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

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 haemoglobinβand lactoperoxidase.Conceivably,the binding of these proteins to the surfaces may contribute to biological processes occurring on them.

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

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 botmd 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 β and lactoperoxidase. Conceivably, the binding of these proteins to the surfaces may contribute to biological processes occurring on them.

Expression of Recombinant Bovine Prion Protein PrP27-30 in CHO-K1 Cells

[Objective] To investigate the possibilities of expressing bovine PrP27-30 gene in CHO-K1 cells. [Method] The purified PCR products of PrP27-30 were digested and ligated to the pCI-neo vector to yield pCI-neo-PrP27-30 that was used as an expression vector. Then CHO-K1 cells were transfected by pCI-neo-PrP27-30, and stable expression clone cells were screened by methotrexate (MTX) at a concentration of 0.1 and 1.0 umol/L. The transient expression was detected by indirect immunofluorescence assay and western blot. [ Result] After drug selection with MTX, the expression of PrP27-30 gene was detected in CHO-K1 cells. [Conclusion] Recombinant protein PrP27-30 expressed in CHO-K1 cells has better immunoreactivity and can be used to study secondary structure and regulation mechanism of pathological isoform of prion protein (prpC).

Expression of Recombinant Protein Bovine Prion pCIp264 in COS-7 Cells and Its Detection

Bovine spongiform encephalopathy （BSE） is thought to be caused by prions initially derived from sheep scrapie, and epidemiological studies suggest that new viriant form of CJD manifest in Great Britain may be caused by BSE prions. PrP^Sc is thought to pathogenic factor of transmissible spongiform encephalopathy （TSE）, which invariably involve a post-translational modification process of PrPc encoded by the host euchromosome PrP gene during the period it converted into the pathogenic form （PrP^Sc）, PrP is nomal cellular protein which has been found in both neuronal and nonneuronal tissues. Since the crucial infectious event in protein-transmitted diseases is an induced misfolding of prion proteins （PrP^c） catalyzed by already misfolded PrP^Sc, it is of high importance that such collisions are enhanced by two-dimensional diffusion in cell membranes is of high importance compared to three-dimensional diffusion in solution. The level of PrP mRNA in brain is higher than other tissue, but purification of PrPc from rodent has been difficult. To understand the formation of PrP^Sc, it seemed useful to develop a system for produced a large quantities of PrPc since there is no nature source of PrP^c. The pCI-neo mammalian expression vector contains the neomycin phosphtransferase gene which serves as a marker for the selection of stable transfected cells with G418. COS-7 cells constitutively express simian viruse 40 （SV40） T-antigen and support replication of expression plasmids containing the SV40 origin of replication, amplifying the introduced expression cassettes, now become important routine of expression a large number of heterologous gene products. In this paper, the authors used pCI-neo vector to construct a recombnant pCIp264 （cotains mPrP, N-signalpeptide and C-GPI anchor） plasmid to express it in the COS-7 cells and meanwhile detect the expression fusion using IN-ELISA, IN-IFA and western blot, and obtain some approximative nature PrPc.

Efficacy of fermented milk and whey proteins in Helicobacter pylori eradication:A review

Helicobacter pylori (H. pylori) eradication is considered a necessary step in the management of peptic ulcer disease, chronic gastritis, gastric adenocarcinoma and mucosa associated lymphoid tissue lymphoma. Standard triple therapy eradication regimens are inconvenient and achieve unpredictable and often poor results. Eradication rates are decreasing over time with increase in antibiotic resistance. Fermented milk and several of its component whey proteins have emerged as candidates for complementary therapy. In this context the current review seeks to summarize the current evidence available on their role in H. pylori eradication. Pertinent narrative/systematic reviews, clinical trials and laboratory studies on individual components including fermented milk, yogurt, whey proteins, lactoferrin, α-lactalbumin (α-LA), glycomacropeptide and immunoglobulin were comprehensively searched and retrieved from Medline, Embase, Scopus, Cochrane Controlled Trials Register and abstracts/proceedings of conferences up to May 2013. A preponderance of the evidence available on fermented milk-based probiotic preparations and bovine lactoferrin suggests a beneficial effect in Helicobacter eradication. Evidence for α-LA and immunoglobulins is promising while that for glycomacropeptide is preliminary and requires substantiation. The magnitude of the potential benefit documented so far is small and the precise clinical settings are ill defined. This restricts the potential use of this group as a complementary therapy in a nutraceutical setting hinging on better patient acceptability/compliance. Further work is necessary to identify the optimal substrate, fermentation process, dose and the ideal clinical setting (prevention/treatment, first line therapy/recurrence, symptomatic/asymptomatic, gastritis/ulcer diseases etc.). The potential of this group in high antibiotic resistance or treatment failure settings presents interesting possibilities and deserves further exploration.

Serum Proteins Stabilized Calcium Phosphate Nanoparticles and Its Effect on Bel-7402 Cells

Hydroxyapatite has a high affinity to biological macromolecules, especially to proteins. Bovine serum proteins were extracted to be used as stablizer to prepare calcium phosphate nanoparticles . 167.7 nm and 87.7 nm particles were respectively prepared by using bovine serum protein fractions at the concentration of 0. 5 mg/mL and 1.0 mg/mL. As the polysaccharide stabilized hydroxyapatite nanoparticles, the protein-stablized nanoparticles also inhibited the proliferation rate of Bel-7402 cells. It suggested that proteins could be applied to prepare calcium phosphate nanoparticles and it also has the anticaneer effect.

APPLICATION OF NONIONIC TEMPERATURE SENSITIVE HYDROGEL FOR CONCENTRATION OF PROTEIN AQUEOUS SOLUTION

Six different N-alkyl substituted acrylarnide nonionic hydrogels were prepared and their swelling characteristics were measured. Poly N-isopropyl acrylamide (PNIPA) and poly N-n-propyl-acrylamide (PNNPA) temperature sensitive hydrogels were chosen as the nonionic temperature sensitive hydrogels for concentration of very dilute aqueous protein solution. The separation properties of PNIPA and PNNPA hydr0gels with different network dimensions were studied and the modification of the hydrogels was surveyed in order to decrease their surface adsorption of protein molecules. The experimental results of the concentration of BSA (Bovin serum albumin) dilute aqueous solution by hydroxylpropyl methacrylate (HPMA) copolymerized PNIPA hydrogel were given. The value and the limitation of concentration of dilute aqueous protein solution by this method was evaluated.

Effects of fabrication of conjugates between different polyphenols and bovine bone proteins on their structural and functional properties

Covalent complexes of bovine bone proteins(BBP)with four polyphenols(chlorogenic acid(CA),quercetin(Q),rutin(R),epigallocatechin gallate(EGCG))with were prepared to increase the high-value utilization degree of bovine bone.The differences in coupling effect and structural and functional properties between the conjugates were also investigated.The results showed that EGCG-BBP had the highest polyphenolic equivalent at 88.05 mg/g protein and the lowest sulfhydryl group content.EGCG-BBP also had the best covalent effect.The SDS-PAGE and fluorescence spectra results revealed that the molecular weight of the conjugates increased significantly as different polyphenols conjugated with the aromatic amino acids in BBP,resulting in significant changes in the structure of EGCG-BBP.Fourier Transform infrared spectroscopy(FTIR)analysis demonstrated that the amide I band of conjugates was slightly shifted,indicating that the polyphenols were covalently bonded with BBP.Based on the results of this study and well-established mechanisms,it could be speculated that the best covalent effect of EGCG may be not only due to the eight phenolic hydroxyl groups but also to its two benzene rings,which are greatly increased in its reaction site.In terms of functional characteristics,EGCG-BBP had the highest 1-diphenyl-2-picryl-hydrazyl radical(DPPH·)scavenging capacity,while Q-BBP had the best solubility.Thus,this study provides a theoretical basis for different polyphenols’structural and functional effects on covalently modified proteins.

Preliminary study of adsorption behavior of bovine serum albumin(BSA) protein and its effect on antibacterial and corrosion property of Ti-3Cu alloy

The adsorption behavior, antibacterial, and corrosion properties of a Ti-3 Cu alloy were studied in a phosphate-buffered saline solution containing 0, 1, 3, and 6 gL^(-1) bovine serum albumin protein at 37℃ and pH = 7.4(±0.2). The protein adsorption behavior was examined via cyclic voltammetry, secondary ions mass spectroscopy(SIMS), and angle-resolved X-ray photoelectron spectroscopy(ARXPS). The corrosion property was analyzed by the open circuit potential(OCP), potentiodynamic polarization(PD),and electrochemical impedance spectroscopy(EIS) examinations. The antibacterial test was conducted according to the GB/T 21510 China Standard. It was observed that the surface charge density(QA DS) was directly proportional to the amount of the adsorbed BSA protein, signifying that the protein adsorption was accompanied by the charge transfer, pointing to chemisorptions phenomena. BSA amino groups and other organic species were observed in the surface analysis examinations. It was shown that the formation of barrier complexes between the TiO_(2) oxide-layer and PBS solution resulted in decreasing the release of Cu-ions, which consequently reduced the antibacterial activity. On the other hand, these barrier complexes improved the corrosion resistance by increasing the charge transfer resistance and double-layer capacitance of the Ti-3 Cu alloy.