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1

Original article Effects of bovine colostrum acid protein on bone  

E-print Network

Original article Effects of bovine colostrum acid protein on bone loss and hemobiochemistry indexes that bovine milk and its basic proteins, and bovine colostrums (BC) and their extracts have positive effects hazard on blood lipids of rats under present experimental condition. bovine colostrum / acid protein

Boyer, Edmond

2

Modification of food contacting surfaces by plasma polymerization technique. Part II: Static and dynamic adsorption behavior of a model protein “bovine serum albumin” on stainless steel surface  

Microsoft Academic Search

The static and dynamic adsorption behaviors of a model protein “Bovine Serum Albumin” on plain (SS 316) and 2-hydroxethylmethacrylate (HEMA) or polyethyleneglycolmethacrylate (PEGMA) plasma polymerization (PlzP) modified stainless steel surfaces were studied. For equilibrium adsorption behavior, Freundlich model was attempted and model parameters for Freundlich (KF and n) were obtained. The values of the KF and n were 24.4, 0.88;

Selma Mutlu; Dilek Çökeliler; Mehmet Mutlu

2007-01-01

3

Proteins of Bovine Ephemeral Fever Virus  

Microsoft Academic Search

The proteins of bovine ephemeral fever virus (BEFV) were examined in purified virions and in infected BHK- 21 cells. Five structural proteins were named L (180K), G (81K), N (52K), M1 (43K) and M2 (29K). The 81KG protein incorporated (3H)glucosamine, was removed from virions by treatment with Triton X-100 and bound monoclonal antibodies which were both neutral- izing and protective.

Peter J. Walker; Keren A. Byrne; Daisy H. Cybinski; Denise L. Doolan; Yonghong Wang

1991-01-01

4

Proteins of bovine ephemeral fever virus.  

PubMed

The proteins of bovine ephemeral fever virus (BEFV) were examined in purified virions and in infected BHK-21 cells. Five structural proteins were named L (180K), G (81K), N (52K), M1 (43K) and M2 (29K). The 81K G protein incorporated [3H]glucosamine, was removed from virions by treatment with Triton X-100 and bound monoclonal antibodies which were both neutralizing and protective. Treatment of virions with Triton X-100 and 0.2 to 1.0 M-NaCl progressively released L, M1 and M2. The N protein remained associated with nucleocapsids in up to 2.5 M-NaCl. The glycoprotein (G), nucleoprotein (N) and matrix protein (M2) were phosphorylated. In BEFV-infected BHK-21 cells, five virus-induced proteins were detected from 12 h post-infection. The L, N, M1 and M2 proteins corresponded to those detected in virions whereas the G protein existed in two forms. In tunicamycin-treated cells these occurred as 67K and 71K non-glycosylated precursors. In the absence of tunicamycin, 77K and 79K glycosylated forms were further modified to produce the 81K virion G protein and a 90K cell-associated form. Five viral proteins were also detected in cells infected with the closely related Berrimah virus; the Berrimah virus G protein was also present in two forms. PMID:1990067

Walker, P J; Byrne, K A; Cybinski, D H; Doolan, D L; Wang, Y H

1991-01-01

5

Nonenzymatic glycosylation of bovine myelin basic protein  

SciTech Connect

In the CNS myelin sheath the nonenzymatic glycosylation reaction (at the early stage of the Amadori product) occurs only with the myelin basic protein and not with the other myelin proteins. This was observed in isolated bovine myelin by in vitro incubation with (/sup 14/C)-galactose and (/sup 14/C)-glucose. The respective in-vitro incorporation rates for purified bovine myelin basic protein with D-galactose, D-glucose and D-mannose were 7.2, 2.4 and 2.4 mmoles/mole myelin basic protein per day at 37/sup 0/C. A more rapid, HPLC method was devised and characterized to specifically analyze for the Amadori product. The HPLC method was correlated to the (/sup 14/C)-sugar incorporation method for myelin basic protein under a set of standard reaction conditions using (/sup 14/C)-glucose and (/sup 14/C)-mannose with HPLC values at 1/6 and 1/5 of the (/sup 14/C)-sugar incorporation method. A novel myelin basic protein purification step has been developed that yields a relativity proteolytic free preparation that is easy to work with, being totally soluble at a neutral pH. Nine new spots appear for a trypsinized glycosylated MBP in the paper peptide map of which eight correspond to positions of the (/sup 3/H)-labeled Amadori product in affinity isolated peptides. These studies provide a general characterization of and a structural basis for investigations on nonenzymatically glycosylated MBP as well as identifying MBP as the only nonenzymatically glycosylated protein in the CNS myelin sheath which may accumulate during aging, diabetes, and demyelinating diseases in general.

Hitz, J.B.

1987-01-01

6

Fatty acid-binding protein in bovine skeletal muscle  

E-print Network

FATTY ACID-BINDING PROTEIN IN BOVINE SKELETAL MUSCLE A Thesis by KIMBERLY KIRBY MOORE Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE... December 1989 Major Subject: Nutrition FATTY ACID-BINDING PROTEIN IN BOVINE SKELETAL MUSCLE A Thesis by KIMBERLY KIRBY MOORE Approved as to style and content by: Ste en B. Smith (Chair of Committee) Karen S. Kubena (Member) Gary C. Smith (Head...

Moore, Kimberly Kirby

2012-06-07

7

Chemical alteration by tooth bleaching of human salivary proteins that infiltrated subsurface enamel lesions -Experimental study with bovine lesion model systems-.  

PubMed

Salivary macromolecules infiltrate white and brown spot enamel lesions and adsorb onto hydroxyapatite. Calcium-binding salivary proteins such as statherin hinder remineralization of these lesions. We assessed whether bleaching agents can remove salivary components that have infiltrated and bound to experimental subsurface lesions in bovine enamel prepared by immersing specimens in acid and then human saliva. Transversal microradiography showed that such demineralized lesions mimicked incipient carious lesions. Bound proteins to the experimental and untreated control specimens were eluted in a stepwise manner with phosphatebuffered saline, 0.4 M phosphate buffer, and 1 M HCl. SDS-PAGE of dialyzed extracts showed that specific salivary proteins bound to the lesions, while virtually no protein bands were detected if the specimens were bleached. Western blotting showed that even statherin, which was more firmly bound than other proteins, was removed. In-office bleaching agent may be useful in treating enamel lesions for removing proteins bound to these lesions. PMID:25273046

Iizuka, Junko; Mukai, Yoshiharu; Taniguchi, Motoe; Mikuni-Takagaki, Yuko; Ten Cate, Jacob Martien; Teranaka, Toshio

2014-01-01

8

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

9

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

10

Binding of bovine prion protein to heparin: A fluorescence polarization study  

Microsoft Academic Search

Glycosaminoglycans (GAGs) are believed to be associated with prion disease pathology and also with metabolism of the prion protein. Fluorescence polarization assay (FPA) of binding between bovine recombinant prion protein (brecPrP) and heparin labelled with AlexaFluor488 was used in model experiments to study glycosaminoglycan–prion protein interaction. Heparin binding to brecPrP was a rapid reversible event which occurred under defined conditions.

Olga Andrievskaia; Zhanna Potetinova; Aru Balachandran; Klaus Nielsen

2007-01-01

11

Bovine immunoglobulin protein isolates for the nutritional management of enteropathy.  

PubMed

The gastrointestinal tract is responsible for a multitude of digestive and immune functions which depend upon the balanced interaction of the intestinal microbiota, diet, gut barrier function, and mucosal immune response. Disruptions in one or more of these factors can lead to intestinal disorders or enteropathies which are characterized by intestinal inflammation, increased gut permeability, and reduced capacity to absorb nutrients. Enteropathy is frequently associated with human immunodeficiency virus (HIV) infection, inflammatory bowel disease, autoimmune enteropathy, radiation enteritis, and irritable bowel syndrome (IBS), where pathologic changes in the intestinal tract lead to abdominal discomfort, bloating, abnormal bowel function (e.g., diarrhea, urgency, constipation and malabsorption). Unfortunately, effective therapies for the management of enteropathy and restoring intestinal health are still not available. An accumulating body of preclinical studies has demonstrated that oral administration of plasma- or serum-derived protein concentrates containing high levels of immunoglobulins can improve weight, normalize gut barrier function, and reduce the severity of enteropathy in animal models. Recent studies in humans, using serum-derived bovine immunoglobulin/protein isolate, demonstrate that such protein preparations are safe and improve symptoms, nutritional status, and various biomarkers associated with enteropathy. Benefits have been shown in patients with HIV infection or diarrhea-predominant IBS. This review summarizes preclinical and clinical studies with plasma/serum protein concentrates and describes the effects on host nutrition, intestinal function, and markers of intestinal inflammation. It supports the concept that immunoglobulin-containing protein preparations may offer a new strategy for restoring functional homeostasis in the intestinal tract of patients with enteropathy. PMID:25206275

Petschow, Bryon W; Blikslager, Anthony T; Weaver, Eric M; Campbell, Joy M; Polo, Javier; Shaw, Audrey L; Burnett, Bruce P; Klein, Gerald L; Rhoads, J Marc

2014-09-01

12

Bovine immunoglobulin protein isolates for the nutritional management of enteropathy  

PubMed Central

The gastrointestinal tract is responsible for a multitude of digestive and immune functions which depend upon the balanced interaction of the intestinal microbiota, diet, gut barrier function, and mucosal immune response. Disruptions in one or more of these factors can lead to intestinal disorders or enteropathies which are characterized by intestinal inflammation, increased gut permeability, and reduced capacity to absorb nutrients. Enteropathy is frequently associated with human immunodeficiency virus (HIV) infection, inflammatory bowel disease, autoimmune enteropathy, radiation enteritis, and irritable bowel syndrome (IBS), where pathologic changes in the intestinal tract lead to abdominal discomfort, bloating, abnormal bowel function (e.g., diarrhea, urgency, constipation and malabsorption). Unfortunately, effective therapies for the management of enteropathy and restoring intestinal health are still not available. An accumulating body of preclinical studies has demonstrated that oral administration of plasma- or serum-derived protein concentrates containing high levels of immunoglobulins can improve weight, normalize gut barrier function, and reduce the severity of enteropathy in animal models. Recent studies in humans, using serum-derived bovine immunoglobulin/protein isolate, demonstrate that such protein preparations are safe and improve symptoms, nutritional status, and various biomarkers associated with enteropathy. Benefits have been shown in patients with HIV infection or diarrhea-predominant IBS. This review summarizes preclinical and clinical studies with plasma/serum protein concentrates and describes the effects on host nutrition, intestinal function, and markers of intestinal inflammation. It supports the concept that immunoglobulin-containing protein preparations may offer a new strategy for restoring functional homeostasis in the intestinal tract of patients with enteropathy. PMID:25206275

Petschow, Bryon W; Blikslager, Anthony T; Weaver, Eric M; Campbell, Joy M; Polo, Javier; Shaw, Audrey L; Burnett, Bruce P; Klein, Gerald L; Rhoads, J Marc

2014-01-01

13

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

14

Identification of highly active flocculant proteins in bovine blood.  

PubMed

Synthetic polymeric flocculants are used extensively for wastewater remediation, soil stabilization, and reduction in water leakage from unlined canals. Sources of highly active, inexpensive, renewable flocculants are needed to replace synthetic flocculants. High kaolin flocculant activity was documented for bovine blood (BB) and blood plasma with several anticoagulant treatments. BB serum also had high flocculant activity. To address the hypothesis that some blood proteins have strong flocculating activity, the BB proteins were separated by SEC. Then, the major proteins of the flocculant-active fractions were separated by SDS-PAGE. Identity of the major protein components was determined by tryptic digestion and peptide analysis by MALDI TOF MS. The sequence of selected peptides was confirmed using TOF/TOF-MS/MS fragmentation. Hemoglobin dimer (subunits ? and ?) was identified as the major protein component of the active fraction in BB; its high flocculation activity was confirmed by testing a commercial sample of hemoglobin. In the same manner, three proteins from blood plasma (fibrinogen, ?-globulin, ?-2-macroglobulin) were found to be highly active flocculants, but bovine serum albumin, ?-globulin, and ?-globulin were not flocculants. On a mass basis, hemoglobin, ?-globulin, ?-2-macroglobulin were as effective as anionic polyacrylamide (PAM), a widely used synthetic flocculant. The blood proteins acted faster than PAM, and unlike PAM, the blood proteins flocculants did not require calcium salts for their activity. PMID:22194055

Piazza, George J; Nuñez, Alberto; Garcia, Rafael A

2012-03-01

15

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

16

Envelope proteins of bovine herpesvirus 1: immunological and biochemical studies  

SciTech Connect

The authors studied immunological and biochemical properties of the bovid herpesvirus 1 (BHV-1) envelope proteins in order to understand the pathogenesis of BHV-1 infection and to provide basic information for the production of effective subunit vaccines against BHV-1. Ten glycoproteins MW 180, 150, 130, 115, 97, 77, 74, 64, 55, and 45 kilodaltons (K), and a single non-glycosylated 108 K protein were quantitatively removed from purified BHV-1 virions by detergent treatment. These glycoproteins were present on the virion envelope and on the surface of BHV-1 infected cells. The quantitative removal from virions by treatment with nonionic detergents and their presence on the surface of infected cells indicate that 180/97, 150/77, and 130/74/55 K are major components of the BHV-1 envelope and are also the targets of virus neutralizing humoral immune response. Envelope glycoproteins of herpes simplex type 1 (HSV-1) bind immunoglobulin by the Fc end and it is suggested this may increase pathogenicity of this virus. They searched for a similar function in BVH-1 by measuring the ability of BHV-1 infected cells and viral envelope proteins to bind radiolabelled rabbit and bovine IgG. Binding activity for rabbit IgG or bovine IgG-Fc could not be demonstrated by BHV-1 infected MDBK cells, whereas, MDBK cells infected with HSV-1 bound rabbit IgG and bovine IgG-Fc. None of the three major envelope proteins of BHV-1 bound to rabbit or bovine IgG. The results of this study indicate that BHV-1, unlike some other herpesviruses, lack Fc binding activity.

Rodriguez Roque, L.L.

1986-01-01

17

Biosynthesis and processing of bovine cartilage link proteins  

SciTech Connect

We have examined posttranslational modifications which are responsible for converting an apparently single precursor to the two major forms of link protein in bovine articular cartilage. Resistance to endoglycosidases H and F suggests that Asn-linked oligosaccharides of link protein secreted by bovine chondrocytes in culture are of the complex or hybrid type. There is no evidence for O-linked oligosaccharides. There is no apparent precursor-product relationship between link protein (LP)1 and LP2, since after a short pulse with (3H)leucine two forms are present, consistent with the existence of two glycosylation sites. An immunoprecipitate of LP1 from pulse-labeled chondrocytes was observed to show a decrease in electrophoretic mobility and increased microheterogeneity during transit through the Golgi, whereas LP2 did not change. During processing both LP1 and LP2 become endoglycosidase H resistant. LP1, but not LP2, can be biosynthetically labeled with (35S)sulfate. Incorporation of (35S)sulfate is inhibited by tunicamycin, indicating that the sulfate is associated with Asn-linked carbohydrate. Sulfation may be important for normal processing, secretion, or degradation of link protein and with sialylation may confer considerable charge heterogeneity upon LP1. We conclude that there are considerable biochemical differences between glycoproteins LP1 and LP2 which may provide a basis for functional differences.

Hering, T.M.; Sandell, L.J. (Univ. of Washington, Seattle (USA))

1990-02-05

18

Mouse and bovine models for human IVF.  

PubMed

It is obvious that the first prerequisite is to define for what purpose a model is needed for humans. There are huge differences in reproductive physiology between the mouse, human and cow. As far as maturation is concerned, the plasticity of the mouse model is not the same in cows and humans. The final stages of oocyte maturation seem to be more finely regulated in cows and humans, where a minimum size of follicle is necessary to complete maturation in vitro. Bovine and human preimplantation embryos seem to be more similar in terms of biochemical and intrinsic paternal and maternal regulatory processes. Once again, interactions between the embryo and the corpus luteum are similar in cows and humans, but mouse and human embryo implantations are closer. Mouse oocytes and embryos should not be overlooked, but excessive generalization between mammalian species must be avoided. PMID:12470581

Ménézo, Yves J R; Hérubel, François

2002-01-01

19

Epitope mapping of bovine viral diarrhea virus nonstructural protein 3.  

PubMed

Six consecutive overlapped coding regions (F1-F6) of whole NS3 molecule of bovine viral diarrhea virus (BVDV) were cloned into pMAL-c2X plasmid vector and expressed in Escherichia coli cells (BL21 strain). The recombinant proteins were then purified by amylose resin to determine the most immunogenic domain(s) of the NS3 molecule. Evaluation of the recombinant proteins was carried out by indirect ELISAs using several bovine sera (previously characterized by virus neutralization test, a commercial ELISA kit, and a newly developed NS3-ELISA) and 6 monoclonal antibodies. The experiments showed that the most immunogenic domain of the NS3 protein was the fourth designed fragment (F4), a 122 amino-acid (AA) region of about 13.5kDa (nucleotide 1003-1368; residue 335-456). Purified recombinant F4 was also evaluated as single ELISA antigen (F4-ELISA) for the detection of anti-BVDV antibodies in sera of infected cattle. Although this small recombinant fragment of NS3 protein was almost completely soluble and expressed more efficient respect to whole NS3 molecule, it did not show enough sensitivity and specificity to be a proper substitute for NS3 as ELISA antigen to detect specific antibodies against BVDV. However, statistical analyses showed a medium correlation between the results of the developed F4-ELISA and virus neutralization test (kappa coefficient=0.63, P<0.001), with the relative sensitivity and specificity of 78.05% and 84.91%, respectively, suggesting the potential use of this fragment as an ELISA antigen along with other antigens or monoclonal antibody(s) in a competitive ELISA. PMID:25205011

Mahmoodi, Pezhman; Seyfi Abad Shapouri, Masoud Reza; Ghorbanpour, Masoud; Ekhtelat, Maryam; Haji Hajikolaei, Mohammad Rahim; Lotfi, Mohsen; Pourmahdi Boroujeni, Mahdi; Daghari, Maryam

2014-10-15

20

Pilot study on binding of bovine salivary proteins to grit silicates and plant phytoliths.  

PubMed

Mostly fed with grass in fresh or conserved form, cattle and other livestock have to cope with silicate defence bodies from plants (phytoliths) and environmental silicates (grit), which abrade tooth enamel and could additionally interact with various salivary proteins. To detect potential candidates for silicate-binding proteins, bovine whole saliva was incubated with grass-derived phytoliths and silicates. Interactions of salivary proteins with pulverized bovine dental enamel and dentine were additionally analysed. After intense washing, the powder fractions were loaded onto 1D-polyacrylamide gels, most prominent adhesive protein bands were cut out and proteins were identified by mass spectrometry within three independent replicates. All materials were mainly bound by bovine odorant-binding protein, bovine salivary protein 30×10(3) and carbonic anhydrase VI. The phytolith/silicate fraction showed additional stronger interaction with haemoglobin ? and lactoperoxidase. Conceivably, the binding of these proteins to the surfaces may contribute to biological processes occurring on them. PMID:23776006

Mau, Marcus; M Kaiser, Thomas; Südekum, Karl-Heinz

2013-06-01

21

Distribution of Proteins Providing Homeostasis of Iron Ions in Bovine Retina  

Microsoft Academic Search

Distribution of proteins providing homeostasis of iron ions in bovine retina was studied by methods of indirect immunohistochemistry, which allowed detection of localization of transferrin, ferritin, and transferrin receptor. In bovine retina, transferrin is revealed in the region of outer and inner segments of photoreceptors and in the external plexiform layer. Distributions of ferritin and transferrin receptor are identical; they

M. G. Yefimova; J.-C. Jeanny; Y. Courtois

2002-01-01

22

Effect of Polyelectrolyte Structure on Protein-Polyelectrolyte Coacervates: Coacervates of Bovine Serum Albumin with  

E-print Network

-polyelectrolyte interactions. We report here on the properties of coacervates obtained for bovine serum albumin (BSA Serum Albumin with Poly(diallyldimethylammonium chloride) versus Chitosan A. Basak Kayitmazer,*,, SabinaEffect of Polyelectrolyte Structure on Protein-Polyelectrolyte Coacervates: Coacervates of Bovine

Dubin, Paul D.

23

Characterization of a putative receptor protein for bovine viral diarrhea virus  

Microsoft Academic Search

In a previous communication, we reported a 50-kDa cell surface protein from Madin-Darby bovine kidney (MDBK) cells as a putative receptor for bovine viral diarrhea virus (BVDV). The present study delineates further characterization of the receptor protein. Protease treatment of cultured MDBK cells adversely affected the receptor, thus abolishing the binding of anti-D89 (BVDV anti-idiotypes) to the cells. However, pretreatment

Wenzhi Xue; Shucheng Zhang; Harish C. Minocha

1997-01-01

24

Molecular weight distribution of hydrolysis products during biodegradation of model macromolecules in suspended and biofilm cultures I. Bovine serum albumin  

Microsoft Academic Search

Macromolecules can comprise a significant portion of dissolved organic carbon in wastewater and affect wastewater treatability by engineered systems, but little information is available about mechanisms of macromolecule degradation. The release of protein hydrolytic fragments was monitored during the degradation of the model protein bovine serum albumin (BSA) in batch and continuous suspended cultures and in fixed-film reactor systems. Three

David R. Confer; Bruce E. Logan

1997-01-01

25

Predictive qualitative risk model of bovine rabies occurrence in Brazil.  

PubMed

Bovine rabies remains endemic in Brazil and despite control efforts, the disease still spreads insidiously. The main vector is the hematophagous bat, Desmodus rotundus. The present work aimed to create a predictive qualitative model of the occurrence of bovine rabies in each municipality in 25 of the 27 Brazilian States. The risk of rabies transmission from bats to bovine was estimated using decision-tree models of receptivity and vulnerability. Questionnaires, which covered a number of questions related to the surveillance of possible risk factors, such as bovine rabies outbreaks in the previous year, the presence of bat roosts, bat rabies positivity and environmental changes, were sent to the local veterinary units of each State. The bovine density and geomorphologic features were obtained from national databases and geographic information systems. Of the 433 municipalities presenting bovine rabies outbreaks in 2010, 178 (41.1%) were classified by the model as high risk, 212 (49.0%) were classified as moderate risk, 25 (5.8%) were classified as low risk, whereas the risk was undetermined in 18 municipalities (4.1%). An ROC curve was built to determine if the risk evaluated by the model could adequately discriminate between municipalities with and without rabies occurrence in future years. The risk estimator for the year 2011 was classified as moderately accurate. In the future, these models could allow the targeting of rabies control efforts, with the adoption of control measures directed to the higher risk locations and the optimization of the field veterinary staff deployment throughout the country. Additionally, efforts must be made to encourage continuous surveillance of risk factors. PMID:24433635

Braga, Guilherme Basseto; Grisi-Filho, José Henrique Hildebrand; Leite, Bruno Meireles; de Sena, Elaine Fátima; Dias, Ricardo Augusto

2014-03-01

26

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

27

A unique property of fetal bovine serum: High levels of protein-glutathione mixed disulfides  

Microsoft Academic Search

Summary  Fetal bovine serum has been reported to delay or inhibit “spontaneous” neoplastic transformation in vitro as compared with\\u000a all other sera tested. The present results indicate that fetal bovine serum is also unique in containing high levels of protein-glutathione\\u000a mixed disulfides (3 to 7 ?g glutathione as mixed disulfide per ml serum). The level of mixed disulfide appears to vary

Edward A. Bump; Donald J. Reed

1977-01-01

28

NMR structure of the bovine prion protein Francisco Lo pez Garcia, Ralph Zahn, Roland Riek, and Kurt Wu thrich*  

E-print Network

NMR structure of the bovine prion protein Francisco Lo´ pez Garci´a, Ralph Zahn, Roland Riek structures of the recombinant 217-residue polypeptide chain of the mature bovine prion protein, bPrP(23 there are characteristic local differences relative to the confor- mations of the murine and Syrian hamster prion proteins

Riek, Roland

29

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

30

31P NMR and AFM studies on the destabilization of cell and model membranes by the major bovine seminal plasma protein, PDC-109.  

PubMed

The effect of PDC-109 binding to dimyristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylglycerol (DPPG) multilamellar vesicles (MLVs) and supported membranes was investigated by (31)P NMR spectroscopy and atomic force microscopy. Additionally, the effect of cholesterol on the binding of PDC-109 to phosphatidylcholine (PC) membranes was studied. Binding of PDC-109 to MLVs of DMPC and DPPG induced the formation of an isotropic signal in their (31)P NMR spectra, which increased with increasing protein/lipid ratio and temperature, consistent with protein-induced disruption of the MLVs and the formation of small unilamellar vesicles or micelles but not inverse hexagonal or cubic phases. Incorporation of cholesterol in the DMPC MLVs afforded a partial stabilization of the lamellar structure, consistent with previous reports of membrane stabilization by cholesterol. AFM results are consistent with the above findings and show that addition of PDC-109 leads to a complete breakdown of PC membranes. The fraction of isotropic signal in (31)P NMR spectra of DPPG in the presence of PDC-109 was less than that of DMPC under similar conditions, suggesting a significantly higher affinity of the protein for PC. Confocal microscopic studies showed that addition of PDC-109 to human erythrocytes results in a disruption of the plasma membrane and release of hemoglobin into the solution, which was dependent on the protein concentration and incubation time. PMID:21117173

Damai, Rajani S; Sankhala, Rajeshwer S; Anbazhagan, Veerappan; Swamy, Musti J

2010-11-01

31

The characterization of DNA methylation-mediated regulation of bovine placental lactogen and bovine prolactin-related protein-1 genes  

PubMed Central

Background Bovine trophoblast binucleate cells (BNC) express a plethora of molecules including bovine placental lactogen (bPL, gene name is bCSH1) and bovine prolactin-related protein-1 (bPRP1). BCSH1 and bPRP1 are members of the growth hormone (GH)/prolactin (PRL) gene family, which are expressed simultaneously in BNC and are central to placentation and the progression of pregnancy in cattle. However, there is a paucity of information on the transcriptional regulatory mechanisms of both the bCSH1 and bPRP1 genes. Recent studies, however, have demonstrated that the expression of a number of genes is controlled by the methylation status of their promoter region. In the present study, we examined the cell-type-specific epigenetic alterations of the 5'-flanking region of the bCSH1 and bPRP1 genes to gain an insight into their regulatory mechanisms. Results Analysis of 5-aza-2'-deoxycytidine treatment demonstrated that bCSH1 expression is moderately induced in fibroblast cultures but enhanced in BT-1 cells. Sodium bisulfite based sequencing revealed that bCSH1 is hypomethylated in the cotyledonary tissue but not in the fetal skin, and this pattern was not altered with the progression of pregnancy. On the other hand, the methylation status of bPRP1 was similar between the cotyledon and fetal skin. The bPRP1 gene was exclusively hypermethylated in a bovine trophoblast cell-derived BT-1 cell-line. While the activity of bCSH1 was similar in both BT-1 and bovine fibroblast cells, that of bPRP1 was specific to BT-1. Treatment with a demethylating agent and luciferase assays provided in vitro evidence of the positive regulation of bCSH1 but not bPRP1. Conclusion This is the first report to identify the differential regulatory mechanisms of the bCSH1 and bPRP1 genes and indicates that bCSH1 might potentially be the only transcript that is subject to DNA methyltransferase regulation. The data indicates the possibility of novel kinetics of induction of the synchronously expressed BNC-specific bCSH1 and bPRP1 transcripts, which may aid the understanding of the intricate regulation and specific role(s) of these important molecules in bovine placentogenesis and the progression of pregnancy. PMID:19261194

Nakaya, Yuki; Kizaki, Keiichiro; Takahashi, Toru; Patel, Osman V; Hashizume, Kazuiyoshi

2009-01-01

32

Comparison of the principal proteins in bovine, caprine, buffalo, equine and camel milk.  

PubMed

Proteomic analysis of bovine, caprine, buffalo, equine and camel milk highlighted significant interspecies differences. Camel milk was found to be devoid of ?-lactoglobulin, whereas ?-lactoglobulin was the major whey protein in bovine, buffalo, caprine, and equine milk. Five different isoforms of ?-casein were found in camel milk, analogous to the micro-heterogeneity observed for bovine ?-casein. Several spots observed in 2D-electrophoretograms of milk of all species could tentatively be identified as polypeptides arising from the enzymatic hydrolysis of caseins. The understanding gained from the proteomic comparison of these milks may be of relevance both in terms of identifying sources of hypoallergenic alternatives to bovine milk and detection of adulteration of milk samples and products. PMID:22365180

Hinz, Katharina; O'Connor, Paula M; Huppertz, Thom; Ross, R Paul; Kelly, Alan L

2012-05-01

33

Effect of Langmuir monolayer of bovine serum albumin protein on the morphology of calcium carbonate  

E-print Network

Bovine serum albumin (BSA) Langmuir monolayer, as a model of biomineralization-associated proteins, was used to study its effect on regulated biomineralization of calcium carbonate. The effects of the BSA Langmuir monolayer and the concentration of the subphase solution on the nucleation and growth processes and morphology of the calcium carbonate crystal were investigated. The morphology and polymorphic phase of the resulting calcium carbonate crystals were characterized by scanning electron microscopy (SEM) and X-ray diffraction analysis (XRD). Moreover, the interaction mechanisms of the subphase solution with the BSA Langmuir monolayer were discussed. It was found that BSA Langmuir monolayer could be used as a template to successfully manipulate the polymorphic phase and crystal morphology of calcium carbonate and had obvious influence on the oriented crystallization and growth. The final morphology or aggregation mode of the calcite crystal was closely dependent on the concentration of calcium bicarbonate...

Xue, Zhong-Hui; Hu, Bin-Bin; Du, Zu-Liang; 10.1016/j.msec.2009.03.016

2011-01-01

34

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

35

Tracer diffusion coefficients of proteins by means of holographic relaxation spectroscopy: application to bovine serum albumin  

SciTech Connect

Holographic relaxation spectroscopy has been used to measure tracer diffusion coefficients for photochromically labeled bovine serum albumin in solutions having total bovine serum albumin concentrations in the range 3.25 to 257 g/liter. In the limit of zero concentration, the diffusion coefficient was found to be 5.9 X 10(-7) cm/sup 2//s and the initial slope was zero. The concentration dependence of the diffusion coefficient was not significantly affected by the fraction of protein molecules which were labeled. Holographic relaxation spectroscopy permits rapid, accurate determination of tracer diffusion coefficients for proteins in mixtures.

Arunyawongsakorn, U.; Johnson, C.S. Jr.; Gabriel, D.A.

1985-04-01

36

Stabilizing effect of saccharides on bovine plasma protein: a calorimetric study.  

PubMed

Bovine plasma proteins provide the needed amino acids for the growth and development of an organism. With the purpose of preserving the native structure, related with the protein functional properties, the oligosaccharide inulin was used as protective agent and was compared with glucose and sucrose, during freeze-drying. In the present study, the thermal stability of protein was investigated as a function of type of saccharide in a concentration range of 5-15% (w/v), and at different pHs. The effect of these variables on phase transition, thermal stability and miscibility was assessed by differential scanning calorimetry (DSC) and scanning electron microscopy (SEM). The results of thermal protein properties (denaturation temperature and enthalpy), demonstrated that endothermic transition shifted to higher temperatures, being the stabilizing effect: inulin>glucose>sucrose. The thermal behavior suggests compatibility or interactions between the components of blends. In this way, the micrographs showed a homogeneous distribution of the different phases, corroborating the miscibility in the matrix. The unfolding process was irreversible and could be adequately described by a two-state model. PMID:22445165

Rodriguez Furlán, Laura T; Lecot, Javier; Pérez Padilla, Antonio; Campderrós, Mercedes E; Zaritzky, Noemi

2012-08-01

37

Insulin regulates milk protein synthesis at multiple levels in the bovine mammary gland  

Microsoft Academic Search

The role of insulin in milk protein synthesis is unresolved in the bovine mammary gland. This study examined the potential\\u000a role of insulin in the presence of two lactogenic hormones, hydrocortisone and prolactin, in milk protein synthesis. Insulin\\u000a was shown to stimulate milk protein gene expression, casein synthesis and 14C-lysine uptake in mammary explants from late pregnant cows. A global

Karensa K. Menzies; Christophe Lefèvre; Keith L. Macmillan; Kevin R. Nicholas

2009-01-01

38

Characterization of a cross-reactive linear epitope in human genogroup I and bovine genogroup III norovirus capsid proteins.  

PubMed

The Southampton norovirus (SV) capsid protein was expressed as VLPs by recombinant baculoviruses in insect cells and was used to immunize mice for the production of monoclonal antibodies (mAbs). One mAb, CM54, showed broad cross-reactivity to genogroup I (GI) noroviruses, but was not reactive to GII capsid proteins. Interestingly mAb CM54 reacted to a bovine norovirus capsid protein. Immunoblot analysis indicated the binding site for CM54 was located in the shell domain between amino acid residues 102-225 of the SV capsid protein. The epitope was mapped to high resolution using a peptide array and was located to the sequence LEDVRN at amino acid residues 162-167. Alignment of norovirus capsid protein sequences confirmed the epitope sequence was common to particular groups of human and bovine noroviruses. Modeling of the epitope onto the recombinant NV capsid protein revealed it was located to the inner surface of the shell domain. PMID:16934306

Batten, Carrie A; Clarke, Ian N; Kempster, Sarah L; Oliver, Stefan L; Bridger, Janice C; Lambden, Paul R

39

Is double C2 protein (DOC2) expressed in bovine adrenal medulla? A commercial anti-DOC2 monoclonal antibody recognizes a major bovine mitochondrial antigen.  

PubMed Central

We have examined the expression in bovine adrenal medulla of double C2 protein (DOC2), a vesicular protein which associates with intracellular phospholipid and Ca(2+) and is implicated in the modulation of regulated exocytosis. Extensive reverse transcription-PCR, Northern blot analyses and in vitro translation reactions have been combined with immunological studies to provide data to suggest that neither DOC2alpha nor DOC2beta is expressed at detectable levels in bovine adrenal chromaffin cells, and that a widely used monoclonal antibody directed against DOC2 also recognizes mitochondrial complex III core protein 2. PMID:10998344

Duncan, R R; Apps, D K; Learmonth, M P; Shipston, M J; Chow, R H

2000-01-01

40

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

41

~ristalsof bovine chyrnotrypsin. Enzymes are proteins specialized to catalyze biological reac-  

E-print Network

Figure 8-1 ~ristalsof bovine chyrnotrypsin. 1 Enzymes are proteins specialized to catalyze of biochemistry is the history of en- zyme research. The name enzyme ("in yeast") was not used until 1877- furic acid. Although Louis Pasteur recognized that fermentation is catalyzed by enzymes, he postulated

Vallino, Joseph J.

42

Oxidative damage of bovine serum albumin and other enzyme proteins by iron-chelate complexes  

Microsoft Academic Search

Direct oxidative protein damage by iron-nitrilotriacetate (NTA), as well as physiological iron complexes, iron-citrate and iron-ADP was studied in the presence or absence of H2O2, using bovine serum albumin (BSA), glucose-6-phosphate dehydrogenase (G-6-PD), glutathione reductase (GSSGRase) and catalase as the target proteins. Both Fe(III)NTA + H2O2 and Fe(II)NTA + H2O2 caused marked BSA fragmentation which accompanied the decrease in the

Tetsuya Ogino; Shigeru Okada

1995-01-01

43

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.

44

Surface-Expressed Mig Protein Protects Streptococcus dysgalactiae against Phagocytosis by Bovine Neutrophils†  

PubMed Central

The mig gene of Streptococcus dysgalactiae, a major bovine mastitis pathogen, encodes two plasma protein-binding receptors, ?2-macroglobulin (?2-M) and immunoglobulin G (IgG). In this study, the mig gene from one S. dysgalactiae isolate was cloned and expressed in Escherichia coli. The IgG receptor region encoded by mig was conserved in 16 S. dysgalactiae strains. An isogenic mig mutant was constructed by allele replacement mutagenesis of the wild-type gene in S. dysgalactiae. The IgG-binding activity was lost in the mig mutant strain, whereas the ?2-M receptor activity was still expressed but was detected only in the culture supernatant. In flow cytometry phagocytosis and bacterial-colony-counting bactericidal assays, the wild-type strain was found to be significantly more resistant to phagocytosis and killing by bovine neutrophils (PMNs) than the mig mutant strain when bacteria were preincubated with bovine serum. We therefore speculate that the Mig protein of S. dysgalactiae plays a role in virulence of the bacteria by binding to the plasma protein ?2-M or IgG and thus preventing phagocytosis by bovine PMNs. PMID:11553540

Song, Xin-Ming; Perez-Casal, Jose; Bolton, Alexandra; Potter, Andrew A.

2001-01-01

45

Mouse and bovine models for human IVF  

Microsoft Academic Search

It is obvious that the first prerequisite is to define for what purpose a model is needed for humans. There are huge differences in reproductive physiology between the mouse, human and cow. As far as maturation is concerned, the plasticity of the mouse model is not the same in cows and humans. The final stages of oocyte maturation seem to

Yves JR Ménézo; François Hérubel

2002-01-01

46

Genomic Heritability of Bovine Growth Using a Mixed Model  

PubMed Central

This study investigated heritability for bovine growth estimated with genomewide single nucleotide polymorphism (SNP) information obtained from a DNA microarray chip. Three hundred sixty seven Korean cattle were genotyped with the Illumina BovineSNP50 BeadChip, and 39,112 SNPs of 364 animals filtered by quality assurance were analyzed to estimate heritability of body weights at 6, 9, 12, 15, 18, 21, and 24 months of age. Restricted maximum likelihood estimate of heritability was obtained using covariance structure of genomic relationships among animals in a mixed model framework. Heritability estimates ranged from 0.58 to 0.76 for body weights at different ages. The heritability estimates using genomic information in this study were larger than those which had been estimated previously using pedigree information. The results revealed a trend that the heritability for body weight increased at a younger age (6 months). This suggests an early genetic evaluation for bovine growth using genomic information to increase genetic merits of animals. PMID:25358309

Ryu, Jihye; Lee, Chaeyoung

2014-01-01

47

Genomic heritability of bovine growth using a mixed model.  

PubMed

This study investigated heritability for bovine growth estimated with genomewide single nucleotide polymorphism (SNP) information obtained from a DNA microarray chip. Three hundred sixty seven Korean cattle were genotyped with the Illumina BovineSNP50 BeadChip, and 39,112 SNPs of 364 animals filtered by quality assurance were analyzed to estimate heritability of body weights at 6, 9, 12, 15, 18, 21, and 24 months of age. Restricted maximum likelihood estimate of heritability was obtained using covariance structure of genomic relationships among animals in a mixed model framework. Heritability estimates ranged from 0.58 to 0.76 for body weights at different ages. The heritability estimates using genomic information in this study were larger than those which had been estimated previously using pedigree information. The results revealed a trend that the heritability for body weight increased at a younger age (6 months). This suggests an early genetic evaluation for bovine growth using genomic information to increase genetic merits of animals. PMID:25358309

Ryu, Jihye; Lee, Chaeyoung

2014-11-01

48

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

49

Effect of Langmuir monolayer of bovine serum albumin protein on the morphology of calcium carbonate  

E-print Network

Bovine serum albumin (BSA) Langmuir monolayer, as a model of biomineralization-associated proteins, was used to study its effect on regulated biomineralization of calcium carbonate. The effects of the BSA Langmuir monolayer and the concentration of the subphase solution on the nucleation and growth processes and morphology of the calcium carbonate crystal were investigated. The morphology and polymorphic phase of the resulting calcium carbonate crystals were characterized by scanning electron microscopy (SEM) and X-ray diffraction analysis (XRD). Moreover, the interaction mechanisms of the subphase solution with the BSA Langmuir monolayer were discussed. It was found that BSA Langmuir monolayer could be used as a template to successfully manipulate the polymorphic phase and crystal morphology of calcium carbonate and had obvious influence on the oriented crystallization and growth. The final morphology or aggregation mode of the calcite crystal was closely dependent on the concentration of calcium bicarbonate solution. It is expected that this research would help to better understand the mechanism of biomineralization by revealing the interactions between protein matrices and crystallization of calcium carbonate crystal.

Zhong-Hui Xue; Shu-Xi Dai; Bin-Bin Hu; Zu-Liang Du

2011-06-14

50

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

2013-04-01

51

The 4.1-like proteins of the bovine lens: spectrin-binding proteins closely related in structure to red blood cell protein 4.1  

Microsoft Academic Search

Abstract. The superficial cortical fiber cells of the bovine lens contain membrane-associated proteins of 150,000, 80,000, and 78,000 D that cross-react with an- tisera prepared,against red blood cell (RBC) protein 4.1 (Aster, J. C., G. J. Brewer, S. M. Hanash, and H. Maisel, 1984, Biochem. J., 224:609-616). To further study their relationship to protein 4.1, these proteins were immunoprecipitated from,detergent

Jon C. Aster; George J. Brewer; Harry Maisel

1986-01-01

52

Improved mass spectrometric characterization of protein glycosylation reveals unusual glycosylation of maize-derived bovine trypsin  

PubMed Central

Although bottom-up proteomics using tryptic digests is widely used to locate posttranslational modifications (PTM) in proteins, there are cases where the protein has several potential modification sites within a tryptic fragment, and MS2 strategies fail to pinpoint the location. We report here a method using two proteolytic enzymes, trypsin and pepsin, in combination followed by tandem mass spectrometric analysis to provide fragments that allow one to locate the modification sites. We used this strategy to find a glycosylation site on bovine trypsin expressed in maize (TrypZean™). Several glycans are present, and all are attached to a nonconsensus N-glycosylation site on the protein. PMID:21077632

Zhang, Hao; Huang, Richard Y-C; Jalili, Pegah R.; Irungu, Janet W.; Nicol, Gordon R.; Ray, Kevin B.; Rohrs, Henry W.; Gross, Michael L.

2010-01-01

53

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

54

Inhibition of the bovine papillomavirus E2 protein activity by peptide nucleic acid  

Microsoft Academic Search

The bovine papillomavirus type-1 E2 protein is the master regulator of the papillomavirus transcription and replication, the activity of which is regulated through sequence-specific DNA binding. Peptide nucleic acid (PNA) is a nucleic acid analogue, which associates with high affinity to complementary DNA, RNA or PNA, yielding in formation of stable complexes. The potential use of PNA as a sequence-specific

Reet Kurg; Ülo Langel; Mart Ustav

2000-01-01

55

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

Microsoft Academic Search

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 FeClâ, 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 μl was

CAROL A. KREADER

1996-01-01

56

Proteomic Analysis of Differentially Expressed Proteins in Bovine Endometrium with Endometritis  

PubMed Central

Endometritis is one of the primary reasons for reproductive failure. In order to investigate endometritis-associated marker proteins, proteomic analysis was performed on bovine endometrium with endometritis. In bovine endometritis, desmin, ?-actin-2, heat-shock protein (HSP) 27, peroxiredoxin-6, luteinizing hormone receptor isoform 1, collectin-43 precursor, deoxyribonuclease-I (DNase-I), and MHC class I heavy chain (MHC-Ih) were up-regulated. In contrast, transferrin, interleukin-2 precursor, hemoglobin ? subunit, and potassium channel tetramerisation domain-containing 11 (KCTD11) were down-regulated in comparison to normal endometrium. The proteomic results were validated by semiquantitative-PCR and immunoblot analysis. The mRNA levels of desmin, transferrin, ?-actin-2, HSP27, KCTD11, and MHC-Ih were up-regulated by over 1.5-fold, and showed a pattern similar to their proteomic profiles. Desmin and ?-actin-2 protein showed positive correlations between proteomic analysis and immunoblot analysis. These results suggest that desmin and ?-actin-2 may play important roles in endometritis-related function, and could be useful markers for the diagnosis of bovine endometritis. PMID:20827334

Choe, Changyong; Park, Jeong-Won; Kim, Eun-Suk; Lee, Sung-Gyu; Park, Sun-Young; Lee, Jeong-Soon; Cho, Myung-Je; Kang, Kee Ryeon; Han, Jaehee

2010-01-01

57

Identification of a High Affinity Nucleocapsid Protein Binding Element from The Bovine Leukemia Virus Genome  

PubMed Central

Retroviral genome recognition is mediated by interactions between the nucleocapsid (NC) domain of the virally encoded Gag polyprotein and cognate RNA packaging elements that, for most retroviruses, appear to reside primarily within the 5?-untranslated region (5?-UTR) of the genome. Recent studies suggest that a major packaging determinant of Bovine Leukemia Virus (BLV), a member of the human T-cell leukemia virus (HTLV)/BLV family and a non-primate animal model for HTLV-induced leukemogenesis, resides within the gag open reading frame. We have prepared and purified the recombinant BLV NC protein and conducted electrophoretic mobility shift and isothermal titration calorimetry studies with RNA fragments corresponding to these proposed packaging elements. The gag-derived RNAs did not exhibit significant affinity for NC, suggesting an alternate role in packaging. However, an 83-nucleotide fragment of the 5?-UTR that resides just upstream of the gag start codon binds NC stoichiometrically and with high affinity (Kd = 136 ± 21 nM). These nucleotides were predicted to form tandem hairpin structures, and studies with smaller fragments indicate that the NC binding site resides exclusively within the distal hairpin (residues G369- U399, Kd = 67 ± 8 nM at physiological ionic strength). Unlike all other structurally characterized retroviral NC binding RNAs, this fragment is not expected to contain exposed guanosines, suggesting that RNA binding may be mediated by a previously uncharacterized mechanism. PMID:22846919

Yildiz, F. Zehra; Babalola, Kathleen; Summers, Michael F.

2012-01-01

58

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

59

Production and Functional Analysis of Recombinant Bovine Morphogenic Protein 15  

E-print Network

kDa protein. Bioactivity and BMP receptor signaling specificity were ascertained using in vitro treatment of HeLa cells and Western blotting for the BMP signaling molecule phosphorylated-SMAD 1/5. Inhibition of this signaling cascade using...

Burns, Gregory Willis

2013-11-21

60

Localization of bovine papillomavirus type 1 E5 protein to transformed basal keratinocytes and permissive differentiated cells in fibropapilloma tissue.  

PubMed Central

We examined expression of the E5 transforming protein of bovine papillomavirus type 1 (BPV-1) in naturally and experimentally infected bovine cells. Bovine conjunctival fibroblasts transformed in vitro by experimental infection with purified BPV-1 virions expressed significantly higher amounts of the 7-kDa E5 protein than BPV-1-transformed murine C127 cells. Indirect immunofluourescence analysis revealed a cytoplasmic, predominantly juxtanuclear, localization of E5 protein in the in vitro virus-transformed bovine cells. In naturally infected bovine skin fibropapilloma tissue, two widely separated sites of E5 protein synthesis were identified within the epithelial layers. Transformed basal layer keratinocytes throughout the tumor tissue expressed cytoplasmic E5 protein at a low uniform level. In addition, abundant amounts of cytoplasmic E5 protein with a granular staining pattern were detected in highly differentiated keratinocytes in close association with sites of viral capsid protein synthesis. These observations imply roles for the viral E5 oncogene in the growth transformation of basal epidermal keratinocytes as well as in the differentiation-linked process of viral maturation. Detection of a papillomavirus protein in the basal cell population of warts lends support to the hypothesis that these cells are maintained in a transformed state by continuous expression of a viral transforming gene. Images PMID:1319069

Burnett, S; Jareborg, N; DiMaio, D

1992-01-01

61

Measurement of protein synthesis and degradation in C 2 C 22 myoblasts using extracts of muscle from hormone treated bovine  

Microsoft Academic Search

A detailed methodology is described for determination of treatment effects on muscle cell protein synthesis and muscle cell protein degradation in a cell culture system. C2C22 mouse myoblasts were treated with growth media containing muscle extracts from bovine treated with different pharmaceutical agents. Radiolabeled amino acids were added to the growth media to determine treatment effects on protein synthesis and

J. L. Montgomery; W. M. Harper; M. F. Miller; K. J. Morrow; J. R. Blanton

2002-01-01

62

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

2014-06-21

63

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

NASA Astrophysics Data System (ADS)

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.

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

2014-11-01

64

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

65

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. PMID:24904221

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

2014-01-01

66

Cloning of the bovine antiapoptotic regulator, BCL2-related protein A1, and its expression in trophoblastic binucleate cells of bovine placenta.  

PubMed

This report studied the identification and sequence of a full-length cDNA for the bovine BCL2 antiapoptotic family member, BCL2-related protein A1 (BCL2A1), and its localized and quantitative expression in the placenta to clarify the regulatory mechanism of trophoblast cell proliferation and differentiation during implantation and placental development. We cloned a full-length bovine BCL2A1 cDNA with 725 nucleotides and an open-reading frame corresponding to a protein of 175 amino acids. The predicted amino acid sequence shared 78% homology with human BCL2A1. All BCL2 homology domains (BH1, BH2, BH3, and BH4) in bovine BCL2A1 were conserved as well as in other mammalian BCL2A1. In the placentomes, in situ hybridization demonstrated that the BCL2A1 was limited in binucleate cells expressing various pregnancy-specific molecules like placental lactogen. BCL2-associated X protein (BAX) was also expressed in binucleate cells. Quantitative real-time RT-PCR detection exhibited a high-level expression of BCL2A1 in the conceptus at Day 21 of gestation, and it was expressed and increased in the extraembryonic membrane, cotyledon, and intercotyledon from implantation to term. BAX expression intensity increased with progression of gestation and remained elevated in postpartum. Caspase-3 protein (CASP3) and mRNA (CASP3) were detected from late gestation to postpartum in placenta as well as in the results of TUNEL detection. We believe that the apoptosis of binucleate cells may be regulated by the balance of the BCL2A1 and BAX. BCL2A1 genes produced a BCL2A1 protein in the mammalian cell-expression system. This molecule is a new candidate for antiapoptotic maintenance of the binucleate cells that support placental functions throughout gestation in bovine. PMID:16221993

Ushizawa, Koichi; Takahashi, Toru; Kaneyama, Kanako; Hosoe, Misa; Hashizume, Kazuyoshi

2006-02-01

67

Association of Bovine Papillomavirus E2 Protein with Nuclear Structures In Vivo  

PubMed Central

Papillomaviruses are small DNA viruses which have the capacity to establish a persistent infection in mammalian epithelial cells. The papillomavirus E2 protein is a central coordinator of viral gene expression, genome replication, and maintenance. We have investigated the distribution of bovine papillomavirus E2 protein in nuclei of proliferating cells and found that E2 is associated with cellular chromatin. This distribution does not change during the entire cell cycle. The N-terminal transactivation domain, but not the C-terminal DNA-binding domain, of the E2 protein is responsible for this association. The majority of the full-length E2 protein can only be detected in chromatin-enriched fractions but not as a free protein in the nucleus. Limited micrococcal nuclease digestion revealed that the E2 protein partitioned to different chromatin regions. A fraction of the E2 protein was located at nuclear sites that are resistant against nuclease attack, whereas the remaining E2 resided on compact chromatin accessible to micrococcal nuclease. These data suggest that there are two pools of E2 in the cell nucleus: one that localizes on transcriptionally inactive compact chromatin and the other, which compartmentalizes to transcriptionally active nuclear structures of the cell. Our data also suggest that E2 associates with chromatin through cellular protein(s), which in turn is released from chromatin at 0.4 M salt. PMID:16051845

Kurg, Reet; Sild, Kristiina; Ilves, Aigi; Sepp, Mari; Ustav, Mart

2005-01-01

68

Function of Bovine CD46 as a Cellular Receptor for Bovine Viral Diarrhea Virus Is Determined by Complement Control Protein 1  

PubMed Central

The pestivirus bovine viral diarrhea virus (BVDV) was shown to bind to the bovine CD46 molecule, which subsequently promotes entry of the virus. To assess the receptor usage of BVDV type 1 (BVDV-1) and BVDV-2, 30 BVDV isolates including clinical samples were assayed for their sensitivity to anti-CD46 antibodies. With a single exception the infectivity of all tested strains of BVDV-1 and BVDV-2 was inhibited by anti-CD46 antibodies, which indicates the general usage of CD46 as a BVDV receptor. Molecular analysis of the interaction between CD46 and the BVD virion was performed by mapping the virus binding site on the CD46 molecule. Single complement control protein modules (CCPs) within the bovine CD46 were either deleted or replaced by analogous CCPs of porcine CD46, which does not bind BVDV. While the epitopes recognized by anti-CD46 monoclonal antibodies which block BVDV infection were attributed to CCP1 and CCP2, in functional assays only CCP1 turned out to be essential for BVDV binding and infection. Within CCP1 two short peptides on antiparallel beta strands were identified as crucial for the binding of BVDV. Exchanges of these two peptide sequences were sufficient for a loss of function in bovine CD46 as well as a gain of function in porcine CD46. Determination of the size constraints of CD46 revealed that a minimum length of four CCPs is essential for receptor function. An increase of the distance between the virus binding domain and the plasma membrane by insertion of one to six CCPs of bovine C4 binding protein exhibited only a minor influence on susceptibility to BVDV. PMID:16571808

Krey, Thomas; Himmelreich, Anke; Heimann, Manuela; Menge, Christian; Thiel, Heinz-Jürgen; Maurer, Karin; Rümenapf, Till

2006-01-01

69

The Protein Model Portal.  

PubMed

Structural Genomics has been successful in determining the structures of many unique proteins in a high throughput manner. Still, the number of known protein sequences is much larger than the number of experimentally solved protein structures. Homology (or comparative) modeling methods make use of experimental protein structures to build models for evolutionary related proteins. Thereby, experimental structure determination efforts and homology modeling complement each other in the exploration of the protein structure space. One of the challenges in using model information effectively has been to access all models available for a specific protein in heterogeneous formats at different sites using various incompatible accession code systems. Often, structure models for hundreds of proteins can be derived from a given experimentally determined structure, using a variety of established methods. This has been done by all of the PSI centers, and by various independent modeling groups. The goal of the Protein Model Portal (PMP) is to provide a single portal which gives access to the various models that can be leveraged from PSI targets and other experimental protein structures. A single interface allows all existing pre-computed models across these various sites to be queried simultaneously, and provides links to interactive services for template selection, target-template alignment, model building, and quality assessment. The current release of the portal consists of 7.6 million model structures provided by different partner resources (CSMP, JCSG, MCSG, NESG, NYSGXRC, JCMM, ModBase, SWISS-MODEL Repository). The PMP is available at http://www.proteinmodelportal.org and from the PSI Structural Genomics Knowledgebase. PMID:19037750

Arnold, Konstantin; Kiefer, Florian; Kopp, Jürgen; Battey, James N D; Podvinec, Michael; Westbrook, John D; Berman, Helen M; Bordoli, Lorenza; Schwede, Torsten

2009-03-01

70

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. PMID:12124257

Baysal, Canan; Atilgan, Ali Rana

2002-01-01

71

A Protein Encoded by the Bovine Herpesvirus 1 Latency-Related Gene Interacts with Specific Cellular Regulatory Proteins, Including CCAAT Enhancer Binding Protein Alpha?  

PubMed Central

Following acute infection, bovine herpesvirus 1 establishes latency in sensory neurons of trigeminal ganglia (TG). Reactivation from latency occurs periodically, resulting in the shedding of infectious virus. The latency-related (LR) RNA is abundantly expressed in TG of latently infected calves, and the expression of LR proteins is necessary for dexamethasone-induced reactivation from latency. Previously published studies also identified an alternatively spliced LR transcript which is abundantly expressed in TG at 7 days after infection and has the potential to encode a novel LR fusion protein. Seven days after infection is when extensive viral gene expression is extinguished in TG and latency is established, suggesting that LR gene products influence the establishment of latency. In this study, we used a bacterial two-hybrid assay to identify cellular proteins that interact with the novel LR fusion protein. The LR fusion protein interacts with two proteins that can induce apoptosis (Bid and Cdc42) and with CCAAT enhancer binding protein alpha (C/EBP-?). Additional studies confirmed that the LR fusion protein interacts with human or insect C/EBP-?. C/EBP-? protein expression is induced in TG neurons of infected calves and after dexamethasone-induced reactivation from latency. Wild-type C/EBP-?, but not a DNA binding mutant of C/EBP-?, enhances plaque formation in bovine cells. We hypothesize that interactions between the LR fusion protein and C/EBP-? promote the establishment of latency. PMID:16987965

Meyer, Florencia; Perez, Sandra; Geiser, Vicki; Sintek, Mark; Inman, Melissa; Jones, Clinton

2007-01-01

72

L233P mutation of the Tax protein strongly correlated with leukemogenicity of bovine leukemia virus.  

PubMed

The bovine leukemia virus (BLV) Tax protein is believed to play a crucial role in leukemogenesis by the virus. BLV usually causes asymptomatic infections in cattle, but only one-third develop persistent lymphocytosis that rarely progress after a long incubation period to lymphoid tumors, namely enzootic bovine leucosis (EBL). In the present study, we demonstrated that the BLV tax genes could be divided into two alleles and developed multiplex PCR detecting an L233P mutation of the Tax protein. Then, in order to define the relationship between the Tax protein and leukemogenicity, we examined 360 tumor samples randomly collected from dairy or breeding cattle in Japan, of which Tax proteins were categorized, for age at the time of diagnosis of EBL. The ages of 288 animals (80.0%) associated with L233-Tax and those of 70 animals (19.4%) with P233-Tax individually followed log-normal distributions. Only the two earliest cases (0.6%) with L233-Tax disobeyed the log-normal distribution. These findings suggest that the animals affected by EBL were infected with the virus at a particular point in life, probably less than a few months after birth. Median age of those with P233-Tax was 22 months older than that with L233-Tax and geometric means exhibited a significant difference (P<0.01). It is also quite unlikely that viruses carrying the particular Tax protein infect older cattle. Here, we conclude that BLV could be divided into two categories on the basis of amino acid at position 233 of the Tax protein, which strongly correlated with leukemogenicity. PMID:24139177

Inoue, Emi; Matsumura, Keiko; Soma, Norihiko; Hirasawa, Shintaro; Wakimoto, Mayuko; Arakaki, Yoshihiro; Yoshida, Takashi; Osawa, Yoshiaki; Okazaki, Katsunori

2013-12-27

73

Fluorescent probing of protein bovine serum albumin stability and denaturation using polarity sensitive spectral response of a charge transfer probe.  

PubMed

The polarity sensitive photo-induced intra-molecular charge transfer (ICT) fluorescence probe (E)-3-(4-methylamino-phenyl)-acrylic acid ethyl ester (MAPAEE) has been used to study the model protein Bovine Serum Albumin (BSA) in its native and thermal and urea induced denatured states. The interaction between BSA and the regular surfactant Sodium Dodecyl Sulphate (SDS) as well as the biologically relevant steroid-based amphiphile Sodium Deoxycholate (NaDC) has also been very keenly followed using this ICT probe. The variation of micellar properties of both SDS and NaDC with increasing ionic strengths and in presence of the chaotrope urea has also been well documemted by the same probe. Steady-state spectroscopy, FRET, and fluorescence anisotropy measurements have been used to gain better insight into these processes and the molecule MAPAEE to be a full-bodied fluorescent probe for studying such intricate biological systems, their properties and interactions. PMID:20922468

Ghosh, Shalini; Jana, Sankar; Nath, Debnarayan; Guchhait, Nikhil

2011-01-01

74

In Vitro and In Vivo Oncogenic Potential of Bovine Leukemia Virus G4 Protein  

PubMed Central

In addition to the genes involved in the structure of the viral particle, the bovine leukemia virus (BLV) genome contains a region called X which contains at least four genes. Among them, the tax and rex genes, respectively, are involved in transcriptional and posttranscriptional regulation of viral transcription. Two other genes, R3 and G4, were identified after cloning of the corresponding mRNAs from BLV-infected lymphocytes. Although the function of the two latter genes is still unknown, they appear to have important roles, since deletion of them restricts viral propagation in vivo. In order to assess the oncogenic potential of the R3 and G4 proteins, we first analyzed their ability to immortalize and/or transform primary rat embryo fibroblasts (Refs). In this assay, the G4 but not the R3 protein cooperated with the Ha-ras oncogene to induce tumors in nude mice. It thus appears that G4 exhibited oncogenic potential in vitro. To extend these observations in vivo, the pathology induced by recombinant viruses with mutations in G4 and in R3 and G4 was next evaluated with the sheep animal model. Viral propagation, as measured by semiquantitative PCR, appeared to be reduced when the R3 and G4 genes were deleted. These observations confirm and extend our previous data underlining the biological function of these genes. In addition, we present the results of a clinical survey that involves 39 sheep infected with six different BLV recombinants. Over a period of 40 months, 83% of the sheep infected with a wild-type virus developed leukemias and/or lymphosarcomas. In contrast, none out of 13 sheep infected with viruses with mutations in G4 or in R3 and G4 developed disease. We conclude that in addition to its oncogenic potential in vitro, G4 is required for pathogenesis in vivo. These observations should help us gain insight into the process of leukemogenesis induced by the related human T-cell leukemia virus type 1. PMID:9499124

Kerkhofs, Pierre; Heremans, Hubertine; Burny, Arsene; Kettmann, Richard; Willems, Luc

1998-01-01

75

Non-oxidative antibacterial activity of bovine neutrophil granule proteins towards mastitis pathogens.  

PubMed

Acid extracts of bovine neutrophil granules displayed potent antibacterial activity towards a number of mastitis pathogens in vitro. Killing of pathogens by acid extractable granule protein was dependent on incubation time, protein concentration, bacterial cell load, pH and ionic strength. Gram-negative and Gram-positive organisms showed variable sensitivity to granule extract. Strains of Staphylococcus aureus were the most resistant of tested organisms to granule extract. Gram-negative organisms were neither consistently more nor less sensitive than Gram-positive organisms. Maximal killing of Gram-positive pathogens, after 30 minutes exposure to granule extract at 37 degrees C, occurred between pH 7.0 and 8.0. The Gram-negative organism Escherichia coli B117 was more sensitive to neutrophil granule extract at pH 5.0. PMID:2816175

Grieve, P A; Mattila, T

1989-09-01

76

Anti-Ebola MAb17A3 Reacts with Bovine and Human Alpha-2-macroglobulin Proteins  

PubMed Central

Monoclonal antibodies (MAbs) were developed against soluble Ebola virus (EBOV) envelope glycoprotein (GP) for the study of the diversity of EBOV envelope and development of diagnostic reagents. Of the three anti-EBOV GP mouse MAbs produced, MAb 15H10 recognized all human EBOV GP species tested (Zaire, Sudan, Ivory Coast), and as well as reacted with the Reston nonhuman primate EBOV GPs. A second MAb, 6D11 recognized EBOV GP species of Sudan and Sudan-Gulu. The third MAb, 17A3, was reported originally in the same article to be EBOV GP-specific has now been found to be specific for bovine and human ?-2 macroglobulin (?-2M) proteins which were contaminants in the Ebola envelope protein preparation. Thus, while MAbs 15H10 and 6D11 are indeed EBOV GP specific, MAb 17A3 is an ?-2 macroglobulin MAb. PMID:20447422

Yu, Jae-Sung; Ma, BenJiang; Scearce, Richard M.; Liao, Hua-Xin; Haynes, Barton F.

2010-01-01

77

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

78

Bone morphogenetic protein 15 in the pro-mature complex form enhances bovine oocyte developmental competence.  

PubMed

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. PMID:25058588

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

2014-01-01

79

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

2014-01-01

80

The Quaternary Structure of the Recombinant Bovine Odorant-Binding Protein Is Modulated by Chemical Denaturants  

PubMed Central

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

81

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. PMID:23376657

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

2013-01-01

82

Evaluation of bovine viral diarrhea virus control using a mathematical model of infection dynamics  

Microsoft Academic Search

A mathematical model for infection with bovine viral diarrhea virus (BVDV) was created comprising a series of coupled differential equations. The model architecture is a development of the traditional model framework using susceptible, infectious and removed animals (the SIR model). The model predicts 1.2% persistent infection (within the range of field estimates) and is fairly insensitive to alterations of structure

B. R Cherry; M. J Reeves; G Smith

1998-01-01

83

Assessing the Susceptibility of Transgenic Mice Overexpressing Deer Prion Protein to Bovine Spongiform Encephalopathy  

PubMed Central

Several transgenic mouse models have been developed which facilitate the transmission of chronic wasting disease (CWD) of cervids and allow prion strain discrimination. The present study was designed to assess the susceptibility of the prototypic mouse line, Tg(CerPrP)1536+/?, to bovine spongiform encephalopathy (BSE) prions, which have the ability to overcome species barriers. Tg(CerPrP)1536+/? mice challenged with red deer-adapted BSE resulted in 90% to 100% attack rates, and BSE from cattle failed to transmit, indicating agent adaptation in the deer. PMID:24257620

Vickery, Christopher M.; Lockey, Richard; Holder, Thomas M.; Thorne, Leigh; Beck, Katy E.; Wilson, Christina; Denyer, Margaret; Sheehan, John; Marsh, Sarah; Webb, Paul R.; Dexter, Ian; Norman, Angela; Popescu, Emma; Schneider, Amanda; Holden, Paul; Griffiths, Peter C.; Plater, Jane M.; Dagleish, Mark P.; Martin, Stuart; Telling, Glenn C.; Simmons, Marion M.

2014-01-01

84

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

85

Induced thermotolerance in bovine two-cell embryos and the role of heat shock protein 70 in embryonic development  

Microsoft Academic Search

Induced thermotolerance is a phenomenon whereby exposure to a mild heat shock can induce heat shock proteins (HSP) and other cellular changes to make cells more resistant to a subsequent, more severe heat shock. Given that the 2-cell bovine embryo is very sensitive to heat shock, but can also produce HSP70 in response to elevat- ed temperature, experiments were conducted

Y. M. Al-Katanani; P. J. Hansen

2002-01-01

86

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. PMID:25072040

Peruffo, Antonella; Cozzi, Bruno

2014-01-01

87

SimulatingWater MOLECULAR DYNAMICS MODEL of bovine pancreatic  

E-print Network

in their native, watery environs. Today, given the great advances in computing technology, we can model proteins of the tetrahedron, hydrogen atoms at two of the four corners and clouds of negative charge at the other two corners. The clouds of negative charge result from the way in which the atomic structures of oxygen and hydrogen

Levitt, Michael

88

4528 J. Phys. Chem. 1993,97, 4528-4534 Electrophoretic and Quasi-ElasticLight Scattering of Soluble Protein-Polyelectrolyte Complexes  

E-print Network

Form: November 30, I992 Complexation between globular proteins (bovine serum albumin, bovine pancreas of bovine serum albumin (BSA) and quaternized poly(4-vinylpyridines), Kabanov22proposed a model in which

Dubin, Paul D.

89

Isothermal Titration Calorimetric Studies on the Interaction of the Major Bovine Seminal Plasma Protein, PDC-109 with Phospholipid Membranes  

PubMed Central

The interaction of the major bovine seminal plasma protein, PDC-109 with lipid membranes was investigated by isothermal titration calorimetry. Binding of the protein to model membranes made up of diacyl phospholipids was found to be endothermic, with positive values of binding enthalpy and entropy, and could be analyzed in terms of a single type of binding sites on the protein. Enthalpies and entropies for binding to diacylphosphatidylcholine membranes increased with increase in temperature, although a clear-cut linear dependence was not observed. The entropically driven binding process indicates that hydrophobic interactions play a major role in the overall binding process. Binding of PDC-109 with dimyristoylphosphatidylcholine membranes containing 25 mol% cholesterol showed an initial increase in the association constant as well as enthalpy and entropy of binding with increase in temperature, whereas the values decreased with further increase in temperature. The affinity of PDC-109 for phosphatidylcholine increased at higher pH, which is physiologically relevant in view of the basic nature of the seminal plasma. Binding of PDC-109 to Lyso-PC could be best analysed in terms of two types of binding interactions, a high affinity interaction with Lyso-PC micelles and a low-affinity interaction with the monomeric lipid. Enthalpy-entropy compensation was observed for the interaction of PDC-109 with phospholipid membranes, suggesting that water structure plays an important role in the binding process. PMID:22022488

Anbazhagan, V.; Sankhala, Rajeshwer S.; Singh, Bhanu Pratap; Swamy, Musti J.

2011-01-01

90

Purification and characterization of a protein-tyrosine kinase from bovine thymus.  

PubMed

A protein-tyrosine kinase has been isolated from a soluble extract of bovine thymus based on its ability to phosphorylate the tyrosine-containing peptide angiotensin I. The purification procedure employs sequential chromatography on columns of DEAE-cellulose, heparin-agarose, casein-agarose, butyl-agarose, and Sephadex G-75. The purified enzyme (p40) is a monomer of Mr = 40,000. The p40 kinase contains an ATP-binding site as determined by photoaffinity labeling experiments and catalyzes an intramolecular autophosphorylation reaction that leads to its modification on tyrosine. Of several proteins tested only the cytoplasmic domain of the erythrocyte band 3 protein serves as a good substrate for p40 (Km = 12 microM). Increasing concentrations of NaCl stimulate the phosphorylation of angiotensin I, inhibit the phosphorylation of band 3, and have no effect on the autophosphorylation of p40. At low concentrations of NaCl, Mn2+ is the preferred divalent cation. Peptide mapping experiments indicate that p40 is distinct from pp60src and from the major phosphotyrosine containing proteins of T and B lymphocyte membranes. PMID:3782080

Zioncheck, T F; Harrison, M L; Geahlen, R L

1986-11-25

91

The bovine basic pancreatic trypsin inhibitor (Kunitz inhibitor): a milestone protein.  

PubMed

The pancreatic Kunitz inhibitor, also known as aprotinin, bovine basic pancreatic trypsin inhibitor (BPTI), and trypsin-kallikrein inhibitor, is one of the most extensively studied globular proteins. It has proved to be a particularly attractive and powerful tool for studying protein conformation as well as molecular bases of protein/protein interaction(s) and (macro)molecular recognition. BPTI has a relatively broad specificity, inhibiting trypsin- as well as chymotrypsin- and elastase-like serine (pro)enzymes endowed with very different primary specificity. BPTI reacts rapidly with serine proteases to form stable complexes, but the enzyme: inhibitor complex formation may involve several intermediates corresponding to discrete reaction steps. Moreover, BPTI inhibits the nitric oxide synthase type-I and -II action and impairs K+ transport by Ca2+-activated K+ channels. Clinically, the use of BPTI in selected surgical interventions, such as cardiopulmonary surgery and orthotopic liver transplantation, is advised, as it significantly reduces hemorrhagic complications and thus blood-transfusion requirements. Here, the structural, inhibition, and bio-medical aspects of BPTI are reported. PMID:12769721

Ascenzi, Paolo; Bocedi, Alessio; Bolognesi, Martino; Spallarossa, Andrea; Coletta, Massimo; De Cristofaro, Raimondo; Menegatti, Enea

2003-06-01

92

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

93

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

PubMed Central

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

2014-01-01

94

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

95

Effects of water quality on in vitro fertilization and development of bovine oocytes in protein-free medium  

Microsoft Academic Search

Examination was made of the effects of water quality in medium preparation on fertilization and early development of bovine in vitro matured (IVM) oocytes in a protein-free medium. The IVM oocytes were inseminated and cultured for 7 d in protein-free media prepared with 4 different types of water preparations: tap, deionized, twice-distilled, and purified water using the Milli-Q system (Milli-Q

Y. Nagao; K. Saeki; M. Hoshi; Y. Takahashi; H. Kanagawa

1995-01-01

96

Aging Phenomena in Spermatozoa. IV. Immunoserologic Characterization of the Deoxyribonucleic Acid-Protein Extracted from Fresh and Aged Bovine Spermatozoa  

Microsoft Academic Search

The deoxyribonucleic acid-protein com- plex was extracted from fresh and aged bovine spermatozoa and characterized by immunoserologic techniques. Seventeen five~ to eight-month-old rabbits were injected with the deoxyribonucleic acid-protein complex, or the deoxyribonucleic acid-pro- tein complex treated with ribonuclease, deoxyribonuclease, or both, respectively. Rabbits injected with washed spermatozoa, seminal plasma, trypsin, ribonuclease, de- oxyribonuelease, or both ribonuclease and deoxyribonuclease, were

R. A. Todorovic; C. N. Graves; G. W. Salisbury

1969-01-01

97

Acetic Acid Activates the AMP-Activated Protein Kinase Signaling Pathway to Regulate Lipid Metabolism in Bovine Hepatocytes  

PubMed Central

The effect of acetic acid on hepatic lipid metabolism in ruminants differs significantly from that in monogastric animals. Therefore, the aim of this study was to investigate the regulation mechanism of acetic acid on the hepatic lipid metabolism in dairy cows. The AMP-activated protein kinase (AMPK) signaling pathway plays a key role in regulating hepatic lipid metabolism. In vitro, bovine hepatocytes were cultured and treated with different concentrations of sodium acetate (neutralized acetic acid) and BML-275 (an AMPK? inhibitor). Acetic acid consumed a large amount of ATP, resulting in an increase in AMPK? phosphorylation. The increase in AMPK? phosphorylation increased the expression and transcriptional activity of peroxisome proliferator-activated receptor ?, which upregulated the expression of lipid oxidation genes, thereby increasing lipid oxidation in bovine hepatocytes. Furthermore, elevated AMPK? phosphorylation reduced the expression and transcriptional activity of the sterol regulatory element-binding protein 1c and the carbohydrate responsive element-binding protein, which reduced the expression of lipogenic genes, thereby decreasing lipid biosynthesis in bovine hepatocytes. In addition, activated AMPK? inhibited the activity of acetyl-CoA carboxylase. Consequently, the triglyceride content in the acetate-treated hepatocytes was significantly decreased. These results indicate that acetic acid activates the AMPK? signaling pathway to increase lipid oxidation and decrease lipid synthesis in bovine hepatocytes, thereby reducing liver fat accumulation in dairy cows. PMID:23861826

Li, Xinwei; Chen, Hui; Guan, Yuan; Li, Xiaobing; Lei, Liancheng; Liu, Juxiong; Yin, Liheng; Liu, Guowen; Wang, Zhe

2013-01-01

98

Prion protein gene (PRNP) variants and evidence for strong purifying selection in functionally important regions of bovine exon 3  

PubMed Central

Amino acid replacements encoded by the prion protein gene (PRNP) have been associated with transmissible and hereditary spongiform encephalopathies in mammalian species. However, an association between bovine spongiform encephalopathy (BSE) and bovine PRNP exon 3 has not been detected. Moreover, little is currently known regarding the mechanisms of evolution influencing the bovine PRNP gene. Therefore, in this study we evaluated the patterns of nucleotide variation associated with PRNP exon 3 for 36 breeds of domestic cattle and representative samples for 10 additional species of Bovinae. The results of our study indicate that strong purifying selection has intensely constrained PRNP over the long-term evolutionary history of the subfamily Bovinae, especially in regions considered to be of functional, structural, and pathogenic importance in humans as well as other mammals. The driving force behind this intense level of purifying selection remains to be explained. PMID:15477588

Seabury, Christopher M.; Honeycutt, Rodney L.; Rooney, Alejandro P.; Halbert, Natalie D.; Derr, James N.

2004-01-01

99

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

100

Segmental differentiation of permeability, protein glycosylation, and morphology of cultured bovine lung vascular endothelium.  

PubMed

The barrier function, surface biochemistry, and morphology of confluent monolayers of endothelial cells isolated from different segments of the bovine lung vasculature [microvessels (BLMVEC), vein (BPVEC) and artery (BPAEC)] were grown in culture and compared. A number of common cell surface proteins were identified along with two proteins of 46 and 48 kDa found exclusively on BPVEC. Lectin affinity chromatography revealed multiple glycosylation differences. The lectins, Arachis hypogaea (AHA) and Lycopersicum esculentum (LEA) agglutinins, interacted with several glycoproteins of BLMVEC but not of BPAEC. Bandeiraea simplicifolia (BS-1) and Caragana arborescens (CAA) agglutinins recognized several glycoproteins of BPVEC and BPAEC but not BLMVEC. Permeabilities were much lower for BLMVEC than BPAEC or BPVEC monolayers, with a range of about 16-fold less for sucrose to 2-fold less for albumin. Electron microscopy revealed that BLMVEC have a greater surface density of plasmalemmal vesicles (approximately 4-fold) and more extensively developed intercellular junctions with more focal membrane adhesion sites per junction (approximately 9-fold) than the other cells. We conclude that: i) BLMVEC monolayers form a much more restrictive barrier to molecular transport as a result of the tighter junctional formation; and ii) endothelial surface glycoproteins may be differentially glycosylated depending on their segmental location within the vasculature. PMID:8123001

Schnitzer, J E; Siflinger-Birnboim, A; Del Vecchio, P J; Malik, A B

1994-02-28

101

Subcellular Localization of the Bovine Leukemia Virus R3 and G4 Accessory Proteins  

PubMed Central

Bovine leukemia virus (BLV) is a complex retrovirus that belongs to the Deltaretrovirus genus, which also includes Human T-cell leukemia virus type 1 (HTLV-1). Both viruses contain an X region coding for at least four proteins: Tax and Rex, which are involved in transcriptional and posttranscriptional regulation, respectively, and the accessory proteins R3 and G4 (for BLV) and p12I, p13II, and p30II (for HTLV-1). The present study was aimed at characterizing the subcellular localization of BLV R3 and G4. The results of immunofluorescence experiments on transfected HeLa Tat cells demonstrated that R3 is located in the nucleus and in cellular membranes, as previously reported for HTLV-1 p12I. In contrast, G4, like p13II, is localized both in the nucleus and in mitochondria. In addition, we have shown that G4 harbors a mitochondrial targeting signal consisting of a hydrophobic region and an amphipathic ?-helix. Thus, despite a lack of significant primary sequence homology, R3 and p12I and G4 and p13II exhibit similar targeting properties, suggesting possible overlap in their functional properties. PMID:12097596

Lefebvre, Laurent; Ciminale, Vincenzo; Vanderplasschen, Alain; D'Agostino, Donna; Burny, Arsene; Willems, Luc; Kettmann, Richard

2002-01-01

102

Comparative modeling of breakthrough curves of bovine serum albumin in anion-exchange chromatography  

Microsoft Academic Search

The experimental results of a previous study of the mass transfer kinetics of bovine serum albumin (BSA) in ion-exchange chromatography, under nonlinear conditions, were reevaluated using the general rate model of chromatography. Solutions of this model were obtained numerically. The influences of axial dispersion, the resistance to mass transfer from the bulk mobile phase to the surface of the packing

Krzysztof Kaczmarski; Dorota Antos; Hong Sajonz; Peter Sajonz; Georges Guiochon

2001-01-01

103

Determination of immunoglobulin G in bovine colostrum and milk powders, and in dietary supplements of bovine origin by protein G affinity liquid chromatography: collaborative study.  

PubMed

An AOAC collaborative study was conducted to evaluate an affinity LC procedure for measuring immunoglobulin G (IgG) in selected dairy powders. The powders were extracted with 0.15 M sodium chloride solution and the pH was adjusted to 4.6 to precipitate caseins, which would otherwise lead to an overestimation of IgG. The analyte was then bound to a commercially available Protein G affinity cartridge and selectively eluted with a glycine buffer at pH 2.5. Detection was at 280 nm and quantification was made against a calibration curve prepared from bovine serum IgG. The samples analyzed included the likely matrixes for which this assay will find commercial use, namely, high- and low-protein-content colostrum powders, tablets containing colostrum powder, and some IgG-containing dairy powders; milk protein isolate, whey protein concentrate, and skim milk powder. Eleven laboratories provided data for the study and assayed blind duplicates of six materials. The repeatability RSD values ranged from 2.1 to 4.2% and the reproducibility RSD values ranged from 6.4 to 18.5%. The Protein G method with casein removal has adequate reproducibility for measuring IgG in colostrum-derived powders that are traded on the basis of IgG content as a colostral marker. PMID:20480910

Abernethy, Grant; Otter, Don; Arnold, K; Austad, J; Christiansen, S; Ferreira, I; Irvine, F; Marsh, C; Massom, L R; Otter, D; Pearce, K; Stevens, J; Szpylka, J; Vyas, P; Woollard, D; Wu, C

2010-01-01

104

Time-resolved fluorescence studies on the single tryptophan in bovine brain S-100a protein  

NASA Astrophysics Data System (ADS)

The fluorescence emission of the single tryptophan in bovine brain S-lOOa protein has been studied by using time-resolved laser fluorescence spectroscopy. The tryptophan fluorescence emission was isolated by a Schott 0-54 cut-off filter at right angles to the excitation direction by a Hamamatsu R955P photomultiplier. With excitation at 295 nm, the fluorescence decay of S-lOOa protein in 25 mM Tris buffer was best represented by a sum of three exponential terms regardless of solvent conditions. At 20°C and pH 7.2, the three components of lifetime for apo-S-lOOa protein were (tau)1~0.43 ns, (tau)2~1.24 ns, and (tau)3~4.05 ns. The corresponding fluorescence contributions of each component were 31%, 31% and 38%, respectively. When the protein was saturated with Mg2, the lifetimes increased slightly and the contribution of the shortest fluorescence component ((tau)1) to the total emission intensity increased slightly at the expense of the other two components. Binding of Ca2+ to S-lOOa protein resulted in a significant decrease of (tau)1, and a substantial increase of (tau)2 and (tau)3, and an increase of the average of the three lifetimes. Under this condition the fractional fluorescence contributions associated with (tau)2 and (tau)3 increased very significantly at the expense of the shortest component. The fluorescence decay behavior of each of he S-lOOa samples was relatively insensitive to the variation of temperature (4~20°C). At pH 8.2 and pH 8.4, the decay behavior of the protein in response to binding of Ca2+ and Mg2+, and changes of temperature was very similar to those observed at pH 7.2. The triple exponential decay kinetics of the single tryptophan in S-lOOa protein could be rationalized by the existence of multiple local conformers.

Wang, Chien-Kao; Mani, Rajam S.; Kay, Cyril M.; Cheung, Herbert C.

1990-05-01

105

Sequence-specific 1H NMR assignments and structural characterization of bovine seminal fluid protein PDC-109 domain b.  

PubMed

Sequence-specific resonance assignments for the isolated second or b domain of the bovine seminal fluid protein PDC-109 have been obtained from analysis of two-dimensional 1H NMR experiments recorded at 500 MHz. These assignments include the identification of all aromatic and most aliphatic amino acid resonances. Stereospecific assignment of resonances stemming from the Val2 CH3 gamma,gamma' groups and from seven CH beta,beta' geminal pairs has been accomplished by analysis of 3J alpha beta coupling constants in conjunction with patterns of cross-peak intensities observed in two-dimensional nuclear Overhauser effect (NOESY) spectra. Analysis of NOESY and 3J alpha NH data reveals a small antiparallel beta-sheet involving stretches containing residues 25-28 and 39-42, a cis-proline residue (Pro4), antiparallel strands consisting of residues 1-3, 5-7, and 10-13, and an aromatic cluster composed of Tyr7, Trp26, and Tyr33. The results of distance geometry and restrained molecular dynamics calculations indicate that the global fold of the PDC-109 b domain, a type II module related to those found in fibronectin, is somewhat different from that predicted by modeling the structure on the basis of homology between type II and kringle units. A shallow depression in the molecular surface which presents a solvent-exposed hydrophobic area--a potential ligand-binding site-is identified in the NMR-based models. PMID:1993183

Constantine, K L; Ramesh, V; Bányai, L; Trexler, M; Patthy, L; Llinás, M

1991-02-12

106

Effects of Protein-Polyelectrolyte Affinity and Polyelectrolyte Molecular Weight on Dynamic Properties of Bovine Serum  

E-print Network

Properties of Bovine Serum Albumin-Poly(diallyldimethylammonium chloride) Coacervates H. Bohidar, P. L. Dubin serum albumin (BSA) and poly(diallyldimethylammonium chloride) (PDADMAC) spontaneously form, over Potsdam-Golm, Germany Received December 28, 2004; Revised Manuscript Received February 21, 2005 Bovine

Dubin, Paul D.

107

Lack of repair of rat skull critical size defect treated with bovine morphometric protein bound to microgranular bioabsorbable hydroxyapatite  

Microsoft Academic Search

The ability of a pool of bovine bone morphogenetic proteins bound to synthetic microgranular hydroxyapatite (BMPb-HA) to stimulate bone repair was determined in rat critical size defects. An 8-mm diameter defect was created in the calvaria of 25 rats. In 15 rats, the defects were filled with BMPb-HA homogenized with blood (experimental group), and in 10 rats the defects were

Gabriel Ramalho Ferreira; Tania Mary Cestari; José Mauro Granjeiro; Rumio Taga

2004-01-01

108

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. PMID:24475205

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

2014-01-01

109

Evaluation of a Bovine Temperament Model for Endophenotypes Associated with Hypothalamic-Pituitary-Adrenal Axis Dysfunction  

E-print Network

with the bovine temperament model have yet to be determined, QTL analysis of temperament traits in beef and dairy cattle continues to point to aspects of the HPA, including: dopamine metabolism (Hiendleder et al., 2003) and the dopamine receptor (Guti?rrez- Gil...

Curley, Kevin

2012-07-16

110

An epidemiological and economic simulation model to evaluate the spread and control of infectious bovine rhinotracheitis in the Netherlands  

Microsoft Academic Search

Bovine herpesvirus type I (BHV1), causing infectious bovine rhinotracheitis (IBR), was introduced in the Netherlands in 1971. In 1993, about 42% of the dairy cows had antibodies against BHV1. In the future, stricter requirements are anticipated regarding the health status of exported breeding cows and material. To support policymakers in their decisions on IBR eradication, a simulation model was developed

A. Vonk Noordegraaf; J. A. A. M. Buijtels; A. A. Dijkhuizen; P. Franken; J. A. Stegeman; J. Verhoeff

1998-01-01

111

Location of neutralizing epitopes on the G protein of bovine ephemeral fever rhabdovirus.  

PubMed

The surface glycoprotein G is the major neutralizing and protective antigen of bovine ephemeral fever rhabdovirus (BEFV). Twelve neutralizing MAbs against BEFV strain BB7721 were used to select 33 neutralization escape mutants. The mutants had been classified previously into three major antigenic sites (G1-G3) based on their cross-neutralization patterns. The nucleotide sequence of the entire extracellular domain of the G protein gene was determined for all mutants. Each contained a single nucleotide change leading to a single amino acid substitution. The 16 mutants assigned to the linear antigenic site G1 mapped to aa 487-503 of the 623 aa G protein. Results of antibody binding to several overlapping octapeptides covering this region mapped the sequence of two common minimal B cell epitopes recognized by the five G1 MAbs to (488)EEDE(491) and (499)NPHE(502). Site G2 mutations mapped either at aa 169 or 187. The 12 mutants representing antigenic site G3 (G3a and G3b) mapped to aa 49, 57, 218, 229 and 265, indicating that this site is likely to combine complex discontinuous epitopes. Comparison of the deduced amino acid sequence from five BEFV field isolates and BB7721 identified aa 218 to be critical for the site G3a neutralization. Alignment of the glycoproteins of rabies virus, vesicular stomatitis Indiana virus, vesicular stomatitis New Jersey virus, infectious haematopoietic necrosis virus and BEFV revealed similarities in the location of the neutralizing epitopes and extensive conservation of cysteine residues, suggesting that basic elements of the folded structure of these glycoproteins are preserved. PMID:9820132

Kongsuwan, K; Cybinski, D H; Cooper, J; Walker, P J

1998-11-01

112

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

PubMed Central

Background In 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 that a mammary stem cell population can be functionally identified and isolated from the bovine mammary gland. We also demonstrate that this stem cell population may be a promising target for manipulating the composition of cow's milk using gene transfer. Methods and Findings We show that the in vitro colony-forming cell assay for detecting normal primitive bipotent and lineage-restricted human mammary clonogenic progenitors are applicable to bovine mammary cells. Similarly, the ability of normal human mammary stem cells to regenerate functional bilayered structures in collagen gels placed under the kidney capsule of immunodeficient mice is shared by a subset of bovine mammary cells that lack aldehyde dehydrogenase activity. We also find that this activity is a distinguishing feature of luminal-restricted bovine progenitors. The regenerated structures recapitulate the organization of bovine mammary tissue, and milk could be readily detected in these structures when they were assessed by immunohistochemical analysis. Transplantation of the bovine cells transduced with a lentivirus encoding human ?-CASEIN led to expression of the transgene and secretion of the product by their progeny regenerated in vivo. Conclusions These findings point to a common developmental hierarchy shared by human and bovine mammary glands, providing strong evidence of common mechanisms regulating the maintenance and differentiation of mammary stem cells from both species. These results highlight the potential of novel engineering and transplant strategies for a variety of commercial applications including the production of modified milk components for human consumption. PMID:20976049

Martignani, Eugenio; Eirew, Peter; Accornero, Paolo

2010-01-01

113

Small-angle neutron scattering study of differences in phase behavior of silica nanoparticles in the presence of lysozyme and bovine serum albumin proteins  

NASA Astrophysics Data System (ADS)

The differences in phase behavior of anionic silica nanoparticles (88 Å) in the presence of two globular proteins [cationic lysozyme (molecular weight (MW) 14.7 kD) and anionic bovine serum albumin (BSA) (MW 66.4 kD)] have been studied by small-angle neutron scattering. The measurements were carried out on a fixed concentration (1 wt %) of Ludox silica nanoparticles with varying concentrations of proteins (0-5 wt %) at pH = 7. It is found that, despite having different natures (opposite charges), both proteins can render to the same kind of aggregation of silica nanoparticles. However, the concentration regions over which the aggregation is observed are widely different for the two proteins. Lysozyme with very small amounts (e.g., 0.01 wt %) leads to the aggregation of silica nanoparticles. On the other hand, silica nanoparticles coexist with BSA as independent entities at low protein concentrations and turn to aggregates at high protein concentrations (>1 wt %). In the case of lysozyme, the charge neutralization by the protein on the nanoparticles gives rise to the protein-mediated aggregation of the nanoparticles. The nanoparticle aggregates coexist with unaggregated nanoparticles at low protein concentrations, whereas, they coexist with a free protein at higher protein concentrations. For BSA, the nonadsorbing nature of the protein produces the depletion force that causes the aggregation of the nanoparticles at higher protein concentrations. The evolution of the interaction is modeled by the two Yukawa potential, taking account of both attractive and repulsive terms of the interaction in these systems. The nanoparticle aggregation is found to be governed by the short-range attraction for lysozyme and the long-range attraction for BSA. The aggregates are characterized by the diffusion limited aggregate type of mass fractal morphology.

Yadav, Indresh; Kumar, Sugam; Aswal, V. K.; Kohlbrecher, J.

2014-03-01

114

Finding biomarkers in non-model species: literature mining of transcription factors involved in bovine embryo development  

PubMed Central

Background Since processes in well-known model organisms have specific features different from those in Bos taurus, the organism under study, a good way to describe gene regulation in ruminant embryos would be a species-specific consideration of closely related species to cattle, sheep and pig. However, as highlighted by a recent report, gene dictionaries in pig are smaller than in cattle, bringing a risk to reduce the gene resources to be mined (and so for sheep dictionaries). Bioinformatics approaches that allow an integration of available information on gene function in model organisms, taking into account their specificity, are thus needed. Besides these closely related and biologically relevant species, there is indeed much more knowledge of (i) trophoblast proliferation and differentiation or (ii) embryogenesis in human and mouse species, which provides opportunities for reconstructing proliferation and/or differentiation processes in other mammalian embryos, including ruminants. The necessary knowledge can be obtained partly from (i) stem cell or cancer research to supply useful information on molecular agents or molecular interactions at work in cell proliferation and (ii) mouse embryogenesis to supply useful information on embryo differentiation. However, the total number of publications for all these topics and species is great and their manual processing would be tedious and time consuming. This is why we used text mining for automated text analysis and automated knowledge extraction. To evaluate the quality of this “mining”, we took advantage of studies that reported gene expression profiles during the elongation of bovine embryos and defined a list of transcription factors (or TF, n?=?64) that we used as biological “gold standard”. When successful, the “mining” approach would identify them all, as well as novel ones. Methods To gain knowledge on molecular-genetic regulations in a non model organism, we offer an approach based on literature-mining and score arrangement of data from model organisms. This approach was applied to identify novel transcription factors during bovine blastocyst elongation, a process that is not observed in rodents and primates. As a result, searching through human and mouse corpuses, we identified numerous bovine homologs, among which 11 to 14% of transcription factors including the gold standard TF as well as novel TF potentially important to gene regulation in ruminant embryo development. The scripts of the workflow are written in Perl and available on demand. They require data input coming from all various databases for any kind of biological issue once the data has been prepared according to keywords for the studied topic and species; we can provide data sample to illustrate the use and functionality of the workflow. Results To do so, we created a workflow that allowed the pipeline processing of literature data and biological data, extracted from Web of Science (WoS) or PubMed but also from Gene Expression Omnibus (GEO), Gene Ontology (GO), Uniprot, HomoloGene, TcoF-DB and TFe (TF encyclopedia). First, the human and mouse homologs of the bovine proteins were selected, filtered by text corpora and arranged by score functions. The score functions were based on the gene name frequencies in corpora. Then, transcription factors were identified using TcoF-DB and double-checked using TFe to characterise TF groups and families. Thus, among a search space of 18,670 bovine homologs, 489 were identified as transcription factors. Among them, 243 were absent from the high-throughput data available at the time of the study. They thus stand so far for putative TF acting during bovine embryo elongation, but might be retrieved from a recent RNA sequencing dataset (Mamo et al. , 2012). Beyond the 246 TF that appeared expressed in bovine elongating tissues, we restricted our interpretation to those occurring within a list of 50 top-ranked genes. Among the transcription factors identified therein, half belonged to the gold standard (ASCL2, c-F

2012-01-01

115

Cholesterol modulates ligand binding and G-protein coupling to serotonin(1A) receptors from bovine hippocampus.  

PubMed

The serotonin(1A) (5-HT(1A)) receptor is an important member of the superfamily of seven-transmembrane domain G-protein-coupled receptors. We have examined the modulatory role of cholesterol on the ligand binding activity and G-protein coupling of the bovine hippocampal 5-HT(1A) receptor by depleting cholesterol from native membranes using methyl-beta-cyclodextrin (MbetaCD). Removal of cholesterol from bovine hippocampal membranes using varying concentrations of MbetaCD results in a concentration-dependent reduction in specific binding of the agonist 8-OH-DPAT to 5-HT(1A) receptors. This is accompanied by alterations in binding affinity and sites obtained from analysis of binding data. Importantly, cholesterol depletion affected G-protein-coupling of the receptor as monitored by the GTP-gamma-S assay. The concomitant changes in membrane order were reported by changes in fluorescence polarization of membrane probes such as DPH and TMA-DPH, which are incorporated at different locations (depths) in the membrane. Replenishment of membranes with cholesterol led to recovery of ligand binding activity as well as membrane order to a considerable extent. Our results provide evidence, for the first time, that cholesterol is necessary for ligand binding and G-protein coupling of this important neurotransmitter receptor. These results could have significant implications in understanding the influence of the membrane lipid environment on the activity and signal transduction of other G-protein-coupled transmembrane receptors. PMID:15157621

Pucadyil, Thomas J; Chattopadhyay, Amitabha

2004-05-27

116

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

Microsoft Academic Search

\\u000a Tryptone (pancreatic digest of casein) is a common ingredient in laboratory and fermentation media for growing wild-type and\\u000a genetically modified microorganisms. Many of the commercially manufactured products such as human growth hormone, antibiotics,\\u000a insulin, etc. are produced by recombinant strains grown on materials derived from bovine sources. With the emergence of Bovine\\u000a Spongiform Encephalopathy (BSE) and the consequent increase in

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

2010-01-01

117

Osmotically unresponsive water fraction on proteins: non-ideal osmotic pressure of bovine serum albumin as a function of pH and salt concentration.  

PubMed

How much does protein-associated water differ in colligative properties (freezing point, boiling point, vapor pressure and osmotic behavior) from pure bulk water? This question was approached by studying the globular protein bovine serum albumin (BSA), using changes in pH and salt concentration to alter its native structural conformation and state of aggregation. BSA osmotic pressure was investigated experimentally and analyzed using the molecular model of Fullerton et al. [Biochem Cell Biol 1992;70(12):1325]. Analysis yielded both the extent of osmotically unresponsive water (OUW) and the effective molecular weight values of the membrane-impermeable BSA solute. Manipulation of BSA conformation and aggregation by membrane-penetrating cosolutes show that alterations in pH and salt concentration change the amount of bulk water that escapes into BSA from a minimum of 1.4 to a maximum of 11.7 g water per g dry mass BSA. PMID:16376113

Fullerton, Gary D; Kanal, Kalpana M; Cameron, Ivan L

2006-01-01

118

Adsorption of bovine serum albumin onto hydroxyapatite  

Microsoft Academic Search

The adsorption of bovine serum albumin (BSA) onto hydroxyapatite (HA) has been studied as a function of protein concentration, pH and ionic strength. Isotherm data (adsorption being a reversible process) have been analysed using the Langmuir model, the adsorption parameters AT (maximum amount of protein adsorbed, mg m?2) and K (affinity constant, L g?1) being calculated for each solution condition

Diana T. Hughes Wassell; Rachel C. Hall; Graham Embery

1995-01-01

119

Comparative Protein Structure Modeling Using Modeller  

PubMed Central

Functional characterization of a protein sequence is one of the most frequent problems in biology. This task is usually facilitated by accurate three-dimensional (3-D) structure of the studied protein. In the absence of an experimentally determined structure, comparative or homology modeling can sometimes provide a useful 3-D model for a protein that is related to at least one known protein structure. Comparative modeling predicts the 3-D structure of a given protein sequence (target) based primarily on its alignment to one or more proteins of known structure (templates). The prediction process consists of fold assignment, target-template alignment, model building, and model evaluation. This unit describes how to calculate comparative models using the program MODELLER and discusses all four steps of comparative modeling, frequently observed errors, and some applications. Modeling lactate dehydrogenase from Trichomonas vaginalis (TvLDH) is described as an example. The download and installation of the MODELLER software is also described. PMID:18428767

Eswar, Narayanan; Marti-Renom, Marc A.; Madhusudhan, M.S.; Eramian, David; Shen, Min-yi; Pieper, Ursula

2014-01-01

120

Evaluation of three serum i-ELISAs using monoclonal antibodies and protein G as peroxidase conjugate for the diagnosis of bovine brucellosis  

Microsoft Academic Search

Three i-ELISAs using LPS, the immunodominant component of Brucella abortus, were developed with three different conjugates: monoclonal antibodies 1C8 (anti-bovine IgG1) and 3H3 (mainly specific for bovine IgG2 but also reacting with IgG1) and protein G (reacts with both bovine IgG subclasses). Using a cut-off value of 2.5U\\/ml, the i-ELISA with 3H3 as conjugate had a specificity (95% CI: 98.32–99.63%)

C Saegerman; L De Waele; D Gilson; J Godfroid; P Thiange; P Michel; B Limbourg; T. K.-O Vo; J Limet; J.-J Letesson; D Berkvens

2004-01-01

121

Phorbol ester receptors in bovine luteal cells: relationship to protein kinase C.  

PubMed

We investigated the binding kinetics of the tumor-promoting phorbol ester, phorbol-12,13-dibutyrate (PBt2) to dispersed total bovine luteal cells, purified small luteal cells, and purified luteal protein kinase C (PKC). Saturation analysis and competitive displacement techniques were used. Binding of [3H]PBt2 to total luteal cell preparations resulted in two distinct affinities. The high affinity component was characterized by a Kd of 4.5 +/- 1.5 nM. Analysis of [3H]PBt2 binding to total cells using competitive displacement demonstrated that the low affinity binding was specific and displaceable but dependent on concentrations of [3H]PBt2 far above the Kd for the high affinity binding. In contrast to the total cell preparations, only high affinity binding was observed in intact purified small luteal cells (Kd = 0.96 +/- 0.04 nM). Partial purification of luteal cytosolic PKC by DEAE-Sephadex chromatography resulted in co-elution of PKC enzyme activity and the [3H]PBt2 binding activity. Under conditions of saturating calcium (0.1 mM) and phosphatidylserine (PS) (100 micrograms/tube) concentrations, binding to the partially purified PKC preparation was found to be of a single high affinity and exhibited a Kd (1.3 +/- 0.2 nM) similar to the high affinity binding observed in intact cells. These results suggest that the primary phorbol ester receptor in luteal cells is PKC. However, a low affinity, high capacity [3H]PBt2 binding site also exists within the corpus luteum, either in the large cells or in the accessory cell fraction which consists mainly of endothelial cells. PMID:2328828

Dowd, J P; Alila, H W; Hansel, W

1990-03-01

122

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 Å 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 Å CRMS from the crystal structure for the ligand atoms. This predicted conformation has the carbonyl O only 2.82 Å from the N of Lys-296. Making this Schiff base bond and minimizing leads to a final conformation only 0.62 Å 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 Å from the N of Lys-296. Making this Schiff base bond and minimizing leads to a final conformation only 2.92 Å 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. PMID:15041637

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

2004-01-01

123

A Bovine Model of Respiratory Chlamydia psittaci Infection: Challenge Dose Titration  

PubMed Central

This study aimed to establish and evaluate a bovine respiratory model of experimentally induced acute C. psittaci infection. Calves are natural hosts and pathogenesis may resemble the situation in humans. Intrabronchial inoculation of C. psittaci strain DC15 was performed in calves aged 2–3 months via bronchoscope at four different challenge doses from 106 to 109 inclusion-forming units (ifu) per animal. Control groups received either UV-inactivated C. psittaci or cell culture medium. While 106 ifu/calf resulted in a mild respiratory infection only, the doses of 107 and 108 induced fever, tachypnea, dry cough, and tachycardia that became apparent 2–3 days post inoculation (dpi) and lasted for about one week. In calves exposed to 109 ifu C. psittaci, the respiratory disease was accompanied by severe systemic illness (apathy, tremor, markedly reduced appetite). At the time point of most pronounced clinical signs (3 dpi) the extent of lung lesions was below 10% of pulmonary tissue in calves inoculated with 106 and 107 ifu, about 15% in calves inoculated with 108 and more than 30% in calves inoculated with 109 ifu C. psittaci. Beside clinical signs and pathologic lesions, the bacterial load of lung tissue and markers of pulmonary inflammation (i.e., cell counts, concentration of proteins and eicosanoids in broncho-alveolar lavage fluid) were positively associated with ifu of viable C. psittaci. While any effect of endotoxin has been ruled out, all effects could be attributed to infection by the replicating bacteria. In conclusion, the calf represents a suitable model of respiratory chlamydial infection. Dose titration revealed that both clinically latent and clinically manifest infection can be reproduced experimentally by either 106 or 108 ifu/calf of C. psittaci DC15 while doses above 108 ifu C. psittaci cannot be recommended for further studies for ethical reasons. This defined model of different clinical expressions of chlamydial infection allows studying host-pathogen interactions. PMID:22299031

Reinhold, Petra; Ostermann, Carola; Liebler-Tenorio, Elisabeth; Berndt, Angela; Vogel, Anette; Lambertz, Jacqueline; Rothe, Michael; Ruttger, Anke; Schubert, Evelyn; Sachse, Konrad

2012-01-01

124

Involvement of the cyclic AMP-responsive element binding protein in bovine leukemia virus expression in vivo.  

PubMed Central

The TAR element (Tax-responsive element; also called TxRE) is a major determinant of the regulation of bovine leukemia virus (BLV) expression. In order to gain insight into the mechanisms of viral expression, complexes formed between proteins and the TAR enhancer DNA were analyzed by gel retardation assays. We report here that nuclear lysates from ex vivo-isolated B lymphocytes contain proteins that specifically bind to TAR. An antibody directed toward the cyclic AMP-responsive element binding (CREB) protein supershifted a complex (C1) present only in BLV-infected B lymphocytes. The CREB protein thus appears to be a major transcription factor involved in BLV expression in vivo. Images PMID:8057465

Adam, E; Kerkhofs, P; Mammerickx, M; Kettmann, R; Burny, A; Droogmans, L; Willems, L

1994-01-01

125

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

126

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

127

Experimental ruminant models for bovine neosporosis: what is known and what is needed.  

PubMed

At present, bovine neosporosis is an important worldwide concern because of its wide geographic distribution and economic impact. Abortion is the main clinical sign of bovine neosporosis in both dairy and beef cattle. Ruminant challenge models are critical to evaluate potential vaccine candidates to help tackle bovine neosporosis and to study pathogenesis and host responses to infection. Several research groups have developed ruminant models of Neospora caninum infection independently of others, resulting in a high degree of variability due to the use of different species of animals, breeds, strains/isolates of N. caninum, doses, routes and times of inoculation. Standardization is greatly needed to advance research in a more collaborative, timely and efficient manner. In the absence of widely accepted international guidelines, this manuscript serves to summarize and discuss the different models and parameters currently in use. Parameters essential for the development of non-pregnant and pregnant ruminant models are outlined and the main knowledge gaps are identified. This information could act as the basis to develop a consensus for international standard guidelines for ruminant models of neosporosis that would be helpful for researchers in this field worldwide. PMID:24926962

Benavides, Julio; Collantes-Fernández, Esther; Ferre, Ignacio; Pérez, Valentín; Campero, Carlos; Mota, Rinaldo; Innes, Elisabeth; Ortega-Mora, Luis M

2014-09-01

128

In Vitro Digestion of Proteins and Growth Factors in a Bovine Whey Protein Extract as Determined Using a Computer-Controlled Dynamic Gastrointestinal System (TIM1)  

Microsoft Academic Search

The digestion of major whey proteins\\/peptides, transforming growth factor-beta2 (TGF-?2) and insulin-like growth factor-I\\u000a (IGF-I) in a bovine whey protein extract (BWPE) was investigated in vitro using a dynamic gastrointestinal digestion system\\u000a (TIM-1). ?-lactoglobulin and glycomacropeptide were the whey components most resistant to 3 h of gastric digestion (pepsin)\\u000a and were also the last to be cleared from the gastric compartment.

Samira Nabil; Sylvie F. Gauthier; Réjean Drouin; Patrice E. Poubelle; Yves Pouliot

129

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

130

Role of Alpha/Beta Interferons in the Attenuation and Immunogenicity of Recombinant Bovine Respiratory Syncytial Viruses Lacking NS Proteins  

PubMed Central

Alpha/beta interferons (IFN-?/?) are not only a powerful first line of defense against pathogens but also have potent immunomodulatory activities. Many viruses have developed mechanisms of subverting the IFN system to enhance their virulence. Previous studies have demonstrated that the nonstructural (NS) genes of bovine respiratory syncytial virus (BRSV) counteract the antiviral effects of IFN-?/?. Here we demonstrate that, in contrast to wild-type BRSVs, recombinant BRSVs (rBRSVs) lacking the NS proteins, and those lacking NS2 in particular, are strong inducers of IFN-?/? in bovine nasal fibroblasts and bronchoalveolar macrophages. Furthermore, whereas the NS deletion mutants replicated to wild-type rBRSV levels in cells lacking a functional IFN-?/? system, their replication was severely attenuated in IFN-competent cells and in young calves. These results suggest that the NS proteins block the induction of IFN-?/? gene expression and thereby increase the virulence of BRSV. Despite their poor replication in the respiratory tract of young calves, prior infection with virus lacking either the NS1 or the NS2 protein induced serum antibodies and protection against challenge with virulent BRSV. The greater level of protection induced by the NS2, than by the NS1, deletion mutant, was associated with higher BRSV-specific antibody titers and greater priming of BRSV-specific, IFN-?-producing CD4+ T cells. Since there were no detectable differences in the ability of these mutants to replicate in the bovine respiratory tract, the greater immunogenicity of the NS2 deletion mutant may be associated with the greater ability of this virus to induce IFN-?/?. PMID:12857912

Valarcher, Jean-Francois; Furze, Julie; Wyld, Sara; Cook, Roy; Conzelmann, Karl-Klaus; Taylor, Geraldine

2003-01-01

131

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

132

Effective vaccination of cattle using the virion G protein of bovine ephemeral fever virus as an antigen.  

PubMed

In a series of experiments, the envelope glycoprotein (G protein) of bovine ephemeral fever virus (BEFV) induced immunity against challenge with virulent virus. Protection correlated with the level of specific serum antibodies to G protein measured by a blocking ELISA test and with the level of neutralizing antibody. The optimum vaccination regimen consisted of two injections given 21 days apart at a dose rate of 0.32 microgram per cow of purified G protein emulsified in the adjuvant Quil A. This schedule conferred immunity for the duration of the preliminary experiment (46 days). Immunity to severe disease, but not to infection, remained for at least 12 months after vaccination, although BEFV could not be reisolated from vaccinated cattle following challenge. Unvaccinated cattle used as controls exhibited typical signs of clinical ephemeral fever and BEFV was recovered from all control animals following challenge. PMID:7975863

Uren, M F; Walker, P J; Zakrzewski, H; St George, T D; Byrne, K A

1994-07-01

133

Fetal bovine serum xenoproteins modulate human monocyte adhesion and protein release on biomaterials in vitro  

Microsoft Academic Search

Monocyte-derived macrophages are critical in the host–foreign body response to biomaterials and have been studied extensively in various culture conditions in vitro, such as medium supplemented with fetal bovine serum (FBS) or autologous human serum (AHS). Since monocyte maturation into macrophages is highly plastic and may vary considerably depending on the surface, isolation procedures and in vitro culture conditions, we

David Schmidt; Evan James Joyce; Weiyuan John Kao

2011-01-01

134

Oncoviral Bovine Leukemia Virus G4 and Human T-Cell Leukemia Virus Type 1 p13II Accessory Proteins Interact with Farnesyl Pyrophosphate Synthetase  

PubMed Central

G4 and p13II are accessory proteins encoded by the X region of bovine leukemia virus and human T-cell leukemia virus type 1 (HTLV-1), respectively. Disruption of the G4 and p13II open reading frames interferes with viral spread in animal model systems, indicating that the corresponding proteins play a key role in viral replication. In addition, G4 is oncogenic in primary cell cultures and is absolutely required for efficient onset of leukemogenesis in sheep. To gain insight into the function of these proteins, we utilized the yeast two-hybrid system to identify protein partners of G4. Results revealed that G4 interacts with farnesyl pyrophosphate synthetase (FPPS), a protein involved in the mevalonate/squalene pathway and in synthesis of FPP, a substrate required for prenylation of Ras. The specificity of the interaction was verified by glutathione S-transferase (GST) pull-down assays and by coimmunoprecipitation experiments. Furthermore, confocal microscopy showed that the subcellular localization of G4 was profoundly affected by FPPS. The G4 protein itself was not prenylated, at least in rabbit reticulocyte lysate-based assays. The domain of G4 required for binding to FPPS was restricted to an amphipathic ?-helix rich in arginine residues. Subtle mutation of this ?-helix abrogated G4 oncogenic potential in vitro, providing a biological relevance for FPPS-G4 complex formation in cells. Finally, HTLV-1 p13II was also found to specifically interact with FPPS (in yeast as well as in GST pull-down assays) and to colocalize with G4 in mitochondria, suggesting a functional analogy between these oncoviral accessory proteins. Identification of FPPS as a molecular partner for p13II and G4 accessory proteins opens new prospects for treatment of retrovirus-induced leukemia. PMID:11773414

Lefebvre, Laurent; Vanderplasschen, Alain; Ciminale, Vincenzo; Heremans, Hubertine; Dangoisse, Olivier; Jauniaux, Jean-Claude; Toussaint, Jean-Francois; Zelnik, Vlado; Burny, Arsene; Kettmann, Richard; Willems, Luc

2002-01-01

135

Empirical solvation models can be used to differentiate native from near-native conformations of bovine pancreatic trypsin inhibitor.  

PubMed

Several hydration models for peptides and proteins based on solvent accessible surface area have been proposed previously. We have evaluated some of these models as well as four new ones in the context of near-native conformations of a protein. In addition, we propose an empirical site-site distance-dependent correction that can be used in conjunction with any of these models. The set of near-native structures consisted of 39 conformations of bovine pancreatic trypsin inhibitor (BPTI) each of which was a local minimum of an empirical energy function (ECEPP) in the absence of solvent. Root-mean-square (rms) deviations from the crystallographically determined structure were in the following ranges: 1.06-1.94 A for all heavy atoms, 0.77-1.36 A for all backbone heavy atoms, 0.68-1.33 A for all alpha-carbon atoms, and 1.41-2.72 A for all side-chain heavy atoms. We have found that there is considerable variation among the solvent models when evaluated in terms of concordance between the solvation free energy and the rms deviations from the crystallographically determined conformation. The solvation model for which the best concordance (0.939) with the rms deviations of the C alpha atoms was found was derived from NMR coupling constants of peptides in water combined with an exponential site-site distance dependence of the potential of mean force. Our results indicate that solvation free energy parameters derived from nonpeptide free energies of hydration may not be transferrable to peptides. Parameters derived from peptide and protein data may be more applicable to conformational analysis of proteins. A general approach to derive parameters for free energy of hydration from ensemble-averaged properties of peptides in solution is described. PMID:1715564

Vila, J; Williams, R L; Vásquez, M; Scheraga, H A

1991-01-01

136

Do-it-yourself histidine-tagged bovine enterokinase: A handy member of the protein engineer's toolbox?  

PubMed Central

Enterokinase, a two-chain duodenal serine protease, activates trypsinogen by removing its N-terminal propeptide. Due to a clean cut after the non-primed site recognition sequence, the enterokinase light chain is frequently employed in biotechnology to separate N-terminal affinity tags from target proteins with authentic N-termini. In order to obtain large quantities of this protease, we adapted an in vitro folding protocol for a pentahistidine-tagged triple mutant of the bovine enterokinase light chain. The purified, highly active enzyme successfully processed recombinant target proteins, while the pentahistidine-tag facilitated post-cleavage removal. Hence, we conclude that producing enterokinase in one's own laboratory is an efficient alternative to the commercial enzyme. PMID:24184090

Skala, Wolfgang; Goettig, Peter; Brandstetter, Hans

2013-01-01

137

Do-it-yourself histidine-tagged bovine enterokinase: a handy member of the protein engineer's toolbox.  

PubMed

Enterokinase, a two-chain duodenal serine protease, activates trypsinogen by removing its N-terminal propeptide. Due to a clean cut after the non-primed site recognition sequence, the enterokinase light chain is frequently employed in biotechnology to separate N-terminal affinity tags from target proteins with authentic N-termini. In order to obtain large quantities of this protease, we adapted an in vitro folding protocol for a pentahistidine-tagged triple mutant of the bovine enterokinase light chain. The purified, highly active enzyme successfully processed recombinant target proteins, while the pentahistidine-tag facilitated post-cleavage removal. Hence, we conclude that producing enterokinase in one's own laboratory is an efficient alternative to the commercial enzyme. PMID:24184090

Skala, Wolfgang; Goettig, Peter; Brandstetter, Hans

2013-12-01

138

Mapping of nuclear import signal and importin {alpha}3 binding regions of 52K protein of bovine adenovirus-3  

SciTech Connect

The L1 region of bovine adenovirus (BAdV)-3 encodes a non-structural protein designated 52K. Anti-52K serum detected a protein of 40 kDa, which localized to the nucleus but not to the nucleolus in BAdV-3-infected or transfected cells. Analysis of mutant 52K proteins suggested that three basic residues ({sup 105}RKR{sup 107}) of the identified domain (amino acids {sup 102}GMPRKRVLT{sup 110}) are essential for nuclear localization of 52K. The nuclear import of a GST-52K fusion protein utilizes the classical importin {alpha}/{beta}-dependent nuclear transport pathway. The 52K protein is preferentially bound to the cellular nuclear import receptor importin {alpha}3. Although deletion of amino acid 102-110 is sufficient to abrogate the nuclear localization of 52K, amino acid 90-133 are required for interaction with importin-{alpha}3 and localizing a cytoplasmic protein to the nucleus. These results suggest that 52K contains a bipartite NLS, which preferentially utilize an importin {alpha}3 nuclear import receptor-mediated pathway to transport 52K to the nucleus.

Paterson, Carolyn P.; Ayalew, Lisanework E. [Vaccine and Infectious Disease Organization-International Vaccine Center (VIDO-InterVac), University of Saskatchewan, Saskatoon, SK S7N 5E3 Canada (Canada) [Vaccine and Infectious Disease Organization-International Vaccine Center (VIDO-InterVac), University of Saskatchewan, Saskatoon, SK S7N 5E3 Canada (Canada); Veterinary Microbiology, University of Saskatchewan, Saskatoon, SK S7N 5E3 S7N 5B4 Canada (Canada); Tikoo, Suresh K., E-mail: suresh.tik@usask.ca [Vaccine and Infectious Disease Organization-International Vaccine Center (VIDO-InterVac), University of Saskatchewan, Saskatoon, SK S7N 5E3 Canada (Canada); Veterinary Microbiology, University of Saskatchewan, Saskatoon, SK S7N 5E3 S7N 5B4 Canada (Canada); School of Public Health, University of Saskatchewan, Saskatoon, SK S7N 5E5 Canada (Canada)

2012-10-10

139

Subunit rearrangement accompanies sequence-specific DNA binding by the bovine papillomavirus-1 E2 protein 1 1 Edited by P. E. Wright  

Microsoft Academic Search

The 2.5 Å crystal structures of the DNA-binding domain of the E2 protein from bovine papillomavirus strain 1 and its complex with DNA are presented. E2 is a transcriptional regulatory protein that is also involved in viral DNA replication. It is the structural prototype for a novel class of DNA-binding proteins: dimeric ?-barrels with surface ?-helices that serve as recognition

Rashmi S Hegde; Ai-Fei Wang; Seung-Sup Kim; Matthieu Schapira

1998-01-01

140

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. PMID:22691138

2012-01-01

141

Location of neutralizing epitopes on the G protein of bovine ephemeral fever rhabdovirus  

Microsoft Academic Search

The surface glycoprotein G is the major neutralizing and protective antigen of bovine ephemeral fever rhabdovirus (BEFV). Twelve neutralizing MAbs against BEFV strain BB7721 were used to select 33 neutralization escape mutants. The mutants had been classified previously into three major antigenic sites (G1-G3) based on their cross-neutralization patterns. The nucleotide sequence of the entire extracellular domain of the G

Kritaya Kongsuwan; Daisy H. Cybinski; Juan Cooper; Peter J. Walker

1998-01-01

142

Cytosolic calcium homeostasis in bovine parathyroid cells and its modulation by protein kinase C.  

PubMed

1. The effects of protein kinase C (PKC) activators and inhibitors on the mechanisms regulating cytosolic Ca2+ homeostasis in dissociated bovine parathyroid cells loaded with fura-2 were examined. 2. Stepwise increases in the concentration of extracellular Ca2+ (from 0.5 to 2 or 3 mM) elicited transient followed by sustained increases in the concentration of intracellular free Ca2+ ([Ca2+]i). Cytosolic Ca2+ transients reflected the mobilization of intracellular Ca2+ and influx of extracellular Ca2+ whereas sustained increases in [Ca2+]i resulted from the influx of extracellular Ca2+. Brief (1-2 min) pretreatment with phorbol myristate acetate (PMA) shifted the concentration-response curve for extracellular Ca(2+)-induced cytosolic Ca2+ transients to the right without affecting the maximal response. Cytosolic Ca2+ transients elicited by extracellular Mg2+ were similarly affected by PMA. 3. These effects of PMA were mimicked by various other activators of PKC with the rank order of potency PMA > phorbol dibutyrate > bryostatin , > (-)indolactam V > mezerein. Isomers or analogues of these compounds that do not alter PKC activity (4 alpha-phorbols and (+)indolactam V) did not alter [Ca2+]i. 4. PKC activators depressed evoked increases in [Ca2+]i when influx of extracellular Ca2+ was blocked with Gd3+. Cytosolic Ca2+ transients elicited by extracellular Mg2+ in the absence of extracellular Ca2+ were similarly inhibited by PKC activators. Activation of PKC thus inhibits the mobilization of intracellular Ca2+ elicited by extracellular divalent cations. 5. Increases in the concentration of extracellular Ca2+ caused corresponding increases in the formation of [3H]inositol 1,4,5-trisphosphate ([3H]InsP3). Pretreatment with PMA shifted the concentration-response curve for extracellular Ca(2+)-induced [3H]InsP3 formation to the right without affecting the maximal response. 6. PKC activators also caused some depression of steady-state increases in [Ca2+]i elicited by extracellular Ca2+. In contrast, PMA did not affect increases in [Ca2+]i elicited by ionomycin or thapsigargin. 7. Ba2+ was used to monitor divalent cation influx. PMA decreased the rate of rise of the fluorescent signal elicited by extracellular Ba2+. 8. All these effects of PKC activators on [Ca2+]i were blocked or reversed by staurosporine at concentrations (30-100 nM) that inhibited PKC activity in parathyroid cells. Staurosporine alone potentiated cytosolic Ca2+ responses evoked by submaximal concentrations of extracellular divalent cations. 9. PKC thus depresses both the mobilization of intracellular Ca2+ and the influx of extracellular Ca2+ in parathyroid cells. The effects on [Ca2+]i provide evidence for a Ca2+ receptor on the surface of parathyroid cells that uses transmembrane signalling mechanisms common to some other Ca(2+)-mobilizing receptors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:8254504

Racke, F K; Nemeth, E F

1993-08-01

143

Cytosolic calcium homeostasis in bovine parathyroid cells and its modulation by protein kinase C.  

PubMed Central

1. The effects of protein kinase C (PKC) activators and inhibitors on the mechanisms regulating cytosolic Ca2+ homeostasis in dissociated bovine parathyroid cells loaded with fura-2 were examined. 2. Stepwise increases in the concentration of extracellular Ca2+ (from 0.5 to 2 or 3 mM) elicited transient followed by sustained increases in the concentration of intracellular free Ca2+ ([Ca2+]i). Cytosolic Ca2+ transients reflected the mobilization of intracellular Ca2+ and influx of extracellular Ca2+ whereas sustained increases in [Ca2+]i resulted from the influx of extracellular Ca2+. Brief (1-2 min) pretreatment with phorbol myristate acetate (PMA) shifted the concentration-response curve for extracellular Ca(2+)-induced cytosolic Ca2+ transients to the right without affecting the maximal response. Cytosolic Ca2+ transients elicited by extracellular Mg2+ were similarly affected by PMA. 3. These effects of PMA were mimicked by various other activators of PKC with the rank order of potency PMA > phorbol dibutyrate > bryostatin , > (-)indolactam V > mezerein. Isomers or analogues of these compounds that do not alter PKC activity (4 alpha-phorbols and (+)indolactam V) did not alter [Ca2+]i. 4. PKC activators depressed evoked increases in [Ca2+]i when influx of extracellular Ca2+ was blocked with Gd3+. Cytosolic Ca2+ transients elicited by extracellular Mg2+ in the absence of extracellular Ca2+ were similarly inhibited by PKC activators. Activation of PKC thus inhibits the mobilization of intracellular Ca2+ elicited by extracellular divalent cations. 5. Increases in the concentration of extracellular Ca2+ caused corresponding increases in the formation of [3H]inositol 1,4,5-trisphosphate ([3H]InsP3). Pretreatment with PMA shifted the concentration-response curve for extracellular Ca(2+)-induced [3H]InsP3 formation to the right without affecting the maximal response. 6. PKC activators also caused some depression of steady-state increases in [Ca2+]i elicited by extracellular Ca2+. In contrast, PMA did not affect increases in [Ca2+]i elicited by ionomycin or thapsigargin. 7. Ba2+ was used to monitor divalent cation influx. PMA decreased the rate of rise of the fluorescent signal elicited by extracellular Ba2+. 8. All these effects of PKC activators on [Ca2+]i were blocked or reversed by staurosporine at concentrations (30-100 nM) that inhibited PKC activity in parathyroid cells. Staurosporine alone potentiated cytosolic Ca2+ responses evoked by submaximal concentrations of extracellular divalent cations. 9. PKC thus depresses both the mobilization of intracellular Ca2+ and the influx of extracellular Ca2+ in parathyroid cells. The effects on [Ca2+]i provide evidence for a Ca2+ receptor on the surface of parathyroid cells that uses transmembrane signalling mechanisms common to some other Ca(2+)-mobilizing receptors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:8254504

Racke, F K; Nemeth, E F

1993-01-01

144

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. PMID:23673274

2013-01-01

145

Expression of a functional influenza viral cap-recognizing protein by using a bovine papilloma virus vector.  

PubMed Central

The gene for the influenza viral PB2 protein, which recognizes and binds the 5'-terminal cap 1 structures (m7GpppNm) on eukaryotic mRNAs, was inserted into a bovine papilloma virus vector under the control of a mouse metallothionein I (MT-I) promoter. After transfection of this vector into mouse NIH 3T3 cells, a cell line, cPB2, was obtained that produces PB2-specific mRNA and authentic PB2 protein. Induction of the MT-I promoter with CdCl2 causes about a 10-fold increase in PB2 mRNA and protein levels. The expressed PB2 protein is functional, as it relieves the block in viral mRNA synthesis exhibited by a temperature-sensitive viral mutant containing a cap-binding defect in the PB2 protein. This demonstrates complementation of a function of a negative-strand RNA virus by a gene product expressed in a cell line from recombinant DNA. Images PMID:2989815

Braam-Markson, J; Jaudon, C; Krug, R M

1985-01-01

146

Agonist-induced conformational changes in bovine rhodopsin: insight into activation of G-protein-coupled receptors.  

PubMed

Activation of G-protein-coupled receptors (GPCRs) is initiated by conformational changes in the transmembrane (TM) helices and the intra- and extracellular loops induced by ligand binding. Understanding the conformational changes in GPCRs leading to activation is imperative in deciphering the role of these receptors in the pathology of diseases. Since the crystal structures of activated GPCRs are not yet available, computational methods and biophysical techniques have been used to predict the structures of GPCR active states. We have recently applied the computational method LITiCon to understand the ligand-induced conformational changes in beta(2)-adrenergic receptor by ligands of varied efficacies. Here we report a study of the conformational changes associated with the activation of bovine rhodopsin for which the crystal structure of the inactive state is known. Starting from the inactive (dark) state, we have predicted the TM conformational changes that are induced by the isomerization of 11-cis retinal to all-trans retinal leading to the fully activated state, metarhodopsin II. The predicted active state of rhodopsin satisfies all of the 30 known experimental distance constraints. The predicted model also correlates well with the experimentally observed conformational switches in rhodopsin and other class A GPCRs, namely, the breaking of the ionic lock between R135(3.50) at the intracellular end of TM3 (part of the DRY motif) and E247(6.30) on TM6, and the rotamer toggle switch on W265(6.48) on TM6. We observe that the toggling of the W265(6.48) rotamer modulates the bend angle of TM6 around the conserved proline. The rotamer toggling is facilitated by the formation of a water wire connecting S298(7.45), W265(6.48) and H211(5.46). As a result, the intracellular ends of TMs 5 and 6 move outward from the protein core, causing large conformational changes at the cytoplasmic interface. The predicted outward movements of TM5 and TM6 are in agreement with the recently published crystal structure of opsin, which is proposed to be close to the active-state structure. In the predicted active state, several residues in the intracellular loops, such as R69, V139(3.54), T229, Q237, Q239, S240, T243 and V250(6.33), become more water exposed compared to the inactive state. These residues may be involved in mediating the conformational signal from the receptor to the G protein. From mutagenesis studies, some of these residues, such as V139(3.54), T229 and V250(6.33), are already implicated in G-protein activation. The predicted active state also leads to the formation of new stabilizing interhelical hydrogen-bond contacts, such as those between W265(6.48) and H211(5.46) and E122(3.37) and C167(4.56). These hydrogen-bond contacts serve as potential conformational switches offering new opportunities for future experimental investigations. The calculated retinal binding energy surface shows that binding of an agonist makes the receptor dynamic and flexible and accessible to many conformations, while binding of an inverse agonist traps the receptor in the inactive state and makes the other conformations inaccessible. PMID:18638482

Bhattacharya, Supriyo; Hall, Spencer E; Vaidehi, Nagarajan

2008-10-01

147

Analyzing models for interactions of aptamers to proteins  

NASA Astrophysics Data System (ADS)

We have devised an experimental and theoretical model, based on fluorescent spectroscopy and molecular modelling, to describe the interaction of aptamer (selected against various protein targets) with proteins and albumins in particular. This model, described in this work, has allowed us to decipher the nature of the interactions between aptamers and albumins, the binding site of the aptamers to albumins, the potential role of primer binding to the albumin and expand to the ability of albumin to carry aptamers in the bloodstream, providing data to better understand the level of free aptamer for target binding. We are presenting the study of a variety of aptamers, including those against the MUC1 tumour marker, heparanase and human kallikrein 6 with bovine and human serum albumins and the effect these interactions may have on the bioavailability of the aptamer for target-specific binding and therapeutic activity.

Silva, Dilson; Missailidis, Sotiris

2014-10-01

148

Glycoinositol phospholipid anchor and protein C-terminus of bovine erythrocyte acetylcholinesterase: analysis by mass spectrometry and by protein and DNA sequencing.  

PubMed Central

Purified bovine erythrocyte acetylcholinesterase (AChE) was radiomethylated on its amine groups and incubated with bacterial phosphatidylinositol-specific phospholipase C to remove the lipid portion of the AChE glycoinositol phospholipid (GPI) anchor, and a C-terminal tryptic fragment that contained the residual GPI glycan was isolated by HPLC. Analysis by electrospray-ionization mass spectrometry revealed a parent ion of m/z 3798. The fragmentation patterns produced by collision-induced dissociation mass spectrometry of the +4 and +5 states of the parent ion indicated a 23-amino acid peptide in amide linkage to ethanolamine-P04-Hex-Hex-Hex(PO4-ethanolamine)(HexNAc)-Hex N(Me)2-inositol phosphate. The glycan structure is completely consistent with that obtained previously for the GPI anchor of human erythrocyte AChE except for the addition of the HexNAc substituent. A nearly complete peptide sequence was deduced from the fragmentation patterns, although four assignments were based only on single fragments of very low abundance. To resolve this uncertainty, a segment of bovine genomic DNA corresponding to the C-terminal AChE sequence was amplified by PCR. DNA sequencing established the 23-amino acid peptide sequence to be FLPKLLSATASEAPCTCSGPAHG, in agreement with the MS data and consistent with results from Edman protein sequencing. Dimerization of AChE polypeptides is mediated by intersubunit disulphide bonding in this C-terminal segment, but the bovine AChE contained two cysteine residues in a ...CTC... motif, in contrast with human AChE which contains only a single cysteine in this segment. Although bovine AChE contained no free thiol groups reactive with iodo[14C]acetamide, partial reduction and alkylation with iodo[14C]acetamide revealed that conversion into monomers occurred with an overall incorporation of only one alkyl group per monomer. An identical level of alkylation was observed when dimeric human AChE was converted into monomers by partial reduction. The question of whether the bovine AChE contains one or two intersubunit disulphide linkages is considered. PMID:8615775

Haas, R; Jackson, B C; Reinhold, B; Foster, J D; Rosenberry, T L

1996-01-01

149

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

150

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. PMID:18513449

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

151

Bovine herpesvirus-1 US3 protein kinase: critical residues and involvement in the phosphorylation of VP22.  

PubMed

The US3 gene product of bovine herpesvirus-1 (BoHV-1) is a protein kinase that is expressed early during infection and capable of autophosphorylation. By examining differentially labelled US3 moieties by co-immunoprecipitation, we demonstrated that the protein kinase interacts with itself in vitro, which supports autophosphorylation by US3. Based on its homology to other serine/threonine protein kinases, we defined two highly conserved lysines in US3, at position 195 within the ATP-binding pocket and at position 282 within the catalytic loop; altering either residue resulted in kinase-dead mutants, demonstrating that these two residues are critical for the catalytic activity of BoHV-1 US3. During immunoprecipitation experiments, US3 interacted weakly with VP22, another tegument protein of BoHV-1. Furthermore, VP22 co-localized with US3 inside the nucleus in BoHV-1-infected cells. In vitro kinase assays demonstrated that VP22 is phosphorylated not only by US3, but also by the cellular casein kinase 2 (CK2) protein. The selective CK2 protein kinase inhibitor, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT) and the less specific CK2 inhibitor Kenpaullone reduced VP22 phosphorylation, while CK1, protein kinase C or protein kinase A inhibitors did not affect phosphorylation. When US3 was included with VP22 in the kinase assay in the presence of DMAT, a low level of VP22 phosphorylation was observed. These data demonstrate that BoHV-1 VP22 interacts with both CK2 and US3, and that CK2 is the major kinase phosphorylating VP22, with US3 playing a minor role. PMID:20016039

Labiuk, Shaunivan L; Lobanov, Vladislav; Lawman, Zoe; Snider, Marlene; Babiuk, Lorne A; van Drunen Littel-van den Hurk, Sylvia

2010-05-01

152

IMMOBILIZATION OF GLUCOSE OXIDASE AND FERROCENE REDOX POLYMER IN CROSS-LINKED POLY (VINYL ALCOHOL) WITH BOVINE SERUM ALBUMIN AS PROTEIN STABILIZER  

Microsoft Academic Search

A method of tethering a mediator to an enzymatic membrane was studied. Glucose oxidase (GOD) and ferrocene redox polymer were immobilized in cross-linked poly (vinyl alcohol) (CLPVA) with bovine serum albumin (BSA) added as a protein stabilizer. Redox hydrogel polyallylamine ferrocene was prepared by cross-linking polyallylamine hydrochloride with glutaraldehyde and attaching the ferrocene covalently. The biosensor response to glucose was

Norhana Jusoh; A Abdul Aziz

153

A pregnant mouse model for bovine Tritrichomonas foetus infection.  

PubMed

The economically important effects of Tritrichomonas foetus infection in cattle are abortion and infertility, yet there has not been an animal model to examine the parasite-host interactions during gestation. In this study, 5- and 7- to 8-week-old BALB/cAnNCr, BALB/cJ, and SCID/NCr mice on a BALB/c background were intravaginally infected with T. foetus. All BALB/cAnNCr and BALB/cJ mice, and 89% of SCID/NCr mice sustained infections for 13 weeks, if inoculated before 5 weeks of age. Infection rates were lower in all mouse strains inoculated at 7 weeks of age, although BALB/cAnNCr mice were significantly more susceptible than BALB/cJ or SCID/NCr mice. Vaginal bacterial flora did not account for the variation in mouse-strain susceptibility, although coagulase-negative staphylococci in vaginal flora were associated with failure of T. foetus to infect. As with infected cattle, T. foetus-specific vaginal immunoglobulin (Ig) G and IgA antibodies were elevated after infection. The number and viability of day-10 fetuses were reduced in mice infected at 5 weeks of age and bred 12 weeks after infection. Lesions in pregnant and nonpregnant infected mice, including suppurative and eosinophilic vaginitis; cervicitis; endometritis with distension of the uterine lumen; endometrial ulceration; and glandular ectasia, with neutrophils in the glandular lumen and loss of gland epithelium, were similar to those in cattle. The decidua and placenta were multifocally necrotic. Immunohistochemistry demonstrated trichomonads in vaginal folds and uterine glands, and adjacent to fetal tissues. In summary, experimentally infected BALB/cAnNCr mice showed many pathologic similarities to cattle and may serve as a model to study host-trichomonad interactions. PMID:18984788

Agnew, D W; Corbeil, L B; Munson, L; Byrne, B A; BonDurant, R H

2008-11-01

154

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

155

Improvement of gluten-free bread properties by the incorporation of bovine plasma proteins and different saccharides into the matrix.  

PubMed

The aim of this work was to improve the quality of gluten-free bread, incorporating plasma bovine proteins concentrated by ultrafiltration and freeze-dried with saccharides (inulin and sucrose). The influence of these compounds on textural properties and final bread quality was assessed. The textural studies revealed that with the addition of proteins and inulin, homogeneous and smaller air cells were achieved improving the textural properties while the bread hardness was comparable with breads with gluten. The volume of gluten-free breads increased with increasing proteins and inulin concentrations, reaching a maximum at a protein concentration of 3.5% (w/w). The addition of the enhancers improved moisture retention of the loaves after cooking and an increase of lightness of crumb with respect to the control was observed. The sensory analysis found no statistically significant difference in sensory attributes evaluated with respect to the control, so these ingredients do not negatively affect the organoleptic properties of bread. PMID:25306343

Rodriguez Furlán, Laura T; Pérez Padilla, Antonio; Campderrós, Mercedes E

2015-03-01

156

Evaluation of Multiplexed Foot-and-Mouth Disease Nonstructural Protein Antibody Assay Against Standardized Bovine Serum Panel  

SciTech Connect

Liquid array technology has previously been used to show proof-of-principle of a multiplexed non structural protein serological assay to differentiate foot-and-mouth infected and vaccinated animals. The current multiplexed assay consists of synthetically produced peptide signatures 3A, 3B and 3D and recombinant protein signature 3ABC in combination with four controls. To determine diagnostic specificity of each signature in the multiplex, the assay was evaluated against a naive population (n = 104) and a vaccinated population (n = 94). Subsequently, the multiplexed assay was assessed using a panel of bovine sera generated by the World Reference Laboratory for foot-and-mouth disease in Pirbright, UK. This sera panel has been used to assess the performance of other singleplex ELISA-based non-structural protein antibody assays. The 3ABC signature in the multiplexed assay showed comparative performance to a commercially available non-structural protein 3ABC ELISA (Cedi test{reg_sign}) and additional information pertaining to the relative diagnostic sensitivity of each signature in the multiplex is acquired in one experiment. The encouraging results of the evaluation of the multiplexed assay against a panel of diagnostically relevant samples promotes further assay development and optimization to generate an assay for routine use in foot-and-mouth disease surveillance.

Perkins, J; Parida, S; Clavijo, A

2007-05-14

157

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

158

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

159

Studies on substantially increased proteins in follicular fluid of bovine ovarian follicular cysts using 2-D PAGE and MALDI-TOF MS  

PubMed Central

Background The objective of this study was to identify substantially increased proteins in bovine cystic follicular fluid (FF) in order to clarify the pathology and etiology of bovine ovarian follicular cysts (BOFC). Methods Proteins in normal and cystic FF samples were subjected to two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and were compared using silver stained gel images with PDQuest image analysis software. Peptides from these increased spots were analyzed by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), and were identified based on the NCBI database by a peptide mass fingerprinting method. Results Comparative proteomic analysis showed 8 increased protein spots present in cystic FF. MS analysis and database searching revealed that the increased proteins in cystic FF were bovine mitochondrial f1-atpase (BMFA), erythroid associated factor (EAF), methionine synthase (MeS), VEGF-receptor, glyceraldehydes 3-phosphate dehydrogenase (GAPDH), heat shock protein 70 (HSP70), ?-lactoglobulin (BLG) and succinate dehydrogenase Ip subunit (SD). Conclusion Our results suggest that these proteins are overexpressed in BOFC, and that they may play important roles in the pathogenesis of BOFC. Furthermore, these proteins in the FF could be useful biomarkers for BOFC. PMID:15941490

Maniwa, Jiro; Izumi, Shunsuke; Isobe, Naoki; Terada, Takato

2005-01-01

160

Bovine Ephemeral Fever Rhabdovirus ?1 Protein Has Viroporin-Like Properties and Binds Importin ?1 and Importin 7  

PubMed Central

ABSTRACT Bovine ephemeral fever virus (BEFV) is an arthropod-borne rhabdovirus that is classified as the type species of the genus Ephemerovirus. In addition to the five canonical rhabdovirus structural proteins (N, P, M, G, and L), the large and complex BEFV genome contains several open reading frames (ORFs) between the G and L genes (?1, ?2/?3, ?, and ?) encoding proteins of unknown function. We show that the 10.5-kDa BEFV ?1 protein is expressed in infected cells and, consistent with previous predictions based on its structure, has the properties of a viroporin. Expression of a BEFV ?1-maltose binding protein (MBP) fusion protein in Escherichia coli was observed to inhibit cell growth and increase membrane permeability to hygromycin B. Increased membrane permeability was also observed in BEFV-infected mammalian cells (but not cells infected with an ?1-deficient BEFV strain) and in cells expressing a BEFV ?1-green fluorescent protein (GFP) fusion protein, which was shown by confocal microscopy to localize to the Golgi complex. Furthermore, the predicted C-terminal cytoplasmic domain of ?1, which contains a strong nuclear localization signal (NLS), was translocated to the nucleus when expressed independently, and in an affinity chromatography assay employing a GFP trap, the full-length ?1 was observed to interact specifically with importin ?1 and importin 7 but not with importin ?3. These data suggest that, in addition to its function as a viroporin, BEFV ?1 may modulate components of nuclear trafficking pathways, but the specific role thereof remains unclear. IMPORTANCE PMID:24257609

Joubert, D. Albert; Blasdell, Kim R.; Audsley, Michelle D.; Trinidad, Lee; Monaghan, Paul; Dave, Keyur A.; Lieu, Kim G.; Amos-Ritchie, Rachel; Jans, David A.; Moseley, Gregory W.; Gorman, Jeffrey J.

2014-01-01

161

Endotoxin-free purification for the isolation of Bovine Viral Diarrhoea Virus E2 protein from insoluble inclusion body aggregates  

PubMed Central

Background Protein expression in Escherichia coli may result in the recombinant protein being expressed as insoluble inclusion bodies. In addition, proteins purified from E. coli contain endotoxins which need to be removed for in vivo applications. The structural protein, E2, from Bovine Viral Diarrhoea Virus (BVDV) is a major immunogenic determinant, and is an ideal candidate as a subunit vaccine. The E2 protein contains 17 cysteine residues creating difficulties in E. coli expression. In this report we outline a procedure for successfully producing soluble and endotoxin-free BVDV E2 protein from inclusion bodies (IB). Results The expression of a truncated form of BVDV-E2 protein (E2-T1) in E. coli resulted in predominantly aggregated insoluble IB. Solubilisation of E2-T1 with high purity and stability from IB aggregates was achieved using a strong reducing buffer containing 100 mM Dithiothreitol. Refolding by dialysis into 50 mM Tris (pH 7.0) containing 0.2% Igepal CA630 resulted in a soluble but aggregated protein solution. The novel application of a two-phase extraction of inclusion body preparations with Triton X-114 reduced endotoxin in solubilised E2-T1 to levels suitable for in vivo use without affecting protein yields. Dynamic light scattering analyses showed 37.5% of the protein was monomeric, the remaining comprised of soluble aggregates. Mice immunised with E2-T1 developed a high titre antibody response by ELISA. Western hybridisation analysis showed E2-T1 was recognised by sera from immunised mice and also by several BVDV-E2 polyclonal and monoclonal antibodies. Conclusion We have developed a procedure using E. coli to produce soluble E2-T1 protein from IB, and due to their insoluble nature we utilised a novel approach using Triton X-114 to efficiently remove endotoxin. The resultant protein is immunogenic and detectable by BVDV-E2 specific antibodies indicating its usefulness for diagnostic applications and as a subunit vaccine. The optimised E. coli expression system for E2-T1 combined with methodologies for solubilisation, refolding and integrated endotoxin removal presented in this study should prove useful for other vaccine applications. PMID:21787435

2011-01-01

162

The use of aqueous two-phase systems to concentrate and purify bovine leukemia virus outer envelope protein gp51.  

PubMed

Enzootic bovine leucosis is a chronic lymphoproliferative disease of cattle. The causative agent, bovine leukemia virus (BLV), is related to the human retroviruses HTLV-I and -II. The external env-protein of BLV, a glycoprotein of 51 kDa, carries neutralizing epitopes and should be an essential component in a vaccine against the virus. Problems have been encountered with the concentration and purification of intact virions of BLV and other retroviruses. During centrifugation procedures the external env-proteins are to a great extent detached and consequently poorly recovered with the virion particles. Therefore, other methods are sought to obtain a high yield of the external glycoproteins. The use of two-phase systems based on water soluble polymers is described for the extraction of BLV-gp51 from culture medium. Several polymer systems were tested and the results showed that some were attractive for large scale application. The classical combination dextran-polyethylene glycol gave promising results; a partition coefficient of about 0.02 was obtained for the distribution of the gp51 between the top and combined inter- and bottom phases. In a single extraction step it was possible to obtain 45% of the glycoprotein in a small volume bottom phase and at the same time about 15-fold purified. That should be compared with a recovery of less than 20% with the conventional centrifugation procedures. It is concluded that extraction in phase systems based on water soluble polymers is a methodology well suited for the concentration and purification of BLV-gp51. PMID:2474306

Hammar, L; Merza, M; Malm, K; Eriksson, S; Morein, B

1989-06-01

163

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

164

Demonstration of membrane estrogen binding proteins in rat brain by ligand blotting using a 17?-estradiol-[ 125I]bovine serum albumin conjugate  

Microsoft Academic Search

This paper describes a ligand blotting procedure to visualize membrane estrogen receptors\\/binding proteins immobilized on nitrocellulose membranes. Using 17?-estradiol covalently linked with [125I]bovine serum albumin (BSA) at the C-6 position (17?-E-6-[125I]BSA) as a ligand, three major binding proteins with molecular masses of approximately 23, 28, and 32 kDa were identified from crude synaptosomal fractions (P2) of female rat brains. The

Jianbiao Zheng; Victor D. Ramirez

1997-01-01

165

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. PMID:24971313

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

166

Heat-induced redistribution of disulfide bonds in milk proteins. 1. Bovine beta-lactoglobulin.  

PubMed

Changes in the structure and chemistry of beta-lactoglobulin (beta-LG) play an important role in the processing and functionality of milk products. In model beta-LG systems, there is evidence that the aggregates of heated beta-LG are held together by a mixture of intermolecular non-covalent association and heat-induced non-native disulfide bonds. Although a number of non-native disulfide bonds have been identified, little is known about the initial inter- and intramolecular disulfide bond rearrangements that occur as a result of heating. These interchange reactions were explored by examining the products of heat treatment to determine the novel disulfide bonds that form in the heated beta-LG aggregates. The native protein and heat-induced aggregates were hydrolyzed by trypsin, and the resulting peptides, before and after reduction with dithiothreitol, were separated by high-performance liquid chromatography and their identities confirmed by electrospray ionization mass spectrometry. Comparisons of these peptide patterns showed that some of the Cys160 was in the reduced form in heated beta-LG aggregates, indicating that the Cys160-Cys66 disulfide bond had been broken during heating. This finding suggests that disulfide bond interchange reactions between beta-LG non-native monomers, or polymers, and other proteins could occur largely via Cys160. PMID:15675818

Creamer, Lawrence K; Bienvenue, Annie; Nilsson, Hanna; Paulsson, Marie; van Wanroij, Miriam; Lowe, Edwin K; Anema, Skelte G; Boland, Michael J; Jiménez-Flores, Rafael

2004-12-15

167

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. PMID:21811561

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

2011-01-01

168

Highly Sensitive Detection of Small Ruminant Bovine Spongiform Encephalopathy within Transmissible Spongiform Encephalopathy Mixes by Serial Protein Misfolding Cyclic Amplification.  

PubMed

It is assumed that sheep and goats consumed the same bovine spongiform encephalopathy (BSE)-contaminated meat and bone meal that was fed to cattle and precipitated the BSE epidemic in the United Kingdom that peaked more than 20 years ago. Despite intensive surveillance for cases of BSE within the small ruminant populations of the United Kingdom and European Union, no instances of BSE have been detected in sheep, and in only two instances has BSE been discovered in goats. If BSE is present within the small ruminant populations, it may be at subclinical levels, may manifest as scrapie, or may be masked by coinfection with scrapie. To determine whether BSE is potentially circulating at low levels within the European small ruminant populations, highly sensitive assays that can specifically detect BSE, even within the presence of scrapie prion protein, are required. Here, we present a novel assay based on the specific amplification of BSE PrP(Sc) using the serial protein misfolding cyclic amplification assay (sPMCA), which specifically amplified small amounts of ovine and caprine BSE agent which had been mixed into a range of scrapie-positive brain homogenates. We detected the BSE prion protein within a large excess of classical, atypical, and CH1641 scrapie isolates. In a blind trial, this sPMCA-based assay specifically amplified BSE PrP(Sc) within brain mixes with 100% specificity and 97% sensitivity when BSE agent was diluted into scrapie-infected brain homogenates at 1% (vol/vol). PMID:25143565

Gough, Kevin C; Bishop, Keith; Maddison, Ben C

2014-11-01

169

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 Muñoz, A

2010-10-01

170

Bovine Herpesvirus 1 Glycoprotein M Forms a Disulfide-Linked Heterodimer with the UL49.5 Protein  

PubMed Central

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 UL10, 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 UL10 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 UL49.5, 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 UL49.5 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.

1998-01-01

171

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

172

In vitro cross-linking of bovine lens proteins photosensitized by promazines  

SciTech Connect

Promazine derivatives induce cross-linking of bovine lens crystallins in vitro by irradiation with near-ultraviolet (UV) light in the presence of O/sub 2/, as revealed by electrophoresis after denaturation. With the five derivatives tested (promazine (PZ), chlorpromazine (CPZ), triflupromazine (TFPZ), methoxypromazine (MTPZ), and acepromazine (ACPZ)), single-hit kinetics are observed. Evidence implicating the cation radicals of the PZ derivatives as the causative agent of this in vitro effect is presented. Hydroxyl radicals do not appear to be involved in the photo-cross-linking reaction. Sodium ascorbate protects against damage induced either by PZ derivatives plus light or by PZ cation radicals in the dark. These findings are discussed with respect to development of cataracts induced by these drugs in vivo.

Merville, M.P.; Decuyper, J.; Piette, J.; Calberg-Bacq, C.M.; Van de Vorst, A.

1984-05-01

173

Major tegument protein VP8 of bovine herpesvirus 1 is phosphorylated by viral US3 and cellular CK2 protein kinases.  

PubMed

The UL47 gene product, VP8, is one of the major tegument proteins of bovine herpesvirus 1 (BoHV-1) and is subject to phosphorylation. Analysis of protein bands co-immunoprecipitated with VP8 from BoHV-1-infected cells by mass spectroscopy suggested that VP8 interacts with two protein kinases: cellular CK2 and viral US3. CK2 is a highly conserved cellular protein, expressed ubiquitously and known to phosphorylate numerous proteins. The US3 gene product is one of the viral kinases produced by BoHV-1 during infection. Interactions of CK2 and US3 with VP8 were confirmed outside the context of infection when FLAG-VP8 was expressed alone or co-expressed with US3-haemagglutinin tag in Cos-7 cells. Furthermore, VP8 and US3 were found to co-localize in the nucleus during viral infection. To explore the significance of these interactions, an in vitro kinase assay was performed, which demonstrated that VP8 is heavily phosphorylated by CK2. In the presence of the highly specific CK2 kinase inhibitor 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT), phosphorylation of VP8 was significantly reduced. Phosphorylation of VP8 was also inhibited by the presence of kenpaullone, a less specific CK2 inhibitor, but not by protein kinase CK1 or protein kinase C inhibitors. When VP8 and US3 were both included in the kinase assay in the presence of DMAT, phosphorylation of VP8 was again observed. Autophosphorylation of US3 was also detected and was not inhibited by DMAT. Based on these results, it is proposed that VP8 interacts with cellular CK2 and viral US3 in BoHV-1-infected cells, and is in turn subject to kinase activities associated with both of these proteins. PMID:19692545

Labiuk, Shaunivan L; Babiuk, Lorne A; van Drunen Littel-van den Hurk, Sylvia

2009-12-01

174

The role of DNA structure and dynamics in the recognition of bovine papillomavirus E2 protein target sequences.  

PubMed

The papillomavirus E2 transcription and replication factors bind to the DNA consensus ACCGN(4)CGGT sequence (E2-BS), through both direct and indirect readout mechanisms. The two symmetric half-sites ACCG.CGGT are highly conserved in the genomes and are hydrogen bound with E2. Although E2 does not contact the N4 spacer, the affinities are modulated by the base composition of this DNA part. Nevertheless, the origin of either the global recognition mechanism or the spacer effect remains unclear, particularly in the case of the bovine papillomavirus type 1 E2 (BPV-1-E2) system, used as model to study the papillomaviruses. We present, herein, studies carried out on oligomers differently recognized by the BPV-1-E2 protein and based on molecular dynamic simulations including counterions and water. The sequences contain the conserved half-sites but three different spacers (CCAT, ACGT and AAAC), resulting in very high, high and low affinity targets for BPV-1-E2. In order to estimate how much the free DNAs resemble the bound conformations, comparisons are made with two DNAs extracted from E2-BS-BPV-1 crystallographic complexes, representative of high and moderate affinity structures. The analysis of 15 ns trajectories reveals that the ACCG/CGGT half-sites, whatever the spacer, have the same behavior and adopt average stable base-pair parameters very close to those of the bound conformations. In contrast, the three different free spacers strongly differ in their BI <--> BII backbone dynamics. The low affinity AAAC spacer exhibits stable BI backbone conformations, the high affinity ACGT spacer is characterized by a dramatic instability of the CpG phosphate groups, and the CpA and GpG backbones in the very high affinity CCAT.ATGG spacer are trapped in BII conformations. All resemble more of the moderate affinity complex DNA than the high affinity one. Nevertheless, the particular behavior of the CCAT and ACGT backbones allows the emergence of BII-rich spacers, a configuration reproducing both local and global helical features of the bound DNA conformation of the high affinity complex and favoring the minor groove curvature required in the complex. In particular, the CCAT-containing site spends almost half of the time in this form that well mimics the bound one. Thus, we propose that the E2 protein could take advantage of the invariant favorable structures of the half-sites to form a pre-complex, but would require a specific spacer intrinsic malleability to lock the interaction. Finally, the backbone conformational states, by their ability to translate information coded in the sequence into structural properties, provide insight into the mechanisms that contribute to fine binding site selection and specific nucleic acid ligand recognition. PMID:15165850

Djuranovic, D; Oguey, C; Hartmann, B

2004-06-11

175

Optimization and characterization of an in vitro bovine mammary cell culture system to study regulation of milk protein synthesis and mammary differentiation  

SciTech Connect

A long term bovine mammary cell culture system that maintains normal mammary cell function was established and optimized to study milk protein synthesis and secretion and mammary differentiation. This culture system used bovine mammary acini isolated from developing or lactating mammary gland by enzymatic dissociation, and cryopreserved until thawed and plated for growth in vitro for these studies. Cells in M199 with lactogenic hormones {plus minus} fetal calf serum (FCS) were cultured on plastic, 100ul and 500ul type I collagen, and Matrigel, or embedded within type I collagen. Cell morphology, cell number, and total TCA-precipitable {sup 35}S-labelled proteins were monitored. Milk protein ({alpha}{sub s,1}-casein, lactoferrin (LF), {alpha}-lactalbumin, and {beta}-lactoglobulin) secretion and intracellular levels were determined by an ELISA assay.

Talhouk, R.S.

1988-01-01

176

Bovine ephemeral fever rhabdovirus ?1 protein has viroporin-like properties and binds importin ?1 and importin 7.  

PubMed

Bovine ephemeral fever virus (BEFV) is an arthropod-borne rhabdovirus that is classified as the type species of the genus Ephemerovirus. In addition to the five canonical rhabdovirus structural proteins (N, P, M, G, and L), the large and complex BEFV genome contains several open reading frames (ORFs) between the G and L genes (?1, ?2/?3, ?, and ?) encoding proteins of unknown function. We show that the 10.5-kDa BEFV ?1 protein is expressed in infected cells and, consistent with previous predictions based on its structure, has the properties of a viroporin. Expression of a BEFV ?1-maltose binding protein (MBP) fusion protein in Escherichia coli was observed to inhibit cell growth and increase membrane permeability to hygromycin B. Increased membrane permeability was also observed in BEFV-infected mammalian cells (but not cells infected with an ?1-deficient BEFV strain) and in cells expressing a BEFV ?1-green fluorescent protein (GFP) fusion protein, which was shown by confocal microscopy to localize to the Golgi complex. Furthermore, the predicted C-terminal cytoplasmic domain of ?1, which contains a strong nuclear localization signal (NLS), was translocated to the nucleus when expressed independently, and in an affinity chromatography assay employing a GFP trap, the full-length ?1 was observed to interact specifically with importin ?1 and importin 7 but not with importin ?3. These data suggest that, in addition to its function as a viroporin, BEFV ?1 may modulate components of nuclear trafficking pathways, but the specific role thereof remains unclear. Although rhabdovirus accessory genes occur commonly among arthropod-borne rhabdoviruses, little is known of their functions. Here, we demonstrate that the BEFV ?1 ORF encodes a protein which has the structural and functional characteristics of a viroporin. We show that ?1 localizes in the Golgi complex and increases cellular permeability. We also show that BEFV ?1 binds importin ?1 and importin 7, suggesting that it may have a yet unknown role in modulating nuclear trafficking. This is the first functional analysis of an ephemerovirus accessory protein and of a rhabdovirus viroporin. PMID:24257609

Joubert, D Albert; Blasdell, Kim R; Audsley, Michelle D; Trinidad, Lee; Monaghan, Paul; Dave, Keyur A; Lieu, Kim G; Amos-Ritchie, Rachel; Jans, David A; Moseley, Gregory W; Gorman, Jeffrey J; Walker, Peter J

2014-02-01

177

Epitope imprinted polymer coating CdTe quantum dots for specific recognition and direct fluorescent quantification of the target protein bovine serum albumin.  

PubMed

A novel epitope molecularly imprinted polymer (EMIP) for specific recognition and direct fluorescent quantification of the target protein bovine serum albumin (BSA) was demonstrated where polymerization was performed on the surface of silica nanospheres embedded CdTe quantum dots (QDs). The synthetic peptide derived from the surface-exposed C-terminus of bovine serum albumin (BSA, residues 599-607) was selected as the template molecule. The resulting EMIP film was able to selectively capture the template peptide and the corresponding target protein BSA via the recognition cavities. Based on the fluorescence quenching, the EMIP-coated QDs (molecular imprinted polymer coating CdTe QDs using epitope as the template) nanospheres were successfully applied to the direct fluorescence quantification of BSA. Compared with BMIP-coated QDs (molecular imprinted polymer coating CdTe QDs using BSA as the template), the imprinting factor and adsorption capacity of EMIP-coated QDs were greatly increased. The prepared EMIP-coated QDs can also discriminate even one mismatched sequences from the original sequences of the epitope of the BSA. The practical analytical performance of the EMIP-coated QDs was examined by evaluating the detection of BSA in the bovine calf serum sample with satisfactory results. In addition, the resulting EMIP-coated QDs nanospheres were also successfully applied to separating BSA from the bovine blood sample. PMID:24287415

Yang, Ya-Qiong; He, Xi-Wen; Wang, Yi-Zhi; Li, Wen-You; Zhang, Yu-Kui

2014-04-15

178

Diffusion models of protein folding  

PubMed Central

In theory and in the analysis of experiments, protein folding is often described as diffusion along a single coordinate. We explore here the application of a one-dimensional diffusion model to interpret simulations of protein folding, where the parameters of a model that “best” describes the simulation trajectories are determined using a Bayesian analysis. We discuss the requirements for such a model to be a good approximation to the global dynamics, and several methods for testing its accuracy. For example, one test considers the effect of an added bias potential on the fitted free energies and diffusion coefficients. Such a bias may also be used to extend our approach to determining parameters for the model to systems which would not normally explore the full coordinate range on accessible time scales. Alternatively, the propagators predicted from the model at different “lag” times may be compared with observations from simulation. We then present some applications of the model to protein folding, including Kramers-like turnover in folding rates of coarse-grained models, the effect of non-native interactions on folding, and the effect of the chosen coordinate on the observed position-dependence of the diffusion coefficients. Lastly, we consider how our results are useful for the interpretation of experiments, and how this type of Bayesian analysis may eventually be applied directly to analyse experimental data. PMID:21842082

2012-01-01

179

Rheology of globular proteins: apparent yield stress, high shear rate viscosity and interfacial viscoelasticity of bovine serum albumin solutions  

E-print Network

viscoelasticity of bovine serum albumin solutions Vivek Sharma,a Aditya Jaishankar,a Ying-Chih Wangb and Gareth H. In this study, we probe the bulk and the interfacial viscoelasticity of surfactant-free bovine serum albumin circulation.1 The concentration of human serum albumin (HSA) in blood plasma is $40 mg mlÃ?1 ($0.6 mM). Bovine

180

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. PMID:24963321

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

2014-01-01

181

Distinctions between bovine herpesvirus 1 and herpes simplex virus type 1 VP22 tegument protein subcellular associations.  

PubMed

The alphaherpesvirus tegument protein VP22 has been characterized with multiple traits including microtubule reorganization, nuclear localization, and nonclassical intercellular trafficking. However, all these data were derived from studies using herpes simplex virus type 1 (HSV-1) and may not apply to VP22 homologs of other alphaherpesviruses. We compared subcellular attributes of HSV-1 VP22 (HVP22) with bovine herpesvirus 1 (BHV-1) VP22 (BVP22) using green fluorescent protein (GFP)-fused VP22 expression vectors. Fluorescence microscopy of cell lines transfected with these constructs revealed differences as well as similarities between the two VP22 homologs. Compared to that of HVP22, the BVP22 microtubule interaction was much less pronounced. The VP22 nuclear interaction varied, with a marbled or halo appearance for BVP22 and a speckled or nucleolus-bound appearance for HVP22. Both VP22 homologs associated with chromatin at various stages of mitosis and could traffic from expressing cells to the nuclei of nonexpressing cells. However, distinct qualitative differences in microtubule, nuclear, and chromatin association as well as trafficking were observed. The differences in VP22 homolog characteristics revealed in this study will help define VP22 function within HSV-1 and BHV-1 infection. PMID:10708447

Harms, J S; Ren, X; Oliveira, S C; Splitter, G A

2000-04-01

182

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 naïve 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-Cañadas, 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

183

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

184

An Acute-phase Protein as a Regulator of Sperm Survival in the Bovine Oviduct: Alpha 1-acid-glycoprotein Impairs Neutrophil Phagocytosis of Sperm In Vitro  

PubMed Central

We have previously shown that polymorphonuclear neutrophils (PMNs) are present in bovine oviduct fluid under physiological conditions, and that the oviduct provides a microenvironment that protects sperm from phagocytosis by PMNs. Alpha 1-acid glycoprotein (AGP) is a major acute-phase protein produced mainly in the liver that has immunomodulatory functions. AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary. Therefore, in this study, we investigated, 1) the local production of AGP in the bovine oviduct, 2) the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production and 3) the impact of AGP desialylation on the PMN phagocytosis of sperm. The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to produce prostaglandin E2 (PGE2) which has been shown to partially contribute to the regulation of sperm phagocytosis in the bovine oviduct. AGP and PGE2 at concentrations detected in the oviducts additively suppressed sperm phagocytosis by PMNs. These results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct. PMID:24931131

LIU, Jinghui; MAREY, Mohamed A.; KOWSAR, Rasoul; HAMBRUCH, Nina; SHIMIZU, Takashi; HANEDA, Shingo; MATSUI, Motozumi; SASAKI, Motoki; HAYAKAWA, Hiroyuki; PFARRER, Christiane; MIYAMOTO, Akio

2014-01-01

185

An Acute-phase Protein as a Regulator of Sperm Survival in the Bovine Oviduct: Alpha 1-acid-glycoprotein Impairs Neutrophil Phagocytosis of Sperm In Vitro.  

PubMed

We have previously shown that polymorphonuclear neutrophils (PMNs) are present in bovine oviduct fluid under physiological conditions, and that the oviduct provides a microenvironment that protects sperm from phagocytosis by PMNs. Alpha 1-acid glycoprotein (AGP) is a major acute-phase protein produced mainly in the liver that has immunomodulatory functions. AGP mRNA is expressed in extrahepatic organs, such as the lung, kidney, spleen, lymph node, uterus, and ovary. Therefore, in this study, we investigated, 1) the local production of AGP in the bovine oviduct, 2) the effect of AGP on the phagocytic activity of PMNs for sperm and superoxide production and 3) the impact of AGP desialylation on the PMN phagocytosis of sperm. The AGP gene was expressed in cultured bovine oviduct epithelial cells (BOECs) and AGP protein was detected in oviduct fluid. Preexposure of PMNs to AGP at physiological levels impaired PMN phagocytosis for sperm and superoxide generation. The desialylation of AGP eliminated these suppressive effects of AGP on PMN. Scanning electron microscopy revealed that AGP drastically reduced the formation of DNA-based neutrophil extracellular traps (NETs) for sperm entanglement. Additionally, AGP dose-dependently stimulated BOECs to produce prostaglandin E2 (PGE2) which has been shown to partially contribute to the regulation of sperm phagocytosis in the bovine oviduct. AGP and PGE2 at concentrations detected in the oviducts additively suppressed sperm phagocytosis by PMNs. These results provide evidence that locally produced AGP may be involved in protecting sperm from phagocytosis by PMNs in the bovine oviduct. PMID:24931131

Liu, Jinghui; Marey, Mohamed A; Kowsar, Rasoul; Hambruch, Nina; Shimizu, Takashi; Haneda, Shingo; Matsui, Motozumi; Sasaki, Motoki; Hayakawa, Hiroyuki; Pfarrer, Christiane; Miyamoto, Akio

2014-10-23

186

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

187

Assignment of the Multifunctional NS3 Protein of Bovine Viral Diarrhea Virus during RNA Replication: an In Vivo and In Vitro Study  

Microsoft Academic Search

Studies on the replication of the pestivirus bovine viral diarrhea virus (BVDV) were considerably facilitated by the recent discovery of an autonomous subgenomic BVDV RNA replicon (DI9c). DI9c comprises mainly the untranslated regions of the viral genome and the coding region of the nonstructural proteins NS3, NS4A, NS4B, NS5A, and NS5B. To assess the significance of the NS3-associated nucleoside triphosphatase\\/helicase

CLAUS W. GRASSMANN; OLAF ISKEN; SVEN-ERIK BEHRENS

1999-01-01

188

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

189

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

190

Microtubule-associated protein tau in bovine retinal photoreceptor rod outer segments: comparison with brain tau  

PubMed Central

Recent studies have suggested a possible involvement of abnormal tau in some retinal degenerative diseases. The common view in these studies is that these retinal diseases share the mechanism of tau-mediated degenerative diseases in brain and that information about these brain diseases may be directly applied to explain these retinal diseases. Here we collectively examine this view by revealing three basic characteristics of tau in the rod outer segment (ROS) of bovine retinal photoreceptors, i.e., its isoforms, its phosphorylation mode and its interaction with microtubules, and by comparing them with those of brain tau. We find that ROS contains at least four isoforms: three are identical to those in brain and one is unique in ROS. All ROS isoforms, like brain isoforms, are modified with multiple phosphate molecules; however, ROS isoforms show their own specific phosphorylation pattern, and these phosphorylation patterns appear not to be identical to those of brain tau. Interestingly, some ROS isoforms, under the normal conditions, are phosphorylated at the sites identical to those in Alzheimer’s patient isoforms. Surprisingly, a large portion of ROS isoforms tightly associates with a membranous component(s) other than microtubules, and this association is independent of their phosphorylation states. These observations strongly suggest that tau plays various roles in ROS and that some of these functions may not be comparable to those of brain tau. We believe that knowledge about tau in the entire retinal network and/or its individual cells are also essential for elucidation of tau-mediated retinal diseases, if any. PMID:23712071

Yamazaki, Akio; Nishizawa, Yuji; Matsuura, Isao; Hayashi, Fumio; Usukura, Jiro; Bondarenko, Vladimir A.

2013-01-01

191

Antithrombin III and its interaction with heparin. Comparison of the human, bovine, and porcine proteins by /sup 1/H NMR spectroscopy  

SciTech Connect

/sup 1/H 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 /sup 1/H NMR spectra. Human antithrombin III has five histidine residues, bovine has six, and porcine has five. The C(2) proton from each of these residues gives a narrow resonance and titrates with pH; the pK/sub a/'s are in the range 5.15-7.25. It is concluded that all histidines in each protein are surface residues with considerable independent mobility. The carbohydrate chains in each protein also give sharp resonances consistent with a surface location and motional flexibility. The /sup 1/H spectra are sensitive to heparin binding. Although heparin resonances obscure protein resonances in the region 3.2-6.0 ppm, difference spectra between antithrombin III with and without heparin show clear perturbation of a small number of aromatic and aliphatic protein protons. For human antithrombin III, it was shown that heparin fragments 8, 10, and 16 sugar residues in length result in almost identical perturbations to the protein. In contrast, tetrasaccharide results in fewer perturbations. Significantly, intact high molecular weight heparin causes the same spectral perturbations as the 16-residue fragment. These data are discussed in terms of requirements for heparin binding.

Gettins, P.

1987-03-10

192

An innovative bovine odorant binding protein-based filtering cartridge for the removal of triazine herbicides from water.  

PubMed

Odorant binding protein (OBP) is a multi-functional scavenger for small hydrophobic molecules dissolved in the mucus lining the nasal epithelia of mammals, characterized by broad ligand binding specificity towards a large number of structurally unrelated natural and synthetic molecules of different chemical classes. Here, we demonstrate for the first time the application of OBP as the active element of an innovative filtering matrix for the removal of environmental pollutants such as triazine herbicides from water samples. The filtering device, obtained by coupling histidine-tagged bovine OBP to a nickel nitrilotriacetic acid (Ni-NTA) agarose resin, was characterized in terms of retention capacity for the herbicides atrazine, simazine, and propazine. Analysis of these herbicides at trace levels with solid-phase microextraction followed by gas chromatography-mass spectrometry using the selected ion monitoring mode proved the capabilities of the proposed device for the decontamination of surface and groundwater samples in the 0.2-2,300 ?g/L concentration range, obtaining a reduction in the triazine content greater than 97 %, thus suggesting its possible use for the potabilization of water. PMID:23104315

Bianchi, Federica; Basini, Giuseppina; Grolli, Stefano; Conti, Virna; Bianchi, Francesco; Grasselli, Francesca; Careri, Maria; Ramoni, Roberto

2013-01-01

193

Identification of alternatively spliced mRNAs encoding potential new regulatory proteins in cattle infected with bovine leukemia virus.  

PubMed Central

The polymerase chain reaction was used to detect and characterize low-abundance bovine leukemia virus (BLV) mRNAs. In infected cattle we could detect spliced mRNA with a splice pattern consistent with a Tax/Rex mRNA, as well as at least four alternatively spliced RNAs. Two of the alternatively spliced mRNAs encoded hitherto unrecognized BLV proteins, designated RIII and GIV. The Tax/Rex and alternatively spliced mRNAs could be detected at their highest levels in BLV-infected cell cultures; the next highest levels were found in samples from calves experimentally infected at 6 weeks postinoculation. Alternatively spliced mRNAs were also expressed, albeit at lower levels, in naturally infected animals; they were detected by a nested polymerase chain reaction. Interestingly, the GIV mRNA was specifically detected in naturally infected cows with persistent lymphocytosis and in two of five calves at 6 months after experimental infection with BLV. Furthermore, the calf with the strongest signal for GIV had the highest lymphocyte counts. These data may suggest a correlation between expression of the GIV product and development of persistent lymphocytosis. Some of the donor and acceptor sites in the alternatively spliced mRNAs were highly unusual. The biological mechanisms and significance of such a choice of unexpected splice sites are currently unknown. Images PMID:8380084

Alexandersen, S; Carpenter, S; Christensen, J; Storgaard, T; Viuff, B; Wannemuehler, Y; Belousov, J; Roth, J A

1993-01-01

194

DNA sequence polymorphisms within the bovine guanine nucleotide-binding protein Gs subunit alpha (Gs?)-encoding (GNAS) genomic imprinting domain are associated with performance traits  

PubMed Central

Background Genes which are epigenetically regulated via genomic imprinting can be potential targets for artificial selection during animal breeding. Indeed, imprinted loci have been shown to underlie some important quantitative traits in domestic mammals, most notably muscle mass and fat deposition. In this candidate gene study, we have identified novel associations between six validated single nucleotide polymorphisms (SNPs) spanning a 97.6 kb region within the bovine guanine nucleotide-binding protein Gs subunit alpha gene (GNAS) domain on bovine chromosome 13 and genetic merit for a range of performance traits in 848 progeny-tested Holstein-Friesian sires. The mammalian GNAS domain consists of a number of reciprocally-imprinted, alternatively-spliced genes which can play a major role in growth, development and disease in mice and humans. Based on the current annotation of the bovine GNAS domain, four of the SNPs analysed (rs43101491, rs43101493, rs43101485 and rs43101486) were located upstream of the GNAS gene, while one SNP (rs41694646) was located in the second intron of the GNAS gene. The final SNP (rs41694656) was located in the first exon of transcripts encoding the putative bovine neuroendocrine-specific protein NESP55, resulting in an aspartic acid-to-asparagine amino acid substitution at amino acid position 192. Results SNP genotype-phenotype association analyses indicate that the single intronic GNAS SNP (rs41694646) is associated (P ? 0.05) with a range of performance traits including milk yield, milk protein yield, the content of fat and protein in milk, culled cow carcass weight and progeny carcass conformation, measures of animal body size, direct calving difficulty (i.e. difficulty in calving due to the size of the calf) and gestation length. Association (P ? 0.01) with direct calving difficulty (i.e. due to calf size) and maternal calving difficulty (i.e. due to the maternal pelvic width size) was also observed at the rs43101491 SNP. Following adjustment for multiple-testing, significant association (q ? 0.05) remained between the rs41694646 SNP and four traits (animal stature, body depth, direct calving difficulty and milk yield) only. Notably, the single SNP in the bovine NESP55 gene (rs41694656) was associated (P ? 0.01) with somatic cell count--an often-cited indicator of resistance to mastitis and overall health status of the mammary system--and previous studies have demonstrated that the chromosomal region to where the GNAS domain maps underlies an important quantitative trait locus for this trait. This association, however, was not significant after adjustment for multiple testing. The three remaining SNPs assayed were not associated with any of the performance traits analysed in this study. Analysis of all pairwise linkage disequilibrium (r2) values suggests that most allele substitution effects for the assayed SNPs observed are independent. Finally, the polymorphic coding SNP in the putative bovine NESP55 gene was used to test the imprinting status of this gene across a range of foetal bovine tissues. Conclusions Previous studies in other mammalian species have shown that DNA sequence variation within the imprinted GNAS gene cluster contributes to several physiological and metabolic disorders, including obesity in humans and mice. Similarly, the results presented here indicate an important role for the imprinted GNAS cluster in underlying complex performance traits in cattle such as animal growth, calving, fertility and health. These findings suggest that GNAS domain-associated polymorphisms may serve as important genetic markers for future livestock breeding programs and support previous studies that candidate imprinted loci may act as molecular targets for the genetic improvement of agricultural populations. In addition, we present new evidence that the bovine NESP55 gene is epigenetically regulated as a maternally expressed imprinted gene in placental and intestinal tissues from 8-10 week old bovine foetuses. PMID:21214909

2011-01-01

195

Polymorphic behavior in protein-surfactant mixtures: the water-bovine serum albumin-sodium taurodeoxycholate system.  

PubMed

Mixtures containing water, bovine serum albumin (BSA), and sodium taurodeoxycholate (NaTDC), a component of the bile in mammals, have been investigated in a wide range of composition and pH. Depending on the concentration of both solutes and the pH, solutions, precipitates, and gels are formed. Under spontaneous pH conditions, the transport properties in dilute solutions indicate the occurrence of significant interactions between BSA and the surfactant. Conversely, acidic media favor the formation of nonsoluble protein-surfactant complexes, with subsequent precipitation. The nucleation kinetics of the protein-surfactant complexes in solid form and the related precipitation processes can be slow or fast, depending on the overall solute content and the mole ratio. At high concentrations, a gel, extending on both sides of the charge neutralization line, and two-phase regions are observed. Gels shrink in open air and swell in the presence of excess water. Depending on concentration and temperature, the gels transform from an essentially liquidlike behavior to that peculiar to true gels (when G' > or = G''). The thermal gelation threshold, the temperature above which G' > or = G'', depends on BSA and NaTDC content and is concomitant to moderate heat effects, inferred by differential scanning calorimetry (DSC). The above data also indicate that the protein thermal denaturation in the gel is shifted to higher temperatures compared to water. Such a stabilizing effect is presumably related to the occurrence of both electrostatic and hydrophobic interactions with NaTDC. Water self-diffusion in the gels is slightly slower than that in the bulk and poorly sensitive to composition: it is about 65% the value of neat H2O in a wide concentration range, irrespective of the BSA, or NaTDC, concentration. A peculiar behavior is also observed in 23Na longitudinal and transverse relaxation rates. The T1 and T2 values, measured at 105.75 MHz on BSA-NaTDC gels, indicate that the motions determining the NMR relaxation of the sodium ions in the hydration layer of the protein-surfactant aggregates are not slow, having frequencies comparable with the Larmor one. The above properties, especially the rheological and the spectroscopic ones, are important for understanding the behavior of gels based on protein-surfactant mixtures. PMID:16800527

Orioni, Barbara; Roversi, Mauro; La Mesa, Camillo; Asaro, Fioretta; Pellizer, Giorgio; D'Errico, Gerardino

2006-06-22

196

The composition and physicochemical properties of bovine nasal-septa protein-polysaccharide complex  

PubMed Central

1. Protein–polysaccharide complexes were prepared in three different ways and the gross stoicheiometry of the complexes compared. 2. The neutral sugar content was ascertained and the possibility of a glycoprotein occurring with chondroitin sulphate and keratosulphate is discussed. 3. Physical data support a molecule of molecular weight 3·2×106–5·8×106 with a roughly spherical domain and an average radius of gyration of 1390Å. Such a particle is highly solvated. The complex is heavily charged with the sulphate groups on the outside. 4. These findings are discussed in the light of the physiological role of protein–polysaccharide light fraction (PPL) in cartilage. PMID:4226525

Luscombe, Mollie; Phelps, C. F.

1967-01-01

197

Microwave Ablation Compared with Radiofrequency Ablation for Breast Tissue in an Ex Vivo Bovine Udder Model  

SciTech Connect

Purpose: To compare the effectiveness of microwave (MW) ablation with radiofrequency (RF) ablation for treating breast tissue in a nonperfused ex vivo model of healthy bovine udder tissue. Materials and Methods: MW ablations were performed at power outputs of 25W, 35W, and 45W using a 915-MHz frequency generator and a 2-cm active tip antenna. RF ablations were performed with a bipolar RF system with 2- and 3-cm active tip electrodes. Tissue temperatures were continuously monitored during ablation. Results: The mean short-axis diameters of the coagulation zones were 1.34 {+-} 0.14, 1.45 {+-} 0.13, and 1.74 {+-} 0.11 cm for MW ablation at outputs of 25W, 35W, and 45W. For RF ablation, the corresponding values were 1.16 {+-} 0.09 and 1.26 {+-} 0.14 cm with electrodes having 2- and 3-cm active tips, respectively. The mean coagulation volumes were 2.27 {+-} 0.65, 2.85 {+-} 0.72, and 4.45 {+-} 0.47 cm{sup 3} for MW ablation at outputs of 25W, 35W, and 45W and 1.18 {+-} 0.30 and 2.29 {+-} 0.55 cm{sup 3} got RF ablation with 2- and 3-cm electrodes, respectively. MW ablations at 35W and 45W achieved significantly longer short-axis diameters than RF ablations (P < 0.05). The highest tissue temperature was achieved with MW ablation at 45W (P < 0.05). On histological examination, the extent of the ablation zone in MW ablations was less affected by tissue heterogeneity than that in RF ablations. Conclusion: MW ablation appears to be advantageous with respect to the volume of ablation and the shape of the margin of necrosis compared with RF ablation in an ex vivo bovine udder.

Tanaka, Toshihiro, E-mail: toshihir@bf6.so-net.ne.jp [RWTH Aachen University, Applied Medical Engineering, Helmholtz-Institute Aachen (Germany); Westphal, Saskia, E-mail: swestphal@ukaachen.de [RWTH Aachen University, Department of Pathology Aachen University Hospital (Germany); Isfort, Peter, E-mail: isfort@hia.rwth-aachen.de [RWTH Aachen University, Applied Medical Engineering, Helmholtz-Institute Aachen (Germany); Braunschweig, Till, E-mail: tbraunschweig@ukaachen.de [RWTH Aachen University, Department of Pathology Aachen University Hospital (Germany); Penzkofer, Tobias, E-mail: penzkofer@hia.rwth-aachen.de; Bruners, Philipp, E-mail: bruners@rad.rwth-aachen.de [RWTH Aachen University, Applied Medical Engineering, Helmholtz-Institute Aachen (Germany); Kichikawa, Kimihiko, E-mail: kkichika@naramed-u.ac.jp [Nara Medical University, Department of Radiology (Japan); Schmitz-Rode, Thomas, E-mail: smiro@hia.rwth-aachen.de; Mahnken, Andreas H., E-mail: mahnken@rad.rwth-aachen.de [RWTH Aachen University, Applied Medical Engineering, Helmholtz-Institute Aachen (Germany)

2012-08-15

198

Ovine prion protein variant A(136)R(154)L(168)Q(171) increases resistance to experimental challenge with bovine spongiform encephalopathy agent.  

PubMed

Susceptibility and incubation periods of transmissible spongiform encephalopathies, such as scrapie in sheep, are modulated by the PrP gene. The standard model of association between ovine PrP genetics and classical scrapie susceptibility is based on PrP genotypes with respect to codons 136, 154 and 171, e.g. alanine-arginine-glutamine (ARQ). It is demonstrated here that a proline to leucine substitution in codon 168 of the ovine PrP protein gene is associated with increased resistance to experimental bovine spongiform encephalopathy (BSE) inoculation. The ARL(168)Q PrP allele was found in heterozygous ARP(168)Q/ARL(168)Q sheep that have so far survived intravenous BSE challenge three times longer than BSE-challenged homozygous ARP(168)Q/ARP(168)Q sheep, which develop disease in around 700 days. In contrast, the L141F polymorphism does not appear to be associated with susceptibility to intravenous BSE challenge. PMID:17098993

Goldmann, Wilfred; Houston, Fiona; Stewart, Paula; Perucchini, Matteo; Foster, James; Hunter, Nora

2006-12-01

199

Bovine Colostrum Increases Pore-Forming Claudin-2 Protein Expression but Paradoxically Not Ion Permeability Possibly by a Change of the Intestinal Cytokine Milieu  

PubMed Central

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

Maletzki, Claudia; Lamprecht, Georg

2013-01-01

200

Presence of ecto-protein tyrosine phosphatase activity is vital for survival of Setaria cervi, a bovine filarial parasite.  

PubMed

The ecto protein tyrosine phosphatases (PTP) are known to play a crucial role in the pathogenesis and survival of the intracellular parasites. However, their presence and role in filarial parasites is still unknown. We found a significant amount of tyrosine phosphatase activity in the surface antigen fraction extracted from Setaria cervi (S. cervi), a bovine filarial parasite. An antibody designed against the conserved catalytic core of human protein tyrosine phosphatases, PTP1B cross reacted with a 63 kDa band in the surface antigen. We detected a significant amount of PTP activity in the intact S. cervi adult parasites as well as microfilariae in this study for the first time. This PTP may be localized on the surface of the parasite with an exposed active site available for the external substrates. The PTP activity was also inhibited by sodium orthovanadate and phenyl arsine oxide, specific inhibitors of PTP in both the life stages. The Km and Vmax for PTP in the adult parasites and microfilariae were determined to be 2.574?±?0.14 mM; 206.3?±?2.75 ?M Pi/h/two parasites and 5.510?±?0.59 mM; 62.27?±?2.27 ?M Pi/h/10(6) parasites respectively using O-P-L-Tyrosine as substrate. Interestingly, a positive correlation was observed between the inhibition in PTP activity and reduction in the motility/ viability of the parasites when they were subjected to the specific PTP inhibitors (Orthovanadate and Phenyl arsine oxide) for 4 h in the KRB maintenance medium. The activity was also significantly inhibited in the parasites exposed to antifilarial drug/compounds for e.g. Diethylcarbamazine, Acetylsalicylic Acid and SK7, a methyl chalcone. Therefore suggesting a possible role played by PTP in the survival of the parasite, its interaction with the host as well as in the screening of newly synthesized antifilarials/drugs. PMID:25028209

Singh, Neetu; Heneberg, Petr; Rathaur, Sushma

2014-10-01

201

Real-Time Trapping of Intact Singly-Charged Bovine Serum Albumin Proteins with a Big Frequency-Adjusted Quadrupole  

SciTech Connect

High-resolution real-time particle mass measurements have not been achievable because the enormous amount of kinetic energy imparted to the particles upon expansion into vacuum competes with and overwhelms the forces applied to the charged particles within the mass spectrometer. It is possible to reduce the kinetic energy of a collimated particulate ion beam through collisions with a buffer gas while radially constraining their motion using a quadrupole guide or trap over a limited mass range. Controlling the pressure drop of the final expansion into a quadrupole trap permits a much broader mass range at the cost of sacrificing collimation. To achieve high-resolution mass analysis of massive particulate ions, an efficient trap with a large tolerance for radial divergence of the injected ions was developed that permits trapping a large range of ions for on-demand injection into an awaiting mass analyzer. The design specifications required that frequency of the trapping potential be adjustable to cover a large mass range and the trap radius be increased to increase the tolerance to divergent ion injection. The large-radius linear quadrupole ion trap was demonstrated by trapping singly-charged bovine serum albumin ions for on-demand injection into a mass analyzer. Additionally, this work demonstrates the ability to measure an electrophoretic mobility cross section (or ion mobility) of singly-charged intact proteins in the low-pressure regime. This work represents a large step toward the goal of high-resolution analysis of intact proteins, RNA, DNA, and viruses.

Koizumi, Hideya [ORNL; Whitten, William B [ORNL; Reilly, Pete [ORNL

2008-01-01

202

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

203

Kinetic properties of bovine brain protein l -isoaspartyl methyltransferase determined using a synthetic isoaspartyl peptide substrate  

Microsoft Academic Search

Proteinl-isoaspartyl methyltransferase, an enzyme enriched in brain, is implicated in the repair of age-damaged proteins containing atypical, isoaspartyl peptide bonds. We have investigated the kinetics of methylation using a synthetic peptide substrate having the structure Trp-Ala-Gly-Gly-isoAsp-Ala-Ser-Gly-Glu. Double-reciprocal plots of initial velocity versus concentration of S-adenosylmethionine (AdoMet) at different fixed concentrations of peptide gave straight lines converging at a positive 1\\/v

Brett A. Johnson; Dana W. Aswad

1993-01-01

204

Hydrogen peroxide-induced alterations of tight junction proteins in bovine brain microvascular endothelial cells  

Microsoft Academic Search

Occludin and zonular occludens (ZO)-1 in tight junctions (TJs) and actin play an important role in maintaining blood–brain barrier (BBB) endothelial ion and solute barriers. Malfunction of BBB by reactive oxygen species (ROS) has been attributed to the disruption of TJs. This study examined H2O2 effects on changes of paracellular permeability, actin, and TJ proteins (occludin and ZO-1) using primary

Hee-Sang Lee; Kee Namkoong; Dong-Hwa Kim; Ki-Jeong Kim; Yoon-Hee Cheong; Sung-Su Kim; Won-Bok Lee; Kyung-Yong Kim

2004-01-01

205

Regulation of a swelling-activated chloride current in bovine endothelium by protein tyrosine phosphorylation and G proteins  

PubMed Central

The role of protein tyrosine phosphorylation and of G proteins in the activation of a swelling-activated Cl? current (ICl,swell) in calf pulmonary artery endothelial (CPAE) cells was studied using the whole-cell patch clamp technique. ICl,swell was activated by reducing the extracellular osmolality by either 12.5% (mild hypotonicity) or 25% (strong hypotonicity).The protein tyrosine kinase (PTK) inhibitors tyrphostin B46, tyrphostin A25 and genistein inhibited ICl,swell with IC50 values of, respectively, 9.2 ± 0.2, 61.4 ± 1.7 and 62.9 ± 1.3?M. Tyrphostin A1, a tyrphostin analogue with little effect on PTK activity, and daidzein, an inactive genistein analogue, were without effect on ICl,swell.The protein tyrosine phosphatase (PTP) inhibitors Na3VO4 (200 ?M) and dephostatin (20 ?M) potentiated ICl,swell activated by mild hypotonicity by 47 ± 9 and 69 ± 15%, respectively.Intracellular perfusion with GTP?S (100 ?M) transiently activated a Cl? current with an identical biophysical and pharmacological profile to ICl,swell. This current was inhibited by the tested PTK inhibitors and potentiated by the PTP inhibitors. Hypertonicity-induced cell shrinkage completely inhibited the GTP?S-activated Cl? current.Intracellular perfusion with GDP?S (1 mM) caused a time-dependent inhibition of ICl,swell, which was more pronounced when the current was activated by mild hypotonicity.Our results demonstrate that the activity of endothelial swelling-activated Cl? channels is dependent on tyrosine phosphorylation and suggest that G proteins regulate the sensitivity to cell swelling. PMID:9490863

Voets, Thomas; Manolopoulos, Vangelis; Eggermont, Jan; Ellory, Clive; Droogmans, Guy; Nilius, Bernd

1998-01-01

206

Preservation of protein clefts in comparative models  

Microsoft Academic Search

BACKGROUND: Comparative, or homology, modelling of protein structures is the most widely used prediction method when the target protein has homologues of known structure. Given that the quality of a model may vary greatly, several studies have been devoted to identifying the factors that influence modelling results. These studies usually consider the protein as a whole, and only a few

David Piedra; Sergi Lois; Xavier de la Cruz

2008-01-01

207

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

PubMed

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-Muñoz, R; González-Halphen, D; Grolli, S; Ramoni, R

2014-07-01

208

Dependence of superoxide anion production on extracellular and intracellular calcium ions and protein kinase C in PMA-stimulated bovine neutrophils.  

PubMed Central

The involvement of both intracellular and extracellular calcium, as well as the activation of protein kinase C (PKC), in phorbol myristate acetate (PMA)-stimulated respiratory burst in bovine neutrophils has been studied. PMA significantly stimulated the superoxide anion production by these cells. The increased production of superoxide anion was inhibited by BAPTA/AM, an intracellular calcium ([Ca2+]i) chelator, but not affected by EGTA, an extracellular calcium ([Ca2+]0) chelator. PMA also induced PKC activation, and a PKC inhibitor, calphostin C, blocked the stimulatory effect of PMA on superoxide anion production by the neutrophils. Therefore, we conclude that PMA-induced respiratory burst in bovine neutrophils is [Ca2+]i- but not [Ca2+]0-dependent, and also requires PKC activation. PMID:9918328

Allard, B; Long, E; Block, E; Zhao, X

1999-01-01

209

Genetic definition of a new bovine papillomavirus type 1 open reading frame, E5B, that encodes a hydrophobic protein involved in altering host-cell protein processing.  

PubMed

We have previously observed that bovine papillomavirus type 1 (BPV-1) induces the appearance of five cellular proteins in C127 mouse fibroblasts, four of which appear to arise by altered processing of resident endoplasmic reticulum proteins. Studies of various cell lines revealed that expression of the 3' end of the BPV early region was sufficient for induction of these changes. To identify the BPV gene responsible, we have utilized the simian virus 40 (SV40)/BPV-1 recombinant virus Pava-1, which expresses the 3' end of the BPV early region behind an SV40 early promoter. C127 cells infected with Pava-1 for 48 h show the expected BPV-associated alterations, as do cells infected with Pava constructs mutated in the E5 or E2 genes. However, a mutation in the start codon of a previously ignored open reading frame extending from nucleotides 4013 to 4170 (E5B) eliminated the BPV-associated changes. Similar results were obtained with COS cells infected with the Pava mutants and C127 cells transformed by full-length mutated BPV. Despite its influence on the processing of cellular endoplasmic reticulum proteins, this mutation in E5B did not alter BPV-transforming efficiency or the ability of transformants to form colonies in soft agar. The E5B open reading frame encodes a hydrophobic 52-amino-acid polypeptide that shares structural similarities with HPV6 E5A and HPV16 E5. Speculations on a role for E5B in the viral life cycle are discussed. PMID:8388507

O'Banion, M K; Winn, V D; Settleman, J; Young, D A

1993-06-01

210

Action of degradative enzymes on the light fraction of bovine septa protein polysaccharide  

PubMed Central

1. Fragments from enzymic degradation of protein–polysaccharide light fraction (PPL) have been analysed. 2. The time-course of action of some proteolytic enzymes and of hyaluronidase on PPL has been followed by viscometric techniques. 3. It is suggested that papain acts to produce single polysaccharide chains, whereas other proteolytic enzymes tried give evidence of twin-chain residues. 4. The molecular weight of the fragments derived from complete enzyme action on PPL supports this postulate. 5. A structure of the PPL complex is suggested. PMID:6033751

Luscombe, Mollie; Phelps, C. F.

1967-01-01

211

Bone temperature during cementation with a heatsink: a bovine model pilot study  

PubMed Central

Background Bone cement is an effective means of supporting implants, but reaches high temperatures while undergoing polymerisation. Bone has been shown to be sensitive to thermal injury with osteonecrosis reported after one minute at 47°C. Necrosis during cementing may lead to loosening of the prosthesis. Some surgeons fill the joint cavity with cool irrigation fluid to provide a heatsink during cementing, but this has not been supported by research. This paper assesses a simple technique to investigate the efficacy of this method. Findings We used a model acetabulum in a bovine humerus to allow measurement of bone temperatures in cementing. Models were prepared with a 50 mm diameter acetabulum and three temperature probe holes; two as close as possible to the acetabular margin at half the depth of the acetabulum and at the full depth of the acetabulum, and one 10 mm from the acetabular rim. Four warmed models were cemented with Palacos RG using a standard mixing system and a 10 mm polyethylene disc to represent an acetabular component. Two of the acetabular models were filled with room temperature water to provide a heatsink. An electronic probe measured temperature at 5 second intervals from the moment of cementing. In the models with no heatsink, peak temperature was 40.3°C. The mean temperature rise was 10.9°C. In the models with a heatsink, there was an average fall in the bone temperature during cementing of 4.4°C. Conclusions These results suggest that using a heatsink while cementing prostheses may reduce the peak bone temperature. This study demonstrates a simple, repeatable technique which may be useful for larger trials. PMID:25099248

2014-01-01

212

Modeling mutations in protein structures  

PubMed Central

We describe an automated method for the modeling of point mutations in protein structures. The protein is represented by all non-hydrogen atoms. The scoring function consists of several types of physical potential energy terms and homology-derived restraints. The optimization method implements a combination of conjugate gradient minimization and molecular dynamics with simulated annealing. The testing set consists of 717 pairs of known protein structures differing by a single mutation. Twelve variations of the scoring function were tested in three different environments of the mutated residue. The best-performing protocol optimizes all the atoms of the mutated residue, with respect to a scoring function that includes molecular mechanics energy terms for bond distances, angles, dihedral angles, peptide bond planarity, and non-bonded atomic contacts represented by Lennard-Jones potential, dihedral angle restraints derived from the aligned homologous structure, and a statistical potential for non-bonded atomic interactions extracted from a large set of known protein structures. The current method compares favorably with other tested approaches, especially when predicting long and flexible side-chains. In addition to the thoroughness of the conformational search, sampled degrees of freedom, and the scoring function type, the accuracy of the method was also evaluated as a function of the flexibility of the mutated side-chain, the relative volume change of the mutated residue, and its residue type. The results suggest that further improvement is likely to be achieved by concentrating on the improvement of the scoring function, in addition to or instead of increasing the variety of sampled conformations. PMID:17766392

Feyfant, Eric; Sali, Andrej; Fiser, Andras

2007-01-01

213

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. PMID:20085727

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

2010-01-01

214

Biopotency of fetal bovine serum, and insulin and insulin-like growth factors I and II in enhancing whole-body protein synthesis of chicken embryos cultured in vitro  

Microsoft Academic Search

Whole-body protein synthesis in chicken embryos was measured to examine the biopotency of fetal bovine serum, bovine insulin, recombinant human insulin, and recombinant human insulin-like growth factors (IGF) I and II. In all experiments chicken embryos at 7 days of incubation age were used and cultured in a synthetic serum-free medium in the presence or absence of the testing substances

Tatsuo Muramatsu; Reinhard Pinontoan; Jun-ichi Okumura

1995-01-01

215

Genomic structure and tissue-specific expression of human and mouse genes encoding homologues of the major bovine seminal plasma proteins.  

PubMed

Sperm capacitation is a maturation event that takes place in the female reproductive tract and is essential for fertilization. A family of phospholipid-binding proteins present in bovine seminal plasma (BSP proteins) binds the sperm membrane at ejaculation and promotes bovine sperm capacitation. Homologues of these proteins have also been isolated from boar, ram, goat, bison and stallion seminal fluid, suggesting that BSP proteins and their homologues are conserved among mammals. However, there have been no reports on BSP-homologous proteins in mice and humans to date. A search of the mouse and human genomes, using the nucleic acid sequences of BSP proteins, revealed the presence of three BSP-like sequences in the mouse genome, named mouse BSP Homologue 1 (mBSPH1), mBSPH2 and mBSPH3, and one sequence in the human genome (hBSPH1). Mouse epididymal expressed sequence tags corresponding to partial sequences of mBSPH1 and mBSPH2 were identified. The entire complementary DNA (cDNA) sequences of mBSPH1 and mBSPH2 from mouse epididymis and hBSPH1 from human epididymis were obtained by 5'-/3'-rapid amplification of cDNA ends (RACE) and encode predicted proteins containing two tandemly repeated fibronectin type II domains, which is the signature of the BSP family of proteins. Using RT-PCR, it was revealed that mBSPH1, mBSPH2 and hBSPH1 mRNA are expressed only in the epididymis. Expression of mBSPH3 was not detected in any tissue and probably represents a pseudogene. This work shows, for the first time, that BSP homologues are expressed in mouse and human and may be involved in sperm capacitation in these species. PMID:17085770

Lefebvre, J; Fan, J; Chevalier, S; Sullivan, R; Carmona, E; Manjunath, P

2007-01-01

216

Modulation of immune function by a modified bovine whey protein concentrate.  

PubMed

The commercial preparation of dairy foodstuffs generates large volumes of by-products, many of which have as yet undocumented effects on mammalian immune function. In the present report, a modified whey protein concentrate (mWPC), derived as a by-product from the commercial manufacture of cheese, was tested for its ability to modulate murine immune function in vitro. The mWPC suppressed T and B lymphocyte proliferative responses to mitogens in a dose-dependent fashion. The mWPC also suppressed alloantigen-induced lymphocyte proliferation during a mixed leucocyte reaction, but showed no suppressive effect against IL-2-sustained proliferation of mitogen-activated T cell blasts. Other indices of lymphocyte activation, such as cytokine secretion and the formation of activated (CD25+) T cell blasts, were suppressed by the mWPC, suggesting that the mode of suppression may be to inhibit the lymphocyte activation process. Enzymatic digestion by pepsin and pancreatin, under physiologically realistic conditions in vitro, ablated the immunomodulatory function of the mWPC. These results are discussed in relation to the potential development of complex-mixture dairy products into health-modulating products. PMID:10457202

Cross, M L; Gill, H S

1999-08-01

217

Use of topical bovine thrombin in an anti-coagulated rat model of hepatic injury.  

PubMed

The need for surgical hemostasis in patients treated with anticoagulant medications is a concern. This study assessed a bovine-derived topical hemostat (FastAct, FA) using a partial hepatectomy hemorrhage model in anticoagulated rats. Ninety rats were randomly assigned to receive preoperative heparin, warfarin, or nothing (n=30/treatment). Within each treatment group, FA, saline, direct pressure (DP), electrocautery, or nothing (n=6/group) was applied to the hepatectomy site. Eight additional rats were used for assessment of the preoperative anticoagulant regimen. Rats that were not anticoagulated and received FA had faster clot times and less hemorrhage than those receiving DP (P<0.05). In warfarin-pretreated rats, FA resulted in faster coagulation times than saline or DP and less hemorrhage than saline (P<0.05). No differences were detected in heparinized rats. Across all groups, rats receiving FA lost less blood and formed clots more frequently than saline (P<0.05). FA may be useful to treat hemorrhage from hepatic lacerations in anticoagulated patients. PMID:22633173

Schmiedt, Chad W; Köhler, Rickard; Brainard, Benjamin M

2012-12-01

218

Identity elements in bovine tRNA(Trp) required for the specific stimulation of gelonin, a plant ribosome-inactivating protein.  

PubMed Central

Ribosome-inactivating proteins (RIPs) are RNA-N-glycosidases widely present in plants that depurinate RNA in ribosomes at a specific universally conserved position, A4324, in the rat 28S rRNA. A small group of RIPs (cofactor-dependent RIPs) require ATP and tRNA to reach maximal activity on isolated ribosomes. Among cofactor-dependent RIPs, gelonin is specifically and uniquely stimulated by tRNA(Trp). The active species are avian (chicken) and mammalian (beef, rat, and rabbit) tRNA(Trp), whereas yeast tRNA(Trp) is completely devoid of stimulating activity. In the present article, bovine and yeast tRNA(Trp) with unmodified bases were prepared by assembly of the corresponding genes from synthetic oligonucleotides followed by PCR and T7 RNA polymerase transcription of the amplified products. The two synthetic tRNAs were fully active (bovine) or inactive (yeast) as the wild-type tRNAs. Construction of chimeric tRNA(Trp) transcripts identified the following bovine nucleotides as recognition elements for gelonin-stimulating activity: G26 and bp G12-C23 in the D arm and G57, A59, and bp G51-C63 and U52-A62 in the T arm. Among single-stranded nucleotides, A59 has a prominent role, but full expression of the gelonin-stimulating activity requires an extensive cooperation between nucleotides in both arms. PMID:10573126

Brigotti, M; Carnicelli, D; Pallanca, A; Rizzi, S; Accorsi, P; Montanaro, L; Sperti, S

1999-01-01

219

Modelling protein-protein interaction networks via a stickiness index  

PubMed Central

What type of connectivity structure are we seeing in protein–protein interaction networks? A number of random graph models have been mooted. After fitting model parameters to real data, the models can be judged by their success in reproducing key network properties. Here, we propose a very simple random graph model that inserts a connection according to the degree, or ‘stickiness’, of the two proteins involved. This model can be regarded as a testable distillation of more sophisticated versions that attempt to account for the presence of interaction surfaces or binding domains. By computing a range of network similarity measures, including relative graphlet frequency distance, we find that our model outperforms other random graph classes. In particular, we show that given the underlying degree information, fitting a stickiness model produces better results than simply choosing a degree-matching graph uniformly at random. Therefore, the results lend support to the basic modelling methodology. PMID:16971339

Przulj, Natasa; Higham, Desmond J

2006-01-01

220

Evaluation of the Recombinant 10-Kilodalton Immunodominant Region of the BP26 Protein of Brucella abortus for Specific Diagnosis of Bovine Brucellosis ?  

PubMed Central

Brucellosis is a disease with worldwide distribution affecting animals and human beings. Brucella abortus is the causative agent of bovine brucellosis. The cross-reactions of currently available diagnostic procedures for B. abortus infection result in false-positive reactions, which make the procedures unreliable. These tests are also unable to differentiate Brucella-infected and -vaccinated animals. The present work is focused on the use of a nonlipopolysaccharide (LPS) diagnostic antigen, a recombinant 10-kDa (r10-kDa) protein of B. abortus, for specific diagnosis of brucellosis. The purified recombinant protein was used as a diagnostic antigen in plate enzyme-linked immunosorbent assay (p-ELISA) format to screen 408 bovine serum samples (70 presumptively negative, 308 random, and 30 vaccinated), and the results were compared with those of the Rose Bengal plate agglutination test (RBPT) and the standard tube agglutination test (STAT). Statistical analysis in presumptive negative samples revealed 100 and 98.41% specificity of p-ELISA with RBPT and STAT, and an agreement of 91.43% with the tests using Cohen's kappa statistics. In random samples, the agreement of p-ELISA was 77.92% and 80.52% with RBPT and STAT, respectively. p-ELISA investigation of vaccinated samples reported no false-positive results, whereas RBPT and STAT reported 30% and 96.6% false-positive results, respectively. The data suggest that p-ELISA with r10-kDa protein may be a useful method for diagnosis of bovine brucellosis. Furthermore, p-ELISA may also be used as a tool for differentiating Brucella-vaccinated and naturally infected animals. PMID:21852548

Tiwari, Arvind Kumar; Kumar, Subodh; Pal, Vijai; Bhardwaj, Bhupendra; Rai, Ganga Prasad

2011-01-01

221

Involvement of different protein kinases and phospholipases A2 in phorbol ester (TPA)-induced arachidonic acid liberation in bovine platelets.  

PubMed

The effect of various phospholipase A2 and protein kinase inhibitors on the arachidonic acid liberation in bovine platelets induced by the protein kinase activator 12-O-tetradecanoylphorbol-13-acetate (TPA) was studied. TPA stimulates arachidonic acid release mainly by activating group IV cytosolic PLA2 (cPLA2), since inhibitors of this enzyme markedly inhibited arachidonic acid formation. However, group VI Ca2+-independent PLA2 (iPLA2) seems to contribute to the arachidonic acid liberation too, since the relatively specific iPLA2 inhibitor bromoenol lactone (BEL) decreased arachidonic acid generation in part. The pronounced inhibition of the TPA-induced arachidonic acid release by the protein kinase C (PKC) inhibitors GF 109203X and Ro 31-82220, respectively, and by the p38 MAP kinase inhibitor SB 202190 suggests that the activation of the PLA2s by TPA is mediated via PKC and p38 MAP kinase. PMID:10877452

Lehr, M; Griessbach, K

2000-01-01

222

Immunoaffinity purification of two major proteins of bovine leukemia virus (gp51 and p24) and their use for discrimination between vaccinated and infected animals.  

PubMed

The outer envelope glycoprotein gp51 and the core protein p24 of bovine leukemia virus (BLV), were purified from culture media of FLK-BLV cells by a single-step procedure, using immunoaffinity chromatography based on monoclonal antibodies to the respective proteins. About 90% of the envelope glycoprotein in the culture medium was recovered as a highly purified product. Both purified protein (gp51 and p24) preparations, were found to be highly specific antigens by ELISA, and did not cross-react with sera raised against the other antigen. The conformational epitopes on the purified gp51 were preserved as judged by their reactions with the corresponding monoclonal antibodies. The p24 ELISA reacted only with sera from naturally infected animals and not with sera from animals immunized with an experimental gp51-iscom vaccine. The p24 antigen is therefore useful for discriminating between BLV-infected animals and those immunized with a gp51 subunit vaccine. PMID:1723735

Merza, M; Sundquist, B; Söber, J; Morein, B

1991-08-01

223

Protein structure homology modeling using SWISS-MODEL workspace  

Microsoft Academic Search

Homology modeling aims to build three-dimensional protein structure models using experimentally determined structures of related family members as templates. SWISS-MODEL workspace is an integrated Web-based modeling expert system. For a given target protein, a library of experimental protein structures is searched to identify suitable templates. On the basis of a sequence alignment between the target protein and the template structure,

Lorenza Bordoli; Florian Kiefer; Konstantin Arnold; Pascal Benkert; James Battey; Torsten Schwede

2008-01-01

224

Protein structure homology modeling using SWISS-MODEL workspace.  

PubMed

Homology modeling aims to build three-dimensional protein structure models using experimentally determined structures of related family members as templates. SWISS-MODEL workspace is an integrated Web-based modeling expert system. For a given target protein, a library of experimental protein structures is searched to identify suitable templates. On the basis of a sequence alignment between the target protein and the template structure, a three-dimensional model for the target protein is generated. Model quality assessment tools are used to estimate the reliability of the resulting models. Homology modeling is currently the most accurate computational method to generate reliable structural models and is routinely used in many biological applications. Typically, the computational effort for a modeling project is less than 2 h. However, this does not include the time required for visualization and interpretation of the model, which may vary depending on personal experience working with protein structures. PMID:19131951

Bordoli, Lorenza; Kiefer, Florian; Arnold, Konstantin; Benkert, Pascal; Battey, James; Schwede, Torsten

2009-01-01

225

The Amino-Terminal Domain of Bovine Viral Diarrhea Virus Npro Protein Is Necessary for Alpha\\/Beta Interferon Antagonism  

Microsoft Academic Search

The alpha\\/beta interferon (IFN-\\/) system is the first line of defense against viral infection and a critical link between the innate and adaptive immune responses. IFN-\\/ secretion is the hallmark of cellular responses to acute RNA virus infections. As part of their survival strategy, many viruses have evolved mechanisms to counteract the host IFN-\\/ response. Bovine viral diarrhea virus (BVDV)

Laura H. V. G. Gil; Israrul H. Ansari; Ventzislav Vassilev; Delin Liang; Vicky C. H. Lai; Weidong Zhong; Zhi Hong; Edward J. Dubovi; Ruben O. Donis

2006-01-01

226

Four Independent Molecular Prion Protein Parameters for Discriminating New Cases of C, L, and H Bovine Spongiform Encephalopathy in Cattle?  

PubMed Central

In anticipation of the emergence of more variants of bovine spongiform encephalopathy (BSE), a semiquantitative display of the following four independent molecular diagnostic prion parameters was designed: N terminus, proteinase K (PK) resistance, glycoprofile, and mixed population. One H BSE case, three L BSE cases, six C BSE cases, and one unusual classical BSE (C BSE) case are reported. PMID:21677067

Langeveld, Jan P. M.; Erkens, Jo H. F.; Rammel, Ines; Jacobs, Jorg G.; Davidse, Aart; van Zijderveld, Fred G.; Bossers, Alex; Schildorfer, Hermann

2011-01-01

227

Four independent molecular prion protein parameters for discriminating new cases of C, L, and h bovine spongiform encephalopathy in cattle.  

PubMed

In anticipation of the emergence of more variants of bovine spongiform encephalopathy (BSE), a semiquantitative display of the following four independent molecular diagnostic prion parameters was designed: N terminus, proteinase K (PK) resistance, glycoprofile, and mixed population. One H BSE case, three L BSE cases, six C BSE cases, and one unusual classical BSE (C BSE) case are reported. PMID:21677067

Langeveld, Jan P M; Erkens, Jo H F; Rammel, Ines; Jacobs, Jorg G; Davidse, Aart; van Zijderveld, Fred G; Bossers, Alex; Schildorfer, Hermann

2011-08-01

228

Intracellular calcium and protein tyrosine phosphorylation during the release of bovine sperm adhering to the fallopian tube epithelium in vitro  

Microsoft Academic Search

In mammals, sperm adhesion to the epithelial cells lining the oviductal isthmus plays a key role in the maintenance of motility and in the selection of superior quality subpopulations. In the bovine species, heparin and other sulfated glycoconjugates powerfully induce the synchronous release of sperm adhering to tubal epithelium in vitro and may represent the signal which triggers release at

Roberto Gualtieri; Raffaele Boni; Elisabetta Tosti; Maria Zagami; Riccardo Talevi

2005-01-01

229

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. PMID:23365203

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

230

Vitamin D signaling in the bovine immune system: a model for understanding human vitamin D requirements.  

PubMed

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. PMID:22666545

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

2012-03-01

231

Native PAGE Protein Expression  

E-print Network

, mg/ml BA Bovine serum albumin Bovine g-globulin Standard curve generation using known standards. A, use Bio-Rad's bovine serum albumin or bovine g-globulin to make your standard curve. For best results and bovine serum albumin standard Catalog # Description 500-0121 RC DC Protein Assay Kit I, includes RC

Lebendiker, Mario

232

Application of systems analysis in modelling the risk of bovine spongiform encephalopathy (BSE)  

Microsoft Academic Search

Bovine spongiform encephalopathy (BSE), widely known as “mad cow disease”, has virtually crippled the British livestock industry. Even though, no cases of BSE have been reported in the United States (US), a similar epidemic in the US would be catastrophic. The added concern for the risk of introduction of the human disease called variant Creutzfeldt-Jacob disease that has been linked

T. Habtemariam; B. Tameru; D. Nganwa; L. Ayanwale; A. Ahmed; D. Oryang; H. AbdelRahman; G. Gray; J. Cohen; S. Kreindel

2002-01-01

233

Bioeconomic modeling of lactational antimicrobial treatment of new bovine subclinical intramammary infections caused by contagious pathogens.  

PubMed

This study determined the direct and indirect epidemiologic and economic effects of lactational treatment of new bovine subclinical intramammary infections (IMI) caused by contagious pathogens using an existing bioeconomic model. The dynamic and stochastic model simulated the dynamics of Staphylococcus aureus, Streptococcus uberis, Streptococcus dysgalactiae, and Escherichia coli during lactation and the dry period in a 100-cow dairy herd during 1 quota year. Input parameters on cure were obtained from recent Dutch field data. The costs of clinical IMI, subclinical IMI, and intervention were calculated into the combined total annual net costs of IMI per herd. The cost effectiveness of 4 scenarios with lactational intervention was determined; scenarios included no intervention, treatment after 1 mo of infection, treatment after 2 mo of infection, and treatment after 1 mo of infection and culling of uncured cows after 2 mo of infection. Model behavior was observed for variation in parameter input values. Compared with no lactational intervention, lactational intervention of new subclinical IMI resulted in fewer clinical flare ups, less transmission within the herd, and much lower combined total annual net costs of IMI in dairy herds. Antimicrobial treatment of IMI after 1 mo of infection and culling of uncured cows after 2 mo of infection resulted in the lowest costs, whereas treatment after 2 mo of infection was associated with the highest costs between the scenarios with intervention. Changing the probability of cure resulted in a nonlinear change in the cumulative incidence of IMI cases and associated costs. Lactational treatment was able to prevent IMI epidemics in dairy herds at high transmission rates of Strep. uberis, Strep. dysgalactiae, and E. coli. Lactational treatment did not limit the spread of Staph. aureus at high transmission rates, although the associated costs were lower compared with no intervention. To improve udder health in a dairy herd, lactational treatment of contagious subclinical IMI must therefore be preceded by management measures that lower the transmission rate. Lactational treatment of environmental subclinical IMI seemed less cost effective. Detection of subclinical IMI needs improvement to be able to most effectively treat subclinical IMI caused by contagious pathogens during lactation. PMID:20723677

van den Borne, B H P; Halasa, T; van Schaik, G; Hogeveen, H; Nielen, M

2010-09-01

234

Impact of external sources of infection on the dynamics of bovine tuberculosis in modelled badger populations  

PubMed Central

Background The persistence of bovine TB (bTB) in various countries throughout the world is enhanced by the existence of wildlife hosts for the infection. In Britain and Ireland, the principal wildlife host for bTB is the badger (Meles meles). The objective of our study was to examine the dynamics of bTB in badgers in relation to both badger-derived infection from within the population and externally-derived, trickle-type, infection, such as could occur from other species or environmental sources, using a spatial stochastic simulation model. Results The presence of external sources of infection can increase mean prevalence and reduce the threshold group size for disease persistence. Above the threshold equilibrium group size of 6–8 individuals predicted by the model for bTB persistence in badgers based on internal infection alone, external sources of infection have relatively little impact on the persistence or level of disease. However, within a critical range of group sizes just below this threshold level, external infection becomes much more important in determining disease dynamics. Within this critical range, external infection increases the ratio of intra- to inter-group infections due to the greater probability of external infections entering fully-susceptible groups. The effect is to enable bTB persistence and increase bTB prevalence in badger populations which would not be able to maintain bTB based on internal infection alone. Conclusions External sources of bTB infection can contribute to the persistence of bTB in badger populations. In high-density badger populations, internal badger-derived infections occur at a sufficient rate that the additional effect of external sources in exacerbating disease is minimal. However, in lower-density populations, external sources of infection are much more important in enhancing bTB prevalence and persistence. In such circumstances, it is particularly important that control strategies to reduce bTB in badgers include efforts to minimise such external sources of infection. PMID:22738118

2012-01-01

235

A soft and transparent handleable protein model  

NASA Astrophysics Data System (ADS)

The field of structural biology currently relies on computer-generated graphical representations of three-dimensional (3D) structures to conceptualize biomolecules. As the size and complexity of the molecular structure increases, model generation and peer discussions become more difficult. It is even more problematic when discussing protein-protein interactions wherein large surface area contact is considered. This report demonstrates the viability of a new handleable protein molecular model with a soft and transparent silicone body similar to the molecule's surface. A full-color printed main chain structure embedded in the silicone body enables users to simultaneously feel the molecular surface, view through the main chain structure, and manually simulate molecular docking. The interactive, hands-on experience deepens the user's intuitive understanding of the complicated 3D protein structure and elucidates ligand binding and protein-protein interactions. This model would be an effective discussion tool for the classroom or laboratory that stimulates inspired learning in this study field.

Kawakami, Masaru

2012-08-01

236

The membrane lateral domain approach in the studies of lipid–protein interaction of GPI-anchored bovine erythrocyte acetylcholinesterase  

Microsoft Academic Search

A novel membrane lateral domain approach was used to test whether the activity of the membrane-bound enzyme acetylcholinesterase (AChE) depends on the local properties (e.g. local lipid ordering) of bovine erythrocyte-ghost membrane. This issue has an additional aspect of interest due to an alternative mode of insertion of AChE molecules into the membrane by the glycosylphosphatidylinositol (GPI) anchor. In our

Zoran Arsov; Milan Schara; Matjaž Zorko; Janez Štrancar

2004-01-01

237

Bovine Leukemia Virus Transmembrane Protein gp30 Physically Associates with the Down-Regulatory Phosphatase SHP1  

Microsoft Academic Search

In B lymphocytes, the down-regulatory phosphatase SHP-1 associates with CD22 and CD32b (also known as Fc?RIIB) and acts as a critical negative regulator of B-cell receptor signaling. Bovine leukemia virus, a retrovirus of the HTLV\\/BLV group, causes persistently increased numbers of peripheral blood B lymphocytes, known as persistent lymphocytosis (PL) and, in some animals, progression to B-cell leukemia and\\/or lymphoma.

Glenn H. Cantor; Suzanne M. Pritchard; Oto Orlik; William C. Davis; Raymond Reeves

1999-01-01

238

Nuclear Import of Bovine Papillomavirus Type 1 E1 Protein Is Mediated by Multiple Alpha Importins and Is Negatively Regulated by Phosphorylation near a Nuclear Localization Signal?  

PubMed Central

Papillomavirus DNA replication occurs in the nucleus of infected cells and requires the viral E1 protein, which enters the nuclei of host epithelial cells and carries out enzymatic functions required for the initiation of viral DNA replication. In this study, we investigated the pathway and regulation of the nuclear import of the E1 protein from bovine papillomavirus type 1 (BPV1). Using an in vitro binding assay, we determined that the E1 protein interacted with importins ?3, ?4, and ?5 via its nuclear localization signal (NLS) sequence. In agreement with this result, purified E1 protein was effectively imported into the nucleus of digitonin-permeabilized HeLa cells after incubation with importin ?3, ?4, or ?5 and other necessary import factors. We also observed that in vitro binding of E1 protein to all three ? importins was significantly decreased by the introduction of pseudophosphorylation mutations in the NLS region. Consistent with the binding defect, pseudophosphorylated E1 protein failed to enter the nucleus of digitonin-permeabilized HeLa cells in vitro. Likewise, the pseudophosphorylation mutant showed aberrant intracellular localization in vivo and accumulated primarily on the nuclear envelope in transfected HeLa cells, while the corresponding alanine replacement mutant displayed the same cellular location pattern as wild-type E1 protein. Collectively, our data demonstrate that BPV1 E1 protein can be transported into the nucleus by more than one importin ? and suggest that E1 phosphorylation by host cell kinases plays a regulatory role in modulating E1 nucleocytoplasmic localization. This phosphoregulation of nuclear E1 protein uptake may contribute to the coordination of viral replication with keratinocyte proliferation and differentiation. PMID:17192311

Bian, Xue-Lin; Rosas-Acosta, German; Wu, Yu-Chieh; Wilson, Van G.

2007-01-01

239

Transdisciplinary habitat models for elk and cattle as a proxy for bovine tuberculosis transmission risk.  

PubMed

Zoonotic diseases such as bovine tuberculosis (TB) that infect wildlife and livestock are particularly difficult to eradicate where wild animals make extensive use of agricultural landscapes. Transmission of TB between cattle (Bos taurus) and wild elk (Cervus elaphus) in southwestern Manitoba, Canada remains poorly understood but there is a risk when commingling occurs on summer pasture. Elk use of cattle summer pastures was assessed using ecological data (187 VHF and 25 GPS collared elk monitored over four years representing 8% of the elk population). Local knowledge was documented by conducting interviews and participatory mapping exercises with 86 cattle producers (98% of those within the study area). Of the 294 cattle pastures mapped by farmers, 13% were used by radio-collared elk, 38% were reported by farmers as being used by elk, and 42% were identified as used by elk when both when all datasets were combined. Cattle pastures that had been used by elk and those that had no elk were compared using binary logistic regression based on each of the three datasets (i.e. farmer observations, radio-collared elk on pasture, and combined dataset). For all three datasets, distance to protected area and proportion of forest cover on the cattle pasture were identified as the most and second most important predictor variables, respectively. There was strong agreement among the relative probabilities of elk occurrence on each pasture derived from the resource selection function (RSF) models developed using farmer interviews and elk collaring data. The farmer interview and collar datasets were then combined to generate a final integrated RSF map summarizing the probability of elk-cattle commingling and were contrasted over each of four cattle grazing seasons (spring, early summer, late summer, and autumn). These predictive maps indicate that use of cattle pastures by elk is extensive, particularly in spring and early summer. Farmer observations indicate that elk and cattle share water sources and livestock mineral supplements on pasture. Local knowledge and conventional ecological data complement and validate one another and help us better understand the temporospatial aspects of shared space use among wildlife and livestock and more generally the risks of disease transmission in agricultural landscapes. PMID:19541377

Brook, Ryan K; McLachlan, Stéphane M

2009-10-01

240

SWISS-MODEL: an automated protein homology-modeling server  

Microsoft Academic Search

SWISS-MODEL (http:\\/\\/swissmodel.expasy.org) is a server for automated comparative modeling of three- dimensional (3D) protein structures. It pioneered the field of automated modeling starting in 1993 and is the most widely-used free web-based automated modeling facility today. In 2002 the server computed 120 000 user requests for 3D protein models. SWISS- MODEL provides several levels of user interaction through its World

Torsten Schwede; Jürgen Kopp; Nicolas Guex; Manuel C. Peitsch

2003-01-01

241

A mathematical model for prokaryotic protein synthesis  

Microsoft Academic Search

A kinetic model for the synthesis of proteins in prokaryotes is presented and analysed. This model is based on a Markov model\\u000a for the state of the DNA strand encoding the protein. The states that the DNA strand can occupy are: ready, repressed, or\\u000a having a mRNA chain of length i in the process of being completed. The case i

Donald A. Drew

2001-01-01

242

The Effect of Molybdenum-Induced Copper Deficiency on Acute-Phase Protein Concentrations, Superoxide Dismutase Activity, Leukocyte Numbers, and Lymphocyte Proliferation in Beef Heifers Inoculated with Bovine Herpesvirus11,2  

Microsoft Academic Search

This study was conducted to deter- mine the effect of Cu deficiency on acute-phase protein concentrations, superoxide dismutase activity, leuko- cyte numbers, and lymphocyte proliferation in heifers inoculated with live bovine herpesvirus-1 (BHV-1). Hereford × Angus heifers were allotted by weight and initial liver Cu concentrations into molybdenum (Mo)-supplemented (n = 6 ) or control (n = 6 ) groups.

J. D. Arthington; L. R. Corah; F. Blecha

2010-01-01

243

Protein hydrogen exchange: Testing current models  

E-print Network

Protein hydrogen exchange: Testing current models John J. Skinner,1 * Woon K. Lim,2 Sabrina Bedard,1 Ben E. Black,1 and S. Walter Englander1 1 Johnson Research Foundation, Department of Biochemistry of protein hydrogen exchange (HX), HX rates of most of the backbone amide hydrogens of Staphylococcal

Englander, S. Walter

244

Expression of a bovine ?-CN cDNA in the mammary gland of transgenic mice utilizing a genomic milk protein gene as an expression cassette  

Microsoft Academic Search

Transgenic mice were produced by microinjection of a DNA construct composed of the bovine ?-casein (?-CN) cDNA under the control of the goat ?-CN 5? promoter elements and 3? flanking regions into pronuclear-stage embryos. The gene construct targeted the expression of bovine ?-CN RNA to the mammary gland and secretion of bovine ?-CN in the milk. In the three lines

Alfonso Gutiérrez; Harry M. Meade; Paul Ditullio; Daniel Pollock; Merry Harvey; Rafael Jiménez-Flores; Gary B. Anderson; James D. Murray; Juan F. Medrano

1996-01-01

245

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

246

A network model to investigate structural and electrical properties of proteins  

E-print Network

One of the main trend in to date research and development is the miniaturization of electronic devices. In this perspective, integrated nanodevices based on proteins or biomolecules are attracting a major interest. In fact, it has been shown that proteins like bacteriorhodopsin and azurin, manifest electrical properties which are promising for the development of active components in the field of molecular electronics. Here we focus on two relevant kinds of proteins: The bovine rhodopsin, prototype of GPCR protein, and the enzyme acetylcholinesterase (AChE), whose inhibition is one of the most qualified treatments of Alzheimer disease. Both these proteins exert their functioning starting with a conformational change of their native structure. Our guess is that such a change should be accompanied with a detectable variation of their electrical properties. To investigate this conjecture, we present an impedance network model of proteins, able to estimate the different electrical response associated with the different configurations. The model resolution of the electrical response is found able to monitor the structure and the conformational change of the given protein. In this respect, rhodopsin exhibits a better differential response than AChE. This result gives room to different interpretations of the degree of conformational change and in particular supports a recent hypothesis on the existence of a mixed state already in the native configuration of the protein.

E. Alfinito; C. Pennetta; L. Reggiani

2007-03-30

247

An economic model to evaluate the mitigation programme for bovine viral diarrhoea in Switzerland.  

PubMed

Economic analyses are indispensable as sources of information to help policy makers make decisions about mitigation resource use. The aim of this study was to conduct an economic evaluation of the Swiss national mitigation programme for bovine viral diarrhoea virus (BVDV), which was implemented in 2008 and concludes in 2017. The eradication phase of the mitigation programme comprised testing and slaughtering of all persistently infected (PI) animals found. First, the whole population was antigen tested and all PI cattle removed. Since October 2008, all newborn calves have been subject to antigen testing to identify and slaughter PI calves. All mothers of PI calves were retested and slaughtered if the test was positive. Antigen testing in calves and elimination of virus-carriers was envisaged to be conducted until the end of 2011. Subsequently, a surveillance programme will document disease freedom or detect disease if it recurs. Four alternative surveillance strategies based on antibody testing in blood from newborn calves and/or milk from primiparous cows were proposed by Federal Veterinary Office servants in charge of the BVDV mitigation programme. A simple economic spreadsheet model was developed to estimate and compare the costs and benefits of the BVDV mitigation programme. In an independent project, the impact of the mitigation programme on the disease dynamics in the population was simulated using a stochastic compartment model. Mitigation costs accrued from materials, labour, and processes such as handling and testing samples, and recording results. Benefits were disease costs avoided by having the mitigation programme in place compared to a baseline of endemic disease equilibrium. Cumulative eradication costs and benefits were estimated to determine the break-even point for the eradication component of the programme. The margin over eradication cost therefore equalled the maximum expenditure potentially available for surveillance without the net benefit from the mitigation programme overall becoming zero. Costs of the four surveillance strategies and the net benefit of the mitigation programme were estimated. Simulations were run for the years 2008-2017 with 20,000 iterations in @Risk for Excel. The mean baseline disease costs were estimated to be 16.04 m CHF (1 Swiss Franc, CHF=0.73 € at the time of analysis) (90% central range, CR: 14.71-17.39 m CHF) in 2008 and 14.89 m CHF (90% CR: 13.72-16.08 m CHF) in 2009. The break-even point was estimated to be reached in 2012 and the margin over eradication cost 63.15m CHF (90% CR: 53.72-72.82 m CHF). The discounted cost for each surveillance strategy was found to be smaller than the margin, so the mitigation programme overall is expected to have a positive net economic benefit irrespective of the strategy adopted. For economic efficiency, the least cost surveillance alternative must be selected. PMID:22402180

Häsler, B; Howe, K S; Presi, P; Stärk, K D C

2012-09-15

248

Quantitative thermodynamic model for globular protein folding  

NASA Astrophysics Data System (ADS)

We present a statistical mechanics formalism for theoretical description of the process of protein folding ? unfolding transition in water environment. The formalism is based on the construction of the partition function of a protein obeying two-stage-like folding kinetics. Using the statistical mechanics model of solvation of hydrophobic hydrocarbons we obtain the partition function of infinitely diluted solution of proteins in water environment. The calculated dependencies of the protein heat capacities upon temperature are compared with the corresponding results of experimental measurements for staphylococcal nuclease and metmyoglobin.

Yakubovich, Alexander V.; Solov'yov, Andrey V.

2014-06-01

249

A minimal model of protein-protein binding affinities.  

PubMed

A minimal model of protein-protein binding affinity that takes into account only two structural features of the complex, the size of its interface, and the amplitude of the conformation change between the free and bound subunits, is tested on the 144 complexes of a structure-affinity benchmark. It yields Kd values that are within two orders of magnitude of the experiment for 67% of the complexes, within three orders for 88%, and fails on 12%, which display either large conformation changes, or a very high or a low affinity. The minimal model lacks the specificity and accuracy needed to make useful affinity predictions, but it should help in assessing the added value of parameters used by more elaborate models, and set a baseline for evaluating their performances. PMID:25270898

Janin, Joël

2014-12-01

250

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. 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; Carratu, Maria Rosaria; Crovace, Antonio

2014-01-01

251

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

252

Human synoviocyte lubricin and bovine synovial fluid lubricin equally improve gliding resistance in a canine model in vitro  

PubMed Central

The lubricating ability of human synoviocyte lubricin and bovine lubricin purified from synovial fluid was investigated and compared using a canine in vitro tendon model. Our null hypothesis was that these two forms of lubricin would have equal lubricating ability. Forty two canine hind-limbs were used. The peroneus longus (PL) tendons were harvested, along with the proximal phalanx and flexor digitorum profundus of the second or fifth digit with its proximal fibro-osseous pulley. Forty PL tendons were randomly assigned to one of four treatment groups. After gliding resistance testing, two intact PL tendons and two tendons in each group were randomly selected for surface observation with scanning electron microscopy (SEM). The variance of the PL saline group mean gliding resistance was significantly different from other groups. There was a significant treatment-cycle interaction effect on the mean gliding resistance. On SEM, the surface of the saline treated PL tendons appeared rough, whereas the other tendon surfaces appeared smooth. Human synoviocyte lubricin functioned as well as bovine synovial fluid lubricin to reduce friction of canine PL tendons in vitro. This data suggest that treatment using the two forms of lubricin are mechanically similar. PMID:22561248

Kohn, Mark D.; Sun, Yu-long; Zhao, Chunfeng; Thoreson, Andrew R.; Jay, Gregory D.; An, Kai-Nan; Amadio, Peter C.

2014-01-01

253

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. PMID:21569635

2011-01-01

254

Characterization and transcript mapping of a bovine herpesvirus type 1 gene encoding a polypeptide homologous to the herpes simplex virus type 1 major tegument proteins VP13\\/14  

Microsoft Academic Search

Using in vitro translation of hybrid-selected mRNA, we have previously shown that bovine herpesvirus type 1 HindlII fragment M encodes an abundant 94K polypeptide. Using immunoprecipitation and sequencing analyses, it has now been shown that the polypeptide is related to the major tegument protein VP8 and is homologous to the herpes simplex virus type 1 major tegument proteins VP13\\/14. The

S. LaBoissiere; Michel Trudel; Claire Simard

1992-01-01

255

Protein structure prediction and model quality assessment  

PubMed Central

Protein structures have proven to be a crucial piece of information for biomedical research. Of the millions of currently sequenced proteins only a small fraction is experimentally solved for structure and the only feasible way to bridge the gap between sequence and structure data is computational modeling. Half a century has passed since it was shown that the amino acid sequence of a protein determines its shape, but a method to translate the sequence code reliably into the 3D structure still remains to be developed. This review summarizes modern protein structure prediction techniques with the emphasis on comparative modeling, and describes the recent advances in methods for theoretical model quality assessment. PMID:19100336

Fidelis, Krzysztof

2009-01-01

256

Analysis of the effect of the bovine adenosine triphosphate-binding cassette transporter G2 single nucleotide polymorphism Y581S on transcellular transport of veterinary drugs using new cell culture models.  

PubMed

In commercial dairy production, the risk of drug residues and environmental pollutants in milk from ruminants has become an outstanding problem. One of the main determinants of active drug secretion into milk is the ATP-binding cassette transporter G2/breast cancer resistance protein (ABCG2/BCRP). It is located in several organs associated with drug absorption, metabolism, and excretion, and its expression is highly induced during lactation in the mammary gland of ruminants, mice, and humans. As a consequence, potential contamination of milk could expose suckling infants to xenotoxins. In cows, a SNP for this protein affecting quality and quantity of milk production has been described previously (Y581S). In this study, our main purpose was to determine whether this polymorphism has an effect on transcellular transport of veterinary drugs because this could alter substrate pharmacokinetics and milk residues. We stably expressed the wild-type bovine ABCG2 and the Y581S variant in Madin-Darby canine kidney epithelial cells (MDCKII) and MEF3.8 cell lines generating cell models in which the functionality of the bovine transporter could be addressed. Functional studies confirmed the greater functional activity in mitoxantrone accumulation assays for the Y581S variant with a greater relative V(MAX) value (P = 0.040) and showed for the first time that the Y581S variant presents greater transcellular transport of the model ABCG2 substrate nitrofurantoin (P = 0.024) and of 3 veterinary antibiotics, the fluoroquinolone agents enrofloxacin (P = 0.035), danofloxacin (P = 0.001), and difloxacin (P = 0.008), identified as new substrates of the bovine ABCG2. In addition, the inhibitory effect of the macrocyclic lactone ivermectin on the activity of wild-type bovine ABCG2 and the Y581S variant was also confirmed, showing a greater inhibitory potency on the wild-type protein at all the concentrations tested (5 ?M, P = 0.017; 10 ?M, P = 0.001; 25 ?M, P = 0.008; and 50 ?M, P = 0.003). Differential transport activity depending on the genotype together with the differential inhibition pattern might have clinical consequences, including changes in substrate pharmacokinetics (and subsequently pharmacodynamics) and more specifically, changes in secretion of ABCG2 substrates into milk, potentially implying important consequences to veterinary therapeutics. PMID:21821808

Real, R; González-Lobato, L; Baro, M F; Valbuena, S; de la Fuente, A; Prieto, J G; Alvarez, A I; Marques, M M; Merino, G

2011-12-01

257

Characterization of gender-specific bovine serum  

Microsoft Academic Search

Animal cell cultures generally require a nutrient-rich medium supplemented with animal serum. Adult bovine serum contains a variety of nutrients including inorganic minerals, vitamins, salts, proteins and lipids as well as growth factors that promote animal cell growth. To evaluate the potential use of gender-specific bovine serum (GSBS) for cell culture, the biochemical properties of male serum (MS), female serum

Jihoe Kim; Minsoo Kim; Sang-Soep Nahm; Dong-Mok Lee; Smritee Pokharel; Inho Choi

2011-01-01

258

A Single Point Mutation in Nonstructural Protein NS2 of Bovine Viral Diarrhea Virus Results in Temperature-Sensitive Attenuation of Viral Cytopathogenicity ?  

PubMed Central

For Bovine viral diarrhea virus (BVDV), the type species of the genus Pestivirus in the family Flaviviridae, cytopathogenic (cp) and noncytopathogenic (ncp) viruses are distinguished according to their effect on cultured cells. It has been established that cytopathogenicity of BVDV correlates with efficient production of viral nonstructural protein NS3 and with enhanced viral RNA synthesis. Here, we describe generation and characterization of a temperature-sensitive (ts) mutant of cp BVDV strain CP7, termed TS2.7. Infection of bovine cells with TS2.7 and the parent CP7 at 33°C resulted in efficient viral replication and a cytopathic effect. In contrast, the ability of TS2.7 to cause cytopathogenicity at 39.5°C was drastically reduced despite production of high titers of infectious virus. Further experiments, including nucleotide sequencing of the TS2.7 genome and reverse genetics, showed that a Y1338H substitution at residue 193 of NS2 resulted in the temperature-dependent attenuation of cytopathogenicity despite high levels of infectious virus production. Interestingly, TS2.7 and the reconstructed mutant CP7-Y1338H produced NS3 in addition to NS2-3 throughout infection. Compared to the parent CP7, NS2-3 processing was slightly decreased at both temperatures. Quantification of viral RNAs that were accumulated at 10 h postinfection demonstrated that attenuation of the cytopathogenicity of the ts mutants at 39.5°C correlated with reduced amounts of viral RNA, while the efficiency of viral RNA synthesis at 33°C was not affected. Taken together, the results of this study show that a mutation in BVDV NS2 attenuates viral RNA replication and suppresses viral cytopathogenicity at high temperature without altering NS3 expression and infectious virus production in a temperature-dependent manner. PMID:19776121

Pankraz, Alexander; Preis, Simone; Thiel, Heinz-Jürgen; Gallei, Andreas; Becher, Paul

2009-01-01

259

Molecular modeling and multispectroscopic studies of the interaction of mesalamine with bovine serum albumin.  

PubMed

The interaction of mesalamine (5-aminosalicylic acid (5-ASA)) with bovine serum albumin (BSA) was investigated by fluorescence quenching, absorption spectroscopy, circular dichroism (CD) techniques, and molecular docking. Thermodynamic parameters (?H<0 and ?S 0) indicated that the hydrogen bond and electrostatic forces played the major role in the binding of 5-ASA to BSA. The results of CD and UV-vis spectroscopy showed that the binding of this drug to BSA induces some conformational changes in BSA. Displacement experiments predicted that the binding of 5-ASA to BSA is located within domain III, Sudlows site 2, that these observations were substantiated by molecular docking studies. In addition, the docking result shows that the 5-ASA in its anionic form mainly interacts with Gln-416 residue through one hydrogen bond between H atom of 5-ASA anion and the adjacent O atom of the hydroxyl group of Gln-416. PMID:24076458

Shahabadi, Nahid; Fili, Soraya Moradi

2014-01-24

260

Molecular modeling and multispectroscopic studies of the interaction of mesalamine with bovine serum albumin  

NASA Astrophysics Data System (ADS)

The interaction of mesalamine (5-aminosalicylic acid (5-ASA)) with bovine serum albumin (BSA) was investigated by fluorescence quenching, absorption spectroscopy, circular dichroism (CD) techniques, and molecular docking. Thermodynamic parameters (?H < 0 and ?S 0) indicated that the hydrogen bond and electrostatic forces played the major role in the binding of 5-ASA to BSA. The results of CD and UV-vis spectroscopy showed that the binding of this drug to BSA induces some conformational changes in BSA. Displacement experiments predicted that the binding of 5-ASA to BSA is located within domain III, Sudlows site 2, that these observations were substantiated by molecular docking studies. In addition, the docking result shows that the 5-ASA in its anionic form mainly interacts with Gln-416 residue through one hydrogen bond between H atom of 5-ASA anion and the adjacent O atom of the hydroxyl group of Gln-416.

Shahabadi, Nahid; Fili, Soraya Moradi

2014-01-01

261

Multiple model approach - dealing wtih alignment ambiguities in protein modeling  

SciTech Connect

Sequence alignments for distantly homologous proteins are often ambiguous, which creates a weak link in structure prediction by homology. We address this problem by using several plausible alignments in a modeling procedure, obtaining many models of the target. All are subsequently evaluated by a threading algorithm. It is shown that this approach can identify best alignments and produce reasonable models, whose quality is now limited only by the extent of the structural similarity between the known and predicted protein. Using a similar approach the structure prediction for the oxidized dimer of S100A1 protein, for which the structure is not known, is presented. 24 refs., 2 figs., 2 tabs.

Pawlowski, K.; Bierzynski, A. [Institute of Biochemistry & Biophysics, Warszawa (Poland); Jaroszewski, L. [Warsaw Univ. (Poland); Godzik, A. [Scripps Research Institute, La Jolla, CA (United States)

1996-12-31

262

Structural modeling of snow flea antifreeze protein.  

PubMed

The glycine-rich antifreeze protein recently discovered in snow fleas exhibits strong freezing point depression activity without significantly changing the melting point of its solution (thermal hysteresis). BLAST searches did not detect any protein with significant similarity in current databases. Based on its circular dichroism spectrum, discontinuities in its tripeptide repeat pattern, and intramolecular disulfide bonding, a detailed theoretical model is proposed for the 6.5-kDa isoform. In the model, the 81-residue protein is organized into a bundle of six short polyproline type II helices connected (with one exception) by proline-containing turns. This structure forms two sheets of three parallel helices, oriented antiparallel to each other. The central helices are particularly rich in glycines that facilitate backbone carbonyl-amide hydrogen bonding to four neighboring helices. The modeled structure has similarities to polyglycine II proposed by Crick and Rich in 1955 and is a close match to the polyproline type II antiparallel sheet structure determined by Traub in 1969 for (Pro-Gly-Gly)(n). Whereas the latter two structures are formed by intermolecular interactions, the snow flea antifreeze is stabilized by intramolecular interactions between the helices facilitated by the regularly spaced turns and disulfide bonds. Like several other antifreeze proteins, this modeled protein is amphipathic with a putative hydrophobic ice-binding face. PMID:17158562

Lin, Feng-Hsu; Graham, Laurie A; Campbell, Robert L; Davies, Peter L

2007-03-01

263

Structural Modeling of Snow Flea Antifreeze Protein  

PubMed Central

The glycine-rich antifreeze protein recently discovered in snow fleas exhibits strong freezing point depression activity without significantly changing the melting point of its solution (thermal hysteresis). BLAST searches did not detect any protein with significant similarity in current databases. Based on its circular dichroism spectrum, discontinuities in its tripeptide repeat pattern, and intramolecular disulfide bonding, a detailed theoretical model is proposed for the 6.5-kDa isoform. In the model, the 81-residue protein is organized into a bundle of six short polyproline type II helices connected (with one exception) by proline-containing turns. This structure forms two sheets of three parallel helices, oriented antiparallel to each other. The central helices are particularly rich in glycines that facilitate backbone carbonyl-amide hydrogen bonding to four neighboring helices. The modeled structure has similarities to polyglycine II proposed by Crick and Rich in 1955 and is a close match to the polyproline type II antiparallel sheet structure determined by Traub in 1969 for (Pro-Gly-Gly)n. Whereas the latter two structures are formed by intermolecular interactions, the snow flea antifreeze is stabilized by intramolecular interactions between the helices facilitated by the regularly spaced turns and disulfide bonds. Like several other antifreeze proteins, this modeled protein is amphipathic with a putative hydrophobic ice-binding face. PMID:17158562

Lin, Feng-Hsu; Graham, Laurie A.; Campbell, Robert L.; Davies, Peter L.

2007-01-01

264

The NS5A Protein of Bovine Viral Diarrhea Virus Contains an Essential Zinc-Binding Site Similar to That of the Hepatitis C Virus NS5A Protein  

PubMed Central

The recent demonstration that the NS5A protein of hepatitis C virus (HCV) contains an unconventional zinc-binding site with the format Cx17CxCx20C and the presence of a similar sequence element in the NS5A proteins of members of the Pestivirus genus has led to the hypothesis that the NS5A protein of the pestivirus bovine viral diarrhea virus (BVDV) is a zinc-binding protein. A method for the expression and partial purification of BVDV NS5A was developed, and the partially purified protein was analyzed for zinc content by atomic absorption spectroscopy. BVDV NS5A was found to coordinate a single zinc atom per protein molecule. Mutation of any of the four cysteines of the predicted zinc-binding motif eliminated zinc coordination. Furthermore, analysis of mutations at these cysteine residues in the context of a BVDV replicon system indicated that these residues were absolutely essential for RNA replication. The recently determined crystal structure of the N-terminal zinc-binding domain of the HCV NS5A protein, combined with secondary structure predictions of the region surrounding the mapped BVDV zinc-binding region, indicates that the BVDV zinc-binding motif fits the general template Cx22CxCx24C and likely comprises a three-stranded antiparallel ?-sheet fold. These data highlight the similarities between the Hepacivirus and Pestivirus NS5A proteins and suggest that both proteins perform a not-yet-defined function in RNA replication that requires coordination of a single zinc atom. PMID:16840325

Tellinghuisen, Timothy L.; Paulson, Matthew S.; Rice, Charles M.

2006-01-01

265

Inhibitor binding changes domain mobility in the iron-sulfur protein of the mitochondrial bc1 complex from bovine heart  

PubMed Central

We have analyzed crystal structures of cytochrome bc1 complexes with electron transfer inhibitors bound to the ubiquinone binding pockets Qi and/or Qo in the cytochrome b subunit. The presence or absence of the Qi inhibitor antimycin A did not affect the binding of the Qo inhibitors. Different subtypes of Qo inhibitors had dramatically different effects on the mobility of the extramembrane domain of the iron–sulfur protein (ISP): Binding of 5-undecyl-6-hydroxy-4,7-dioxobenzothiazol and stigmatellin (subtype Qo–II and Qo–III, respectively) led to a fixation of the ISP domain on the surface of cytochrome b, whereas binding of myxothiazol and methoxyacrylate-stilbene (subtype Qo–I) favored release of this domain. The native structure has an empty Qo pocket and is intermediate between these extremes. On the basis of these observations we propose a model of quinone oxidation in the bc1 complex, which incorporates fixed and loose states of the ISP as features important for electron transfer and, possibly, also proton transport. PMID:9653134

Kim, Hoeon; Xia, Di; Yu, Chang-An; Xia, Jia-Zhi; Kachurin, Anatoly M.; Zhang, Li; Yu, Linda; Deisenhofer, Johann

1998-01-01

266

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

267

Substitution of lysine with glutamic acid at position 193 in bovine CYP11A1 significantly affects protein oligomerization and solubility but not enzymatic activity.  

PubMed

CYP11A1, a mitochondrial cytochrome P450, catalyzes the conversion from cholesterol to pregnenolone, the crucial step in the steroid hormone biosynthesis of mammals. It was shown in prior investigations, that the putative F-G loop of this enzyme is involved in membrane attachment. We produced different bovine CYP11A1 variants by rational protein design and could show that a deletion of 20 amino acids comprising parts of the F-G loop results in an enzyme with a three-fold increased solubility, the highest solubility of a CYP11A1 variant obtained so far. Furthermore, a single amino acid mutation, K193E, could be identified which leads not only to a higher solubility of CYP11A1 as well as a 4-fold improved expression rate, but also lowers the oligomerization of the protein while its activity is only slightly decreased. Therefore, this mutant has many advantages for the biotechnological application of CYP11A1 and is an important step towards crystallization of this mitochondrial P450. PMID:20538078

Janocha, Simon; Bichet, Andreas; Zöllner, Andy; Bernhardt, Rita

2011-01-01

268

Protein Modelling: What Happened to the "Protein Structure Gap"?  

PubMed Central

Computational modeling and prediction of three-dimensional macromolecular structures and complexes from their sequence has been a long standing vision in structural biology as it holds the promise to bypass part of the laborious process of experimental structure solution. Over the last two decades, a paradigm shift has occurred: starting from a situation where the “structure knowledge gap” between the huge number of protein sequences and small number of known structures has hampered the widespread use of structure-based approaches in life science research, today some form of structural information – either experimental or computational – is available for the majority of amino acids encoded by common model organism genomes. Template based homology modeling techniques have matured to a point where they are now routinely used to complement experimental techniques. With the scientific focus of interest moving towards larger macromolecular complexes and dynamic networks of interactions, the integration of computational modeling methods with low-resolution experimental techniques allows studying large and complex molecular machines. Computational modeling and prediction techniques are still facing a number of challenges which hamper the more widespread use by the non-expert scientist. For example, it is often difficult to convey the underlying assumptions of a computational technique, as well as the expected accuracy and structural variability of a specific model. However, these aspects are crucial to understand the limitations of a model, and to decide which interpretations and conclusions can be supported. PMID:24010712

Schwede, Torsten

2013-01-01

269

Models of Crk Adaptor Proteins in Cancer  

PubMed Central

The Crk family of adaptor proteins (CrkI, CrkII, and CrkL), originally discovered as the oncogene fusion product, v-Crk, of the CT10 chicken retrovirus, lacks catalytic activity but engages with multiple signaling pathways through their SH2 and SH3 domains. Crk proteins link upstream tyrosine kinase and integrin-dependent signals to downstream effectors, acting as adaptors in diverse signaling pathways and cellular processes. Crk proteins are now recognized to play a role in the malignancy of many human cancers, stimulating renewed interest in their mechanism of action in cancer progression. The contribution of Crk signaling to malignancy has been predominantly studied in fibroblasts and in hematopoietic models and more recently in epithelial models. A mechanistic understanding of Crk proteins in cancer progression in vivo is still poorly understood in part due to the highly pleiotropic nature of Crk signaling. Recent advances in the structural organization of Crk domains, new roles in kinase regulation, and increased knowledge of the mechanisms and frequency of Crk overexpression in human cancers have provided an incentive for further study in in vivo models. An understanding of the mechanisms through which Crk proteins act as oncogenic drivers could have important implications in therapeutic targeting. PMID:23226572

Bell, Emily S.

2012-01-01

270

Adsorption of bovine serum albumin on to titanium powder  

Microsoft Academic Search

The adsorption of bovine serum albumin (BSA) on to titanium powder has been studied as a function of protein concentration and pH, and in the presence of calcium and phosphate ions. Isotherm data have shown that the adsorption process does not follow the Langmuir model (inflection points). The maximum adsorption (Admax) at pH 6.8 is 1.13 ± 0.21 mg m?2.

Diana T. Hughes Wassell; Graham Embery

1996-01-01

271

Denaturation of bovine spleen galectin-1 in guanidine hydrochloride and fluoroalcohols: structural characterization and implications for protein folding.  

PubMed

Guanidine hydrochloride (GdnHCl)-induced unfolding of bovine spleen galectin-1 (Gal-1) exhibits three-state mechanism involving exclusive, structured tertiary monomer in 0.5 M GdnHCl. Gal-1 has one tryptophan residue (Trp 68) per subunit. Phosphorescence spectra of both Gal-1 dimer and intermediate monomer at 77 K show single (0,0) band at 405.6 nm, characteristics of free tryptophan environment as of N-acetyl-l-tryptophanamide. Unfolded Gal-1 in 4 M GdnHCl gives (0,0) band at longer wavelength (408.6 nm) signifying that Trp 68 is less solvent exposed, being localized in an environment of residual structure. Trifluoroethanol (TFE)- and hexafluoroisopropanol (HFIP)-induced structural changes of Gal-1 dimer and monomer have been investigated by far-UV CD and FTIR. CD results show reversible nature of ?-sheet to ?-helix transition, with 30% helix in 80% TFE or 40% HFIP for Gal-1 dimer. Temperature-dependent studies show that induced helix entails reduced thermal stability. FTIR results reveal partial ?-sheet to ?-helix conversion but with quantitative yield. At intermediate TFE concentration, both Gal-1 dimer and monomer aggregate as evidenced by FTIR band at ?1617/cm, however, the onset of aggregation and stability of aggregates for monomer differ from those of dimer. The results may provide important insights into perturbed folding problem of Gal-1. PMID:24037640

Mandal, Pritha; Molla, Anisur R; Mandal, Dipak K

2013-12-01

272

Mathematical analysis of a model for the growth of the bovine corpus luteum.  

PubMed

The corpus luteum (CL) is an ovarian tissue that grows in the wound space created by follicular rupture. It produces the progesterone needed in the uterus to maintain pregnancy. Rapid growth of the CL and progesterone transport to the uterus require angiogenesis, the creation of new blood vessels from pre-existing ones, a process which is regulated by proteins that include fibroblast growth factor 2 (FGF2). In this paper we develop a system of time-dependent ordinary differential equations to model CL growth. The dependent variables represent FGF2, endothelial cells (ECs), luteal cells, and stromal cells (like pericytes), by assuming that the CL volume is a continuum of the three cell types. We assume that if the CL volume exceeds that of the ovulated follicle, then growth is inhibited. This threshold volume partitions the system dynamics into two regimes, so that the model may be classified as a Filippov (piecewise smooth) system. We show that normal CL growth requires an appropriate balance between the growth rates of luteal and stromal cells. We investigate how angiogenesis influences CL growth by considering how the system dynamics depend on the dimensionless EC proliferation rate, [Formula: see text]. We find that weak (low [Formula: see text]) or strong (high [Formula: see text]) angiogenesis leads to 'pathological' CL growth, since the loss of CL constituents compromises progesterone production or delivery. However, for intermediate values of [Formula: see text], normal CL growth is predicted. The implications of these results for cow fertility are also discussed. For example, inadequate angiogenesis has been linked to infertility in dairy cows. PMID:24337679

Prokopiou, Sotiris A; Byrne, Helen M; Jeffrey, Mike R; Robinson, Robert S; Mann, George E; Owen, Markus R

2014-12-01

273

Annotation of novel transcripts putatively relevant for bovine fat metabolism.  

PubMed

Two bovine transcripts encoded by the interleukin-1 receptor-associated kinase 1 (IRAK1) gene and the locus LOC618944 predicted as similar to human chromosome 6 open reading frame 52 (C6orf52) gene had indicated divergent expression in bovine skeletal muscle containing different amount of intramuscular fat in a pilot screening experiment. However, for both loci any role in the regulation of energy or fat metabolism is not yet described. In this study, we validated and refined gene structure, screened for mRNA splice variants and analyzed the tissue-specific gene expression patterns of both loci as a prerequisite to elucidate their potential physiological function. Based on comparative sequence analysis, a new full-length gene model for the bovine IRAK1 gene was developed and confirmed experimentally. Expression of IRAK1 mRNA was found in a variety of tissues, and a splice variant was identified in skeletal muscle caused by an in-frame deleted segment of 210 bp affecting regions of intrinsic disorder in the respective protein. For the locus LOC618944, our data contributed to a revised gene model and its assignment to BTA23 (bovine chromosome 23) on the current bovine genome assembly supported by comparative similarity analysis between the bovine and human genomes and experimental data. Furthermore, we identified several splice variants in mammary gland, fat and skeletal muscle tissue and detected a highly similar processed pseudogene on BTA26. All transcript variants of LOC618944 detected in the analyzed tissues represent noncoding RNAs. For both loci, our results suggest yet undetected physiological functions in tissues relevant for fat or energy metabolism in cattle. PMID:20127178

Eberlein, Annett; Kalbe, Claudia; Goldammer, Tom; Brunner, Ronald M; Kuehn, Christa; Weikard, Rosemarie

2011-06-01

274

Fold assessment for comparative protein structure modeling  

PubMed Central

Accurate and automated assessment of both geometrical errors and incompleteness of comparative protein structure models is necessary for an adequate use of the models. Here, we describe a composite score for discriminating between models with the correct and incorrect fold. To find an accurate composite score, we designed and applied a genetic algorithm method that searched for a most informative subset of 21 input model features as well as their optimized nonlinear transformation into the composite score. The 21 input features included various statistical potential scores, stereochemistry quality descriptors, sequence alignment scores, geometrical descriptors, and measures of protein packing. The optimized composite score was found to depend on (1) a statistical potential z-score for residue accessibilities and distances, (2) model compactness, and (3) percentage sequence identity of the alignment used to build the model. The accuracy of the composite score was compared with the accuracy of assessment by single and combined features as well as by other commonly used assessment methods. The testing set was representative of models produced by automated comparative modeling on a genomic scale. The composite score performed better than any other tested score in terms of the maximum correct classification rate (i.e., 3.3% false positives and 2.5% false negatives) as well as the sensitivity and specificity across the whole range of thresholds. The composite score was implemented in our program MODELLER-8 and was used to assess models in the MODBASE database that contains comparative models for domains in approximately 1.3 million protein sequences. PMID:17905832

Melo, Francisco; Sali, Andrej

2007-01-01

275

Evaluation of Multiplexed Foot-and-Mouth Disease Nonstructural Protein Antibody Assay Against Standardized Bovine Serum Panel.  

National Technical Information Service (NTIS)

Liquid array technology has previously been used to show proof-of-principle of a multiplexed non structural protein serological assay to differentiate foot-and-mouth infected and vaccinated animals. The current multiplexed assay consists of synthetically ...

A. Clavijo, J. Perkins, S. Parida

2007-01-01

276

Evaluation of Multiplexed Foot-and-Mouth Disease Nonstructural Protein Antibody Assay Against Standardized Bovine Serum Panel  

Microsoft Academic Search

Liquid array technology has previously been used to show proof-of-principle of a multiplexed non structural protein serological assay to differentiate foot-and-mouth infected and vaccinated animals. The current multiplexed assay consists of synthetically produced peptide signatures 3A, 3B and 3D and recombinant protein signature 3ABC in combination with four controls. To determine diagnostic specificity of each signature in the multiplex, the

J Perkins; S Parida; A Clavijo

2007-01-01

277

Is the pre-Tg DSC endotherm observed with solid state proteins associated with the protein internal dynamics? Investigation of bovine serum albumin by solid state hydrogen/deuterium exchange.  

PubMed

DSC thermograms of solid state pure proteins often show a distinct endotherm at a temperature far below the glass transition temperature of the system (Tg). We hypothesized this endotherm represents enthalpy recovery associated with an internal mobility transition of the protein molecule. Although the existence of an internal transition has been postulated, whether this endotherm is associated with such a transition has not previously been discussed. The purpose of this study was to investigate the origin of the pre-Tg endotherm in lyophilized bovine serum albumin (BSA). Due to strong glass behavior, the system Tg was determined by extrapolating Tg data of disaccharide/BSA formulations to zero saccharide. A small pre-Tg endotherm around 40-60 °C was observed in amorphous BSA equilibrated at 11%RH. The apparent activation energy suggested the endotherm was "?-mobility"-related. A solid state hydrogen/deuterium exchange study using FTIR was conducted over a temperature range spanning the endotherm. We found a fast phase, followed by essentially a plateau level which is highly temperature dependent in the 40-60 °C range, suggesting enhanced internal protein motion as the system passes through the temperature range of the endotherm. These results suggest the pre-Tg endotherm is associated with a protein internal mobility transition. PMID:23669417

Mizuno, Masayasu; Pikal, Michael J

2013-10-01

278

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

279

Coordination modes of tyrosinate-ligated catalase-type heme enzymes: Magnetic circular dichroism studies of Plexaura homomalla allene oxide synthase, Mycobacterium avium ssp . Paratuberculosis protein-2744c, and bovine liver catalase in their ferric and ferrous states  

Microsoft Academic Search

Bovine liver catalase (BLC), catalase-related allene oxide synthase (cAOS) from Plexaura homomalla, and a recently isolated protein from the cattle pathogen Mycobacterium avium ssp. paratuberculosis (MAP-2744c (MAP)) are all tyrosinate-ligated heme enzymes whose crystal structures have been reported. cAOS and MAP have low (<20%) sequence similarity to, and significantly different catalytic functions from, BLC. cAOS transforms 8R-hydroperoxy-eicosatetraenoic acid to an

D. M. Indika Bandara; Masanori Sono; Grant S. Bruce; Alan R. Brash; John H. Dawson

280

Conformational changes involved in thermal aggregation processes of bovine serum albumin  

Microsoft Academic Search

We report a kinetic study on thermal aggregation process of the model protein bovine serum albumin (BSA) in low concentration regime. Aim of this study is to provide information on relationship between conformational changes and initial step of aggregation. The experimental approach is based on steady-state fluorescence spectra of the two tryptophans located in two different domains, in way to

Valeria Militello; Valeria Vetri; Maurizio Leone

2003-01-01

281

Enantiomeric separation by ultrafiltration: complexation mechanism of tryptophan analogs to bovine serum albumin  

Microsoft Academic Search

The separation of racemic tryptophan and its analogs can be performed by ultrafiltration in a solution system using bovine serum albumin (BSA) as a complexing agent. Three models have been tested to simulate the complexation mechanism of tryptophan and kynurenine enantiomers to BSA protein. In the pH range from 7 to 11, the most probable complexation mechanism was a competitive

F. Garnier; J. Randon; J. L. Rocca

1999-01-01

282

Effect of retinoic acid on protein synthesis by foetal bovine chondrocytes in high-density culture: down-regulation of the glucose-regulated protein, GRP-78, and type II collagen.  

PubMed Central

The effect of 0.1-10 microM retinoic acid (RA) on foetal bovine chondrocytes was investigated in high-density cultures (0.6 x 10(6) cells/cm2). After 5 days of culture in ascorbate-free medium, control chondrocytes presented a typical rounded shape and synthesized type II, IX, XI and III collagens. After RA treatment on days 2-5 of culture, the cells exhibited a fibroblast-like shape and decreased synthesis of total protein (48%) and pepsinresistant proteins (60%) as determined by [35S]methionine labelling. Addition of RA was not followed by the expression of type I collagen, but induced quantitative changes in the synthesis of cartilage-specific collagens (II, IX and XI) as measured by direct autoradiography of the corresponding bands after SDS/PAGE. The main change was in type II collagen synthesis, with a 80% decrease in the cell-layer fraction and a 89% decrease in culture-medium fraction; inhibition of type IX and XI collagen synthesis was limited to 25 and 31% respectively. Modifications to intracellular proteins induced by RA were determined by using two-dimensional electrophoresis associated with a computerized imaging system. Synthesis of one of the more abundant proteins (pI 4.8; 78 kDa) was decreased by 75% after RA treatment. This protein was characterized by micro-sequencing as the glucose-regulated protein 78 (GRP 78). It was reported previously to bind denatured collagen and mutated type I procollagen molecule and to function as a molecular chaperone for collagen molecules. It remains to demonstrate whether the parallel down-regulation of GRP 78 and type II collagen observed here corresponds to a co-ordinate regulation of these two proteins. Images Figure 1 Figure 2 Figure 3 PMID:7832751

Freyria, A M; Ronzière, M C; Boutillon, M M; Herbage, D

1995-01-01

283

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

284

Molecular modelling indicates that the pathological conformations of prion proteins might be beta-helical.  

PubMed Central

Creutzfeldt-Jakob disease, kuru, scrapie and bovine spongiform encephalopathy are diseases of the mammalian central nervous system that involve the conversion of a cellular protein into an insoluble extracellular isoform. Spectroscopic studies have shown that the precursor protein contains mainly alpha-helical and random-coil conformations, whereas the prion isoform is largely in the beta conformation. The pathogenic prion is resistant to denaturation and protease digestion and can promote the conversion of the precursor protein to the pathogenic form. These properties have yet to be explained in terms of the structural conformations of the proteins. In the present study, molecular modelling showed that prion proteins could adopt the beta-helical conformation, which has been established for a number of fibrous proteins and has been suggested previously as the basis of amyloid fibrils. The beta-helical conformation provides explanations for the biophysical and biochemical stability of prions, their ability to form templates for the transmission of pathological conformation, and the existence of phenotypical strains of the prion diseases. PMID:10510313

Downing, D T; Lazo, N D

1999-01-01

285

Bovine spongiform encephalopathy associated insertion/deletion polymorphisms of the prion protein gene in the four beef cattle breeds from North China.  

PubMed

Two insertion/deletion (indel) polymorphisms of the prion protein gene (PRNP), a 23-bp indel in the putative promoter region and a 12-bp indel within intron I, are associated with the susceptibility to bovine spongiform encephalopathy (BSE) in cattle. In the present study, the polymorphism frequencies of the two indels in four main beef cattle breeds (Hereford, Simmental, Black Angus, and Mongolian) from North China were studied. The results showed that the frequencies of deletion genotypes and alleles of 23- and 12-bp indels were lower, whereas the frequencies of insertion genotypes and alleles of the two indels were higher in Mongolian cattle than in the other three cattle breeds. In Mongolian cattle, the 23-bp insertion / 12-bp insertion was the major haplotype, whereas in Hereford, Simmental, and Black Angus cattle, the 23-bp deletion / 12-bp deletion was the major haplotype. These results demonstrated that Mongolian cattle could be more resistant to BSE, compared with the other three cattle breeds, because of its relatively low frequencies of deletion genotypes and alleles of 23- and 12-bp indel polymorphisms. Thus, this race could be important for selective breeding to improve resistance against BSE in this area. PMID:21923635

Zhu, Xiang-Yuan; Feng, Fu-Ying; Xue, Su-Yuan; Hou, Ting; Liu, Hui-Rong

2011-10-01

286

Time-of-flight neutron diffraction study of bovine [gamma]-chymotrypsin at the Protein Crystallography Station  

SciTech Connect

The overarching goal of this research project is to determine, for a subset of proteins, exact hydrogen positions using neutron diffraction, thereby improving H-atom placement in proteins so that they may be better used in various computational methods that are critically dependent upon said placement. In order to be considered applicable for neutron diffraction studies, the protein of choice must be amenable to ultrahigh-resolution X-ray crystallography, be able to form large crystals (1 mm{sup 3} or greater) and have a modestly sized unit cell (no dimension longer than 100 {angstrom}). As such, {gamma}-chymotrypsin is a perfect candidate for neutron diffraction. To understand and probe the role of specific active-site residues and hydrogen-bonding patterns in {gamma}-chymotrypsin, neutron diffraction studies were initiated at the Protein Crystallography Station (PCS) at Los Alamos Neutron Science Center (LANSCE). A large single crystal was subjected to H/D exchange prior to data collection. Time-of-flight neutron diffraction data were collected to 2.0 {angstrom} resolution at the PCS with 85% completeness. Here, the first time-of-flight neutron data collection from {gamma}-chymotrypsin is reported.

Lazar, Louis M.; Fisher, S. Zoe; Moulin, Aaron G.; Kovalevsky, Andrey; Novak, Walter R.P.; Langan, Paul; Petsko, Gregory A.; Ringe, Dagmar (Brandeis); (LANL)

2012-02-06

287

Characterization of cholate-solubilized dopamine D/sub 2/ receptors coupled to guanine nucleotide regulatory protein in bovine striatum  

SciTech Connect

Considerable evidence suggest that membrane-bound dopamine D/sub 2/ receptor exhibits high and low affinity states for agonists and the high affinity sites are coupled to guanine nucleotide regulatory protein (G-protein). Since discrepant findings were reported for the retention of functional dopamine - G-protein complexes following solubilization, they carried out the detailed characterization of cholic acid - NaCl solubilized receptors using (/sup 3/H)-spiperone, (/sup 3/H)-N-propylnorapomorphine (NPA) and (/sup 35/S)-GTP..gamma..S binding. The solubilized receptor not only retained all of its pharmacological specificities, but also exhibited high affinity (/sup 35/S)-GTP..gamma..S and (/sup 3/H)-NPA binding and the agonist (NPA) interaction was completely abolished by 100 ..mu..M Gpp(NH)p. Further, Gpp(NH)p significantly reduced the affinity of agonists in (/sup 3/H)-spiperone/agonists competition studies. When the solubilized preparation was subjected to DEAE-chromatography, the receptors eluted in two distinct peaks. Several evidence indicate that the two peaks represent different subpopulation of dopamine receptors with reference to high affinity agonist binding and sensitivity to guanine nucleotides. These findings suggest that the D/sub 2/ receptor-G-protein complex is retained following solubilization and the agonist high affinity component can be resolved chromatographically.

Kazmi, S.M.I.; Ramwani, J.; Mishra, R.K.

1986-03-05

288

Divergent antibody isotype responses induced in mice by systemic exposure to proteins: a comparison of ovalbumin with bovine serum albumin  

Microsoft Academic Search

Whereas many foreign proteins are immunogenic, only a proportion is associated commonly with allergy, having the potential to induce the quality of immune response necessary for IgE antibody production and the development of immediate type hypersensitivity reactions in the gastrointestinal and\\/or respiratory tracts. In the context of toxicological evaluations there is a need to identify those properties that confer on

R. J Dearman; H Caddick; D. A Basketter; I Kimber

2000-01-01

289

Mitogen-activated protein kinase signaling in bovine articular chondrocytes in response to fluid flow does not require calcium mobilization  

Microsoft Academic Search

In the present study, the role of mitogen-activated protein kinases (MAPKs) in chondrocyte mechanotransduction was investigated. We hypothesized that MAPKs participate in fluid flow-induced chondrocyte mechanotransduction. To test our hypothesis, we studied cultured chondrocytes subjected to a well-defined mechanical stimulus generated with a laminar flow chamber. The extracellular signal-regulated kinases 1 and 2 (ERK1\\/2) were activated 1.6–3-fold after 5–15min of

Clark T. Hung; D. Ross Henshaw; Christopher C.-B. Wang; Robert L. Mauck; Frank Raia; Glyn Palmer; Pen-Hsiu Grace Chao; Van C. Mow; Anthony Ratcliffe; Wilmot B. Valhmu

2000-01-01

290

Encapsulation of bovine serum albumin within ?-cyclodextrin\\/gelatin-based polymeric hydrogel for controlled protein drug release  

Microsoft Academic Search

Novel biodegradable pH-responsive polymeric hydrogels based on ?-cyclodextrin (?-CD) and gelatin (G) were prepared for controlled protein drug delivery studies. The polymeric hydrogels, G-g-poly(GMA) were obtained by grafting copolymerization of glycidyl methacrylate monomer (GMA) onto gelatin in presence of ammonium ceric nitrate (CAN) as initiator. Immobilization of ?-CD onto G-g-poly(GMA) hydrogel was then carried out. The prepared polymeric hydrogels were

A. A. Haroun; N. R. El-Halawany

2010-01-01

291

Protein Simulation Data in the Relational Model  

PubMed Central

High performance computing is leading to unprecedented volumes of data. Relational databases offer a robust and scalable model for storing and analyzing scientific data. However, these features do not come without a cost—significant design effort is required to build a functional and efficient repository. Modeling protein simulation data in a relational database presents several challenges: the data captured from individual simulations are large, multi-dimensional, and must integrate with both simulation software and external data sites. Here we present the dimensional design and relational implementation of a comprehensive data warehouse for storing and analyzing molecular dynamics simulations using SQL Server. PMID:23204646

Simms, Andrew M.; Daggett, Valerie

2011-01-01

292

Detection of silver protein complex injections in the bovine udder using x-ray fluorescence: A preliminary investigation  

PubMed Central

Abstract To determine the feasibility of using x-ray fluorescence (XRF) to detect the presence of silver in the mammary gland of dairy cows injected with mild silver protein suspension. The XRF spectroscopy was conducted on cadaver udders with and without mild silver protein injected. Spectral analysis was performed in order to determine the amplitude of the silver K-alpha peak that was detected. By comparing the amplitude of the K-alpha peak to the background, a minimum time of collection was determined, as a measure of the time required to observe a silver signal that is significantly non-zero. The minimum detection time required for evidence of injected silver suspension was calculated to be 2.8 ± 0.2 s. Even with an additional requirement that the net signal exceed 50 counts, the clear indication of the presence of silver will be observed within 4 min of interrogation. X-ray fluorescence spectroscopy was shown to be a viable method for the detection of injected silver protein in cadaver mammary glands of dairy cows. While these findings are promising, further studies must be conducted to investigate the time dependence of the silver signal when diffusion, absorption, and redistribution are involved, under conditions that better mimic those encountered at an exhibition. This technique, used in conjunction with screening ultrasound examinations, has the potential to confirm sites of injection of silver compounds. PMID:15971676

2005-01-01

293

Inlet protein aggregation: a new mechanism for lubricating film formation with model synovial fluids  

PubMed Central

This paper reports a fundamental study of lubricant film formation with model synovial fluid components (proteins) and bovine serum (BS). The objective was to investigate the role of proteins in the lubrication process. Film thickness was measured by optical interferometry in a ball-on-disc device (mean speed range of 2–60?mm/s). A commercial cobalt–chromium (CoCrMo) metal femoral head was used as the stationary component. The results for BS showed complex time-dependent behaviour, which was not representative of a simple fluid. After a few minutes sliding BS formed a thin adherent film of 10–20?nm, which was attributed to protein absorbance at the surface. This layer was augmented by a hydrodynamic film, which often increased at slow speeds. At the end of the test deposited surface layers of 20–50?nm were measured. Imaging of the contact showed that at slow speeds an apparent ‘phase boundary’ formed in the inlet just in front of the Hertzian zone. This was associated with the formation of a reservoir of high-viscosity material that periodically moved through the contact forming a much thicker film. The study shows that proteins play an important role in the film-forming process and current lubrication models do not capture these mechanisms. PMID:21870377

Fan, J; Myant, C W; Underwood, R; Cann, P M; Hart, A

2011-01-01

294

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

295

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

296

Ca(2+) signaling mechanisms in bovine adrenal chromaffin cells.  

PubMed

Calcium (Ca(2+)) is a crucial intracellular messenger in physiological aspects of cell signaling. Adrenal chromaffin cells are the secretory cells from the adrenal gland medulla that secrete catecholamines, which include epinephrine and norepinephrine important in the 'fight or flight' response. Bovine adrenal chromaffin cells have long been used as an important model for secretion -(exocytosis) not only due to their importance in the short-term stress response, but also as a neuroendocrine model of neurotransmtter release, as they have all the same exocytotic proteins as neurons but are easier to prepare, culture and use in functional assays. The components of the Ca(2+) signal transduction cascade and it role in secretion has been extensively characterized in bovine adrenal chromaffin cells. The Ca(2+) sources, signaling molecules and how this relates to the short-term stress response are reviewed in this book chapter in an endeavor to generally -overview these mechanisms in a concise and uncomplicated manner. PMID:22453973

Weiss, Jamie L

2012-01-01

297

Characterisation of the conformational and quaternary structure-dependent heparin-binding region of bovine seminal plasma protein PDC-109.  

PubMed

PDC-109, the major heparin-binding protein of bull seminal plasma, binds to sperm choline lipids at ejaculation and modulates capacitation mediated by heparin. Affinity chromatography on heparin-Sepharose showed that polydisperse, but not monomeric, PDC-109 displayed heparin-binding capability. We sought to characterise the surface topology of the quaternary structure-dependent heparin-binding region of PDC-109 by comparing the arginine- and lysine-selective chemical modification patterns of the free and the heparin-bound protein. A combination of reversed-phase peptide mapping of endoproteinase Lys-C-digested PDC-109 derivatives and mass spectrometry was employed to identify modified and heparin-protected residues. PDC-109 contains two tandemly arranged fibronectin type II domains (a, Cys24-Cys61; b, Cys69-Cys109). The results show that six basic residues (Lys34, Arg57, Lys59, Arg64, Lys68, and Arg104) were shielded from reaction with acetic anhydride and 1,2-cyclohexanedione in heparin-bound PDC-109 oligomers. In the 1H-NMR solution structures of single fibronectin type II domains, residues topologically equivalent to PDC-109 Arg57 (Arg104) and Lys59 lay around beta-strand D on the same face of the domain. In full-length PDC-109, Arg64 and Lys68 are both located in the intervening polypeptide between domains a and b. Our data suggest possible quaternary structure arrangements of PDC-109 molecules to form a heparin-binding oligomer. PMID:10050771

Calvete, J J; Campanero-Rhodes, M A; Raida, M; Sanz, L

1999-02-12

298

Bovine Leukemia Virus SU Protein Interacts with Zinc, and Mutations within Two Interacting Regions Differently Affect Viral Fusion and Infectivity In Vivo  

PubMed Central

Bovine leukemia virus (BLV) and human T-cell lymphotropic virus type 1 (HTLV-1) belong to the genus of deltaretroviruses. Their entry into the host cell is supposed to be mediated by interactions of the extracellular (SU) envelope glycoproteins with cellular receptors. To gain insight into the mechanisms governing this process, we investigated the ability of SU proteins to interact with specific ligands. In particular, by affinity chromatography, we have shown that BLV SU protein specifically interacted with zinc ions. To identify the protein domains involved in binding, 16 peptides distributed along the sequence were tested. Two of them appeared to be able to interact with zinc. To unravel the role of these SU regions in the biology of the virus, mutations were introduced into the env gene of a BLV molecular clone in order to modify residues potentially interacting with zinc. The fusogenic capacity of envelope mutated within the first zinc-binding region (104 to 123) was completely abolished. Furthermore, the integrity of this domain was also required for in vivo infectivity. In contrast, mutations within the second zinc-binding region (218 to 237) did not hamper the fusogenic capacity; indeed, the syncytia were even larger. In sheep, mutations in region 218 to 237 did not alter infectivity or viral spread. Finally, we demonstrated that the envelope of the related HTLV-1 was also able to bind zinc. Interestingly, zinc ions were found to be associated with the receptor-binding domain (RBD) of Friend murine leukemia virus (Fr-MLV) SU glycoprotein, further supporting their relevance in SU structure. Based on the sequence similarities shared with the Fr-MLV RBD, whose three-dimensional structure has been experimentally determined, we located the BLV zinc-binding peptide 104-123 on the opposite side of the potential receptor-binding surface. This observation supports the hypothesis that zinc ions could mediate interactions of the SU RBD either with the C-terminal part of SU, thereby contributing to the SU structural integrity, or with a partner(s) different from the receptor. PMID:12134000

Gatot, Jean-Stephane; Callebaut, Isabelle; Van Lint, Carine; Demonte, Dominique; Kerkhofs, Pierre; Portetelle, Daniel; Burny, Arsene; Willems, Luc; Kettmann, Richard

2002-01-01

299

Crystal structure of the cytochrome bc⁠complex from bovine heart mitochondria  

Microsoft Academic Search

On the basis of x-ray diffraction data to a resolution of 2.9 angstroms, atomic models of most protein components of the bovine cytochrome bc⁠complex were built, including core 1, core 2, cytochrome b, subunit 6, and subunit 7, a carboxyl-terminal fragment of cytochrome câ, and an amino-terminal fragment of the iron-sulfur protein. The positions of the four iron centers

Di Xia; Hoeon Kim; J. Deisenhofer; Li Zhang

1997-01-01

300

PreImplantation factor (PIF) detection in maternal circulation in early pregnancy correlates with live birth (bovine model)  

PubMed Central

Background Early identification of viable pregnancy is paramount for successful reproduction. Detection of specific signals from pre-implantation viable embryos in normal pregnancy circulation would indicate initiation of embryo-maternal interaction and create a continuum to accurately reflect embryo/fetal well-being post-implantation. Viable mammalian embryos secrete PreImplantation Factor (PIF), a biomarker which plays key, multi-targeted roles to promote implantation, trophoblast invasion and modulate maternal innate and adaptive immunity toward acceptance. Anti-PIF monoclonal antibody (mAb-based chemiluminescent ELISA) accurately detects PIF in singly cultured embryos media and its increased levels correlate with embryo development up to the blastocyst stage. Herein reported that PIF levels (ELISA) in early maternal serum correlate with pregnancy outcome. Methods Artificially inseminated (AI) blind-coded Angus cattle (N?=?21-23) serum samples (day10,15 & 20 post-AI) with known calf birth were blindly tested, using both non-pregnant heifers (N?=?30) and steer serum as negative controls. Assay properties and anti-PIF monoclonal antibody specificity were determined by examining linearity, spike and recovery experiments and testing the antibody against 234 different circulating proteins by microarray. Endogenous PIF was detected using <3 kDa filter separation followed by anti-PIF mAb-based affinity chromatography and confirmed by ELISA and HPLC. PIF expression was established in placenta using anti-PIF mAb-based IHC. Results PIF detects viable pregnancy at day 10 post-AI with 91.3% sensitivity, reaching 100% by day 20 and correlating with live calf birth. All non-pregnant samples were PIF negative. PIF level in pregnant samples was a stringent 3?+?SD higher as compared to heifers and steer sera. Assay is linear and spike and recovery data demonstrates lack of serum interference. Anti-PIF mAb is specific and does not interact with circulating proteins. Anti-PIF based affinity purification demonstrates that endogenous PIF is what ELISA detects. The early bovine placenta expresses PIF in the trophoblast layer. Conclusion Data herein documents that PIF is a specific, reliable embryo-derived biomarker conveniently detectable in early maternal circulation. PIF ELISA emerges as practical tool to detect viable early pregnancy from day 20 post-AI. PMID:24238492

2013-01-01

301

Detection of bovine spongiform encephalopathy, ovine scrapie prion-related protein (PrPSc) and normal PrPc by monoclonal antibodies raised to copper-refolded prion protein.  

PubMed Central

Prion-related protein (PrP) is a glycosylphosphatidylinositol-linked cell-surface protein expressed by a wide variety of cells, including those of the nervous system and the immune system. Several functions of normal cellular PrP (PrPc) have been proposed that may be associated with the capacity of this protein to bind copper. In the present study, we describe the generation of a panel of monoclonal antibodies raised to copper-refolded PrP, which may be used to analyse the normal and disease-associated forms of this protein. The anti-PrP monoclonal antibodies were reactive by Western blot and ELISA with recombinant murine PrPc refolded in the presence or absence of either copper or manganese, and with the disease-susceptible allelic form V136R154Q171 ('VRQ'; where single-letter amino-acid notation has been used) and disease-resistant allelic form A136R154R171 ('ARR') of recombinant ovine PrPc. FACS analysis of lymphoid cells using these monoclonal antibodies showed that wild-type non-activated mouse lymphocytes expressed little, if any, PrPc. These monoclonal antibodies were shown to react with the unglycosylated and monoglycosylated forms of PrPSc (abnormal disease-specific conformation of PrP) in prion-infected tissue samples from all of the different species tested by Western blot. In addition, this analysis allowed one to make a distinction between bovine spongiform encephalopathy ('BSE') and scrapie PrPSc) isolates from experimentally infected sheep on the basis of their different electrophoretic mobilities. PMID:12429022

Thackray, Alana M; Madec, Jean-Yves; Wong, Edmond; Morgan-Warren, Robert; Brown, David R; Baron, Thierry; Bujdoso, Raymond

2003-01-01

302

Targeting the Human Papillomavirus E6 and E7 Oncogenes through Expression of the Bovine Papillomavirus Type 1 E2 Protein Stimulates Cellular Motility?†  

PubMed Central

Expression of the high-risk human papillomavirus (HPV) E6 and E7 oncogenes is essential for the initiation and maintenance of cervical cancer. The repression of both was previously shown to result in activation of their respective tumor suppressor targets, p53 and pRb, and subsequent senescence induction in cervical cancer cells. Consequently, viral oncogene suppression is a promising approach for the treatment of HPV-positive tumors. One well-established method of E6/E7 repression involves the reexpression of the viral E2 protein which is usually deleted in HPV-positive cancer cells. Here, we show that, surprisingly, bovine papillomavirus type 1 (BPV1) E2 but not RNA interference-mediated E6/E7 repression in HPV-positive cervical cancer cells stimulates cellular motility and invasion. Migration correlated with the dynamic formation of cellular protrusions and was dependent upon cell-to-cell contact. While E2-expressing migratory cells were senescent, migration was not a general feature of cellular senescence or cell cycle arrest and was specifically observed in HPV-positive cervical cancer cells. Interestingly, E2-expressing cells not only were themselves motile but also conferred increased motility to admixed HeLa cervical cancer cells. Together, our data suggest that repression of the viral oncogenes by E2 stimulates the motility of E6/E7-targeted cells as well as adjacent nontargeted cancer cells, thus raising the possibility that E2 expression may unfavorably increase the local invasiveness of HPV-positive tumors. PMID:21835799

Morrison, Monique A.; Morreale, Richard J.; Akunuru, Shailaja; Kofron, Matthew; Zheng, Yi; Wells, Susanne I.

2011-01-01

303

Silica vesicles as nanocarriers and adjuvants for generating both antibody and T-cell mediated immune resposes to Bovine Viral Diarrhoea Virus E2 protein.  

PubMed

Bovine Viral Diarrhoea Virus (BVDV) is widely distributed in cattle industries and causes significant economic losses worldwide annually. A limiting factor in the development of subunit vaccines for BVDV is the need to elicit both antibody and T-cell-mediated immunity as well as addressing the toxicity of adjuvants. In this study, we have prepared novel silica vesicles (SV) as the new generation antigen carriers and adjuvants. With small particle size of 50 nm, thin wall (?6 nm), large cavity (?40 nm) and large entrance size (5.9 nm for SV-100 and 16 nm for SV-140), the SV showed high loading capacity (? 250 ?g/mg) and controlled release of codon-optimised E2 (oE2) protein, a major immunogenic determinant of BVDV. The in vivo functionality of the system was validated in mice immunisation trials comparing oE2 plus Quil A (50 ?g of oE2 plus 10 ?g of Quil A, a conventional adjuvant) to the oE2/SV-140 (50 ?g of oE2 adsorbed to 250 ?g of SV-140) or oE2/SV-140 together with 10 ?g of Quil A. Compared to the oE2 plus Quil A, which generated BVDV specific antibody responses at a titre of 10(4), the oE2/SV-140 group induced a 10 times higher antibody response. In addition, the cell-mediated response, which is essential to recognise and eliminate the invading pathogens, was also found to be higher [1954-2628 spot forming units (SFU)/million cells] in mice immunised with oE2/SV-140 in comparison to oE2 plus Quil A (512-1369 SFU/million cells). Our study has demonstrated that SV can be used as the next-generation nanocarriers and adjuvants for enhanced veterinary vaccine delivery. PMID:25239045

Mody, Karishma T; Mahony, Donna; Zhang, Jun; Cavallaro, Antonino S; Zhang, Bing; Popat, Amirali; Mahony, Timothy J; Yu, Chengzhong; Mitter, Neena

2014-12-01

304

The skin compatibility of distilled tall oils: evaluation with the bovine udder skin in vitro model system.  

PubMed

Distilled tall oil (DTO) is a natural product, often added as an emulsifying ingredient in cutting fluids used as lubricants and coolants in metal working. The in vitro model used to test the skin compatibility of these substances, was the isolated perfused ex vivo bovine udder skin (BUS) model. After three exposure periods (0.5, 1, and 5 hours), cytotoxic effects were determined by using the 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and tissue levels of the pre-inflammatory mediator prostaglandin E2 (PGE2) in treated whole skin biopsies were assessed by using an enzyme immunoassay. The BUS standard study design, involving a single application, was previously developed to investigate the skin irritation potential of cosmetics and chemicals. In the current study, four different batches of undiluted DTO, and tall oil fatty acids as a reference compound, were applied both singly and repeatedly (three times), under open conditions which were in line with the potential usage conditions in the work place. Under the standardised single application conditions, no major differences in cytotoxic effects or PGE2 levels between the samples were apparent, so no indication of a skin irritation potential could be concluded. This result is in accordance with prior in vivo studies for acute dermal toxicity. Under repeated application conditions, signs of cytotoxicity were observed after the application of one of the DTO samples, which was known to be derived from different raw materials. Therefore, it was concluded that, generally, the presence of DTO at a concentration of up to 10% in cutting fluids, is not expected to result in any DTO-related deterioration of the skin. PMID:19292577

Pittermann, Wolfgang; Hopfgarten, Fredrik; Kietzmann, Manfred

2009-02-01

305

The effects of bovine serum albumin and fetal bovine serum on the development of pre- and postcleavage-stage bovine embryos cultured in modified CR2 and M199 media  

Microsoft Academic Search

The effects of protein supplementation on bovine embryo development in vitro was evaluated using a 4 × 2 factorial arrangement with ten replications. A total of 6438 oocytes collected from abattoir ovaries were used. Bovine serum albumin (BSA) and fetal bovine serum (FBS) were added in various combinations to simple (modified CR2) and complex (M199) media during culture of precleavage-stage

S. Wang; Y. Liu; G. R. Holyoak; T. D. Bunch

1997-01-01

306

Diffuse X-ray scattering to model protein motions.  

PubMed

Problems in biology increasingly need models of protein flexibility to understand and control protein function. At the same time, as they improve, crystallographic methods are marching closer to the limits of what can be learned from Bragg data in isolation. It is thus inevitable that mainstream protein crystallography will turn to diffuse scattering to model protein motions and improve crystallographic models. The time is ripe to make it happen. PMID:24507780

Wall, Michael E; Adams, Paul D; Fraser, James S; Sauter, Nicholas K

2014-02-01

307

Modeling of loops in protein structures.  

PubMed Central

Comparative protein structure prediction is limited mostly by the errors in alignment and loop modeling. We describe here a new automated modeling technique that significantly improves the accuracy of loop predictions in protein structures. The positions of all nonhydrogen atoms of the loop are optimized in a fixed environment with respect to a pseudo energy function. The energy is a sum of many spatial restraints that include the bond length, bond angle, and improper dihedral angle terms from the CHARMM-22 force field, statistical preferences for the main-chain and side-chain dihedral angles, and statistical preferences for nonbonded atomic contacts that depend on the two atom types, their distance through space, and separation in sequence. The energy function is optimized with the method of conjugate gradients combined with molecular dynamics and simulated annealing. Typically, the predicted loop conformation corresponds to the lowest energy conformation among 500 independent optimizations. Predictions were made for 40 loops of known structure at each length from 1 to 14 residues. The accuracy of loop predictions is evaluated as a function of thoroughness of conformational sampling, loop length, and structural properties of native loops. When accuracy is measured by local superposition of the model on the native loop, 100, 90, and 30% of 4-, 8-, and 12-residue loop predictions, respectively, had <2 A RMSD error for the mainchain N, C(alpha), C, and O atoms; the average accuracies were 0.59 +/- 0.05, 1.16 +/- 0.10, and 2.61 +/- 0.16 A, respectively. To simulate real comparative modeling problems, the method was also evaluated by predicting loops of known structure in only approximately correct environments with errors typical of comparative modeling without misalignment. When the RMSD distortion of the main-chain stem atoms is 2.5 A, the average loop prediction error increased by 180, 25, and 3% for 4-, 8-, and 12-residue loops, respectively. The accuracy of the lowest energy prediction for a given loop can be estimated from the structural variability among a number of low energy predictions. The relative value of the present method is gauged by (1) comparing it with one of the most successful previously described methods, and (2) describing its accuracy in recent blind predictions of protein structure. Finally, it is shown that the average accuracy of prediction is limited primarily by the accuracy of the energy function rather than by the extent of conformational sampling. PMID:11045621

Fiser, A.; Do, R. K.; Sali, A.

2000-01-01

308

Mitogen-activated protein kinase signaling in bovine articular chondrocytes in response to fluid flow does not require calcium mobilization.  

PubMed

In the present study, the role of mitogen-activated protein kinases (MAPKs) in chondrocyte mechanotransduction was investigated. We hypothesized that MAPKs participate in fluid flow-induced chondrocyte mechanotransduction. To test our hypothesis, we studied cultured chondrocytes subjected to a well-defined mechanical stimulus generated with a laminar flow chamber. The extracellular signal-regulated kinases 1 and 2 (ERK1/2) were activated 1.6-3-fold after 5-15 min of fluid flow exposure corresponding to a chamber wall shear stress of 1.6 Pa. Activation of ERK1/2 was observed in the presence of both 10% FBS and 0.1% BSA, suggesting that the flow effects do not require serum agonists. Treatment with thapsigargin or EGTA had no significant effect on the ERK1/2 activation response to flow, suggesting that Ca2+ mobilization is not required for this response. To assess downstream effects of the activated MAPKs on transcription, flow studies were performed using chondrocytes transfected with a chimeric luciferase construct containing 2.4 kb of the promoter region along with exon 1 of the human aggrecan gene. Two-hour exposure of transfected chondrocytes to fluid flow significantly decreased aggrecan promoter activity by 40%. This response was blocked by treatment of chondrocytes with the MEK-1 inhibitor PD98059. These findings demonstrate that, under the conditions of the present study, fluid flow-induced signals activate the MEK-1/ERK signaling pathway in articular chondrocytes, leading to down-regulation of expression of the aggrecan gene. PMID:10609520

Hung, C T; Henshaw, D R; Wang, C C; Mauck, R L; Raia, F; Palmer, G; Chao, P H; Mow, V C; Ratcliffe, A; Valhmu, W B

2000-01-01

309

Modeling Intrinsically Disordered Proteins with Bayesian Statistics  

PubMed Central

The characterization of intrinsically disordered proteins is challenging because accurate models of these systems require a description of both their thermally accessible conformers and the associated relative stabilities or weights. These structures and weights are typically chosen such that calculated ensemble averages agree with some set of prespecified experimental measurements; however, the large number of degrees of freedom in these systems typically leads to multiple conformational ensembles that are degenerate with respect to any given set of experimental observables. In this work we demonstrate that estimates of the relative stabilities of conformers within an ensemble are often incorrect when one does not account for the underlying uncertainty in the estimates themselves. Therefore, we present a method for modeling the conformational properties of disordered proteins that estimates the uncertainty in the weights of each conformer. The Bayesian weighting (BW) formalism incorporates information from both experimental data and theoretical predictions to calculate a probability density over all possible ways of weighting the conformers in the ensemble. This probability density is then used to estimate the values of the weights. A unique and powerful feature of the approach is that it provides a built-in error measure that allows one to assess the accuracy of the ensemble. We validate the approach using reference ensembles constructed from the five-residue peptide met-enkephalin and then apply the BW method to construct an ensemble of the K18 isoform of the tau protein. Using this ensemble, we indentify a specific pattern of long-range contacts in K18 that correlates with the known aggregation properties of the sequence. PMID:20925316

2010-01-01

310

Molecular Dynamics Studies on Free and Bound Targets of the Bovine Papillomavirus Type I E2 Protein: The Protein Binding Effect on DNA and the Recognition Mechanism  

Microsoft Academic Search

Molecular dynamics simulations of a total duration of 30ns in explicit solvent were carried out on the BPV-1-E2 protein complexed to a high-affinity DNA target containing the two hydrogen-bonded ACCG.CGGT half-sites separated by the noncontacted ACGT sequence. The analysis of the trajectories focuses on the DNA structure and on the dynamics. The data are compared to those issued from recent

D. Djuranovic; B. Hartmann

2005-01-01

311

Lactoferrin from bovine colostrum regulates prolyl hydroxylase 2 activity and prevents prion protein-mediated neuronal cell damage via cellular prion protein.  

PubMed

Prion disorders are associated with the conversion of normal cellular prion protein (PrPc) to the abnormal scrapie isoform of prion protein (PrPsc). Recent studies have shown that expression of normal PrPc is regulated by hypoxia-inducible factor-1 alpha (HIF-1?), and that lactoferrin increases full-length PrPc on the cell surface. Lactoferrin is an 80-kDa iron-binding glycoprotein with various biological activities, including iron-chelating ability. HIF-1? and the associated ubiquitin-proteasome pathway are regulated by HIF prolyl-hydroxylases 2 (PHD2). We hypothesized that lactoferrin regulates PHD2 expression and enzymatic activity, and the PHD2 regulation promotes HIF-1? stability and prevention of neuronal cell death mediated by prion protein (PrP) residues (106-126). Lactoferrin prevented PrP (106-126)-induced neurotoxicity by the induction of PrPc expression via promoting HIF-1? stability in neuronal cells. Our results demonstrated that lactoferrin prevented PrP (106-126)-induced neurotoxicity via the up-regulation of HIF-1? stability determined by PHD2 expression and enzymatic activity. These findings suggest that possible therapies such as PHD2 inhibition, or promotion of lactoferrin secretion, may have clinical benefits in neurodegenerative diseases, including prion disease. PMID:24875174

Park, Y-G; Moon, J-H; Park, S-Y

2014-08-22

312

Reconstitution of Membrane Proteins into Model Membranes: Seeking Better Ways to Retain Protein Activities  

PubMed Central

The function of any given biological membrane is determined largely by the specific set of integral membrane proteins embedded in it, and the peripheral membrane proteins attached to the membrane surface. The activity of these proteins, in turn, can be modulated by the phospholipid composition of the membrane. The reconstitution of membrane proteins into a model membrane allows investigation of individual features and activities of a given cell membrane component. However, the activity of membrane proteins is often difficult to sustain following reconstitution, since the composition of the model phospholipid bilayer differs from that of the native cell membrane. This review will discuss the reconstitution of membrane protein activities in four different types of model membrane—monolayers, supported lipid bilayers, liposomes and nanodiscs, comparing their advantages in membrane protein reconstitution. Variation in the surrounding model environments for these four different types of membrane layer can affect the three-dimensional structure of reconstituted proteins and may possibly lead to loss of the proteins activity. We also discuss examples where the same membrane proteins have been successfully reconstituted into two or more model membrane systems with comparison of the observed activity in each system. Understanding of the behavioral changes for proteins in model membrane systems after membrane reconstitution is often a prerequisite to protein research. It is essential to find better solutions for retaining membrane protein activities for measurement and characterization in vitro. PMID:23344058

Shen, Hsin-Hui; Lithgow, Trevor; Martin, Lisandra L.

2013-01-01

313

The variable and conserved interfaces of modeled olfactory receptor proteins  

E-print Network

The variable and conserved interfaces of modeled olfactory receptor proteins YITZHAK PILPEL models of other G-protein-coupled receptors, allows us to analyze the OR amino acid variability patterns postions in other G-protein-coupled receptors, the rest are suggested to form an olfactory-unique aspect

Church, George M.

314

Investigation of a dual fetal infection model with bovine viral diarrhoea viruses (BVDV)-1 and BVDV-2.  

PubMed

Two studies were performed in pregnant heifers to determine whether inoculation with two bovine viral diarrhoea viruses (BVDV), one BVDV-1 and one BVDV-2, inoculated separately into either nostril, results in fetal infection with both viruses. Dual transplacental infection of the fetus with BVDV-1 and BVDV-2 was observed in one case, but not consistently. PMID:21597952

Makoschey, B; Janssen, M G J

2011-10-01

315

MODBASE, a database of annotated comparative protein structure models  

Microsoft Academic Search

MODBASE (http:\\/\\/guitar.rockefeller.edu\\/modbase) is a relational database of annotated comparative protein structure models for all available protein sequences matched to at least one known protein structure. The models are calculated by MODPIPE, an automated modeling pipeline that relies on PSI-BLAST, IMPALA and MODELLER. MODBASE uses the MySQL relational database management system for flexible and efficient querying, and the MODVIEW Netscape plugin

Ursula Pieper; Narayanan Eswar; Ashley C. Stuart; Valentin A. Ilyin; Andrej Sali

2002-01-01

316

Bayesian Modeling of MPSS Data: Gene Expression Analysis of Bovine Salmonella Infection  

PubMed Central

Massively Parallel Signature Sequencing (MPSS) is a high-throughput counting-based technology available for gene expression profiling. It produces output that is similar to Serial Analysis of Gene Expression (SAGE) and is ideal for building complex relational databases for gene expression. Our goal is to compare the in vivo global gene expression profiles of tissues infected with different strains of Salmonella obtained using the MPSS technology. In this article, we develop an exact ANOVA type model for this count data using a zero-inflated Poisson (ZIP) distribution, different from existing methods that assume continuous densities. We adopt two Bayesian hierarchical models—one parametric and the other semiparametric with a Dirichlet process prior that has the ability to “borrow strength” across related signatures, where a signature is a specific arrangement of the nucleotides, usually 16-21 base-pairs long. We utilize the discreteness of Dirichlet process prior to cluster signatures that exhibit similar differential expression profiles. Tests for differential expression are carried out using non-parametric approaches, while controlling the false discovery rate. We identify several differentially expressed genes that have important biological significance and conclude with a summary of the biological discoveries. PMID:21165171

Dhavala, Soma S.; Datta, Sujay; Mallick, Bani K.; Carroll, Raymond J.; Khare, Sangeeta; Lawhon, Sara D.; Adams, L. Garry

2010-01-01

317

Modeling colloid deposition on a protein layer adsorbed to iron-oxide-coated sand  

NASA Astrophysics Data System (ADS)

Our recent study reported that conformation change of granule-associated Bovine Serum Albumin (BSA) may influence the role of the protein controlling colloid deposition in porous media (Flynn et al., 2012). The present study conceptualized the observed phenomena with an ellipsoid morphology model, describing BSA as an ellipsoid taking a side-on or end-on conformation on granular surface, and identified the following processes: (1) at low adsorbed concentrations, BSA exhibited a side-on conformation blocking colloid deposition; (2) at high adsorbed concentrations, BSA adapted to an end-on conformation promoted colloid deposition; and (3) colloid deposition on the BSA layer may progressively generate end-on molecules (sites) by conformation change of side-on BSA, resulting in sustained increasing deposition rates. Generally, the protein layer lowered colloid attenuation by the porous medium, suggesting the overall effect of BSA was inhibitory at the experimental time scale. A mathematical model was developed to interpret the ripening curves. Modeling analysis identified the site generation efficiency of colloid as a control on the ripening rate (declining rate in colloid concentrations), and this efficiency was higher for BSA adsorbed from a more dilute BSA solution.

Yang, X.; Flynn, R.; von der Kammer, F.; Hofmann, T.

2012-11-01

318

Homology Modeling and Analysis of Structure Predictions of the Bovine Rhinitis B Virus RNA Dependent RNA Polymerase (RdRp).  

PubMed

Bovine Rhinitis B Virus (BRBV) is a picornavirus responsible for mild respiratory infection of cattle. It is probably the least characterized among the aphthoviruses. BRBV is the closest relative known to Foot and Mouth Disease virus (FMDV) with a ~43% identical polyprotein sequence and as much as 67% identical sequence for the RNA dependent RNA polymerase (RdRp), which is also known as 3D polymerase (3D(pol)). In the present study we carried out phylogenetic analysis, structure based sequence alignment and prediction of three-dimensional structure of BRBV 3D(pol) using a combination of different computational tools. Model structures of BRBV 3D(pol) were verified for their stereochemical quality and accuracy. The BRBV 3D(pol) structure predicted by SWISS-MODEL exhibited highest scores in terms of stereochemical quality and accuracy, which were in the range of 2Å resolution crystal structures. The active site, nucleic acid binding site and overall structure were observed to be in agreement with the crystal structure of unliganded as well as template/primer (T/P), nucleotide tri-phosphate (NTP) and pyrophosphate (PPi) bound FMDV 3D(pol) (PDB, 1U09 and 2E9Z). The closest proximity of BRBV and FMDV 3D(pol) as compared to human rhinovirus type 16 (HRV-16) and rabbit hemorrhagic disease virus (RHDV) 3D(pols) is also substantiated by phylogeny analysis and root-mean square deviation (RMSD) between C-? traces of the polymerase structures. The absence of positively charged ?-helix at C terminal, significant differences in non-covalent interactions especially salt bridges and CH-pi interactions around T/P channel of BRBV 3D(pol) compared to FMDV 3D(pol), indicate that despite a very high homology to FMDV 3D(pol), BRBV 3D(pol) may adopt a different mechanism for handling its substrates and adapting to physiological requirements. Our findings will be valuable in the design of structure-function interventions and identification of molecular targets for drug design applicable to Aphthovirus RdRps. PMID:22942748

Rai, Devendra K; Rieder, Elizabeth

2012-01-01

319

Homology Modeling and Analysis of Structure Predictions of the Bovine Rhinitis B Virus RNA Dependent RNA Polymerase (RdRp)  

PubMed Central

Bovine Rhinitis B Virus (BRBV) is a picornavirus responsible for mild respiratory infection of cattle. It is probably the least characterized among the aphthoviruses. BRBV is the closest relative known to Foot and Mouth Disease virus (FMDV) with a ~43% identical polyprotein sequence and as much as 67% identical sequence for the RNA dependent RNA polymerase (RdRp), which is also known as 3D polymerase (3Dpol). In the present study we carried out phylogenetic analysis, structure based sequence alignment and prediction of three-dimensional structure of BRBV 3Dpol using a combination of different computational tools. Model structures of BRBV 3Dpol were verified for their stereochemical quality and accuracy. The BRBV 3Dpol structure predicted by SWISS-MODEL exhibited highest scores in terms of stereochemical quality and accuracy, which were in the range of 2Å resolution crystal structures. The active site, nucleic acid binding site and overall structure were observed to be in agreement with the crystal structure of unliganded as well as template/primer (T/P), nucleotide tri-phosphate (NTP) and pyrophosphate (PPi) bound FMDV 3Dpol (PDB, 1U09 and 2E9Z). The closest proximity of BRBV and FMDV 3Dpol as compared to human rhinovirus type 16 (HRV-16) and rabbit hemorrhagic disease virus (RHDV) 3Dpols is also substantiated by phylogeny analysis and root-mean square deviation (RMSD) between C-? traces of the polymerase structures. The absence of positively charged ?-helix at C terminal, significant differences in non-covalent interactions especially salt bridges and CH-pi interactions around T/P channel of BRBV 3Dpol compared to FMDV 3Dpol, indicate that despite a very high homology to FMDV 3Dpol, BRBV 3Dpol may adopt a different mechanism for handling its substrates and adapting to physiological requirements. Our findings will be valuable in the design of structure-function interventions and identification of molecular targets for drug design applicable to Aphthovirus RdRps. PMID:22942748

Rai, Devendra K.; Rieder, Elizabeth

2012-01-01

320

Recombinant Bovine Dihydrofolate Reductase Produced by Mutagenesis and Nested PCR of Murine Dihydrofolate Reductase cDNA  

PubMed Central

Recent reports of the slow-tight binding inhibition of bovine liver dihydrofolate reductase (bDHFR) in the presence of polyphenols isolated from green tea leaves has spurred renewed interest in the biochemical properties of bDHFR. Earlier studies were done with native bDHFR but in order to validate models of polyphenol binding to bDHFR, larger quntities of bDHFR are necessary to support structural studies. Bovine DHFR differs from its closest sequence homologue, murine DHFR, by 19 amino acids. To obtain the bDHFR cDNA, murineDHFR cDNA was transformed by a series of nested PCR reactions to reproduce the amino acid coding sequence for bovine DHFR. The bovine liver DHFR cDNA has an open reading frame of 561 base pairs encoding a protein of 187 amino acids that has a high level of conservation at the primary sequence level with other DHFR enzymes, and more so for the amino acid residues in the active site of the mammalian DHFR enzymes. Expression of the bovine DHFR cDNA in bacterial cells produced a stable recombinant protein with high enzymatic activity and kinetic properties similar to those previously reported for the native protein. PMID:18672067

Cody, Vivian; Mao, Qilong; Queener, Sherry F.

2008-01-01

321

High bovine blastocyst development in a static in vitro production system using sofaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins  

Microsoft Academic Search

We describe a bovine embryo culture system that supports repeatable high development in the presence of serum or BSA as well as under defined conditions in the absence of those components. In the first experiment, embryo development in SOF with amino acids (SOFaa), sodium citrate (SOFaac) and myo-inositol (SOFaaci) and with BSA or polyvinyl alcohol (PVA) was compared with that

P. Holm; P. J. Booth; M. H. Schmidt; T. Greve; H. Callesen

1999-01-01

322

Monoclonal antibodies to the E2 protein of a new genotype (type 2) of bovine viral diarrhea virus define three antigenic domains involved in neutralization  

Microsoft Academic Search

Bovine viral diarrhea virus (BVDV) has recently been segregated into two genotypes, namely, BVDV 1 and BVDV 2. Viruses of the BVDV 2 genotype are a cause of hemorrhagic and acute fatal disease in cattle in the US and Canada. In this study, monoclonal antibodies (mAbs) to the newly described BVDV 2 were produced after immunization with virus or a

Dirk Deregt; Piet A. van Rijn; Tania Y. Wiens; Jan van den Hurk

1998-01-01

323

Isolation from Fetal Bovine Serum of a Fragment b of Complement Factor B-like Protein Improving a Long-Term Survival of Human Endothelial Cells  

Microsoft Academic Search

It is known that serum is a most important factor supporting cell survival and growth. Particularly, the deprivation of serum would result in the death of human endothelial cell. Our previous paper reported an endothelial cell-viability maintaining factor (EC-VMFa) purified from fetal bovine serum and identified as an apolipoprotein. In the present further study, it is demonstrated that another potent

Guoping Cai; Takeshi Satoh; Hiroyoshi Hoshi

1997-01-01

324

Sequences outside that of residues 93-102 of 3A protein can contribute to the ability of foot-and-mouth disease virus (FMDV) to replicate in bovine-derived cells.  

PubMed

Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease of cloven-hoofed animals. During 2010 and 2011, there was an epidemic of the Mya-98 lineage of the Southeast Asia (SEA) topotype in East Asia, including China. Changes in the FMDV 3A protein have been previously reported to be associated with the inability of FMDV to grow in bovine cells and cause disease in cattle. In this paper, we report the generation of a full-length infectious cDNA clone of FMDV O/SEA/Mya-98 strain O/GZSB/2011 for the first time along with two genetically modified viruses with deletion at positions 93-102 and 133-143 in 3A based on the established infectious clone. All the recombinant viruses grew well and displayed growth properties and plaque phenotypes similar to those of the parental virus in baby hamster kidney (BHK-21) cells, porcine kidney (PK-15) cells, and primary fetal porcine kidney (FPK) cells. While the recombinant viruses rvGZSB and rvSB?133-143 exhibited similar growth properties and plaque phenotypes with the parental virus in primary fetal bovine kidney (FBK) cells, the recombinant virus rvSB?93-102, containing deletion at positions 93-102 in 3A, grew at a slower rate and had a smaller plaque size phenotype in FBK cells than that of the parental virus. Therefore, the results suggest that the deletion at positions 93-102 of 3A protein does not affect FMDV replication efficiency in BHK-21, PK-15 and FPK cells, but affects virus replication efficiency in FBK cells, although, cannot alone account for the inability to replicate in bovine cells. PMID:25116389

Ma, Xueqing; Li, Pinghua; Bai, Xingwen; Sun, Pu; Bao, Huifang; Lu, Zengjun; Cao, Yimei; Li, Dong; Chen, Yingli; Qiao, Zilin; Liu, Zaixin

2014-10-13

325

Tactile Teaching: Exploring Protein Structure/Function Using Physical Models  

ERIC Educational Resources Information Center

The technology now exists to construct physical models of proteins based on atomic coordinates of solved structures. We review here our recent experiences in using physical models to teach concepts of protein structure and function at both the high school and the undergraduate levels. At the high school level, physical models are used in a…

Herman, Tim; Morris, Jennifer; Colton, Shannon; Batiza, Ann; Patrick, Michael; Franzen, Margaret; Goodsell, David S.

2006-01-01

326

Bioinorganic Chemical Modeling of Dioxygen-Activating Copper Proteins.  

ERIC Educational Resources Information Center

Discusses studies done in modeling the copper centers in the proteins hemocyanin (a dioxygen carrier), tyrosinase, and dopamine beta-hydroxylase. Copper proteins, model approach in copper bioinorganic chemistry, characterization of reversible oxygen carriers and dioxygen-metal complexes, a copper mono-oxygenase model reaction, and other topics are…

Karlin, Kenneth D.; Gultneh, Yilma

1985-01-01

327

ISOLATION AND PROPERTIES OF BOVINE COLOSTRAL TRYPSIN INHIBITOR  

E-print Network

COLOSTRUM DES BOVINS. ― L'inhibiteur de la trypsine du colostrum des bovins a été isolé par chromato colostrum (CTI) is a heat- and acid-resistant protein, which was originally isolated by Laskowski the dynamics of the secretion of CTI and those of colostral immunoglobulins. Materials and Methods 1. COLOSTRUM

Paris-Sud XI, Université de

328

Mechanisms of m-cresol-induced Protein Aggregation Studied Using a Model Protein Cytochrome c  

E-print Network

Mechanisms of m-cresol-induced Protein Aggregation Studied Using a Model Protein Cytochrome c) to inhibit microbial growth during long-term storage of unused formulations. m-cresol (CR) is one such AP; proteins; spectroscopy; formulation; cytochrome c; cresol INTRODUCTION Approximately one

Mallela, Krishna M. G.

329

Modeling and mitigating winter hay bale damage by elk in a low prevalence bovine tuberculosis endemic zone.  

PubMed

Wildlife causes extensive crop damage throughout much of North America and these shared feeds are a key risk factor in the transmission of diseases between wildlife and livestock, including bovine tuberculosis (TB). Predicting wildlife use of agricultural crops can provide insight directed toward targeted disease mitigation at areas of potential indirect interaction. In this study, we quantified use of hay bales by elk (Cervus canadensis) during the winter in southwestern Manitoba, Canada using a database of 952 damage claims paid compensation from 1994 to 2012. We evaluated environmental factors predicted to determine risk of hay bale damage on each quarter section by elk using a Resource Selection Probability Function (RSPF) model. The most important variables (as measured for each quarter section and based on cumulative Akaike weights that scale from 0 to 1) were distance to protected areas (1.00), forest including a buffer around the quarter section (1.00), forage crop including a buffer around the quarter section (1.00), distance from streams (0.99), forage crop (0.92), cereal and oilseed crop cover including a buffer (0.85), and forest cover (0.82). We then developed an RSPF-based predictive map of damage to hay bales by elk that identified key areas with high probability of damage (RSPF?0.6), accounting for 3.5% of the study area. We then multiplied the RSPF values by the inverse of the proximity to known cases of TB positive elk and determined that 0.51% of the study area had an overall high combined probability of hay bale damage and proximity to TB positive elk (i.e. adjusted probability of ?0.6). In the southern half of the study area where 164 hay yard barrier fences have been implemented since 2002, there has been a significant decrease in the number of annual claims. Barrier fencing around Riding Mountain National Park has been successful at reducing elk damage where it has been implemented. In our study area, prevalence of TB in both cattle (0.003%) and elk (0.89%) is very low, precluding conventional epidemiological analyses. In the absence of clear evidence of specific routes of TB transmission, we advocate that on-farm risk assessments and mitigation efforts should continue to address areas where elk cause damage to hay bales in winter using barrier fencing. Mitigation effort should especially focus on areas where TB positive elk have been identified, as these sites provide potential for indirect interaction between cattle and elk. PMID:24486094

Gooding, R M; Brook, R K

2014-05-01

330

Characterization of Carbohydrate Structures of Bovine MUC15 and Distribution of the Mucin in Bovine Milk  

Microsoft Academic Search

The present work reports the characterization of car- bohydrate structures and the distribution of the newly identified mucin MUC15, a highly glycosylated protein associated with the bovine milk fat globule membrane (MFGM). Distribution of MUC15 was investigated in various fractions of bovine milk by densitometric scan- ning of Western blots. In raw milk, MUC15 was shown to constitute 0.08% (wt)

L. T. Pallesen; L. R. L. Pedersen; T. E. Petersen; J. T. Rasmussen

2007-01-01

331

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

Microsoft Academic Search

  \\u000a Incisional hernia is a frequent complication of abdominal surgery developing in 11–20 % of patients undergoing an abdominal\\u000a operation. Regarding morbidity and loss of manpower, incisional hernias continue to be a fundamental problem for surgeons.\\u000a In this experimental study, three commonly used mesh materials (Goretex PTFE; Tutoplast Fascia lata; Tutopatch Pericardium\\u000a bovine) were compared according to effectiveness, strength, adhesion

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

2003-01-01

332

A Model of Consumers' Risk Perceptions Toward Recombinant Bovine Growth Hormone (rbGH): The Impact of Risk Characteristics  

Microsoft Academic Search

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

Deana Grobe; Robin Douthitt; Lydia Zepeda

1999-01-01

333

Tissue engineering approach to repair abdominal wall defects using cell-seeded bovine tunica vaginalis in a rabbit model.  

PubMed

The aim of this study was to engineer skeletal muscle tissue for repair abdominal wall defects. Myoblast were seeded onto the scaffolds and cultivated in vitro for 5 days. Full thickness abdominal wall defects (3 x 4 cm) were created in 18 male New Zealand white rabbits and randomly divided into two equal groups. The defects of the first group were repaired with myoblast-seeded-bovine tunica vaginalis whereas the second group repaired with non-seeded-bovine tunica vaginalis and function as a control. Three animals were sacrificed at 7th, 14th, and 30th days of post-implantation from each group and the explanted specimens were subjected to macroscopic and microscopic analysis. In every case, seeded scaffolds have better deposition of newly formed collagen with neo-vascularisation than control group. Interestingly, multinucleated myotubes and myofibers were only detected in cell-seeded group. This study demonstrated that myoblast-seeded-bovine tunica vaginalis can be used as an effective scaffold to repair severe and large abdominal wall defects with regeneration of skeletal muscle tissue. PMID:20135201

Ayele, T; Zuki, A B Z; Noorjahan, B M A; Noordin, M M

2010-05-01

334

ASPDock: protein-protein docking algorithm using atomic solvation parameters model  

Microsoft Academic Search

BACKGROUND: Atomic Solvation Parameters (ASP) model has been proven to be a very successful method of calculating the binding free energy of protein complexes. This suggests that incorporating it into docking algorithms should improve the accuracy of prediction. In this paper we propose an FFT-based algorithm to calculate ASP scores of protein complexes and develop an ASP-based protein-protein docking method

Lin Li; Dachuan Guo; Yangyu Huang; Shiyong Liu; Yi Xiao

2011-01-01

335

Molecular modelling of the mass density of single proteins  

Microsoft Academic Search

Using molecular dynamics (MD) simulations, the density of single proteins and its temperature dependence was modelled starting from the experimentally determined protein structure and a generic, transferable force field, without the need of prior parameterization. Although all proteins consist of the same 20 amino acids, their density in aqueous solution varies up to 10% and the thermal expansion coefficient up

Meike Hutt; Tobias Kulschewski; Jürgen Pleiss

2012-01-01

336

Effects of exposure to calves persistently infected with bovine viral diarrhea virus type 1b and subsequent infection with Mannheima haemolytica on clinical signs and immune variables: model for bovine respiratory disease via viral and bacterial interaction.  

PubMed

The objective was to determine effects of an intratracheal Mannheimia haemolytica challenge after 72-h exposure to bovine viral diarrhea virus type 1b (BVDV1b) persistently infected (PI) calves on serum antibody production, white blood cell count (WBC), cytokine concentrations, and blood gases in feedlot steers. Twenty-four steers (initial BW = 314 +/- 31 kg) were randomly allocated to 1 of 4 treatments (6 steers/treatment) arranged as a 2 x 2 factorial. Treatments were 1) steers not exposed to steers PI with BVDV nor challenged with M. haemolytica (control; CON); 2) steers exposed to 2 steers PI with BVDV for 72 h (BVD); 3) steers intratracheally challenged with M. haemolytica (MH); and 4) steers exposed to 2 steers PI with BVDV for 72 h and challenged with M. haemolytica (BVD+MH). There were 12 h between exposure to PI steers and challenge with M. haemolytica. Rectal temperature was increased (P < 0.001) for MH and BVD+MH during the initial 24 h after the M. haemolytica challenge. For MH and BVD+MH, total WBC count was increased (P < 0.01) at 36 h post M. haemolytica challenge compared with CON, whereas in BVD steers, WBC count was decreased (P < 0.01). Total lymphocyte count was increased (P = 0.004) during the initial 72 h post BVDV exposure for the BVD and BVD+MH groups compared with MH and CON, and this difference remained at 96 h post M. haemolytica challenge. An increased (P < 0.001) total neutrophil count was observed during the initial 36 h for the MH group and at 72 h for the BVD+MH challenge group. Interleukin 1beta, IL-6, and tumor necrosis factor alpha (TNFalpha) concentrations were greater (P model was successful at inducing bovine respiratory disease (BRD) associated with BVDV and M. haemolytica. Understanding the physiological changes in morbid animals will lead to improved strategies for decreasing severity and economic losses associated with BRD. PMID:20154164

Burciaga-Robles, L O; Step, D L; Krehbiel, C R; Holland, B P; Richards, C J; Montelongo, M A; Confer, A W; Fulton, R W

2010-06-01

337

Virtual Ligand Screening Against Comparative Protein Structure Models  

PubMed Central

Virtual ligand screening uses computation to discover new ligands of a protein by screening one or more of its structural models against a database of potential ligands. Comparative protein structure modeling extends the applicability of virtual screening beyond the atomic structures determined by X-ray crystallography or NMR spectroscopy. Here, we describe an integrated modeling and docking protocol, combining comparative modeling by MODELLER and virtual ligand screening by DOCK. PMID:22183533

Fan, Hao; Irwin, John J.; Sali, Andrej

2012-01-01

338

Weak self-interactions of globular proteins studied by small-angle X-ray scattering and structure-based modeling  

E-print Network

We investigate protein-protein interactions in solution by small-angle X-ray scattering (SAXS) and theoretical modeling. The structure factor for solutions of bovine pancreatic trypsin inhibitor (BPTI), myoglobin (Mb), and intestinal fatty acid-binding protein (IFABP) is determined from SAXS measurements at multiple concentrations, from Monte Carlo simulations with a coarse-grained structure-based interaction model, and from analytic approximate solutions of two idealized colloidal interaction models without adjustable parameters. By combining these approaches, we find that the structure factor is essentially determined by hard-core and screened electrostatic interactions. Other soft short-ranged interactions (van der Waals and solvation-related) are either individually insignificant or tend to cancel out. The structure factor is also not significantly affected by charge fluctuations. For Mb and IFABP, with small net charge and relatively symmetric charge distribution, the structure factor is well described b...

Kaieda, Shuji; Plivelic, Tomás S; Halle, Bertil

2014-01-01

339

Characterization of the keratan sulphate proteoglycans from bovine corneal stroma.  

PubMed Central

The keratan sulphate proteoglycans that can be prepared from bovine corneal stroma [Axelsson & Heinegård (1975) Biochem. J. 145, 491-500] were characterized by gel chromatography, gel electrophoresis and analytical ultracentrifugation in associative (0.6 M-NaCl) and dissociative (6M-guanidinum chloride) solvents. The proteoglycans aggreagated at low salt concentrations and pH. The weight-average molecular weight of the monomer proteoglycans was established. Keratan sulphate peptides and oligosaccharide peptides were isolated after proteolysis. Their composition indicated that both are linked to protein via asparagine residues. A tentative model for corneal keratan sulphate proteoglycans is suggested. PMID:646789

Axelsson, I; Heinegard, D

1978-01-01

340

Proteolysis of Specific Muscle Structural Proteins by m-Calpain at Low pH and Temperature Is Similar to Degradation in Postmortem Bovine Muscle1,2  

Microsoft Academic Search

Postmortem (PM) and m-calpain- induced degradation of specific skeletal muscle pro- teins was monitored by SDS-PAGE and Western blotting. Samples were removed from bovine longissi- mus thoracis (LT) at approximately 45 min PM for the preparation of at-death (0-d) myofibrils (MF). The LT was excised at 1 d PM, vacuum-packaged, and stored at 2°C. Samples were removed for Warner- Bratzler

Elisabeth Huff-Lonergan; Tomiko Mitsuhashi; Dirk D. Beekman; F. C. Parrish; Dennis G. Olson; Richard M. Robson

2010-01-01

341

Modelling the trend of bovine spongiform encephalopathy prevalence in France: Use of restricted cubic spline regression in age–period–cohort models to estimate the efficiency of control measures  

Microsoft Academic Search

An age–period–cohort (APC) analysis was used to assess the trend in prevalence of bovine spongiform encephalopathy (BSE) in France over time in relation to the control measures adopted since onset of the epidemic. Restricted cubic regression splines were used to model the functional forms of the non-linear effects of age at screening, birth cohort and date of diagnosis of the

Carole Sala; Eric Morignat; Christian Ducrot; Didier Calavas

2009-01-01

342

Secretory activity of bovine ovarian granulosa cells transfected with sense and antisense insulin-like growth factor (IGF) binding protein-3 and the response to IGF-I, GH, LH, oxytocin and oestradiol.  

PubMed

The aim of our in vitro experiments was to examine if IGF binding protein (IGFBP)-3 is involved in control of bovine ovarian secretory activity. For this purpose we performed the transfection of bovine granulosa cells with cDNA sense and antisense constructs increasing or inhibiting IGFBP-3 synthesis. The release of IGFBP-3, progesterone, oxytocin, IGF-I and prostaglandins F (PGF) and E (PGE) by control and transfected cells was compared. The transfected ovarian cells were cultured with and without bLH (100 ng/ml), bGH (100 ng/ml), IGF-I (10 ng/ml), oxytocin (10 ng/ml) and oestradiol-17beta (100 ng/ml). The concentration of IGFBP-3 produced was assessed using ligand and western blotting and secretion of progesterone, oxytocin, IGF-I, PGF and PGE was evaluated using RIA/IRMA techniques. Transfection of cells with the sense IGFBP-3 cDNA construct resulted in the expected increase in IGFBP-3 release, whereas the antisense IGFBP-3 construct induced the expected reduction in IGFBP-3 output. The granulosa cells transfected to overexpress IGFBP-3 had an increase in IGF-I, PGF and PGE release, and a decrease in basal and hormone- or growth factor-induced accumulation of progesterone and oxytocin. The granulosa cells transfected to have reduced IGFBP-3 expression gave primarily significant opposite findings. The present results suggest the involvement of IGFBP-3 in control of bovine ovarian steroid, peptide hormone, growth factor and prostaglandin release. IGFBP-3 is a physiological stimulator of IGF-I and prostaglandin release and an inhibitor of steroid and peptide hormone output. PMID:11719285

Sirotkin, A V; Makarevich, A V; Corkins, M R; Kotwica, J; Kwon, H B; Bulla, J; Hetényi, L

2001-12-01

343

Virtual Ligand Screening Against Comparative Protein Structure Models  

E-print Network

Chapter 8 Virtual Ligand Screening Against Comparative Protein Structure Models Hao Fan, John J. Irwin, and Andrej Sali Abstract Virtual ligand screening uses computation to discover new ligands of a protein by screening one or more of its structural models against a database of potential ligands

Sali, Andrej

344

Automated protein model building combined with iterative structure refinement  

Microsoft Academic Search

In protein crystallography, much time and effort are often required to trace an initial model from an interpretable electron density map and to refine it until it best agrees with the crystallographic data. Here, we present a method to build and refine a protein model automatically and without user intervention, starting from diffraction data extending to resolution higher than 2.3

Richard Morris; Victor S. Lamzin; Anastassis Perrakis

1999-01-01

345

Expression and localization of progesterone receptor membrane component 1 and 2 and serpine mRNA binding protein 1 in the bovine corpus luteum during the estrous cycle and the first trimester of pregnancy.  

PubMed

The aim of this study was to evaluate the mRNA and protein expression and the localization of progesterone receptor membrane component 1 (PGRMC1), PGRMC2, and the PGRMC1 partner serpine mRNA binding protein 1 (SERBP1) in the bovine CL on Days 2 to 5, 6 to 10, 11 to 16, and 17 to 20 of the estrous cycle as well as during Weeks 3 to 5, 6 to 8, and 9 to 12 of pregnancy (n = 5-6 per each period). The highest levels of PGRMC1 and PGRMC2 mRNA expression were found on Days 6 to 16 (P < 0.05) and 11 to 16, respectively, of the estrous cycle and during pregnancy (P < 0.001). The level of PGRMC1 protein was the highest (P < 0.05) on Days 11 to 16 of the estrous cycle compared with the other stages of the estrous cycle and pregnancy, whereas PGRMC2 protein expression (P < 0.001) was the highest on Days 17 to 20 and also during pregnancy. The mRNA expression of SERBP1 was increased (P < 0.05) on Days 11 to 16, whereas the level of its protein product was decreased (P < 0.05) on Days 6 to 10 of the estrous cycle and was at its lowest (P < 0.001) on Days 17 to 20. In pregnant cows, the patterns of SERBP1 mRNA and protein expression remained constant and were comparable with those observed during the estrous cycle. Progesterone receptor membrane component 1 and PGRMC2 localized to both large and small luteal cells, whereas SERBP1 was observed mainly in small luteal cells and much less frequently in large luteal cells. All proteins were also localized in the endothelial cells of blood vessels. The data obtained indicate the variable expression of PGRMC1, PGRMC2, and SERBP1 mRNA and protein in the bovine CL and suggest that progesterone may regulate CL function via its membrane receptors during both the estrous cycle and pregnancy. PMID:25168721

Kowalik, Magdalena K; Rekawiecki, Robert; Kotwica, Jan

2014-11-01

346

Chronic low protein intake reduces tissue protein synthesis in a pig model of protein malnutrition.  

PubMed

To determine the effect of severe chronic protein deficiency on protein synthesis in different tissues and total protein in plasma, and on plasma biochemical constituents involved in amino acid metabolism, we fed diets containing either 20 or 3% protein to two groups of four age-matched piglets. After consuming the diets for 8 wk, the pigs received a primed-constant infusion of 2 H3-leucine for 8 h to measure the fractional synthesis rates (FSR) of tissue protein and total protein in plasma. Plasma urea and amino acid concentrations, particularly indispensable amino acids, were significantly lower in protein-deficient pigs. Fractional protein synthesis rates were lower in skin by 66% (P < 0.01), in jejunal mucosa by 50% (P < 0.05), in masseter muscle by 40% (P < 0.05), and in liver by 25% (P < 0.02); the fractional synthesis rate of the longissimus muscle was not different than controls. Although plasma protein concentration was significantly (P < 0.01) lower in protein-deficient pigs, the fractional synthesis rate of the total intravascular plasma protein pool was not different. We conclude that adaptation to a low protein diet involves a reduction in the rate of protein synthesis in most body tissues, with the most marked changes occurring in skin and intestine, two tissues which frequently exhibit severe functional impairment in protein malnutrition. PMID:8618147

Wykes, L J; Fiorotto, M; Burrin, D G; Del Rosario, M; Frazer, M E; Pond, W G; Jahoor, F

1996-05-01

347

Human activated protein C variants in a rat model of arterial thrombosis  

PubMed Central

Background Activated protein C (APC) inhibits coagulation by degrading activated factor V (FVa) and factor VIII (FVIIIa), protein S (PS) functioning as a cofactor to APC. Methods By mutagenesis of the vitamin K-dependent Gla domain of APC, we have recently created an APC variant having enhanced anticoagulant activity due to increased affinity for negatively charged phospholipid membranes. In the present study, the potential antithrombotic effects of this APC variant, and of a variant APC that is additionally mutated in the serine protease domain, have been evaluated in a blind randomized study in a rat model of arterial thrombosis. In this model, we have previously found the combination of bovine APC and PS to be highly antithrombotic. Four treatment groups each containing 10 rats were, in a blind random fashion, given intravenous bolus injections of wild-type or mutant variants of APC (0.8 mg/kg) together with human PS (0.6 mg/kg) or human PS (0.6 mg/kg) alone. A control group with 20 animals where given vehicle only. Results A trend to increased patency rates was noted in a group receiving one of the APC variants, but it did not reach statistical significance. Conclusion In conclusion, administration of human APC variants having enhanced anticoagulant efficacy together with human PS in a rat model of arterial thrombosis did not give an efficient antithrombotic effect. The lack of effect may be due to species-specific differences between the human protein C system and the rat hemostatic system. PMID:18957140

Malm, Karl; Arnljots, Bjorn; Dahlback, Bjorn

2008-01-01

348

Tracking Membrane Protein Association in Model Membranes  

PubMed Central

Membrane proteins are essential in the exchange processes of cells. In spite of great breakthrough in soluble proteins studies, membrane proteins structures, functions and interactions are still a challenge because of the difficulties related to their hydrophobic properties. Most of the experiments are performed with detergent-solubilized membrane proteins. However widely used micellar systems are far from the biological two-dimensions membrane. The development of new biomimetic membrane systems is fundamental to tackle this issue. We present an original approach that combines the Fluorescence Recovery After fringe Pattern Photobleaching technique and the use of a versatile sponge phase that makes it possible to extract crucial informations about interactions between membrane proteins embedded in the bilayers of a sponge phase. The clear advantage lies in the ability to adjust at will the spacing between two adjacent bilayers. When the membranes are far apart, the only possible interactions occur laterally between proteins embedded within the same bilayer, whereas when membranes get closer to each other, interactions between proteins embedded in facing membranes may occur as well. After validating our approach on the streptavidin-biotinylated peptide complex, we study the interactions between two membrane proteins, MexA and OprM, from a Pseudomonas aeruginosa efflux pump. The mode of interaction, the size of the protein complex and its potential stoichiometry are determined. In particular, we demonstrate that: MexA is effectively embedded in the bilayer; MexA and OprM do not interact laterally but can form a complex if they are embedded in opposite bilayers; the population of bound proteins is at its maximum for bilayers separated by a distance of about 200 Å, which is the periplasmic thickness of Pseudomonas aeruginosa. We also show that the MexA-OprM association is enhanced when the position and orientation of the protein is restricted by the bilayers. We extract a stoichiometry for the complex that exhibits a strong pH dependance: from 2 to 6 MexA per OprM trimer when the pH decreases from 7.5 to 5.5. Our technique allows to study membrane protein associations in a membrane environment. It provides some challenging information about complexes such as geometry and stoichiometry. PMID:19337368

Reffay, Myriam; Gambin, Yann; Benabdelhak, Houssain; Phan, Gilles; Taulier, Nicolas; Ducruix, Arnaud; Hodges, Robert S.; Urbach, Wladimir

2009-01-01

349

Ensemble modeling of [beta]-sheet proteins  

E-print Network

Our ability to characterize protein structure and dynamics is vastly outpaced by the speed of modern genetic sequencing, creating a growing divide between our knowledge of biological sequence and structure. Structural ...

O'Donnell, Charles William

2011-01-01

350

Parmodel: a web server for automated comparative modeling of proteins.  

PubMed

Parmodel is a web server for automated comparative modeling and evaluation of protein structures. The aim of this tool is to help inexperienced users to perform modeling, assessment, visualization, and optimization of protein models as well as crystallographers to evaluate structures solved experimentally. It is subdivided in four modules: Parmodel Modeling, Parmodel Assessment, Parmodel Visualization, and Parmodel Optimization. The main module is the Parmodel Modeling that allows the building of several models for a same protein in a reduced time, through the distribution of modeling processes on a Beowulf cluster. Parmodel automates and integrates the main softwares used in comparative modeling as MODELLER, Whatcheck, Procheck, Raster3D, Molscript, and Gromacs. This web server is freely accessible at . PMID:15555595

Uchôa, Hugo Brandão; Jorge, Guilherme Eberhart; Freitas Da Silveira, Nelson José; Camera, João Carlos; Canduri, Fernanda; De Azevedo, Walter Filgueira

2004-12-24

351

BRUCELLOSE BOVINE EXPRIMENTALE VII. —  

E-print Network

BRUCELLOSE BOVINE EXPÃ?RIMENTALE VII. — INFLUENCE SUR L'Ã?VOLUTION DE L'INFECTION D'UNE FAIBLE à partir du mucus vaginal (avant et après mise bas), du colostrum, du lait, des organes et ganglions'Homme, enparticulier par les tétracyclines, l'usage de ces antibiotiques dans le traitement de la Brucellose bovine s

Paris-Sud XI, Université de

352

A conditional neural fields model for protein threading  

PubMed Central

Motivation: Alignment errors are still the main bottleneck for current template-based protein modeling (TM) methods, including protein threading and homology modeling, especially when the sequence identity between two proteins under consideration is low (<30%). Results: We present a novel protein threading method, CNFpred, which achieves much more accurate sequence–template alignment by employing a probabilistic graphical model called a Conditional Neural Field (CNF), which aligns one protein sequence to its remote template using a non-linear scoring function. This scoring function accounts for correlation among a variety of protein sequence and structure features, makes use of information in the neighborhood of two residues to be aligned, and is thus much more sensitive than the widely used linear or profile-based scoring function. To train this CNF threading model, we employ a novel quality-sensitive method, instead of the standard maximum-likelihood method, to maximize directly the expected quality of the training set. Experimental results show that CNFpred generates significantly better alignments than the best profile-based and threading methods on several public (but small) benchmarks as well as our own large dataset. CNFpred outperforms others regardless of the lengths or classes of proteins, and works particularly well for proteins with sparse sequence profiles due to the effective utilization of structure information. Our methodology can also be adapted to protein sequence alignment. Contact: j3xu@ttic.edu Supplementary information: Supplementary data are available at Bioinformatics online. PMID:22689779

Ma, Jianzhu; Peng, Jian; Wang, Sheng; Xu, Jinbo

2012-01-01

353

Increasing Antiviral Activity of Surfactant Protein D Trimers By Introducing Residues From Bovine Serum Collectins: Dissociation of mannan-binding and antiviral activity  

PubMed Central

Collectins contribute to host defense through interactions with glycoconjugates on pathogen surfaces. We have prepared recombinant trimeric neck and carbohydrate recognition domains (NCRDs) of collectins and we now show that the NCRDs of bovine conglutinin and CL-46 (like that of CL-43) have greater intrinsic antiviral activity for influenza A virus (IAV) than the human SP-D NCRD (hSP-D-NCRD). The three serum collectins differ from SP-D by having insertions adjacent to amino acid 325 and substitution of hydrophobic residues for arginine 343. We previously showed that a 3 amino acid (RAK) insertion, as found in CL-43, increases antiviral activity and mannan binding activity of the hSP-D-NCRD, while the substitution of valine at 343, as in conglutinin, more strongly increased these activities. Mannan binding activity of collectins has been considered to predict for ability to bind to high mannose glycans on viruses or other pathogens. We now show, however, that combined mutants containing the RAK insertion and R343V or R343I substitutions have greatly increased mannan binding ability, but lower IAV binding or inhibiting activity than mutants containing R343V or R343I substitutions only. These findings indicate differences in the recognition of glycan structures of mannan and IAV by the NCRDs and emphasize the importance of the flanking sequences in determining the differing interactions of human SP-D and bovine serum collectins with mannose-rich glycoconjugates on IAV and other pathogens. Of interest, we show conservation of some monoclonal antibody binding epitopes between bovine collectin NCRDs and hSP-D, suggesting shared structural motifs. PMID:20591072

Hartshorn, Kevan L.; White, Mitchell R.; Smith, Kelly; Sorensen, Grith; Kuroki, Yoshio; Holmskov, Uffe; Head, James; Crouch, Erika C.

2012-01-01

354

Interfacial protein-protein associations.  

PubMed

While traditional models of protein adsorption focus primarily on direct protein-surface interactions, recent findings suggest that protein-protein interactions may play a central role. Using high-throughput intermolecular resonance energy transfer (RET) tracking, we directly observed dynamic, protein-protein associations of bovine serum albumin on polyethylene glycol modified surfaces. The associations were heterogeneous and reversible, and associating molecules resided on the surface for longer times. The appearance of three distinct RET states suggested a spatially heterogeneous surface - with areas of high protein density (i.e., strongly interacting clusters) coexisting with mobile monomers. Distinct association states exhibited characteristic behavior, i.e., partial-RET (monomer-monomer) associations were shorter-lived than complete-RET (protein-cluster) associations. While the fractional surface area covered by regions with high protein density (i.e., clusters) increased with increasing concentration, the distribution of contact times between monomers and clusters was independent of solution concentration, suggesting that associations were a local phenomenon, and independent of the global surface coverage. PMID:24274729

Langdon, Blake B; Kastantin, Mark; Walder, Robert; Schwartz, Daniel K

2014-01-13

355

Using hidden markov models to discover new protein transport machines.  

PubMed

Protein import and export pathways are driven by protein translocases, often comprised of multiple subunits, and usually conserved across a range of organisms. Protein import into mitochondria is fundamental to eukaryotic organisms and is initiated when substrate proteins are translocated across the mitochondrial outer membrane by the TOM complex. The essential subunit of this complex is a protein called Tom40, which is probably a beta-barrel in structure and serves as the translocation pore. We describe a hidden Markov model search designed to find the Tom40 sequence in the amoeba Entamoeba histolytica. This organism has a highly reduced "mitosome", an organelle whose relationship to mitochondria has been the subject of controversy. The Tom40 sequence could not be found with BLAST-based searches, but a hidden Markov model search identified a likely candidate to form the protein import pore in the outer mitosomal membrane in E. histolytica. PMID:20419416

Likic, Vladimir A; Dolezal, Pavel; Celik, Nermin; Dagley, Michael; Lithgow, Trevor

2010-01-01

356

Developing algorithms for predicting protein-protein interactions of homology modeled proteins.  

SciTech Connect

The goal of this project was to examine the protein-protein docking problem, especially as it relates to homology-based structures, identify the key bottlenecks in current software tools, and evaluate and prototype new algorithms that may be developed to improve these bottlenecks. This report describes the current challenges in the protein-protein docking problem: correctly predicting the binding site for the protein-protein interaction and correctly placing the sidechains. Two different and complementary approaches are taken that can help with the protein-protein docking problem. The first approach is to predict interaction sites prior to docking, and uses bioinformatics studies of protein-protein interactions to predict theses interaction site. The second approach is to improve validation of predicted complexes after docking, and uses an improved scoring function for evaluating proposed docked poses, incorporating a solvation term. This scoring function demonstrates significant improvement over current state-of-the art functions. Initial studies on both these approaches are promising, and argue for full development of these algorithms.

Martin, Shawn Bryan; Sale, Kenneth L.; Faulon, Jean-Loup Michel; Roe, Diana C.

2006-01-01

357

Bovine sperm raft membrane associated Glioma Pathogenesis-Related 1-like protein 1 (GliPr1L1) is modified during the epididymal transit and is potentially involved in sperm binding to the zona pellucida.  

PubMed

Glioma pathogenesis-related 1-like protein1 (GliPr1L1) was identified by liquid chromatography-tandem mass spectrometry analyses of proteins associated to bovine sperm lipid raft membrane domains. This protein belongs to the CAP superfamily including cysteine-rich secretory proteins, Antigen 5 and pathogenesis-related 1 protein. PCR analysis revealed that GliPr1L1 is expressed in testis and, at a much lower level, all along the epididymis. Western blotting showed a similar distribution of GliPr1L1 in testicular and epididymal tissue extracts. In the epididymal lumen, GliPr1L1 was associated with the maturing spermatozoa and epididymosomes all along the excurrent duct but was undetectable in the soluble fraction of epididymal fluid. The protein was detectable as multiple isoforms with a higher MW form in the testis and proximal caput. Treatments with PNGase F revealed that N-glycosylation was responsible of multiple bands detected on Western blots. These results suggest that the N-glycosylation moiety of GliPr1L1 is processed during the transit in the caput. Western blots demonstrated that GliPr1L1 was associated with the sperm plasma membrane preparation. GliPr1L1 is glycosyl phosphatidyl inositol (GPI) anchored to caput and cauda spermatozoa as demonstrated by the ability of phosphatidylinositol specific phospholipase C to release GliPr1L1 from intact sperm cells. Lipid raft membrane domains were separated from caput and cauda epididymal spermatozoa. GliPr1L1 was immunodetectable in the low buoyant density fractions where lipid rafts are distributed. GliPr1L1 was localized on sperm equatorial segment and neck. In vitro fertilization performed in presence of anti-GliPr1L1 showed that this protein is involved in sperm-zona pellucida interaction. PMID:22552861

Caballero, Julieta; Frenette, Gilles; D'Amours, Olivier; Belleannée, Clémence; Lacroix-Pepin, Nicolas; Robert, Claude; Sullivan, Robert

2012-12-01

358

Quantum Mechanical Model for Information Transfer from DNA to Protein  

E-print Network

A model for the information transfer from DNA to protein using quantum information and computation techniques is presented. DNA is modeled as the sender and proteins are modeled as the receiver of this information. On the DNA side, a 64-dimensional Hilbert space is used to describe the information stored in DNA triplets (codons). A Hamiltonian matrix is constructed for this space, using the 64 possible codons as base states. The eigenvalues of this matrix are not degenerate. The genetic code is degenerate and proteins comprise only 20 different amino acids. Since information is conserved, the information on the protein side is also described by a 64-dimensional Hilbert space, but the eigenvalues of the corresponding Hamiltonian matrix are degenerate. Each amino acid is described by a Hilbert subspace. This change in Hilbert space structure reflects the nature of the processes involved in information transfer from DNA to protein.

Ioannis G. Karafyllidis

2008-01-18

359

GRAPHICAL MODELS FOR PROTEIN FUNCTION AND  

E-print Network

of bioinformatics can be viewed, in one sense, as the history of adopting and applying new algorithms to biological fashion and thus allows more sensitive searches of similarities in proteins. HMM has been widely applied describe the MRF and the CRF in comparison with the HMM. What follows are applications of the two graphical

Kihara, Daisuke

360

ANIMAL MODELS FOR PROTEIN RESPIRATORY SENSITIZERS  

EPA Science Inventory

Protein induced respiratory hypersensitivity, particularly atopic disease in general, and allergic asthma in particular, has increased dramatically over the last several decades in the U.S. and other industrialized nations as a result of ill-defined changes in living conditions i...

361

Structured Probabilistic Models of Proteins across Spatial and Fitness Landscapes  

E-print Network

Games, Protein Structure, Protein Design, Drug Design #12;Dedicated to the memory of my uncle, P interactions quickly and accurately. We then develop a method of learning generative models of amino acid, this dissertation develops a game-theoretic ap- proach to drug design (GAMUT). GAMUT determines the affects

362

Accuracy of functional surfaces on comparatively modeled protein structures  

E-print Network

. Keywords Protein binding surface Á Comparative model Á Signatures of binding pockets Á Amylase Abbreviation Signature of local active regions Introduction Proteins carry out their biological functions through bind- oped a method using active-site profiles to identify resi- dues located in the spatial environment

Dai, Yang

363

Assessment of Bdellovibrio bacteriovorus 109J killing of Moraxella bovis in an in vitro model of infectious bovine keratoconjunctivitis  

PubMed Central

The objective of this study was to determine the potential of Bdellovibrio bacteriovorus 109J as an alternative non-chemotherapeutic treatment of infectious bovine keratoconjunctivitis (IBK). To accomplish this, various parameters of B. bacteriovorus predation of Moraxella bovis were determined in vitro. Initial passage of B. bacteriovorus using M. bovis as prey required 10 d for active cultures to develop compared with 2 d for culture on normal Escherichia coli prey; however by the 5th passage, time to active predatory morphology was reduced to 2 d. This high passage B. bacteriovorus culture [1 × 1010 plaque forming units (PFU)/mL] killed 76% of M. bovis [1 × 107 colony forming units (CFU)/mL] present in suspension broth in a 4 h assay. The minimal level of M. bovis supporting B. bacteriovorus predation was 1 × 104 CFU/mL. To assess the ability of B. bacteriovorus to kill M. bovis on an epithelial surface mimicking IBK, an in vitro assay with Madin-Darby bovine kidney (MDBK) cells inoculated with 4 × 107 CFU/mL M. bovis was used. Treatment with a B. bacteriovorus suspension (1.6 × 1011 PFU/mL) decreased adherence of M. bovis to MDBK cells by 6-fold at 12 h of treatment, as well as decreased the number of unattached M. bovis cells by 1.4-fold. This study demonstrates that B. bacteriovorus has potential as an effective biological control of M. bovis at levels likely present in IBK-infected corneal epithelia and ocular secretions. PMID:22468026

Boileau, Melanie J.; Clinkenbeard, Kenneth D.; Iandolo, John J.

2011-01-01

364

MicroRNA Regulation of Bovine Monocyte Inflammatory and Metabolic Networks in an In Vivo Infection Model  

PubMed Central

Bovine mastitis is an inflammation-driven disease of the bovine mammary gland that costs the global dairy industry several billion dollars per year. Because disease susceptibility is a multifactorial complex phenotype, an integrative biology approach is required to dissect the molecular networks involved. Here, we report such an approach using next-generation sequencing combined with advanced network and pathway biology methods to simultaneously profile mRNA and miRNA expression at multiple time points (0, 12, 24, 36 and 48 hr) in milk and blood FACS-isolated CD14+ monocytes from animals infected in vivo with Streptococcus uberis. More than 3700 differentially expressed (DE) genes were identified in milk-isolated monocytes (MIMs), a key immune cell recruited to the site of infection during mastitis. Upregulated genes were significantly enriched for inflammatory pathways, whereas downregulated genes were enriched for nonglycolytic metabolic pathways. Monocyte transcriptional changes in the blood, however, were more subtle but highlighted the impact of this infection systemically. Genes upregulated in blood-isolated monocytes (BIMs) showed a significant association with interferon and chemokine signaling. Furthermore, 26 miRNAs were DE in MIMs and three were DE in BIMs. Pathway analysis revealed that predicted targets of downregulated miRNAs were highly enriched for roles in innate immunity (FDR < 3.4E?8), particularly TLR signaling, whereas upregulated miRNAs preferentially targeted genes involved in metabolism. We conclude that during S. uberis infection miRNAs are key amplifiers of monocyte inflammatory response networks and repressors of several metabolic pathways. PMID:24470219

Lawless, Nathan; Reinhardt, Timothy A.; Bryan, Kenneth; Baker, Mike; Pesch, Bruce; Zimmerman, Duane; Zuelke, Kurt; Sonstegard, Tad; O'Farrelly, Cliona; Lippolis, John D.; Lynn, David J.

2014-01-01

365

fast_protein_cluster: parallel and optimized clustering of large-scale protein modeling data  

PubMed Central

Motivation: fast_protein_cluster is a fast, parallel and memory efficient package used to cluster 60 000 sets of protein models (with up to 550 000 models per set) generated by the Nutritious Rice for the World project. Results: fast_protein_cluster is an optimized and extensible toolkit that supports Root Mean Square Deviation after optimal superposition (RMSD) and Template Modeling score (TM-score) as metrics. RMSD calculations using a laptop CPU are 60× faster than qcprot and 3× faster than current graphics processing unit (GPU) implementations. New GPU code further increases the speed of RMSD and TM-score calculations. fast_protein_cluster provides novel k-means and hierarchical clustering methods that are up to 250× and 2000× faster, respectively, than Clusco, and identify significantly more accurate models than Spicker and Clusco. Availability and implementation: fast_protein_cluster is written in C++ using OpenMP for multi-threading support. Custom streaming Single Instruction Multiple Data (SIMD) extensions and advanced vector extension intrinsics code accelerate CPU calculations, and OpenCL kernels support AMD and Nvidia GPUs. fast_protein_cluster is available under the M.I.T. license. (http://software.compbio.washington.edu/fast_protein_cluster) Contact: lhhung@compbio.washington.edu Supplementary information: Supplementary data are available at Bioinformatics online. PMID:24532722

Hung, Ling-Hong; Samudrala, Ram

2014-01-01

366

A Least Square Method Based Model for Identifying Protein Complexes in Protein-Protein Interaction Network  

PubMed Central

Protein complex formed by a group of physical interacting proteins plays a crucial role in cell activities. Great effort has been made to computationally identify protein complexes from protein-protein interaction (PPI) network. However, the accuracy of the prediction is still far from being satisfactory, because the topological structures of protein complexes in the PPI network are too complicated. This paper proposes a novel optimization framework to detect complexes from PPI network, named PLSMC. The method is on the basis of the fact that if two proteins are in a common complex, they are likely to be interacting. PLSMC employs this relation to determine complexes by a penalized least squares method. PLSMC is applied to several public yeast PPI networks, and compared with several state-of-the-art methods. The results indicate that PLSMC outperforms other methods. In particular, complexes predicted by PLSMC can match known complexes with a higher accuracy than other methods. Furthermore, the predicted complexes have high functional homogeneity.

Dai, Qiguo; Guo, Maozu; Guo, Yingjie; Liu, Xiaoyan; Liu, Yang; Teng, Zhixia

2014-01-01

367

Roles of ?-Turns in Protein Folding: From Peptide Models to Protein Engineering  

PubMed Central

Reverse turns are a major class of protein secondary structure; they represent sites of chain reversal and thus sites where the globular character of a protein is created. It has been speculated for many years that turns may nucleate the formation of structure in protein folding, as their propensity to occur will favor the approximation of their flanking regions and their general tendency to be hydrophilic will favor their disposition at the solvent-accessible surface. Reverse turns are local features, and it is therefore not surprising that their structural properties have been extensively studied using peptide models. In this article, we review research on peptide models of turns to test the hypothesis that the propensities of turns to form in short peptides will relate to the roles of corresponding sequences in protein folding. Turns with significant stability as isolated entities should actively promote the folding of a protein, and by contrast, turn sequences that merely allow the chain to adopt conformations required for chain reversal are predicted to be passive in the folding mechanism. We discuss results of protein engineering studies of the roles of turn residues in folding mechanisms. Factors that correlate with the importance of turns in folding indeed include their intrinsic stability, as well as their topological context and their participation in hydrophobic networks within the protein’s structure. PMID:18275088

Marcelino, Anna Marie C.; Gierasch, Lila M.

2010-01-01

368

Differential Virulence of Clinical and Bovine-Biased Enterohemorrhagic Escherichia coli O157:H7 Genotypes in Piglet and Dutch Belted Rabbit Models  

PubMed Central

Enterohemorrhagic Escherichia coli O157:H7 (EHEC O157) is an important cause of food and waterborne illness in the developed countries. Cattle are a reservoir host of EHEC O157 and a major source of human exposure through contaminated meat products. Shiga toxins (Stxs) are an important pathogenicity trait of EHEC O157. The insertion sites of the Stx-encoding bacteriophages differentiate EHEC O157 isolates into genogroups commonly isolated from cattle but rarely from sick humans (bovine-biased genotypes [BBG]) and those commonly isolated from both cattle and human patients (clinical genotypes [CG]). Since BBG and CG share the cardinal virulence factors of EHEC O157 and are carried by cattle at similar prevalences, the infrequent occurrence of BBG among human disease isolates suggests that they may be less virulent than CG. We compared the virulence potentials of human and bovine isolates of CG and BBG in newborn conventional pig and weaned Dutch Belted rabbit models. CG-challenged piglets experienced severe disease accompanied by early and high mortality compared to BBG-challenged piglets. Similarly, CG-challenged rabbits were likely to develop lesions in kidney and intestine compared with the BBG-challenged rabbits. The CG strains used in this study carried stx2 and produced significantly higher amounts of Stx, whereas the BBG strains carried the stx2c gene variant only. These results suggest that BBG are less virulent than CG and that this difference in virulence potential is associated with the Stx2 subtype(s) carried and/or the amount of Stx produced. PMID:22025512

Shringi, Smriti; Garcia, Alexis; Lahmers, Kevin K.; Potter, Kathleen A.; Muthupalani, Sureshkumar; Swennes, Alton G.; Hovde, Carolyn J.; Call, Douglas R.; Fox, James G.

2012-01-01

369

Food proteins and gut mucosal barrier. IV. Effects of acute and chronic ethanol administration on handling and uptake of bovine serum albumin by rat small intestine  

SciTech Connect

The effects of ethanol exposure on small intestinal handling and uptake of radiolabeled bovine serum albumin were investigated using everted gut sacs. There was less breakdown of BSA after acute ethanol administration in vitro and after acute and chronic in vivo exposure. Thus, the vascular compartment of the small intestine was confronted with more complete and potentially more antigenic material after ethanol. Changes in BSA binding and uptake after acute exposure were shown to be reversible after 4-6 hr. In all groups, there was more BSA binding when the small intestine was exposed to ethanol. This difference was most pronounced after chronic exposure. In the same group, uptake of BSA was correlated with binding and significantly increased. Combined effects of ethanol on the gut mucosal barrier may account for changes in food antigen handling and uptake.

Stern, M.; Carter, E.A.; Walker, W.A.

1986-11-01

370

Infectious Bovine Rhinotracheitis  

E-print Network

Infectious bovine rhinotracheitis (IBR) is a complex of disease syndromes occuring throughout the United States and the other major cattle-producing areas of the world. It affects cattle and some wild ruminants. This publication describes...

Sprott, L. R.

1998-11-30

371

Short Communication The effect of bovine serum albumin on batch and continuous enzymatic  

E-print Network

Short Communication The effect of bovine serum albumin on batch and continuous enzymatic cellulose Enzyme stability Cellulose a b s t r a c t Bovine serum albumin (BSA) was applied as a model non- teins such as bovine serum albumin (BSA) has been shown to en- hance cellulose hydrolysis by enzymes

California at Riverside, University of

372

Progression to persistent lymphocytosis and tumor development in bovine leukemia virus (BLV)-infected cattle correlates with impaired proliferation of CD4+ T cells in response to gag- and env-encoded BLV proteins.  

PubMed Central

The mechanism of leukemogenesis and persistent lymphocytosis (PL; benign expansion of B lymphocytes) in cattle infected with bovine leukemia virus (BLV; a retrovirus closely related to human T-cell leukemia virus type 1) is unknown; however, the immune system likely plays an important role in controlling the outcome of infection. In this study, we compared T-cell competence in serologically positive alymphocytotic (AL) animals with T-cell functions in animals with progressive stages of infection, PL and tumor bearing (TB). Dramatic differences were observed in lymphocyte proliferation to recombinant proteins encoded by BLV gag (p12, p15, and p24) and env (gp30, and gp51) genes in different disease stages. Lymphocytes from AL cattle recognized an average of three of five recombinant proteins per animal. Expansion of antigen pulsed lymphocytes in interleukin-2 increased protein recognition to almost five per animal. In contrast, lymphocytes from PL and TB animals failed to recognize any BLV recombinant proteins. Short-term T-cell cultures from the PL group expanded in interleukin-2, as well as the PL and TB cells cultured in indomethacin (3 to 6 microg/ml), increased the average of recognized proteins per animal to one. Cells proliferating to BLV antigens were CD4+ T lymphocytes, as shown by cell depletion studies. The positive effect of indomethacin suggests involvement of prostaglandin E2 as a negative regulatory factor in the later stages of disease. Thus, for the first time, advancing stages of BLV infection were correlated with decreased T-cell competence, providing deeper insight into pathogenesis of retroviral infections. PMID:8892878

Orlik, O; Splitter, G A

1996-01-01

373

A Computational Model of the LGI1 Protein Suggests a Common Binding Site for ADAM Proteins  

PubMed Central

Mutations of human leucine-rich glioma inactivated (LGI1) gene encoding the epitempin protein cause autosomal dominant temporal lateral epilepsy (ADTLE), a rare familial partial epileptic syndrome. The LGI1 gene seems to have a role on the transmission of neuronal messages but the exact molecular mechanism remains unclear. In contrast to other genes involved in epileptic disorders, epitempin shows no homology with known ion channel genes but contains two domains, composed of repeated structural units, known to mediate protein-protein interactions. A three dimensional in silico model of the two epitempin domains was built to predict the structure-function relationship and propose a functional model integrating previous experimental findings. Conserved and electrostatic charged regions of the model surface suggest a possible arrangement between the two domains and identifies a possible ADAM protein binding site in the ?-propeller domain and another protein binding site in the leucine-rich repeat domain. The functional model indicates that epitempin could mediate the interaction between proteins localized to different synaptic sides in a static way, by forming a dimer, or in a dynamic way, by binding proteins at different times. The model was also used to predict effects of known disease-causing missense mutations. Most of the variants are predicted to alter protein folding while several other map to functional surface regions. In agreement with experimental evidence, this suggests that non-secreted LGI1 mutants could be retained within the cell by quality control mechanisms or by altering interactions required for the secretion process. PMID:21479274

Leonardi, Emanuela; Andreazza, Simonetta; Vanin, Stefano; Busolin, Giorgia; Nobile, Carlo; Tosatto, Silvio C. E.

2011-01-01

374

Probing protein aggregation using simplified models and discrete molecular dynamics  

PubMed Central

Understanding the role of biomolecular dynamics in cellular processes leading to human diseases and the ability to rationally manipulate these processes is of fundamental importance in scientific research. The last decade has witnessed significant progress in probing biophysical behavior of proteins. However, we are still limited in understanding how changes in protein dynamics and inter-protein interactions occurring in short length- and time-scales lead to aberrations in their biological function. Bridging this gap in biology probed using computer simulations marks a challenging frontier in computational biology. Here we examine hypothesis-driven simplified protein models in conjunction with discrete molecular dynamics in the study of protein aggregation, implicated in series of neurodegenerative diseases, such as Alzheimer's and Huntington's diseases. Discrete molecular dynamics simulations of simplified protein models have emerged as a powerful methodology with its ability to bridge the gap in time and length scales from protein dynamics to aggregation, and provide an indispensable tool for probing protein aggregation. PMID:18508545

Sharma, Shantanu; Ding, Feng; Dokholyan, Nikolay V.

2008-01-01

375

COARSE-GRAINED MODELING OF PROTEIN UNFOLDING DYNAMICS*  

PubMed Central

We present a new dynamic elastic network model (DENM) that describes the unfolding process of a force-loaded protein. The protein interaction network and its potentials are constructed based on information of its native-state structure obtained from the Protein Data Bank, with network nodes positioned at the C? coordinates of the protein backbone. Specifically, to mimic the unfolding process, i.e., to simulate the process of overcoming the local energy barrier on the free energy landscape with force loading, the noncovalent protein network bonds (i.e., hydrogen bonds, salt bridges, hydrophobic contacts, etc.) are broken one-by-one with a certain probability, while the strong covalent bonds along the backbone (i.e., peptide bonds, disulfide bonds, etc.) are kept intact. The jumping event from local energy minima (bonds breaking rate) are chosen according to Kramer’s theory and the Bell model. Moreover, we exploit the self-similar structure of proteins at different scales to design an effective coarse-graining procedure for DENM with optimal parameter selection. The robustness of DENM is validated by coarse-grained molecular dynamics (MD) simulation against atomistic MD simulation of force-extension processes of the Fibrinogen and Titin Immunoglobulin proteins. We observe that the native structure of the proteins determines the total unfolding dynamics (including large deviations) and not just the fluctuations around the native state.

DENG, MINGGE

2014-01-01

376

A CONTINUUM HARD-SPHERE MODEL OF PROTEIN ADSORPTION.  

PubMed

Protein adsorption plays a significant role in biological phenomena such as cell-surface interactions and the coagulation of blood. Two-dimensional random sequential adsorption (RSA) models are widely used to model the adsorption of proteins on solid surfaces. Continuum equations have been developed so that the results of RSA simulations can be used to predict the kinetics of adsorption. Recently, Brownian dynamics simulations have become popular for modeling protein adsorption. In this work a continuum model was developed to allow the results from a Brownian dynamics simulation to be used as the boundary condition in a computational fluid dynamics (CFD) simulation. Brownian dynamics simulations were used to model the diffusive transport of hard-sphere particles in a liquid and the adsorption of the particles onto a solid surface. The configuration of the adsorbed particles was analyzed to quantify the chemical potential near the surface, which was found to be a function of the distance from the surface and the fractional surface coverage. The near-surface chemical potential was used to derive a continuum model of adsorption that incorporates the results from the Brownian dynamics simulations. The equations of the continuum model were discretized and coupled to a CFD simulation of diffusive transport to the surface. The kinetics of adsorption predicted by the continuum model closely matched the results from the Brownian dynamics simulation. This new model allows the results from mesoscale simulations to be incorporated into micro- or macro-scale CFD transport simulations of protein adsorption in practical devices. PMID:23729843

Finch, Craig; Clarke, Thomas; Hickman, James J

2013-07-01

377

Protein-protein interaction networks: probing disease mechanisms using model systems  

PubMed Central

Protein-protein interactions (PPIs) and multi-protein complexes perform central roles in the cellular systems of all living organisms. In humans, disruptions of the normal patterns of PPIs and protein complexes can be causative or indicative of a disease state. Recent developments in the biological applications of mass spectrometry (MS)-based proteomics have expanded the horizon for the application of systematic large-scale mapping of physical interactions to probe disease mechanisms. In this review, we examine the application of MS-based approaches for the experimental analysis of PPI networks and protein complexes, focusing on the different model systems (including human cells) used to study the molecular basis of common diseases such as cancer, cardiomyopathies, diabetes, microbial infections, and genetic and neurodegenerative disorders. PMID:23635424

2013-01-01

378

Weak self-interactions of globular proteins studied by small-angle X-ray scattering and structure-based modeling.  

PubMed

We investigate protein-protein interactions in solution by small-angle X-ray scattering (SAXS) and theoretical modeling. The structure factor for solutions of bovine pancreatic trypsin inhibitor (BPTI), myoglobin (Mb), and intestinal fatty acid-binding protein (IFABP) is determined from SAXS measurements at multiple concentrations, from Monte Carlo simulations with a coarse-grained structure-based interaction model, and from analytic approximate solutions of two idealized colloidal interaction models without adjustable parameters. By combining these approaches, we find that the structure factor is essentially determined by hard-core and screened electrostatic interactions. Other soft short-ranged interactions (van der Waals and solvation-related) are either individually insignificant or tend to cancel out. The structure factor is also not significantly affected by charge fluctuations. For Mb and IFABP, with a small net charge and relatively symmetric charge distribution, the structure factor is well described by a hard-sphere model. For BPTI, with a larger net charge, screened electrostatic repulsion is also important, but the asymmetry of the charge distribution reduces the repulsion from that predicted by a charged hard-sphere model with the same net charge. Such charge asymmetry may also amplify the effect of shape asymmetry on the protein-protein potential of mean force. PMID:25117055

Kaieda, Shuji; Lund, Mikael; Plivelic, Tomás S; Halle, Bertil

2014-08-28

379

Effects of wortmannin on the kinetics of GVBD and the activities of the maturation-promoting factor and mitogen-activated protein kinase during bovine oocyte maturation in vitro.  

PubMed

The present study was conducted with the objective of examining the effect of wortmanin, a specific PI 3-kinase inhibitor, on the kinetic of GVBD, and on the activities of the maturation-promoting factor (MPF) and mitogen-activated protein (MAP) kinase during bovine oocyte maturation. The time sequence for GVBD was not different between oocytes cultured with or without wortmannin. Most of the cultured oocytes were at the filamentous bivalents stage after 4 h of culture. Six hours after the start of culture, most of the oocytes possessed germinal vesicles with condensed bivalent, and by 10 h of culture nearly all of the cultured oocytes underwent GVBD. A gradual increase in MPF activity until 12 h of culture was observed in the presence and absence of wortmannin. A sharp decrease in MPF activity in oocytes cultured without wortmannin treatment was recorded at 14 h of culture. Thereafter, MPF regained activity, reaching a maximum level at 20 to 24 h of culture. For oocytes cultured with wortmannin, no decline in the activity of MPF was observed during the interval from 12 to 24 h of culture. For these oocytes the MPF activity remained nearly stable during this transition until the end of incubation. The presence of wortmannin in the maturation medium did not alter MAP kinase activity. Taken together, these observations indicate that inhibition of PI 3-kinase does not modulate the time sequence of GVBD or the pattern of MAP kinase activity in bovine oocytes. However, PI 3-kinase might be one of the molecules that regulate the sharp reduction in the activity of MPF during the MI/MII transition. PMID:10968422

Anas, M K; Shojo, A; Shimada, M; Terada, T

2000-06-01

380

Evaluation of innate immunity and vector toxicity following inoculation of bovine, porcine or human adenoviral vectors in a mouse model  

PubMed Central

Nonhuman adenovirus (Ad) vectors derived from bovine Ad serotype 3 (BAd3) or porcine Ad serotype 3 (PAd3) can circumvent pre-existing immunity against human Ad (HAd). We have previously reported differential transduction of human and nonhuman cells by these Ad vectors, and their distinct receptor usage and biodistribution. To compare the induction of innate immunity, vector toxicity and vector uptake by Kupffer cells (KCs) following intravenous administration of PAd3, BAd3, or HAd5 vectors in mice, we determined mRNA expression levels of proinflammatory chemokines and cytokines, and Toll-like receptors (TLRs) in the liver and spleen. Tissue toxicity of these vectors was assessed by comparing serum levels of liver-specific enzymes, histopathology and Kupffer cell (KC) depletion. Compared to the HAd5 vector, PAd3 and BAd3 vectors were more potent stimulators of innate immune responses as indicated by enhanced mRNA expression of TLRs and proinflammatory chemokines and cytokine genes. Histopathological changes in the liver were most pronounced in HAd5-inoculated mice while BAd3- or PAd3-inoculated mice revealed mild histologic changes that were confined to early time points. Inoculation with HAd5 or PAd3 vectors resulted in a significant (P <0.05) decline of the number of KCs in the liver. Together, these results extend our previous observations regarding distinct in vivo biology of nonhuman and human Ad vectors. PMID:20659505

Sharma, Anurag; Bangari, Dinesh S.; Tandon, Manish; HogenEsch, Harm; Mittal, Suresh K.

2010-01-01

381

Modeling of chemical inhibition from amyloid protein aggregation kinetics  

PubMed Central

Backgrounds The process of amyloid proteins aggregation causes several human neuropathologies. In some cases, e.g. fibrillar deposits of insulin, the problems are generated in the processes of production and purification of protein and in the pump devices or injectable preparations for diabetics. Experimental kinetics and adequate modelling of chemical inhibition from amyloid aggregation are of practical importance in order to study the viable processing, formulation and storage as well as to predict and optimize the best conditions to reduce the effect of protein nucleation. Results In this manuscript, experimental data of insulin, A?42 amyloid protein and apomyoglobin fibrillation from recent bibliography were selected to evaluate the capability of a bivariate sigmoid equation to model them. The mathematical functions (logistic combined with Weibull equation) were used in reparameterized form and the effect of inhibitor concentrations on kinetic parameters from logistic equation were perfectly defined and explained. The surfaces of data were accurately described by proposed model and the presented analysis characterized the inhibitory influence on the protein aggregation by several chemicals. Discrimination between true and apparent inhibitors was also confirmed by the bivariate equation. EGCG for insulin (working at pH?=?7.4/T?=?37°C) and taiwaniaflavone for A?42 were the compounds studied that shown the greatest inhibition capacity. Conclusions An accurate, simple and effective model to investigate the inhibition of chemicals on amyloid protein aggregation has been developed. The equation could be useful for the clear quantification of inhibitor potential of chemicals and rigorous comparison among them. PMID:24572069

2014-01-01

382

Stabilization and Controlled Release of Bovine Serum Albumin Encapsulated in Poly(D, L-lactide) and Poly(ethylene glycol) Microsphere Blends  

Microsoft Academic Search

Purpose. The acidic microclimate in poly(D, L-lactide-co-glycolide) 50\\/50 microspheres has been previously demonstrated by our group as the primary instability source of encapsulated bovine serum albumin (BSA). The objectives of this study were to stabilize the encapsulated model protein, BSA, and to achieve continuous protein release by using a blend of: slowly degrading poly(D, L-lactide) (PLA), to reduce the production

Wenlei Jiang; Steven P. Schwendeman

2001-01-01

383

Photolysis of Bovine Serum Albumin by near UV irradiation Sarah Foley1, Angela Staicu2, Alexandru Pascu2 & Mironel Enescu1  

E-print Network

Photolysis of Bovine Serum Albumin by near UV irradiation Sarah Foley1, Angela Staicu2, Alexandru August - 3rd September 2011, Coimbra, Portugal Bovine Serum Albumin (BSA) is the most abundant protein

Boyer, Edmond

384

Protein carbonylation, protein aggregation and neuronal cell death in a murine model of multiple sclerosis  

NASA Astrophysics Data System (ADS)

Many studies have suggested that oxidative stress plays an important role in the pathophysiology of both multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). Yet, the mechanism by which oxidative stress leads to tissue damage in these