spot Z(1) and Z(2) (pattern A) (b) one of spot Z(3) (pattern B) (c) five of spot Z(2) and Z(3) (pattern C) (d) one of spot Z(1) (pattern D) and (e) six of spot Z(2) (pattern E). Identification of spot Z(1), Z(2) and Z(3) by LC MS/MS had a similar result as matched to the tektin-4 protein of Bos taurus with a respective score of 171, 557 and 591. The statistical analysis of the 56 kDa protein patterns, tektin-4, indicated a significant effect on spermatozoa motility (P < 0.05) albeit non-significant on spermatozoa morphology. The bulls which showed pattern A had a higher percentage of spermatozoa motility than pattern E (P < 0.05) and not different from pattern C (P > 0.05). The statistical analysis also revealed that the presence of spot Z(1) had an effect on the percentage of spermatozoa motility (P < 0.01), whereas the presence of spot Z(2) and Z(3) had no effect (P > 0.05). The correlation coefficient between the relative protein content of spot Z(1) and the percentage of spermatozoa motility was 0.49. Our study demonstrates that the expression patterns of tektin-4 were a proxy for an effect on spermatozoa motility and consequently bull infertility. It may be that these protein patterns can be used as markers for improving bovine reproduction. PMID:22444558

Thepparat, T; Katawatin, S; Vongpralub, T; Duangjinda, M; Thammasirirak, S; Utha, A

2012-06-01

357

Predicting protein structure using hidden Markov models  

Microsoft Academic Search

We discuss how methods based on hidden Markov models performed in the fold-recognition sectionof the CASP2 experiment. Hidden Markov models were built for a representative set of just over onethousand structures from the Protein Data Bank (pdb). Each CASP2 target sequence was scored againstthis library of hmms. In addition, an hmm was built for each of the target sequences, and

Kevin Karplus; Kimmen Sjölander; Christian Barrett; Melissa Cline; David Haussler; Richard Hughey; Liisa Holm; Chris Sander

1997-01-01

358

Estradiol and Progesterone Exhibit Similar Patterns of Hepatic Gene Expression Regulation in the Bovine Model  

PubMed Central

Female sex steroid hormones, estradiol-17? (E2-17?) and progesterone (P4) regulate reproductive function and gene expression in a broad range of tissues. Given the central role of the liver in regulating homeostasis including steroid hormone metabolism, we sought to understand how E2-17? and P4 interact to affect global gene expression in liver. Ovariectomized cows (n?=?8) were randomly assigned to 4 treatment groups applied in a replicated Latin Square design: 1) No hormone supplementation, 2) E2-17? treatment (ear implant), 3) P4 treatment (intravaginal inserts), and 4) E2-17? combined with P4. After 14 d of treatment, liver biopsies were collected, allowing 28 d intervals between periods. Changes in gene expression in the liver biopsies were monitored using bovine-specific arrays. Treatment with E2-17? altered expression of 479 genes, P4 472 genes, and combined treatment significantly altered expression of 468 genes. In total, 578 genes exhibited altered expression including a remarkable number (346 genes) that responded similarly to E2-17?, P4, or combined treatment. Additional evidence for similar gene expression actions of E2-17ß and/or P4 were: principal component analysis placed almost every treatment array at a substantial distance from controls; Venn diagrams indicated overall treatment effects for most regulated genes; clustering analysis indicated the two major clusters had all treatments up-regulating (172 genes) or down-regulating (173 genes) expression. Thus, unexpectedly, common biological pathways were regulated by E2-17? and/or P4 in liver. This indicates that the mechanism of action of these steroid hormones in the liver might be either indirect or might occur through non-genomic pathways. This unusual pattern of gene expression in response to steroid hormones is consistent with the idea that there are classical and non-classical tissue-specific responses to steroid hormone actions. Future studies are needed to elucidate putative mechanism(s) responsible for overlapping actions of E2-17? and P4 on the liver transcriptome.

Piccinato, Carla A.; Rosa, Guilherme J. M.; N'Jai, Alhaji U.; Jefcoate, Colin R.; Wiltbank, Milo C.

2013-01-01

359

Fertility control as a means of controlling bovine tuberculosis in badger (Meles meles) populations in south-west England: predictions from a spatial stochastic simulation model  

Microsoft Academic Search

SUMMARY A spatial stochastic simulation model was used to assess the potential of fertility control, based on a yet-to- be-developed oral bait-delivered contraceptive directed at females, for the control of bovine tuberculosis in badger populations in south-west England.The contraceptive had a lifelong eˇect so that females rendered sterile in any particular year remained so for the rest of their lives.

PIRAN C. L. WHITE; ALEX J. G. LEWIS; STEPHEN HARRIS

1997-01-01

360

Fertility Control as a Means of Controlling Bovine Tuberculosis in Badger (Meles meles) Populations in South-West England: Predictions from a Spatial Stochastic Simulation Model  

Microsoft Academic Search

A spatial stochastic simulation model was used to assess the potential of fertility control, based on a yet-to-be-developed oral bait-delivered contraceptive directed at females, for the control of bovine tuberculosis in badger populations in south-west England. The contraceptive had a lifelong effect so that females rendered sterile in any particular year remained so for the rest of their lives. The

Piran C. L. White; Alex J. G. Lewis; Stephen Harris

1997-01-01

361

Bovine Tuberculosis in Badger (Meles meles) Populations in Southwest England: An Assessment of Past, Present and Possible Future Control Strategies Using Simulation Modelling  

Microsoft Academic Search

A spatial stochastic simulation model was used to compare the efficacy of different badger control policies and to determine the theoretical requirements for the control of endemic bovine tuberculosis in badger populations in southwest England. Culling-based strategies for controlling endemic disease were compared with strategies employing a yet-to-be-developed oral vaccine which would provide uninfected badgers with immunity to the infection.

Piran C. L. White; Stephen Harris

1995-01-01

362

Modeling the "glass" transition in proteins.  

PubMed

A model of a protein as a disordered system of identical spherical particles (which imitate protein side chains) interacting with each other via a repulsive soft sphere potential U(r) infinity r(-beta) is constructed. The particles undergo the conformational motion (CM) within their own harmonic conformational potentials around some mean equilibrium positions ascribed by the tertiary structure of the protein. A first principles calculation of the positional correlation functions for the CM is carried out. The general analysis is exemplified by the case in which the mean equilibrium positions of the particles form a cubic tightly-packed (face- centered) lattice (each particle has 12 nearest neighbors) with the step b(hydr) =6.6 A (the average distance between the centers of mass of hydrated protein subunits). The model yields dramatic slowing down of the relaxation with the decrease of temperature followed by a sharp glass transition at some crossover temperature T(c) < 200 K. At the transition the liquid-like dynamic behavior (the correlation functions tend to zero with time) is altered by the glass-like one (the correlation functions tend with time to some non-zero limit). In the liquid-like region above the crossover temperature the relaxation exhibits distinct alpha-process following the beta-one. The glass transition results from the interaction of the particles. Thus the model suggests that namely direct interactions of the fragments of protein structure rather than protein-solvent interactions are the origin of the phenomenon of the glass transition. The known increase of T(c) up to 300 K at dehydration of the protein is attributed to the known concomitant compression of the globule upon drying by about 4-6% so that positions of individual atoms displace by about 0.6 A (modeled by the decrease of the step of the lattice b by 0.6 A so that b(dehydr)=6 A). The model suggests that the solvent influences the phenomenon of the glass transition indirectly determining the tertiary structure of the protein rather than via own freezing. In the model the transition from the liquid-like dynamic behavior to the glass-like one can be obtained even in a cluster containing a few particles. Thus the results of the model can be considered as an argument in favor of the point of view that the transition to the glassy behavior can take place for a very small domains of the protein comprising only several constituting fragments of its structure. The model predicts that for the dehydrated protein the alpha-relaxation process is strongly repressed. PMID:11843621

Sitnitsky, A E

2002-02-01

363

Relationship between stearoyl-CoA desaturase 1 gene expression, relative protein abundance, and its fatty acid products in bovine tissues.  

PubMed

Stearoyl-CoA desaturase 1 (SCD1) greatly contributes to the unsaturated fatty acids present in milk and meat of cattle. The SCD1 enzyme introduces a double bond into certain saturated fatty acyl-CoAs producing monounsaturated fatty acids (MUFA). The SCD1 enzyme also has been shown to be active in the bovine mammary gland converting t11 18 : 1 (vaccenic acid) to c9 t11 conjugated linoleic acid (CLA). The objective of this study was to determine any association between the gene expression of SCD1 and occurrence of its products (c9 14 : 1, c9 16 : 1, c9 18 : 1, and c9 t11 18 : 2) in various bovine tissues. Tissue samples were obtained from lactating Holstein cows (n=28) at slaughter, frozen in liquid nitrogen and stored at -80 °C. Total RNA was extracted and converted to complementary DNA for quantitative real time polymerase chain reaction (PCR) analysis of the SCD1 gene. Extracted lipid was converted to fatty acid methyl esters and analysed by GC. Tissues varied in expression of SCD1 gene with mammary, cardiac, intestinal adipose, and skeletal muscle expressing greater copy number as compared with lung, large intestine, small intestine and liver (371, 369, 328, 286, 257, 145, 73, and 21 copies/ng RNA, respectively). Tissues with high mRNA expression of SCD1 contained greater SCD1 protein whereas detection of SCD1 protein in tissues with low SCD1 mRNA expression was very faint or absent. Across tissues, the desaturase indices for c9 18 : 1 (r=0·24) and sum of SCD products (r=0·20) were positively correlated with SCD1 gene expression (P<0·01 for both). Within each tissue, the relationship between SCD1 gene expression and the desaturase indices varied. No correlation was detected between SCD1 expression and desaturase indices in the liver, large and small intestines, lung, cardiac or skeletal muscles. Positive correlations, however, were detected between SCD1 expression and the desaturase indices in intestinal adipose tissue (P<0·02 for all) except 14 : 1, whereas only c9 18 : 1, c9 t11 18 : 2 and sum of all desaturase indices were positively correlated with SCD1 expression in mammary tissue (P?0·03). Overall, the relationship between SCD1 gene expression and occurrence of its products seems to be tissue specific. PMID:24904960

Rezamand, Pedram; Watts, Jason S; Yavah, Katherine M; Mosley, Erin E; Ma, Liying; Corl, Benjamin A; McGuire, Mark A

2014-08-01

364

Administration of bovine, porcine and equine growth hormone to the horse: effect on insulin-like growth factor-I and selected IGF binding proteins  

Microsoft Academic Search

This study investigated the biochemical effects of admin- istration of three types of recombinant growth hormone (GH; somatotropin) to the Thoroughbred horse. Equine or bovine or porcine GH was administered at a recom- mended dosage to 3-5-year old Thoroughbred geldings, for up to 21 days. It was shown that, in addition to equine GH, bovine and porcine GH were active

S S De Kock; J P Rodgers; B C Swanepoel; A J Guthrie

2001-01-01

365

Biological activities and physicochemical properties of Maillard reaction products in sugar-bovine casein peptide model systems.  

PubMed

The purpose of this study was to evaluate the biological activities and physicochemical properties of Maillard reaction products (MRPs), derived from aqueous reducing sugar (ribose, galactose and lactose) and bovine casein peptide (BCP) model systems. The fluorescence intensity of ribose-BCP MRPs reached the maximum value within 1h, while it took 3h for galactose-BCP MRPs. Size exclusion chromatography of all the MRPs indicated molecular rearrangements and production of new smaller molecules, as a function of the heating time. The consumption of ribose and amino groups was the highest in the ribose-BCP MRPs. BCP lost its known angiotensin-I-converting enzyme (ACE) inhibitory activity by the Maillard reaction with reducing sugars. Ribose-BCP MRPs had the lowest ACE inhibitory activity, but they showed the highest 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging activity and ferrous reducing power among all the MRPs. Galactose-BCP MRPs inhibited, slightly the growth of Caco-2 cells, while ribose-BCPand lactose-BCP MRPs had no cytotoxicity. PMID:23993556

Jiang, Zhanmei; Wang, Lizhe; Wu, Wei; Wang, Yu

2013-12-15

366

The development and characterization of a competitive ELISA for measuring active ADAMTS-4 in a bovine cartilage ex vivo model.  

PubMed

ADAMTS-4 (aggrecanase1) is believed to play an important role in the degradation of aggrecan during the progression of joint diseases. ADAMTS-4 is synthesized as a latent pro-enzyme that requires the removal of the pro-domain, exposing the N-terminal neoepitope, to achieve activity. We developed a monoclonal antibody against this neoepitope of active ADAMTS-4. Furthermore, we established and characterized a competitive ELISA for measuring active ADAMTS-4 form applying the specific antibody. We used this assay to profile the presence of active ADAMTS-4 and its aggrecan degradation product (NITEGE(373)) in a bovine cartilage ex vivo model. We found that after stimulation with catabolic factors, the cartilage initially released high levels of aggrecanase-derived aggrecan fragments into supernatant but subsequently decreased to background levels. The level of active ADAMTS-4 released into the supernatant and retained in the cartilage matrix increased continuously throughout the 21days of the study. The activity of ADAMTS-4 on the last day of catabolic stimulation was verified in vitro by adding deglycosylated or native aggrecan to the conditioned medium. Samples of human cartilage affected by varying degrees of osteoarthritis stained strongly for active ADAMTS-4 where surface fibrillation and clustered chondrocytes were observed. This assay could be an effective tool for studying ADAMTS-4 activity and for screening drugs regulating ADAMTS-4 activation. Moreover, it could be a potential biomarker for degenerative joint disease. PMID:23295731

He, Yi; Zheng, Qinlong; Simonsen, Ole; Petersen, Kristian K; Christiansen, Thorbjřrn G; Karsdal, Morten A; Bay-Jensen, Anne C

2013-04-24

367

Binding of teicoplanin and vancomycin to bovine serum albumin in vitro: a multispectroscopic approach and molecular modeling.  

PubMed

In this paper, the binding properties of teicoplanin and vancomycin to bovine serum albumin (BSA) were investigated using fluorescence quenching, synchronous fluorescence, Fourier transform infrared (FTIR), circular dichroism (CD) and UV-vis spectroscopic techniques and molecular docking under simulative physiological conditions. The results obtained from fluorescence quenching data revealed that the drug-BSA interaction altered the conformational structure of BSA. Meanwhile, the 3D fluorescence, CD, FTIR and UV-vis data demonstrated that the conformation of BSA was slightly altered in the presence of teicoplanin and vancomycin, with different reduced ?-helical contents. The binding distances for the drug-BSA system were provided by the efficiency of fluorescence resonance energy transfer (FRET). Furthermore, the thermodynamic analysis implied that hydrogen bond and van der Waals' forces were the main interaction for the drug-BSA systems, which agreed well with the results from the molecular modeling study. The results obtained herein will be of biological significance in future toxicological and pharmacological investigation. PMID:23606567

Lin, Yongxin; Jiao, Genlong; Sun, Guodong; Zhang, Lili; Wang, Shilong; Liu, Hanchao; Li, Zhizhong

2014-03-01

368

A unified model of protein dynamics  

PubMed Central

Protein functions require conformational motions. We show here that the dominant conformational motions are slaved by the hydration shell and the bulk solvent. The protein contributes the structure necessary for function. We formulate a model that is based on experiments, insights from the physics of glass-forming liquids, and the concepts of a hierarchically organized energy landscape. To explore the effect of external fluctuations on protein dynamics, we measure the fluctuations in the bulk solvent and the hydration shell with broadband dielectric spectroscopy and compare them with internal fluctuations measured with the Mössbauer effect and neutron scattering. The result is clear. Large-scale protein motions are slaved to the fluctuations in the bulk solvent. They are controlled by the solvent viscosity, and are absent in a solid environment. Internal protein motions are slaved to the beta fluctuations of the hydration shell, are controlled by hydration, and are absent in a dehydrated protein. The model quantitatively predicts the rapid increase of the mean-square displacement above ?200 K, shows that the external beta fluctuations determine the temperature- and time-dependence of the passage of carbon monoxide through myoglobin, and explains the nonexponential time dependence of the protein relaxation after photodissociation.

Frauenfelder, Hans; Chen, Guo; Berendzen, Joel; Fenimore, Paul W.; Jansson, Helen; McMahon, Benjamin H.; Stroe, Izabela R.; Swenson, Jan; Young, Robert D.

2009-01-01

369

In Vivo Protein Binding and Functional Analysis of cis-Acting Elements in the U3 Region of the Bovine Leukemia Virus Long Terminal Repeat  

PubMed Central

Bovine leukemia virus (BLV) is a member of the human T-cell leukemia virus (HTLV)/BLV group of retroviruses. These viruses regulate their own transcription by producing Tax, a protein which activates the virus promoter region, the long terminal repeat (LTR). To explore the molecular mechanisms involved in the transactivation, we identified protein binding elements by in vivo footprinting and analyzed their function by site-?directed mutagenesis. We used in vivo dimethyl sulfate footprinting by ligation-mediated PCR to detect constitutive in vivo protein-DNA interactions in a BLV-producing cell line, Bat2Cl6. The U3 region and part of the R region of the LTR were footprinted. In addition to the cis-acting elements (three cyclic AMP-responsive elements [CREs] and two AP4 sites) reported by others to be important for Tax-mediated activation of the BLV LTR, we found footprints in regions flanking these elements and in the core promoter region. The importance of these sites for transcriptional activation was studied by site-directed mutagenesis followed by promoter function analysis of the mutants with a chloramphenicol acetyltransferase reporter system. Our data corroborate those of others showing that the CREs are necessary for transactivation of the LTR, and they identify two new functional sites not previously reported by others. We show that the middle region of the BLV U3 contains multiple dual-functioning cis-acting elements which act as either positive or negative regulatory elements depending on the cell type tested. This is the first report of a functional mapping of the cis-acting elements of a virus of the HTLV/BLV group.

Xiao, Jianqiao; Buehring, Gertrude C.

1998-01-01

370

Effect of polyelectrolyte structure on protein-polyelectrolyte coacervates: coacervates of bovine serum albumin with poly(diallyldimethylammonium chloride) versus chitosan.  

PubMed

Electrostatic interactions between synthetic polyelectrolytes and proteins can lead to the formation of dense, macroion-rich liquid phases, with equilibrium microheterogeneities on length scales up to hundreds of nanometers. The effects of pH and ionic strength on the rheological and optical properties of these coacervates indicate microstructures sensitive to protein-polyelectrolyte interactions. We report here on the properties of coacervates obtained for bovine serum albumin (BSA) with the biopolyelectrolyte chitosan and find remarkable differences relative to coacervates obtained for BSA with poly(diallyldimethylammonium chloride) (PDADMAC). Coacervation with chitosan occurs more readily than with PDADMAC. Viscosities of coacervates obtained with chitosan are more than an order of magnitude larger and, unlike those with PDADMAC, show temperature and shear rate dependence. For the coacervates with chitosan, a fast relaxation time in dynamic light scattering, attributable to relatively unrestricted protein diffusion in both systems, is diminished in intensity by a factor of 3-4, and the consequent dominance by slow modes is accompanied by a more heterogeneous array of slow apparent diffusivities. In place of a small-angle neutron scattering Guinier region in the vicinity of 0.004 A-1, a 10-fold increase in scattering intensity is observed at lower q. Taken together, these results confirm the presence of dense domains on length scales of hundreds of nanometers to micrometers, which in coacervates prepared with chitosan are less solidlike, more interconnected, and occupy a larger volume fraction. The differences in properties are thus correlated with differences in mesophase structure. PMID:17892297

Kayitmazer, A Basak; Strand, Sabina P; Tribet, Christophe; Jaeger, Werner; Dubin, Paul L

2007-11-01

371

The solution structure of the bovine S100B protein dimer in the calcium-free state  

Microsoft Academic Search

Background S100B (S100?) is a member of the S100 family of small calcium-binding proteins: members of this family contain two helix-loop-helix calcium-binding motifs and interact with a wide range of proteins involved mainly in the cytoskeleton and cell proliferation. S100B is a neurite-extension factor and levels of S100B are elevated in the brains of patients with Alzheimer's disease or Down's

Peter M Kilby; Linda J Van Eldik; Gordon CK Roberts

1996-01-01

372

Color modeling of protein optical probes.  

PubMed

We present a strategy for modeling optical probes within heterogeneous environments of restricted dimension. The method is based on a multiphysics approach comprising sequential structure modeling by means of hybrid Car-Parrinello molecular dynamics and property modeling by means of quantum mechanics/molecular mechanics response theory. For demonstration we address the structural and optical properties of nile red within the ?-lacto globulin protein. We consider the cases with the probe situated on the surface or within the cavity of the protein, or embedded in a water solvent. We find the absorption properties of the probe to be highly dependent on its position relative to the protein. Structural rearrangements of the optical probe are found to contribute significantly to these environmental effects. PMID:22134524

Murugan, N Arul; Kongsted, Jacob; Rinkevicius, Zilvinas; Ĺgren, Hans

2012-01-21

373

Modelling of DNA-protein recognition  

NASA Technical Reports Server (NTRS)

Computer model-building procedures using stereochemical principles together with theoretical energy calculations appear to be, at this stage, the most promising route toward the elucidation of DNA-protein binding schemes and recognition principles. A review of models and bonding principles is conducted and approaches to modeling are considered, taking into account possible di-hydrogen-bonding schemes between a peptide and a base (or a base pair) of a double-stranded nucleic acid in the major groove, aspects of computer graphic modeling, and a search for isogeometric helices. The energetics of recognition complexes is discussed and several models for peptide DNA recognition are presented.

Rein, R.; Garduno, R.; Colombano, S.; Nir, S.; Haydock, K.; Macelroy, R. D.

1980-01-01

374

In vitro capacitation of bovine spermatozoa: Role of intracellular calcium  

Microsoft Academic Search

The development of successful methods of in vitro fertilization for bovine oocytes has advanced the bovine as a model for reproductive technology. The discovery of heparin as a capacitating agent has made it possible for investigators to have an inexpensive, readily available supply of bovine gametes for experimentation in reproductive biotechnologies such as gene transfer and cloning. The central event

J. J. Parrish; J. L. Susko-Parrish; J. K. Graham

1999-01-01