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Sample records for include bovine serum

  1. Rotavirus-specific antibodies in fetal bovine serum and commercial preparations of serum albumin.

    OpenAIRE

    Offit, P A; Clark, H F; Taylor, A H; Hess, R G; Bachmann, P A; Plotkin, S A

    1984-01-01

    Rotavirus-specific antibodies were detected in fetal bovine serum, bovine serum albumin, and human serum albumin by radioimmunoprecipitation with the NCDV strain of bovine rotavirus as the detecting antigen. Fetal bovine sera neutralized bovine rotavirus in a plaque reduction neutralization test to titers of 1:20 or greater. Immunoglobulins purified from fetal bovine serum by protein A-agarose affinity chromatography precipitated rotavirus antigens but did not neutralize bovine rotavirus. Rot...

  2. Photodynamically generated bovine serum albumin radicals

    DEFF Research Database (Denmark)

    Silvester, J A; Timmins, G S; Davies, Michael Jonathan

    1998-01-01

    Porphyrin-sensitized photoxidation of bovine serum albumin (BSA) results in oxidation of the protein at (at least) two different, specific sites: the Cys-34 residue giving rise to a thiyl radical (RS.); and one or both of the tryptophan residues (Trp-134 and Trp-214) resulting in the formation...... of tertiary carbon-centred radicals and disruption of the tryptophan ring system. In the case of porphyrins such as hematoporphyrin, which bind at specific sites on BSA, these species appear to arise via long-range transfer of damage within the protein structure, as the binding site is some distance from...... the ultimate site of radical formation. This transfer of damage is shown to depend on a number of factors including the conformation of the protein, the presence of blocking groups and pH. Alteration of the protein conformation results in radical formation at additional (or alternative) sites, as does blocking...

  3. Aggregation and fibrillation of bovine serum albumin

    DEFF Research Database (Denmark)

    Holm, NK; Jespersen, SK; Thomassen, LV

    2007-01-01

    The all-alpha helix multi-domain protein bovine serum albumin (BSA) aggregates at elevated temperatures. Here we show that these thermal aggregates have amyloid properties. They bind the fibril-specific dyes Thioflavin T and Congo Red, show elongated although somewhat worm-like morphology...

  4. ADSORPTION OF BOVINE SERUM ALBUMIN ONTO ACTIVATED ...

    African Journals Online (AJOL)

    ABSTRACT. In this study, the adsorption of bovine serum albumin was studied using the activated carbon prepared from Elaeagnus stone with chemical activation. Elaeagnus stone activated carbon was characterized using the point of zero charge, Fourier transform infrared spectra, Brunauer-Emmet-Teller method, ...

  5. On the possible involvement of bovine serum albumin precursor in ...

    Indian Academy of Sciences (India)

    biotin affinity chromatography and mass finger printing analysis technique, herein we report the identification of a 70 kDa size protein (bovine serum albumin precursor, BSAP) which binds strongly with lipoplexes and may play role in lipofection ...

  6. Studies on the interaction between triptolide and Bovine Serum ...

    African Journals Online (AJOL)

    Studies on the interaction between triptolide and Bovine Serum Albumin (BSA) by spectroscopic and molecular modeling methods. Haidong Wang, Hailang Shi, Jie Pang, Xingfa Song, Caiyun Xu, Zengxian Sun ...

  7. Binding of anandamide to bovine serum albumin

    DEFF Research Database (Denmark)

    Bojesen, I.N.; Hansen, Harald S.

    2003-01-01

    The endocannabinoid anandamide is of lipid nature and may thus bind to albumin in the vascular system, as do fatty acids. The knowledge of the free water-phase concentration of anandamide is essential for the investigations of its transfer from the binding protein to cellular membranes, because a...... in aqueous compartments. - Bojesen, I. N., and H. S. Hansen. Binding of anandamide to bovine serum albumin.......The endocannabinoid anandamide is of lipid nature and may thus bind to albumin in the vascular system, as do fatty acids. The knowledge of the free water-phase concentration of anandamide is essential for the investigations of its transfer from the binding protein to cellular membranes, because...... a water-phase shuttle of monomers mediates such transfers. We have used our method based upon the use of albumin-filled red cell ghosts as a dispersed biological "reference binder" to measure the water-phase concentrations of anandamide. These concentrations were measured in buffer (pH 7.3) in equilibrium...

  8. Structural changes during the unfolding of Bovine serum albumin in ...

    Indian Academy of Sciences (India)

    The native form of serum albumin is the most important soluble protein in the body plasma. In order to investigate the structural changes of Bovine serum albumin (BSA) during its unfolding in the presence of urea, a small-angle neutron scattering (SANS) study was performed. The scattering curves of dilute solutions of BSA ...

  9. Interaction of Tannin with Bovine Serum Albumin by Fluorescence Spectrometry

    OpenAIRE

    Dong-Il, Kim; Son-Ae, Choe; Kye-Ryong, Sin

    2016-01-01

    Interaction between tannin and bovine serum albumin (BSA) was examined by the fluorescent quenching. The process of elimination between BSA and tannin was the one of a stationary state, and the coupling coefficient was one. The working strength between the tannin and the beef serum was hydrophobic one.

  10. A Novel Genetic Group of Bovine Hepacivirus in Archival Serum Samples from Brazilian Cattle

    Directory of Open Access Journals (Sweden)

    Cláudio W. Canal

    2017-01-01

    Full Text Available Hepatitis C virus (HCV (genus Hepacivirus; family Flaviviridae is a major human pathogen causing persistent infection and hepatic injury. Recently, emerging HCV-like viruses were described infecting wild animals, such as bats and rodents, and domestic animals, including dogs, horses, and cattle. Using degenerate primers for detecting bovine pestiviruses in a 1996 survey three bovine serum samples showed a low identity with the genus Pestivirus of the Flaviviridae family. A virus could not be isolated in cell culture. The description of bovine hepaciviruses (BovHepV in 2015 allowed us to retrospectively identify the sequences as BovHepV, with a 88.9% nucleotide identity. In a reconstructed phylogenetic tree, the Brazilian BovHepV samples grouped within the bovine HCV-like cluster in a separated terminal node that was more closely related to the putative bovine Hepacivirus common ancestor than to bovine hepaciviruses detected in Europe and Africa.

  11. Control of declared origin of bovine serum, a pilot study

    Science.gov (United States)

    Horacek, M.; Papesch, W.

    2009-04-01

    Bovine serum is the essential culture medium for cell cultures. Therefore it is highly demanded and the quality of the serum, e.g.: absence of bacteria, viruses certain antibodies, etc.., are important criteria. as some cattle diseases are endemic in certain regions, the origin of bovine serum is an important quality measure for its value. Thus the need to control the declared origins is present. Bovine serum was measured for d2H, d13C, d15N and d34S of proteine (dry residue) and d2H and d18O of the serum water. The hydrogen and oxygen are mainly depending by the isotopic composition of the water ingested by the cattle, and thus usually influenced by the isotopic signal of the precipitation. The carbon isotope signal is reflecting the diet of the cattle, whether it mainly feed on C3- or C4-plants. The nitrogen and sulphur isotope ratio is transferred from the ground/soil into the plant material and into the animal tissue, with some offset for nitrogen and without any significant offset for sulphur. Bovine serum samples from Canada, USA, Mexico, Brazil, Australia and New Zealand have been analysed. Due to the variations in the environmental conditions in different countries and regions which influence the isotope signatures of the serum samples it is possible to discriminate samples of different origin. Main discriminating parameters are d2H and d18O, d13C and d34S.

  12. Interaction of amphiphilic drugs with human and bovine serum albumins.

    Science.gov (United States)

    Khan, Abbul Bashar; Khan, Javed Masood; Ali, Mohd Sajid; Khan, Rizwan Hasan; Kabir-Ud-Din

    2012-11-01

    To know the interaction of amphiphilic drugs nortriptyline hydrochloride (NOT) and promazine hydrochloride (PMZ) with serum albumins (i.e., human serum albumin (HSA) and bovine serum albumin (BSA)), techniques of UV-visible, fluorescence, and circular dichroism (CD) spectroscopies are used. The binding affinity is more in case of PMZ with both the serum albumins. The quenching rate constant (k(q)) values suggest a static quenching process for all the drug-serum albumin interactions. The UV-visible results show that the change in protein conformation of PMZ-serum albumin interactions are more prominent as compared to NOT-serum albumin interactions. The CD results also explain the conformational changes in the serum albumins on binding with the drugs. The increment in %α-helical structure is slightly more for drug-BSA complexes as compared to drug-HSA complexes. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Sucrose/bovine serum albumin mediated biomimetic crystallization ...

    Indian Academy of Sciences (India)

    Administrator

    to animal phyla, algae, and in mollusk shells. 5. It has been found that supersaturation is critical in determining the calcium carbonate polymorph pre- ... in figure 1c: s (spherical), r (rhomboidal), f (filamentous). pure. The bovine serum albumin were extra pure. (more than 99%), and purchased from Sigma Chemi- cals.

  14. Serum antibody response to human and bovine IRBP in uveitis

    NARCIS (Netherlands)

    Hoekzema, R.; Hwan, S. B.; Rothova, A.; van Haren, M. A.; Donoso, L. A.; Kijlstra, A.

    1990-01-01

    Interphotoreceptor retinoid binding protein (IRBP) is a 136,000 molecular weight photoreceptor cell protein capable of inducing an experimental autoimmune uveitis (EAU) in susceptible animal strains. The occurrence of serum antibodies against human (Hu) or bovine (Bo) IRBP was investigated in

  15. Bovine Serum Albumin Metal Complexes for Mimic of SOD

    Indian Academy of Sciences (India)

    consequence, it can be considered as a bio-functional mimic of enzyme SOD and has a promising application prospect in antioxidant drug field. Keywords. Bovine serum albumin; biopolymer metal complexes; superoxide; free radical; scavenging activity. 1. Introduction. Reactive oxygen species (ROS), natural byproducts.

  16. Isolation of bovine serum albumin from whey using affinity chromatography

    NARCIS (Netherlands)

    Besselink, T.; Janssen, A.E.M.; Boom, R.M.

    2015-01-01

    The adsorption of bovine serum albumin (BSA) to a chromatography resin with immobilised llama antibody fragments as affinity ligands was investigated. The maximum adsorption capacity of the affinity resin was 21.6 mg mL-1 with a Langmuir equilibrium constant of 20.4 mg mg-1. Using packed bed

  17. Investigation of Bovine Serum Albumin (BSA) Attachment onto Self-Assembled Monolayers (SAMs) Using Combinatorial Quartz Crystal Microbalance with Dissipation (QCM-D) and Spectroscopic Ellipsometry (SE)

    Science.gov (United States)

    Phan, Hanh T. M.; Bartelt-Hunt, Shannon; Rodenhausen, Keith B.; Schubert, Mathias; Bartz, Jason C.

    2015-01-01

    Understanding protein adsorption kinetics to surfaces is of importance for various environmental and biomedical applications. Adsorption of bovine serum albumin to various self-assembled monolayer surfaces including neutral and charged hydrophilic and hydrophobic surfaces was investigated using in-situ combinatorial quartz crystal microbalance with dissipation and spectroscopic ellipsometry. Adsorption of bovine serum albumin varied as a function of surface properties, bovine serum albumin concentration and pH value. Charged surfaces exhibited a greater quantity of bovine serum albumin adsorption, a larger bovine serum albumin layer thickness, and increased density of bovine serum albumin protein compared to neutral surfaces at neutral pH value. The quantity of adsorbed bovine serum albumin protein increased with increasing bovine serum albumin concentration. After equilibrium sorption was reached at pH 7.0, desorption of bovine serum albumin occurred when pH was lowered to 2.0, which is below the isoelectric point of bovine serum albumin. Our data provide further evidence that combinatorial quartz crystal microbalance with dissipation and spectroscopic ellipsometry is a sensitive analytical tool to evaluate attachment and detachment of adsorbed proteins in systems with environmental implications. PMID:26505481

  18. Investigation of Bovine Serum Albumin (BSA Attachment onto Self-Assembled Monolayers (SAMs Using Combinatorial Quartz Crystal Microbalance with Dissipation (QCM-D and Spectroscopic Ellipsometry (SE.

    Directory of Open Access Journals (Sweden)

    Hanh T M Phan

    Full Text Available Understanding protein adsorption kinetics to surfaces is of importance for various environmental and biomedical applications. Adsorption of bovine serum albumin to various self-assembled monolayer surfaces including neutral and charged hydrophilic and hydrophobic surfaces was investigated using in-situ combinatorial quartz crystal microbalance with dissipation and spectroscopic ellipsometry. Adsorption of bovine serum albumin varied as a function of surface properties, bovine serum albumin concentration and pH value. Charged surfaces exhibited a greater quantity of bovine serum albumin adsorption, a larger bovine serum albumin layer thickness, and increased density of bovine serum albumin protein compared to neutral surfaces at neutral pH value. The quantity of adsorbed bovine serum albumin protein increased with increasing bovine serum albumin concentration. After equilibrium sorption was reached at pH 7.0, desorption of bovine serum albumin occurred when pH was lowered to 2.0, which is below the isoelectric point of bovine serum albumin. Our data provide further evidence that combinatorial quartz crystal microbalance with dissipation and spectroscopic ellipsometry is a sensitive analytical tool to evaluate attachment and detachment of adsorbed proteins in systems with environmental implications.

  19. Detection and identification of the atypical bovine pestiviruses in commercial foetal bovine serum batches.

    Directory of Open Access Journals (Sweden)

    Hongyan Xia

    Full Text Available The recently emerging atypical bovine pestiviruses have been detected in commercial foetal bovine serum (FBS of mainly South American origin so far. It is unclear how widely the viruses are presented in commercial FBS of different geographic origins. To further investigate the possible pestivirus contamination of commercially available FBS batches, 33 batches of FBS were obtained from ten suppliers and analysed in this study for the presence of both the recognised and the atypical bovine pestiviruses. All 33 batches of FBS were positive by real-time RT-PCR assays for at least one species of bovine pestiviruses. According to the certificate of analysis that the suppliers claimed for each batch of FBS, BVDV-1 was detected in all 11 countries and BVDV-2 was detected exclusively in the America Continent. The atypical pestiviruses were detected in 13 batches claimed to originate from five countries. Analysis of partial 5'UTR sequences showed a high similarity among these atypical bovine pestiviruses. This study has demonstrated, for the first time that commercial FBS batches of different geographic origins are contaminated not only with the recognised species BVDV-1 and BVDV-2, but also with the emerging atypical bovine pestiviruses.

  20. Affinity extraction of emerging contaminants from water based on bovine serum albumin as a binding agent.

    Science.gov (United States)

    Papastavros, Efthimia; Remmers, Rachael A; Snow, Daniel D; Cassada, David A; Hage, David S

    2018-03-01

    Affinity sorbents using bovine serum albumin as a binding agent were developed and tested for the extraction of environmental contaminants from water. Computer simulations based on a countercurrent distribution model were also used to study the behavior of these sorbents. Several model drugs, pesticides, and hormones of interest as emerging contaminants were considered in this work, with carbamazepine being used as a representative analyte when coupling the albumin column on-line with liquid chromatography and tandem mass spectrometry. The albumin column was found to be capable of extracting carbamazepine from aqueous solutions that contained trace levels of this analyte. Further studies of the bovine serum albumin sorbent indicated that it had higher retention under aqueous conditions than a traditional C 18 support for most of the tested emerging contaminants. Potential advantages of using these protein-based sorbents included the low cost of bovine serum albumin and its ability to bind to a relatively wide range of drugs and related compounds. It was also shown how simulations could be used to describe the elution behavior of the model compounds on the bovine serum albumin sorbents as an aid in optimizing the retention and selectivity of these supports for use with liquid chromatography or methods such as liquid chromatography with tandem mass spectrometry. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Bovine serum albumin: survival and osmolarity effect in bovine spermatozoa stored above freezing point.

    Science.gov (United States)

    Nang, C F; Osman, K; Budin, S B; Ismail, M I; Jaffar, F H F; Mohamad, S F S; Ibrahim, S F

    2012-05-01

    Liquid nitrogen preservation in remote farms is a limitation. The goal of this study was to determine optimum temperature above freezing point for bovine spermatozoa preservation using bovine serum albumin (BSA) as a supplementation. Pooled semen sample from three ejaculates was subjected to various BSA concentration (1, 4, 8 and 12 mg ml(-1)), before incubation in different above freezing point temperatures (4, 25 and 37 °C). Viability assessment was carried out against time from day 0 (fresh sample) until all spermatozoa become nonviable. Optimal condition for bovine spermatozoa storage was at 4 °C with 1 mg ml(-1) BSA for almost 7 days. BSA improved bovine spermatozoa viability declining rate to 44.28% at day 4 and 57.59% at day 7 compared to control, with 80.54% and 98.57% at day 4 and 7 respectively. Increase in BSA concentration did not improve sperm viability. Our results also confirmed that there was a strong negative correlation between media osmolarity and bovine spermatozoa survival rate with r = 0.885, P freezing point. © 2011 Blackwell Verlag GmbH.

  2. Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum

    OpenAIRE

    Ziegler, Anja; Everett, Helen; Hamza, Eman; Garbani, Mattia; Gerber, Vinzenz; Marti, Eliane; Steinbach, Falko

    2016-01-01

    BACKGROUND: Dendritic cells are professional antigen-presenting cells that play an essential role in the initiation and modulation of T cell responses. They have been studied widely for their potential clinical applications, but for clinical use to be successful, alternatives to xenogeneic substances like fetal bovine serum (FBS) in cell culture need to be found. Protocols for the generation of dendritic cells ex vivo from monocytes are well established for several species, including hor...

  3. Inactivation of viral agents in bovine serum by gamma irradiation.

    Science.gov (United States)

    House, C; House, J A; Yedloutschnig, R J

    1990-10-01

    Cell culture origin or suckling mouse brain origin viruses of Akabane disease, Aino, bovine ephemeral fever, swine vesicular disease, hog cholera, bluetongue, and minute virus of mice were each suspended in bovine serum. Aliquots (1 mL) were exposed to various doses of gamma radiation from a 60Co source while at -68 degrees C. Aliquots (100-mL) of serum from a steer experimentally infected with foot-and-mouth disease virus were similarly irradiated. The samples were assayed for infectivity in cell culture systems before and after irradiation, and the data points were analyzed by linear regression. The irradiation doses (in megarads) necessary to inactivate one log10 of viral infectivity (D10) was calculated for each virus. D10 is otherwise known as the slope of the regression line. The r2 value, a measure of association with 1.0 = perfect fit, was also calculated for each regression line. The values (D10, r2) for each virus were as follows: Akabane, 0.25, 0.998; Aino, 0.35, 0.997; bovine ephemeral fever, 0.29, 0.961; swine vesicular disease, 0.50, 0.969; foot-and-mouth disease, 0.53, 0.978; hog cholera, 0.55, 0.974; bluetongue, 0.83, 0.958; and minute virus of mice, 1.07, 0.935.

  4. Genome analysis of an atypical bovine pestivirus from fetal bovine serum.

    Science.gov (United States)

    Gao, Shandian; Du, Junzheng; Tian, Zhancheng; Xing, Shanshan; Chang, Huiyun; Liu, Guangyuan; Luo, Jianxun; Yin, Hong

    2016-08-01

    We report the complete genome sequence of a bovine pestivirus LVRI/cont-1 originated from a commercial batch of fetal bovine serum. Its complete genome consists of 12,282 nucleotides (nt), which contain an open reading frame (ORF) of 11,700 bp flanked by 5' and 3' untranslated regions (383 and 199 bp). The size of the 5'UTR and the individual protein coding region of LVRI/cont-1 are identical to those of the reference virus Th/04_KhonKaen, but it has a deletion of the first 56 nt in the 3'UTR. Alignment of the complete nucleotide sequence and phylogenetic analysis indicate that this viral isolate belongs to atypical pestiviruses.

  5. Improvement of the Shami goat semen quality by adding bovine serum albumin

    Directory of Open Access Journals (Sweden)

    O.I. Azawi

    2014-06-01

    Full Text Available The present study was aimed to improve the quality of Shami goat semen diluted with Tris diluent by adding bovine serum albumin. In the current study, six male goats were used. Semen was collected using artificial vagina of one ejaculate per week of every male included in this study. This study was performed during the breeding season from 1 \\ 10 \\ 2012 to 1 \\ 12 \\ 2012. In this study, two semen diluents were use first; Tris- fructose- egg yolk 2.5% and second Tris - fructose - 2.5% egg yolk with 1% of bovine serum albumin. Diluted semen samples were cooled gradually and stored at 5 ° C. Cooled diluted semen samples were examined every 24 h of storage to 144 h. These tests includes the proportion of live sperm and the percentage of secondary abnormalities of the sperm, the percentage of sperm acrosomal defects and percentage of progressive motility using a computer-aided sperm analysis. These results showed that the addition of bovine serum albumin with egg yolk to semen of male goats led to improved qualities of semen significantly (P<0.05 including the proportion of live sperm and the percentage of secondary abnormalities of the sperm, the percentage of sperm acrosomal defects and percentage of progressive motility. It could be concluded from the results of the current study, the possibility of storing goat semen for more than six days with alive sperm of more than 50% and the percentage of the progressive motility of more than 40% when adding bovine albumin serum to dilute goat semen at 1% level and this result has not reached by any previous study.

  6. 3-hydroxyflavone-bovine serum albumin interaction in Dextran medium

    Directory of Open Access Journals (Sweden)

    Voicescu Mariana

    2015-01-01

    Full Text Available Spectroscopic analysis of a bioactive flavonol, 3-Hydroxyflavone (3-HF, in systems based on Dextran 70 (Dx70 (an important bio-relevant polysacharide and Bovine Serum Albumin (BSA (a carrier protein, have been studied by fluorescence and circular dichroism. Changes produced by different concentrations of Dx70 on the fluorescent characteristics of 3-HF, and on the excited - state intramolecular proton transfer (ESIPT process were studied. The influence of 3-HF binding and of Dx70 on the secondary structure of BSA were investigated by circular dichroism spectroscopy. The influence of temperature (30-80°C range on the intrinsic Tryptophan fluorescence in 3-HF/BSA/Dx70 systems, was investigated. The results are discussed with relevance to 3-HF as a sensitive fluorescence probe for exploring flavone-protein interaction in plasma expander media and also for its biological evaluation.

  7. A low percentage of autologous serum can replace bovine serum to engineer human nasal cartilage

    Directory of Open Access Journals (Sweden)

    F Wolf

    2008-02-01

    Full Text Available For the generation of cell-based therapeutic products, it would be preferable to avoid the use of animal-derived components. Our study thus aimed at investigating the possibility to replace foetal bovine serum (FBS with autologous serum (AS for the engineering of cartilage grafts using expanded human nasal chondrocytes (HNC. HNC isolated from 7 donors were expanded in medium containing 10% FBS or AS at different concentrations (2%, 5% and 10% and cultured in pellets using serum-free medium or in Hyaff®-11 meshes using medium containing FBS or AS. Tissue forming capacity was assessed histologically (Safranin O, immunohistochemically (type II collagen and biochemically (glycosaminoglycans -GAG- and DNA. Differences among experimental groups were assessed by Mann Whitney tests. HNC expanded under the different serum conditions proliferated at comparable rates and generated cartilaginous pellets with similar histological appearance and amounts of GAG. Tissues generated by HNC from different donors cultured in Hyaff®-11 had variable quality, but the accumulated GAG amounts were comparable among the different serum conditions. Staining intensity for collagen type II was consistent with GAG deposition. Among the different serum conditions tested, the use of 2% AS resulted in the lowest variability in the GAG contents of generated tissues. In conclusion, a low percentage of AS can replace FBS both during the expansion and differentiation of HNC and reduce the variability in the quality of the resulting engineered cartilage tissues.

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

    Directory of Open Access Journals (Sweden)

    Petschow BW

    2014-05-01

    Full Text Available Bryon W Petschow, Bruce Burnett, Audrey L Shaw, Eric M Weaver, Gerald L Klein Entera Health, Inc., Cary, NC, USA Abstract: 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. Keywords: bovine immunoglobulins, nutrient, gut barrier, microbiota

  9. Investigation of interactions between dendrimer-coated magnetite nanoparticles and bovine serum albumin

    International Nuclear Information System (INIS)

    Pan Bifeng; Gao Feng; Ao Limei

    2005-01-01

    We investigated the interactions between dendrimer-coated magnetite nanoparticles (MNPs) and the protein serum albumin. The investigation was based on the fluorescence quenching of tryptophan residue of serum albumin after binding with the dendrimer-coated magnetite nanoparticles. The extent of the interactions between bovine serum albumin and dendrimer-coated MNPs strongly depends on their surface groups and pH value

  10. Contact angle studies on PDMS surfaces fouled by bovine serum albumin

    CSIR Research Space (South Africa)

    Windvoel, VT

    2010-01-01

    Full Text Available proteins. This is a limitation to microfluidic applications that require hydrophobic surfaces where proteins are involved. This study determines the change in wetting of PDMS after fouling by a protein, bovine serum albumin (BSA), by measuring contact...

  11. Formation of complexes between tannic acid with bovine serum albumin, egg ovalbumin and bovine beta-lactoglobulin.

    Science.gov (United States)

    Xie, Liyang; Wehling, Randy L; Ciftci, Ozan; Zhang, Yue

    2017-12-01

    Tannic acid (TA) shows strong interactions with proteins and the resulting complexes can be utilized as delivery systems for oral drugs. The complexation of TA with three proteins including bovine serum albumin (BSA), egg ovalbumin (EA) and bovine beta-lactoglobulin (BLG) at pH 7.4 was studied. The tryptophan (Trp) fluorescence of all three proteins was quenched by TA in a static quenching mechanism. BLG showed the highest binding affinity and a smallest binding distance with TA which may suggest that BLG-TA is the most stable complex. The results of circular dichroism, synchronous and three-dimensional fluorescence spectra suggested that the protein structures have been changed at different levels and helix structure was affected more significant than β-strand. Zeta-potential of all three proteins was more negative after binding with TA, which is favorable for the stabilization of protein based nanoparticles. Information derived from this work could be important to potentially use TA-protein complexes as nanoencapsulation systems for oral drug delivery. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Study of the interaction of kaempferol with bovine serum albumin

    Science.gov (United States)

    Tian, Jianniao; Liu, Jiaqin; Tian, Xuan; Hu, Zhide; Chen, Xingguo

    2004-03-01

    The binding of kaempferol with bovine serum albumin (BSA) was investigated at three temperatures, 296, 310 and 318 K, by the fluorescence, circular dichroism (CD) and Fourier transform infrared spectroscopy (FT-IR) at pH 7.40. The CD and FT-IR studies indicate that kaempferol binds strongly to BSA. The association constant K was determined by Stern-Volmer equation based on the quenching of the fluorescence BSA in the presence of kaempferol. The thermodynamic parameters were calculated according to the dependence of enthalpy change on the temperature as follows: Δ H0 and Δ S0 possess small negative (-1.694 kJ/mol) and positive values (88.814 J/mol K), respectively. According to the displacement experimental and the thermodynamic results, it is considered that kaempferol binding site II (subdomain III) mainly by hydrophobic interaction. The results studied by FT-IR and CD experiments indicate that the secondary structures of the protein have been changed by the interaction of kaempferol with BSA. The distance between the tryptophan residues in BSA and kaempferol bound to site II was estimated to be 2.78 nm using Foster's equation on the basis of fluorescence energy transfer.

  13. Bovine serum albumin adsorption on functionalized porous silicon surfaces

    Science.gov (United States)

    Tay, Li-Lin; Rowell, Nelson L.; Lockwood, David J.; Boukherroub, Rabah

    2004-10-01

    The large surface area within porous Si (pSi) and its strong room temperature photoluminescence (PL) make it an ideal host for biological sensors. In particular, the development of pSi-based optical sensors for DNA, enzyme and other biochemical molecules have become of great interest. Here, we demonstrate that the in-situ monitoring of the pSi PL behaviour can be used as a positive identification of bovine serum albumin (BSA) protein adsorption inside the porous matrix. Electrochemically prepared pSi films were first functionalized with undecylenic acid to produce an organic monolayer covalently attached to the porous silicon surfaces. The acid terminal group also provided favourable BSA binding sites on the pSi matrix sidewalls. In-situ PL spectra showed a gradual red shift (up to 12 meV) in the PL peak energy due to the protein incorporation into the porous matrix. The PL then exhibited a continuous blue shift after saturation of the protein molecules in the pores. This blue shift of the PL peak frequency and a steady increase in the PL intensity is evidence of surface oxidation. Comparing the specular reflectance obtained by Fourier transform infrared spectroscopy (FTIR) before and after BSA incubation confirmed the adsorption of protein in the pSi matrix.

  14. Bovine serum albumin adsorption on passivated porous silicon layers

    Science.gov (United States)

    Lockwood, David; Boukherroub, Rabah

    2005-03-01

    Hydrogen-terminated porous silicon (pSi) films were fabricated through electrochemical anodization of crystalline Si in HF-based solutions. The pSi-H surface was chemically functionalized by thermal reaction with undecylenic acid to produce an organic monolayer covalently attached to the silicon surface through Si-C bonds and bearing an acid terminal group. Bovine serum albumin (BSA) was then adsorbed onto the modified surface. SEM showed that the porous films were damaged and partially lifted off the Si substrate after a prolonged BSA adsorption. Ellipsometry revealed that the BSA had penetrated ˜ 1.3 micrometers into the porous structure. The film damage results from BSA anchoring itself tightly through strong electrostatic interactions to the acid-covered Si sidewalls. A change in surface tension during BSA film formation then causes the pSi layer to buckle and lift-off the underlying Si substrate. FTIR results from the modified pSi surfaces showed the presence of strong characteristic Amide I, II and III vibrational bands after BSA adsorption.

  15. Polyacrylic acids–bovine serum albumin complexation: Structure and dynamics

    International Nuclear Information System (INIS)

    Othman, Mohamed; Aschi, Adel; Gharbi, Abdelhafidh

    2016-01-01

    The study of the mixture of BSA with polyacrylic acids at different masses versus pH allowed highlighting the existence of two regimes of weak and strong complexation. These complexes were studied in diluted regime concentration, by turbidimetry, dynamic light scattering (DLS), zeta-potential measurements and nuclear magnetic resonance (NMR). We have followed the pH effect on the structure and properties of the complex. This allowed refining the interpretation of the phase diagram and understanding the observed phenomena. The NMR measurements allowed probing the dynamics of the constituents versus the pH. The computational method was used to precisely determine the electrostatic potential of BSA and how the polyelectrolyte binds to it at different pH. - Highlights: • Influence of physico-chemical parameters on the electrostatic interactions in the complex system (polyelectrolyte/protein). • Stabilization and encapsulation of biological macromolecules solution by mean of polyelectrolyte. • Properties and structure of mixture obtained by screening the charges of globular protein and at different masses of polyacrylic acids. • Dynamic of the constituents formed by complexes particles. • Evaluation of the electrostatic properties of bovine serum albumin versus pH through solution of the Poisson-Boltzmann equation.

  16. Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum.

    Science.gov (United States)

    Ziegler, Anja; Everett, Helen; Hamza, Eman; Garbani, Mattia; Gerber, Vinzenz; Marti, Eliane; Steinbach, Falko

    2016-11-15

    Dendritic cells are professional antigen-presenting cells that play an essential role in the initiation and modulation of T cell responses. They have been studied widely for their potential clinical applications, but for clinical use to be successful, alternatives to xenogeneic substances like fetal bovine serum (FBS) in cell culture need to be found. Protocols for the generation of dendritic cells ex vivo from monocytes are well established for several species, including horses. Currently, the gold standard protocol for generating dendritic cells from monocytes across various species relies upon a combination of GM-CSF and IL-4 added to cell culture medium which is supplemented with FBS. The aim of this study was to substitute FBS with heterologous horse serum. For this purpose, equine monocyte-derived dendritic cells (eqMoDC) were generated in the presence of horse serum or FBS and analysed for the effect on morphology, phenotype and immunological properties. Changes in the expression of phenotypic markers (CD14, CD86, CD206) were assessed during dendritic cell maturation by flow cytometry. To obtain a more complete picture of the eqMoDC differentiation and assess possible differences between FBS- and horse serum-driven cultures, a transcriptomic microarray analysis was performed. Lastly, immature eqMoDC were primed with a primary antigen (ovalbumin) or a recall antigen (tetanus toxoid) and, after maturation, were co-cultured with freshly isolated autologous CD5 + T lymphocytes to assess their T cell stimulatory capacity. The microarray analysis demonstrated that eqMoDC generated with horse serum were indistinguishable from those generated with FBS. However, eqMoDC incubated with horse serum-supplemented medium exhibited a more characteristic dendritic cell morphology during differentiation from monocytes. A significant increase in cell viability was also observed in eqMoDC cultured with horse serum. Furthermore, eqMoDC generated in the presence of horse serum

  17. Interactions between imazethapyr and bovine serum albumin: Spectrofluorimetric study

    Energy Technology Data Exchange (ETDEWEB)

    Pacheco, Maria E. [Division Quimica Analitica, Departamento de Quimica, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires (Argentina); Bruzzone, Liliana, E-mail: bruzzone@quimica.unlp.edu.ar [Division Quimica Analitica, Departamento de Quimica, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Buenos Aires (Argentina)

    2012-10-15

    The interaction between imazethapyr (IMA) and bovine serum albumin (BSA) was investigated by fluorescence spectroscopy. The Stern-Volmer quenching constant (K{sub SV}) at three temperatures was evaluated in order to determine the quenching mechanism. The dependence of fluorescence quenching on viscosity was also evaluated for this purpose. The results showed that IMA quenches the fluorescence intensity of BSA through a static quenching process. The values of the binding constant for the formed BSA-IMA complex and the number of binding sites were found to be 1.51 Multiplication-Sign 10{sup 5} M{sup -1} and 0.77, respectively, at room temperature. Based on the calculated thermodynamic parameters, the forces that dominate the binding process are hydrogen bonds and van der Waals forces, and the binding process is spontaneous and exothermic. The quenching of protein fluorescence by iodide ion was used to probe the accessibility of tryptophan residues in BSA and the change in accessibility induced by the presence of IMA. According to the obtained results, the BSA-IMA complex is formed in the site where the Trp-134 is located, causing it to become less exposed to the solvent. - Highlights: Black-Right-Pointing-Pointer Fluorescence spectroscopy helps to understand protein binding mechanisms. Black-Right-Pointing-Pointer Quenching measurements reveal the nature of the binding process involved. Black-Right-Pointing-Pointer Iodine ion can be used to study the change in accessibility of tryptophan residues. Black-Right-Pointing-Pointer Thermodynamic parameters for the binding reaction confirm binding modes.

  18. Kinetics of Thermal Denaturation and Aggregation of Bovine Serum Albumin.

    Directory of Open Access Journals (Sweden)

    Vera A Borzova

    Full Text Available Thermal aggregation of bovine serum albumin (BSA has been studied using dynamic light scattering, asymmetric flow field-flow fractionation and analytical ultracentrifugation. The studies were carried out at fixed temperatures (60°C, 65°C, 70°C and 80°C in 0.1 M phosphate buffer, pH 7.0, at BSA concentration of 1 mg/ml. Thermal denaturation of the protein was studied by differential scanning calorimetry. Analysis of the experimental data shows that at 65°C the stage of protein unfolding and individual stages of protein aggregation are markedly separated in time. This circumstance allowed us to propose the following mechanism of thermal aggregation of BSA. Protein unfolding results in the formation of two forms of the non-native protein with different propensity to aggregation. One of the forms (highly reactive unfolded form, Uhr is characterized by a high rate of aggregation. Aggregation of Uhr leads to the formation of primary aggregates with the hydrodynamic radius (Rh,1 of 10.3 nm. The second form (low reactive unfolded form, Ulr participates in the aggregation process by its attachment to the primary aggregates produced by the Uhr form and possesses ability for self-aggregation with formation of stable small-sized aggregates (Ast. At complete exhaustion of Ulr, secondary aggregates with the hydrodynamic radius (Rh,2 of 12.8 nm are formed. At 60°C the rates of unfolding and aggregation are commensurate, at 70°C the rates of formation of the primary and secondary aggregates are commensurate, at 80°C the registration of the initial stages of aggregation is complicated by formation of large-sized aggregates.

  19. Fluorescence lifetime measurements of native and glycated human serum albumin and bovine serum albumin

    Science.gov (United States)

    Joshi, Narahari V.; Joshi, Virgina O. d.; Contreras, Silvia; Gil, Herminia; Medina, Honorio; Siemiarczuk, Aleksander

    1999-05-01

    Nonenzymatic glycation, also known as Maillard reaction, plays an important role in the secondary complications of the diabetic pathology and aging, therefore, human serum albumin (HSA) and bovine serum albumin (BSA) were glycated by a conventional method in our laboratory using glucose as the glycating agent. Fluorescence lifetime measurements were carried out with a laser strobe fluorometer equipped with a nitrogen/dye laser and a frequency doubler as a pulsed excitation source. The samples were excited at 295 nm and the emission spectra were recorded at 345 nm. The obtained decay curves were tried for double and triple exponential functions. It has been found that the shorter lifetime increases for glycated proteins as compared with that of the native ones. For example, in the case of glycated BSA the lifetime increased from 1.36 ns to 2.30 ns. Similarly, for HSA, the lifetime increases from 1.58 ns to 2.26 ns. Meanwhile, the longer lifetime changed very slightly for both proteins (from 6.52 ns to 6.72 ns). The increase in the lifetime can be associated with the environmental effect; originated from the attachment of glucose to some lysine residues. A good example is Trp 214 which is in the cage of Lys 225, Lys 212, Lys 233, Lys 205, Lys 500, Lys 199 and Lys 195. If fluorescence lifetime technique is calibrated and properly used it could be employed for assessing glycation of proteins.

  20. Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis

    Directory of Open Access Journals (Sweden)

    Jae Eun Lee

    2015-06-01

    Full Text Available Two dimensional-fluorescence difference gel electrophoresis (2D DIGE is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

  1. Proteomic Analysis of Bovine Pregnancy-specific Serum Proteins by 2D Fluorescence Difference Gel Electrophoresis

    Science.gov (United States)

    Lee, Jae Eun; Lee, Jae Young; Kim, Hong Rye; Shin, Hyun Young; Lin, Tao; Jin, Dong Il

    2015-01-01

    Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum. PMID:25925056

  2. Human autologous serum as a substitute for fetal bovine serum in human Schwann cell culture.

    Directory of Open Access Journals (Sweden)

    Parisa Goodarzi

    2014-04-01

    Full Text Available Nowadays, cell -based and tissue engineered products have opened new horizons in treatment of incurable nervous system disorders. The number of studies on the role of Schwann cells (SC in treating nervous disorders is higher than other cell types. Different protocols have been suggested for isolation and expansion of SC which most of them have used multiple growth factors, mitogens and fetal bovine sera (FBS in culture medium. Because of potential hazards of animal-derived reagents, this study was designed to evaluate the effect of replacing FBS with human autologous serum (HAS on SC's yield and culture parameters. Samples from 10 peripheral nerve biopsies were retrieved and processed under aseptic condition. The isolated cells cultured in FBS (1st group or autologous serum (2nd group. After primary culture the cells were seeded at 10000 cell/cm2 in a 12 wells cell culture plate for each group. At 100% confluency, the cell culture parameters (count, viability, purity and culture duration of 2 groups were compared using paired t-test. The average donors' age was 35.80 (SD=13.35 and except for 1 sample the others cultured successfully. In first group, the averages of cell purity, viability and culture duration were 97% (SD=1.32, 97/33% (SD=1.22 and 11.77 (SD=2.58 days respectively. This parameters were 97.33% (SD=1.00, 97.55% (SD=1.33 and 10.33 days (SD=1.65 in second group. The difference of cell count, purity and viability were not significant between 2 groups (P>0.05. The cells of second group reached to 100% confluency in shorter period of time (P=0.03. The results of this study showed that autologous serum can be a good substitute for FBS in human SC culture. This can reduce the costs and improve the safety of cell product for clinical application.

  3. Interaction of Water-Soluble CdTe Quantum Dots with Bovine Serum Albumin

    Science.gov (United States)

    2011-01-01

    Semiconductor nanoparticles (quantum dots) are promising fluorescent markers, but it is very little known about interaction of quantum dots with biological molecules. In this study, interaction of CdTe quantum dots coated with thioglycolic acid (TGA) with bovine serum albumin was investigated. Steady state spectroscopy, atomic force microscopy, electron microscopy and dynamic light scattering methods were used. It was explored how bovine serum albumin affects stability and spectral properties of quantum dots in aqueous media. CdTe–TGA quantum dots in aqueous solution appeared to be not stable and precipitated. Interaction with bovine serum albumin significantly enhanced stability and photoluminescence quantum yield of quantum dots and prevented quantum dots from aggregating. PMID:27502633

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

    Science.gov (United States)

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

    2013-10-01

    Because the use of serum in the embryo cryopreservation increases the probability of animal health problems such as bovine spongiform encephalopathy (BSE) and viral infections, this study was conducted to examine the effects of sericin supplementation for serum-free freezing medium on the survival and development of bovine embryos after freezing-thawing and direct transfer to recipients. When in vitro-produced bovine embryos were frozen conventionally in the freezing medium supplemented with various concentrations (0.1%, 0.5%, and 1.0%) of sericin, the percentages of damaged zona pellucida, survival, and development of frozen-thawed embryos were similar to those of embryos frozen in freezing medium supplemented with 0.4% bovine serum albumin (BSA) and 20% fetal bovine serum (FBS) (0.4BSA/20F; control). When in vivo-derived embryos were frozen with 0.4BSA/20F (control), 0.5% sericin +20% FBS (0.5S/20F) or 0.5% sericin (0.5S) and were subsequently transferred directly to recipients, the percentages of recipients with pregnancy and normal calving in the 0.5S/20F group were higher than those in the control group (47.3% vs. 40.1% and 94.6% vs. 87.3%, respectively). Moreover, the percentages of recipients with pregnancy and normal calving (42.2% and 92.4%, respectively) in the 0.5S group were similar with those of other groups. In conclusion, these results indicate that serum-free freezing medium supplemented with sericin is available for the cryopreservation of bovine embryos and that it is beneficial for the elimination of a potential source of biological contamination by serum or BSA. Copyright © 2013. Published by Elsevier Inc.

  5. Cell Type-Specific Modulation of Cobalamin Uptake by Bovine Serum.

    Directory of Open Access Journals (Sweden)

    Hua Zhao

    Full Text Available Tracking cellular 57Co-labelled cobalamin (57Co-Cbl uptake is a well-established method for studying Cbl homeostasis. Previous studies established that bovine serum is not generally permissive for cellular Cbl uptake when used as a supplement in cell culture medium, whereas supplementation with human serum promotes cellular Cbl uptake. The underlying reasons for these differences are not fully defined. In the current study we address this question. We extend earlier observations by showing that fetal calf serum inhibits cellular 57Co-Cbl uptake by HT1080 cells (a fibrosarcoma-derived fibroblast cell line. Furthermore, we discovered that a simple heat-treatment protocol (95°C for 10 min ameliorates this inhibitory activity for HT1080 cell 57Co-Cbl uptake. We provide evidence that the very high level of haptocorrin in bovine serum (as compared to human serum is responsible for this inhibitory activity. We suggest that bovine haptocorrin competes with cell-derived transcobalamin for Cbl binding, and that cellular Cbl uptake may be minimised in the presence of large amounts of bovine haptocorrin that are present under routine in vitro cell culture conditions. In experiments conducted with AG01518 cells (a neonatal foreskin-derived fibroblast cell line, overall cellular 57Co-Cbl uptake was 86% lower than for HT1080 cells, cellular TC production was below levels detectable by western blotting, and heat treatment of fetal calf serum resulted in only a modest increase in cellular 57Co-Cbl uptake. We recommend a careful assessment of cell culture protocols should be conducted in order to determine the potential benefits that heat-treated bovine serum may provide for in vitro studies of mammalian cell lines.

  6. Binding of ethyl pyruvate to bovine serum albumin: Calorimetric, spectroscopic and molecular docking studies

    Energy Technology Data Exchange (ETDEWEB)

    Pathak, Mallika [Department of Chemistry, Miranda House, University of Delhi, Delhi 11007 (India); Mishra, Rashmi; Agarwala, Paban K. [Department of Radiation Genetics and Epigenetics, Division of Radioprotective Drug Development Research, Institute of Nuclear Medicine and Allied Sciences, Delhi 110054 (India); Ojha, Himanshu, E-mail: himanshu.drdo@gmail.com [Department of Radiation Genetics and Epigenetics, Division of Radioprotective Drug Development Research, Institute of Nuclear Medicine and Allied Sciences, Delhi 110054 (India); Singh, Bhawna [Department of Radiation Genetics and Epigenetics, Division of Radioprotective Drug Development Research, Institute of Nuclear Medicine and Allied Sciences, Delhi 110054 (India); Singh, Anju; Kukreti, Shrikant [Nucleic Acid Research Laboratory, Department of Chemistry, University of Delhi, Delhi 11007 (India)

    2016-06-10

    Highlights: • ITC study showed binding of ethyl pyruvate with BSA with high binding affinity. • Ethyl pyruvate binding caused conformation alteration of BSA. • Fluorescence quenching mechanism is static in nature. • Electrostatic, hydrogen bonding and hydrophobic forces involved in binding. • Docking confirmed role of electrostatic, hydrogen bonding and hydrophobic forces. - Abstract: Various in vitro and in vivo studies have shown the anti-inflammatory and anticancer potential role of ethyl pyruvate. Bio-distribution of drugs is significantly influenced by the drug-serum protein binding. Therefore, the binding mechanism of the ethyl pyruvate with bovine serum albumin was investigated using UV–vis absorption, fluorescence, circular dichroism, isothermal titration calorimetry and molecular docking techniques. Absorption and fluorescence quenching studies indicated the binding of ethyl pyruvate with protein. Circular dichroism spectra of bovine serum albumin confirmed significant change in the conformation of protein upon binding. Thermodynamic data confirmed that ethyl pyruvate binds to bovine serum albumin at the two different sites with high affinity. Binding of ethyl pyruvate to bovine serum albumin involves hydrogen bonding, van der Waal and hydrophobic interactions. Further, docking studies indicated that ethyl pyruvate could bind significantly at the three binding sites. The results will definitely contribute to the development of ethyl pyruvate as drug.

  7. Bovine Serum Albumin Metal Complexes for Mimic of SOD

    Indian Academy of Sciences (India)

    Proteins are well-known for efficiency and selecti- vity that few other natural or artificial molecules can match,24,25 whether in catalysis or molecular recogni- tion. Owing to 35 cysteine residues, human serum albu- min (HSA) is facile to be combined with metal ions. The serum albumin is readily available, non-antigenic.

  8. Biological Interaction of Molybdenocene Dichloride with Bovine Serum Albumin Using Fluorescence Spectroscopy

    Science.gov (United States)

    Domínguez, Moralba; Cortes-Figueroa, Jose´ E.; Meléndez, Enrique

    2018-01-01

    Bioinorganic topics are ubiquitous in the inorganic chemistry curriculum; however, experiments to enhance understanding of related topics are scarce. In this proposed laboratory, upper undergraduate students assess the biological interaction of molybdenocene dichloride (Cp2MoCl2) with bovine serum albumin (BSA) by fluorescence spectroscopy.…

  9. Reversibility of structural rearrangements in bovine serum albumin during homomolecular exchange from AgI particles

    NARCIS (Netherlands)

    Vermonden, T; Giacomelli, CE; Norde, W

    2001-01-01

    The reversibility of the homomolecular exchange of bovine serum albumin (BSA) from AgI particles was studied by differential scanning calorimetry, the binding of 8-anilino-1-naphthaIene-sulfonic acid, and circular dichroism spectroscopy. The structure of BSA in solution before adsorption, in the

  10. Reversibility of structural rearrangements in Bovine Serum Albumin during homomolecular exchange from AgI particles

    NARCIS (Netherlands)

    Vermonden, T.; Giacomelli, C.E.; Norde, W.

    2001-01-01

    The reversibility of the homomolecular exchange of bovine serum albumin (BSA) from AgI particles was studied by differential scanning calorimetry, the binding of 8-anilino-1-naphthalene-sulfonic acid, and circular dichroism spectroscopy. The structure of BSA in solution before adsorption, in the

  11. Nanoshaving of bovine serum albumin films adsorbed on monocrystalline surfaces and interfaces

    Czech Academy of Sciences Publication Activity Database

    Kolivoška, Viliam; Gál, Miroslav; Lachmanová, Štěpánka; Janda, Pavel; Sokolová, Romana; Hromadová, Magdaléna

    2011-01-01

    Roč. 76, č. 9 (2011), s. 1075-1087 ISSN 0010-0765 R&D Projects: GA ČR GP203/09/P502; GA ČR GA203/08/1157 Institutional research plan: CEZ:AV0Z40400503 Keywords : bovine serum albumin * nanoshaving * proteins Subject RIV: CG - Electrochemistry Impact factor: 1.283, year: 2011

  12. Platelet-rich plasma can replace fetal bovine serum in human meniscus cell cultures

    NARCIS (Netherlands)

    Gonzales, V.K.; Mulder, E.L.W. de; Boer, T. den; Hannink, G.; Tienen, T.G. van; Heerde, W.L. van; Buma, P.

    2013-01-01

    Concerns over fetal bovine serum (FBS) limit the clinical application of cultured tissue-engineered constructs. Therefore, we investigated if platelet-rich plasma (PRP) can fully replace FBS for meniscus tissue engineering purposes. Human PRP and platelet-poor plasma (PPP) were isolated from three

  13. Kinetics of the Adsorption of Bovine Serum Albumin of White Wine ...

    African Journals Online (AJOL)

    This study investigates the kinetics of adsorption of bovine serum albumin, BSA, in white wine model solutions onto activated carbon, AC, and alumina, AL. Pseudo-first order and pseudo-second order models were applied to determine the rate and mechanism of adsorption of the white wine protein during the haze removal ...

  14. Biomimetic coprecipitation of calcium phosphate and bovine serum albumin on titanium alloy

    NARCIS (Netherlands)

    Liu, Yuelian; Layrolle, Pierre; de Bruijn, Joost Dick; van Blitterswijk, Clemens; de Groot, K.

    2001-01-01

    Titanium alloy implants were precoated biomimetically with a thin and dense layer of calcium phosphate and then incubated either in a supersaturated solution of calcium phosphate or in phosphate-buffered saline, each containing bovine serum albumin (BSA) at various concentrations, under

  15. Recovery of bovine Babesia spp. after long-term cryostorage and comparison of bovine donor erythrocytes and serum.

    Science.gov (United States)

    Alhaboubi, Amer; He, Lan; McCormack, Kimberly; Gustafson, Amber; Holman, Patricia J

    2017-08-30

    Cultured Babesia bovis and Babesia bigemina were recovered from liquid nitrogen storage nearly 30 years after they were cryopreserved. Four cattle were compared as donors of erythrocytes and serum for microaerophilous stationary phase (MASP) cultures for recovery of B. bigemina. Erythrocytes and serum from only one (#913) of the four animals supported growth of B. bigemina. Two B. bigemina (frozen in 1986 and 1987) and two B. bovis (both frozen in 1986) cryostocks were recovered from liquid nitrogen storage and all four recovered and thrived in #913 erythrocytes and serum. In the third passage after recovery, B. bovis cultures were cryopreserved. Six months later they were successfully recovered using #913 erythrocytes and serum. This study shows that B. bovis and B. bigemina stored nearly 30 years in liquid nitrogen can be successfully recovered in the MASP system. This study also confirms previous observations that selection of a suitable bovine donor of erythrocytes and serum is critical to the success of the culture. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Determination of testosterone esters and estradiol esters in bovine and porcine blood serum.

    Science.gov (United States)

    Rejtharová, Martina; Rejthar, Libor; Čačková, Katarína

    2017-04-01

    Monitoring of steroid esters in blood serum is desirable in order to detect the possible illegal use of natural hormones as growth promoters. A method for the determination of testosterone propionate, testosterone benzoate, testosterone isocaproate, testosterone decanoate and estradiol benzoate in bovine and porcine blood serum was developed. The procedure consists of protein precipitation and removal of phospholipids using a HybridSPE®-Phospholipid column followed by clean-up on a hydrophilic modified styrene polymer Supel TM -Select HLB column and LC-MS/MS measurement. The method was validated according to Commission Decision 2002/657/EC. Decision limits for all analytes were observed in the range 5-30 pg ml - 1 . The method was shown to be robust for bovine and porcine serum analyses and can be applied for both screening and confirmatory determination in routine residue monitoring.

  17. Bovine milk intake and xanthine oxidase activity in blood serum.

    Science.gov (United States)

    McCarthy, R D; Long, C A

    1976-06-01

    Xanthine oxidase activity in blood serum was measured by a sensitive radio-enzymatic assay. Pigs receiving 7.6 liters of milk daily for 100 days did not show any detectable enzymatic activity in their blood Xanthine oxidase activity in blood serum of 25 human volunteers had an average of 6.7 milliunits per liter with a range of 0 to 34.6 milliunits per liter. Neither a causal nor statistically significant relationship existed between xanthine oxidase activity in blood and average daily milk consumption, age, or sex.

  18. Structural changes during the unfolding of Bovine serum albumin in ...

    Indian Academy of Sciences (India)

    400Da [9]. Low-angle X-ray scattering [10] and neutron scattering [11] also in- dicated serum albumin to be a prolate ellipsoid. However, studies using 1H NMR indicated that a prolate structure was unlikely; rather a heart-shaped structure was proposed [12]. The shape of albumin reveals a heart-shaped molecule that can.

  19. Immobilization of Bovine Serum Albumin Upon Multiwall Carbon Nanotube for High Speed Humidity Sensing Application.

    Science.gov (United States)

    Bhattacharya, Sankhya; Sasmal, Milan

    2016-01-01

    We present a high-speed humidity sensor based on immobilization of bovine serum albumin upon multiwall carbon nanotube (IBC). A simple and versatile drop casting technique was employed to make the humidity sensor using novel material IBC at room temperature. IBC was synthesized using easy solution process technique. The working principle of the IBC humidity sensor depends upon the variation of output current or conductance with the exposure of different humidity level. Humidity sensing properties of our device is explained on the basis of charge transfer from water molecules to IBC and bovine serum albumin to multiwall carbon nanotube (MWCNT). Our sensor exhibits faster response time around 1.2 s and recovery time 1.5 s respectively.

  20. Residual bovine serum albumin (BSA) quantitation in vaccines using automated Capillary Western technology.

    Science.gov (United States)

    Loughney, John W; Lancaster, Catherine; Ha, Sha; Rustandi, Richard R

    2014-09-15

    Bovine serum albumin (BSA) is a major component of fetal bovine serum (FBS), which is commonly used as a culture medium during vaccine production. Because BSA can cause allergic reactions in humans the World Health Organization (WHO) has set a guidance of 50 ng or less residual BSA per vaccine dose. Vaccine manufacturers are expected to develop sensitive assays to detect residual BSA. Generally, sandwich enzyme-linked immunosorbent assays (ELISA) are used in the industry to detect these low levels of BSA. We report the development of a new improved method for residual BSA detection using the SimpleWestern technology to analyze residual BSA in an attenuated virus vaccine. The method is based on automated Capillary Western and has linearity of two logs, >80% spike recovery (accuracy), intermediate precision of CV BSA in four lots of bulk vaccine products and was used to monitor BSA clearance during vaccine process purification. Copyright © 2014 Elsevier Inc. All rights reserved.

  1. Synthesis and Characterization of Bovine Serum Albumin-Conjugated Copper Sulfide Nanocomposites

    Directory of Open Access Journals (Sweden)

    Peng Huang

    2010-01-01

    Full Text Available A simple biomolecule-assisted solution route was developed to synthesize Bovine Serum Albumin-conjugated copper sulfide (CuS/BSA nanocomposites, directly using copper salts and thioacetamide (TAA as the starting materials with a zwitterionic surfactant Bovine Serum Albumin (BSA as foaming and stabilizing agent. The CuS/BSA nanocomposites have been characterized by UV, TEM, Zeta, DLS, XRD, and FTIR. The results indicate that the as-prepared CuS/BSA nanocomposites are approximate sphere with a size distribution from 10 to 35 nm in diameter and good dispersibility, depending highly on concentration of BSA concentration. These protein-assisted synthesized nanocomposites have a great potential application in biomedical engineering and microelectronics.

  2. Interactions of nanobubbles with bovine serum albumin and papain films on gold surfaces

    Czech Academy of Sciences Publication Activity Database

    Kolivoška, Viliam; Gál, Miroslav; Hromadová, Magdaléna; Lachmanová, Štěpánka; Pospíšil, Lubomír

    2011-01-01

    Roč. 6, č. 4 (2011), s. 164-170 ISSN 1559-4106 R&D Projects: GA ČR GP203/09/P502; GA ČR GA203/09/0705; GA ČR GA203/08/1157; GA AV ČR IAA400400802 Institutional research plan: CEZ:AV0Z40400503 Keywords : nanobubbles * bovine serum albumin * gold surfaces Subject RIV: CG - Electrochemistry Impact factor: 3.118, year: 2010

  3. Native and denatured bovine serum albumin. D.c. polarography, stripping voltammetry and constant current chronopotentiometry

    Czech Academy of Sciences Publication Activity Database

    Ostatná, Veronika; Uslu, B.; Dogan, B.; Ozkan, S.; Paleček, Emil

    2006-01-01

    Roč. 593, č. 1-2 (2006), s. 172-178 ISSN 0022-0728 R&D Projects: GA AV ČR(CZ) IAA500040513; GA MŠk(CZ) LC06035 Institutional research plan: CEZ:AV0Z50040507 Keywords : protein electrochemistry * bovine serum albumin * native and denatured proteins Subject RIV: BO - Biophysics Impact factor: 2.339, year: 2006

  4. Thermodynamic Study of the Interaction of Bovine Serum Albumin and Amino Acids with Cellulose Nanocrystals

    OpenAIRE

    Lombardo, Salvatore; Eyley, Sam; Schütz, Christina; Van Gorp, Hans; Rosenfeldt, Sabine; Van den Mooter, Guy; Thielemans, Wim

    2017-01-01

    The interaction of bovine serum albumin (BSA) with sulfated, carboxylated, and pyridinium-grafted cellulose nanocrystals (CNCs) was studied as a function of the degree of substitution by determining the adsorption isotherm and by directly measuring the thermodynamics of interaction. The adsorption of BSA onto positively charged pyridinium-grafted cellulose nanocrystals followed Langmuirian adsorption with the maximum amount of adsorbed protein increasing linearly with increasing degree of sub...

  5. [Study of interaction between sorbitol and bovine serum albumin by fluorescence spectrometry].

    Science.gov (United States)

    Li, Dan; Jiang, Xin-min; Yan, Zheng-yu

    2008-06-01

    The interaction of sorbitol and bovine serum album (BSA) was studied by fluorescence and ultraviolet absorption spectra. As it is well known to us, the interactional analysis between small molecular drug and biology macromolecule (such as protein, DNA, etc) is one of the important interactional analyses, which can not only offer new biological view but also supply chance for chemist and biochemist to synthesize new drug capable of regulating the biology process effectively. In the present paper, fluorescence spectrophotometry was first employed to study the interaction between BSA and sorbitol. At the same time, the synchronous fluorescence spectroscopy was adopted to review the configuration of BSA influenced by sorbitol, which provides important significance for clinical medication. The results show that sorbitol has strongly quenched the fluorescence of bovine serum albumin in natural physiological condition, the quenching mechanism is a static quenching procedure at different temperatures and drug concentration, and the variational absorption spectra also proves this deduction. At the same time, this article has also examined the influences of sorbitol on the fluorescence quenching of bovine serum albumin at different temperatures and drug concentration. The binding constants and the number of binding sites between sorbitol and BSA were calculated at different temperatures. Furthermore, the enthalpy and entropy changes in the interaction of sorbitol and bovine serum album were also obtained by the equations of Stern-Volmer and Lineweaver-Burk. From the thermodynamic parameters, it can be judged that the primary binding power between sorbitol and BSA is electrostatic force. Moreover, the synchronous fluorescence spectroscopy was applied to examine the effect of sorbitol on the configuration of BSA. The alterative configuration of BSA may be induced by the hydrophobicity environment of tyrosine with the increase in drug concentration. In conclusion, the

  6. Influence of the fetal bovine serum proteins on the growth of human osteoblast cells on graphene

    Czech Academy of Sciences Publication Activity Database

    Kalbáčová, M.; Brož, A.; Kalbáč, Martin

    100A, č. 11 (2012), s. 3001-3007 ISSN 1549-3296 R&D Projects: GA AV ČR IAA400400911; GA AV ČR KAN200100801; GA ČR GAP204/10/1677; GA ČR(CZ) GAP208/12/1062; GA MŠk ME09060 Institutional support: RVO:61388955 Keywords : human osteoblast * graphene * fetal bovine serum Subject RIV: CG - Electrochemistry Impact factor: 2.834, year: 2012

  7. The application of two-dimensional fluorescence correlation spectroscopy on the interaction between bovine serum albumin and prulifloxacin

    OpenAIRE

    Xue, Mao-Yun; Yang, Ai-Ping; Ma, Mei-Hua; Li, Xiao-Hua

    2009-01-01

    The interaction between bovine serum albumin (BSA) and prulifloxacin was investigated by ultraviolet spectrophotometer (UV) and fluorescence spectroscopy in this paper. Two-dimensional (2D) correlation spectroscopy was applied to the analysis of fluorescence spectra. The results of spectroscopic measurements suggested that prulifloxacin (PL) have a strong ability to quench the intrinsic fluorescence of bovine serum albumin through static quenching procedure. Thermodynamic parameter enthalpy c...

  8. Use of Rat Estrus Serum for in Vitro Maturation of Bovine Oocytes

    Directory of Open Access Journals (Sweden)

    AR Rafati

    2007-04-01

    Full Text Available Introduction: Superovulation produces complications in some patients, so invitro maturation of oocytes is used to decrease or eliminate these complications and improve IVF. Moreover, IVM is used for different aspects of reproductive researches. Slaughterhouse ovaries are the main source of oocytes for IVM and IVF studies. Different media has been introduced and experimented for in vitro maturation of oocytes. Animal's serum at estrus stage contains different hormones and proteins which are essential for oocyte maturation. The aim of this study was to compare three culture media for in vitro maturation (IVM of bovine oocytes; 1(controlTCM-199, 2HCG and follicular fluid (FF and 3 antibiotic. Methods: Rat estrus serum (RSS or fetal bovine serum (FBS was added to control medium. Total of 1789 compact cumulus oocyte complexes (COCs were aspirated from ovaries of slaughtered animals. Oocytes were randomly cultured in mentioned media and incubated in 38.5◦c, 5% CO2 and 95% humidity for 24 hours. The maturation of oocytes was judged according to cumulus cell expansion or randomly orcein stained oocytes and observation of polar bodies. Results: The results showed that maturation rate was significantly higher in second and third group (90.2%, 78.7% as compared to the control group (p<0.001. There was no significant difference between second and third groups (90.2 % vs. 86.6%. Conclusion: RSS is as effective as FBS for IVM of bovine oocytes and can be used as an alternative.

  9. Iron and ferritin levels in the serum and milk of bovine leukemia virus-infected dairy cows

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    KOICHI eORINO

    2015-05-01

    Full Text Available Iron metabolism was examined in 15 bovine leukemia virus (BLV -infected dairy cows (2.6-7.8 years old. BLV infection was detected by measuring serum antibody titer against BLV virus antigen (gp51. The anti-BLV antibody titers of the BLV-infected cows were significantly higher in serum than in milk; a single serum-positive animal lacked detectable anti-BLV antibodies in its milk. Iron and ferritin concentrations also were significantly higher in serum than in milk. Although most of the BLV-infected dairy cows had past or present anamneses (such as inflammatory diseases, including intramammary infection, the milk ferritin concentrations of the infected cows were significantly lower than those of normal cows; serum ferritin concentrations did not differ significantly between these two groups. The anti-BLV antibody titers in milk samples showed significant correlation with serum iron concentrations. These results suggest that BLV infection affects iron homeostasis through iron metabolism in the dairy cow mammary gland.

  10. Interaction of phosphatidylcholine with bovine serum albumin. Specificity and properties of the complexes.

    Science.gov (United States)

    Jonas, A

    1976-03-18

    Phosphatidylcholine dispersed on Celite was rapidly solubilized by neutral bovine serum albumin solutions. Stable protein-lipid complexes were isolated by Agrose gel filtration or by ultracentrifugal flotation in high density solvents, and the physicochemical properties of the complexes were investigated in terms of the stoichiometry of binding, effect of fatty acid ligands on phosphatidylcholine binding, effect of high ionic strength on the stability of the complexes, intrinsic fluorescence and circular dichroism spectra, and sedimentation velocity coefficients. Complexes containing from 2 to 30 phosphatidylcholine molecules per protein molecule were observed; however, no saturation of binding sites could be detected in this range of molar ratios. Oleic acid binding by serum albumin prevents interaction of the protein with phosphatidylcholine, indicating possible competition of these ligands at low contents of the phospholipid. For molar ratios of up to 10 phosphatidylcholine molecules per serum albumin, binding is primarily due to hydrophobic interactions that have no effect on the overall shape and secondary structure of the native protein except for local modifications at tryptophan residues, whose fluorescence becomes quenched and blue shifted on phosphatidylcholine binding. Similar phosphatidylcholine uptake experiments performed with a series of globular proteins indicated that the lipid extraction from Celite surfaces is a non-specific process, accelerated by several other proteins (e.g. aldolase, egg albumin, chymotrypsinogen, soybean trypsin inhibitor, and the major apolipoprotein from bovine serum high density lipoprotein). Formation of stable protein-lipid complexes, however, was only observed with bovine serum albumin, which in contrast to the other proteins is known to have affinity binding sites for anions with hydrophobic side chains.

  11. Making the switch: alternatives to foetal bovine serum for adipose-derived stromal cell expansion

    Directory of Open Access Journals (Sweden)

    Carla Dessels

    2016-10-01

    Full Text Available Adipose-derived stromal cells (ASCs are being used extensively in clinical trials. These trials require that ASCs are prepared using good manufacturing procedures (GMPs and are safe for use in humans. The majority of clinical trials in which ASCs are expanded make use of fetal bovine serum (FBS. While FBS is used traditionally in the research setting for in vitro expansion, it does carry the risk of xenoimmunization and zoonotic transmission when used for expanding cells destined for therapeutic purposes. In order to ensure a GMP quality product for cellular therapy, in vitro expansion of ASCs has been undertaken using xeno-free (XF, chemically-defined, and human blood-derived alternatives. These investigations usually include the criteria proposed by the International Society of Cellular Therapy (ISCT and International Fat Applied Technology Society (IFATS. The majority of studies use these criteria to compare plastic-adherence, morphology, the immunophenotype and the trilineage differentiation of ASCs under the different medium supplemented conditions. Based on these studies, all of the alternatives to FBS seem to be suitable replacements; however, each has its own advantages and drawbacks. Very few studies have investigated the effects of the supplements on the immunomodulation of ASCs; the transcriptome, proteome and secretome; and the ultimate effects in appropriate animal models. The selection of medium supplementation will depend on the downstream application of the ASCs and their efficacy and safety in preclinical studies.

  12. Microglial cell death induced by glycated bovine serum albumin: nitric oxide involvement.

    Science.gov (United States)

    Khazaei, Mohammad R; Habibi-Rezaei, Mehran; Karimzadeh, Fereshteh; Moosavi-Movahedi, Ali Akbar; Sarrafnejhad, Abdo Alfattah; Sabouni, Farzaneh; Bakhti, Mostafa

    2008-08-01

    Nonenzymatic glycation results in the formation of advanced glycation end products (AGEs) through a nonenzymatic multistep reaction of reducing sugars with proteins. AGEs have been suspected to be involved in the pathogenesis of several chronic clinical neurodegenerative complications including Alzheimer's disease, which is characterized with the activation of microglial cells in neuritic plaques. To find out the consequence of this activation on microglial cells, we treated the cultured microglial cells with different glycation levels of Bovine Serum Albumin (BSA) which were prepared in vitro. Extent of glycation of protein has been characterized during 16 weeks of incubation with glucose. Treatment of microglial cells with various levels of glycated albumin induced nitric oxide (NO) production and consequently cell death. We also tried to find out the mode of death in AGE-activated microglial cells. Altogether, our results suggest that AGE treatment causes microglia to undergo NO-mediated apoptotic and necrotic cell death in short term and long term, respectively. NO production is a consequence of iNOS expression in a JNK dependent RAGE signalling after activation of RAGE by AGE-BSA.

  13. Comprehensive spectroscopic probing the interaction and conformation impairment of bovine serum albumin (BSA) by herbicide butachlor.

    Science.gov (United States)

    Liu, Xiaoyi; Ling, Zhaoxing; Zhou, Xing; Ahmad, Farooq; Zhou, Ying

    2016-09-01

    Butachlor is an effective herbicide to deal with undesired weeds selectively and is used at high levels in Asian countries. However, its interaction and impairment effect on BSA was still not clear. In this study, we investigated the interaction between butachlor and bovine serum albumin (BSA) by multi-spectroscopic methods including UV absorption, circular dichroism (CD) spectra, Fourier transform infrared (FTIR) spectra and fluorescence spectra under physiological conditions (pH=7.4). The results revealed that there was a static quenching of BSA induced by butachlor stemmed from the formation of complex. Based on thermodynamic data, the interaction of butachlor with BSA was due to happen, and van der Waals force as well as hydrogen bond were the major forces contributed to the interaction. The binding constant Kb and number of binding site of butachlor with BSA were 5.158×10(5) and 1.372 at 303K, respectively. The distance r between donor (BSA) and acceptor (butachlor) was 0.113nm, obtained according to the Förster theory. The results revealed that butachlor induced conformational changes in BSA but the secondary structure of BSA was still retained. In addition, the microenvironment around chromophore residues of BSA, for example, tryptophan, changed as well, resulting from the formation of more hydrogen bonds. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Severe anaphylactic reaction to bovine serum albumin at the first attempt of artificial insemination.

    Science.gov (United States)

    Wüthrich, B; Stern, A; Johansson, S G

    1995-02-01

    A 33-year-old woman without history of previous atopic diseases or drug allergies developed a severe anaphylactic reaction with asthma, vomiting, itching, generalized urticaria, and angioedema during artificial insemination with her husband's sperm. The sperm-processing medium contained bovine serum albumin (BSA). Skin prick test and RAST demonstrated an IgE-mediated hypersensitivity to BSA as well as a polyvalent atopic sensitization to pollens, animal danders, cow's milk, beef, pork, and mutton. SDS-PAGE studies indicated serum albumin to be the appropriate allergen with a high degree of cross-reactivity between serum albumin from different animal species. Artificial insemination with fluid containing potential allergens can, therefore, represent an unnecessary risk for atopic females, even in the absence of prior clinical symptoms of allergic diseases. Preoperative testing with the medium is recommended.

  15. [Bovine serum albumin in the presence of zinc (II) by fluorescence method].

    Science.gov (United States)

    Wu, Gen-Hua; Wang, Jie; Chen, Jin-Long; Guo, Chang; Wang, Pei-San; Wang, Dong-Xiang; Wang, Zhu-Qing

    2008-04-01

    The interaction between norfloxacin and bovine serum albumin, and the influence of Zinc (II) on the system of norfloxacin and bovine serum albumin was studied under physiological condition by fluorescence method. It was shown that norfloxacin has a powerful ability to quench the BSA fluorescence via a nonradiative energy transfer mechanism. The fluorescence quenching data were analyzed according to Stern-Volmer equation and double-reciprocal equation, and the binding constant (K) and the binding sites (n) were obtained. In the system of binary complex of NFLX and BSA, K = 6.80 x 10(5) and n = 1.21. There is a strong combination between NFLX and BSA, which offers the condition for the serum protein to be deposited and transported in vivo. Besides, the combination between NFLX and BSA becomes stronger in the presence of Zinc (II). According to Stern-Volmer equation and double-reciprocal equation, the concentration of Znic (II) is denser, and the binding constant (K) and the binding sites (n) are bigger. By studying the binding interaction between Zinc (II), norfloxacin and BSA, the mechanism of the interaction among norfloxacin, Zinc (II) and protein in organism, is furtherly discussed.

  16. Common epitopes in LPS of different Enterobacteriaceae are associated with an immune response against Escherichia coli O157 in bovine serum samples.

    Science.gov (United States)

    Navarro, Armando; Eslava, Carlos; García de la Torre, Guadalupe; León, Luis Antonio; Licona, Delia; León, Lemuel; Zarco, Luis Alberto; Cravioto, Alejandro

    2007-11-01

    Epidemiological studies in both humans and animals conducted in Mexico have shown that the isolation frequency of Escherichia coli O157 : H7 is low. In a previous study, IgG antibodies against E. coli O157, O7 and O116 LPS were found in serum samples from children and adults with no previous history of E. coli O157 : H7 infection. The present study was designed to determine whether a similar immune response against E. coli O157 : H7 and other antigenically related bacteria was present in bovine serum samples. A total of 310 serum samples from different herds in Mexico was analysed by microagglutination assays against different enterobacterial antigens, including E. coli O157. Microagglutination assays were positive against E. coli O7 (55 %), O116 (76 %) and O157 (36 %), Escherichia hermannii (15 %), Salmonella enterica serotype Urbana (14 %) and Salmonella enterica subsp. arizonae (40 %). These results were confirmed using a specific ELISA with purified LPS. A positive reaction was observed against the LPS of E. coli O7 (29 %), O116 (12 %) and O157 (22 %), E. hermannii (4 %), Salmonella Urbana (13 %) and S. enterica subsp. arizonae (12 %). Serum absorption studies of positive serum samples indicated the existence of at least three common epitopes shared by the LPS of E. coli O7, O116 and O157, and two others between E. coli O157 and Salmonella Urbana and S. enterica subsp. arizonae. A bactericidal assay against E. coli O157 : H7 using 31 bovine serum samples was performed, and 22 (71 %) of these serum samples gave positive results. The data demonstrated that bovine serum showed a response against different enterobacteria, including E. coli O157, and that this response could be due to the presence of shared epitopes in the LPS of these organisms.

  17. Identification of reference genes for relative quantification of circulating microRNAs in bovine serum.

    Directory of Open Access Journals (Sweden)

    In-Seon Bae

    Full Text Available Circulating microRNAs in body fluids have been implicated as promising biomarkers for physiopathology disorders. Currently, the expression levels of circulating microRNAs are estimated by reverse transcription quantitative real-time polymerase chain reaction. Use of appropriate reference microRNAs for normalization is critical for accurate microRNA expression analysis. However, no study has systematically investigated reference genes for evaluating circulating microRNA expression in cattle. In this study, we describe the identification and characterization of appropriate reference microRNAs for use in the normalization of circulating microRNA levels in bovine serum. We evaluated the expression stability of ten candidate reference genes in bovine serum by using reverse transcription quantitative real-time polymerase chain reaction. Data were analyzed using geNorm, NormFinder, and BestKeeper statistical algorithms. The results consistently showed that a combination of miR-93 and miR-127 provided the most stably expressed reference. The suitability of these microRNAs was validated, and even when compared among different genders or breeds, the combination of miR-93 and miR-127 was ranked as the most stable microRNA reference. Therefore, we conclude that this combination is the optimal endogenous reference for reverse transcription quantitative real-time polymerase chain reaction-based detection of microRNAs in bovine serum. The data presented in this study are crucial to successful biomarker discovery and validation for the diagnosis of physiopathological conditions in cattle.

  18. The use of bovine serum protein as an oral support therapy following coronavirus challenge in calves.

    Science.gov (United States)

    Arthington, J D; Jaynes, C A; Tyler, H D; Kapil, S; Quigley, J D

    2002-05-01

    The objective of this experiment was to investigate the therapeutic efficacy of a supplemental bovine serum protein blend fed to calves challenged with virulent coronavirus. Twelve Holstein bull calves (approximately 3 wk of age) were allocated by initial body weight to Control (n = 5) and treated (n = 7) groups. On d 0, all calves were orally challenged with 1 x 10(7) plaque-forming units of virulent coronavirus isolate. Infection was allowed to progress for 24 h before treatment was started. On d 1, treated calves began receiving 160 g of dry bovine serum powder (16 g IgG) mixed into milk replacer powder (67 g) at both an a.m. and p.m. feeding. Control calves received only milk replacer powder (227 g) at both feedings. Response to coronavirus challenge and dietary treatment was monitored prior to a.m. and p.m. feeding by the collection of multiple clinical measures. Fecal consistency was decreased by coronavirus challenge but was not affected by dietary treatment. Mean daily rectal temperature and heart rate were not affected by dietary treatment. Average packed cell volume was higher in treated calves than in control (35.0 and 27.0%). Coronavirus challenge resulted in an immediate increase in respiration rate, decreasing by d 7. Control calves tended to have a greater average respiration rate compared with treated (28.7 vs. 26.8 breaths/min). Treated calves had a higher average feed intake than control (0.57 vs. 0.44 kg/d). These data suggest that bovine-serum supplemented milk replacer may decrease the severity of disease in young calves exposed to coronavirus.

  19. Study on the thermodynamic characteristics between fluoroquinolone and bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Guo Ming [Department of Chemistry, Zhejiang Forestry University, LinAn 311300 (China)]. E-mail: guoming@zjfc.edu.cn; Lu Weijun [Department of Chemistry, Zhejiang Forestry University, LinAn 311300 (China); Yi Pinggui [Department of Chemistry, Zhejiang University, Hangzhou 310027 (China); Yu Qingsen [Department of Chemistry, Zhejiang University, Hangzhou 310027 (China)

    2007-03-15

    The binding reactions of the fluoroquinolone with bovine serum albumin (BSA) were investigated by microcalorimetry. The thermodynamic parameters were measured with the help of spectroscopy in a Tris-HCl buffer solution (pH 7.0, made isotonic with sodium chloride) at T = 298 K. Microcalorimetric measurements show that the molar change of enthalpy {delta}{sub r} H {sub m} is insignificant for the reaction, which may suggest that the interaction is governed mainly by entropy, and the interaction between the protein and the drugs is stronger. The results also reveal an entropy-enthalpy compensation relationship of the interaction.

  20. Layer-by-layer films from tartrazine dye with bovine serum albumin

    Science.gov (United States)

    de Souza, Nara C.; Flores, Júlio C. Johner; Silva, Josmary R.

    2009-12-01

    We report on the preparation and study of the adsorption process of layer-by-layer films of tartrazine alternated with bovine serum albumin. UV-Vis spectroscopy indicated that the films form J-aggregates of tartrazine. Adsorption kinetics was fitted by the Johnson-Mehl-Avrami equation and surface morphological analyses by atomic force microscopy suggested that the J-aggregates were column-shaped, which was attributed to the column-like symmetry of the tartrazine molecules. The columnar structures that formed probably arose from the juxtaposition of smaller aggregates that were already present at the beginning of film growth.

  1. Inhibitory effect of gold nanoparticles on the D-ribose glycation of bovine serum albumin

    Directory of Open Access Journals (Sweden)

    Liu W

    2014-11-01

    Full Text Available Weixi Liu,1 Menashi A Cohenford,1–3 Leslie Frost,3 Champika Seneviratne,4 Joel A Dain1 1Department of Chemistry, University of Rhode Island, Kingston, RI, USA; 2Department of Integrated Science and Technology, 3Department of Chemistry, Marshall University, Huntington, WV, USA; 4Department of Chemistry, College of the North Atlantic, Labrador, NL, Canada Abstract: Formation of advanced glycation end products (AGEs by nonenzymatic glycation of proteins is a major contributory factor to the pathophysiology of diabetic conditions including senile dementia and atherosclerosis. This study describes the inhibitory effect of gold nanoparticles (GNPs on the D-ribose glycation of bovine serum albumin (BSA. A combination of analytical methods including ultraviolet–visible spectrometry, high performance liquid chromatography, circular dichroism, and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF mass spectrometry were used to determine the extent of BSA glycation in the presence of citrate reduced spherical GNPs of various sizes and concentrations. GNPs of particle diameters ranging from 2 nm to 20 nm inhibited BSA’s AGE formation. The extent of inhibition correlated with the total surface area of the nanoparticles. GNPs of highest total surface area yielded the most inhibition whereas those with the lowest total surface area inhibited the formation of AGEs the least. Additionally, when GNPs’ total surface areas were set the same, their antiglycation activities were similar. This inhibitory effect of GNPs on BSA’s glycation by D-ribose suggests that colloidal particles may have a therapeutic application for the treatment of diabetes and conditions that promote hyperglycemia. Keywords: gold nanoparticles, glycation, AGEs, GNPs, BSA

  2. Establishing presence of antibodies against bovine respiratory syncytial virus (BRSV, parainfluenza virus 3 (PI3 and bovine herpesvirus 1 (BHV 1 in blood serum of cattle using indirect immunoenzyme probe

    Directory of Open Access Journals (Sweden)

    Šamanc Horea

    2009-01-01

    Full Text Available A total of 92 samples of bovine blood serum were examined for the presence of antibodies against the bovine respiratory syncytial virus using indirect immunoenzyme probe - iELISA. Specific antibodies against the bovine respiratory syncytial virus (BRSV were established in 46, or 50% blood serum samples. Investigations of the 92 blood serum samples of cattle for the presence of antibodies against the parainfluenza virus 3 (PI 3, revealed their presence in 77, or 83.69% of the samples, and the presence of antibodies against the bovine herpesvirus 1 (BHV 1 was established in 19, or 20.65% of the samples.

  3. Fetal bovine serum-free cryopreservation methods for clinical banking of human adipose-derived stem cells.

    Science.gov (United States)

    Park, Seah; Lee, Dong Ryul; Nam, Ji Sun; Ahn, Chul Woo; Kim, Haekwon

    2018-02-13

    The use of fetal bovine serum (FBS) as a cryopreservation supplement is not suitable for the banking of mesenchymal stem cells (MSCs) due to the risk of transmission of disease as well as xenogeneic immune reactions in the transplanted host. Here, we investigated if human serum albumin (HSA), human serum (HS), or knockout serum replacement (KSR) can replace FBS for the cryopreservation of MSCs. In addition, we examined the characteristics of MSCs after multiple rounds of cryopreservation. Human adipose-derived stem cells (ASCs) cryopreserved with three FBS replacements, 9% HSA, 90% HS, or 90% KSR, in combination with 10% dimethyl sulfoxide (Me 2 SO) maintained stem cell properties including growth, immunophenotypes, gene expression patterns, and the potential to differentiate into adipogenic, osteogenic, and chondrogenic lineages, similar to ASCs frozen with FBS. Moreover, the immunophenotype, gene expression, and differentiation capabilities of ASCs were not altered by up to four freeze-thaw cycles. However, the performance of three or four freeze-thaw cycles significantly reduced the proliferation ability of ASCs, as indicated by the longer population doubling time and reduced colony-forming unit-fibroblast frequency. Together, our results suggest that HSA, HS, or KSR can replace FBS for the cryopreservation of ASCs, without altering their stemness, and should be processed with no more than two freeze-thaw cycles for clinical approaches. Copyright © 2018 Elsevier Inc. All rights reserved.

  4. β-Carotene and astaxanthin with human and bovine serum albumins.

    Science.gov (United States)

    Li, Xiangrong; Wang, Gongke; Chen, Dejun; Lu, Yan

    2015-07-15

    β-Carotene and astaxanthin are two carotenoids with powerful antioxidant properties. In this study, the interaction of these two carotenoids with human serum albumin (HSA) and bovine serum albumin (BSA) under physiological conditions was investigated using several spectroscopic techniques. The experimental results indicate the quenching mechanism of HSA/BSA, by the two carotenoids, is a static process. The binding constants and number of binding sites were evaluated at different temperatures. Thermodynamic investigations revealed the interaction between the two carotenoids and HSA/BSA is synergistically driven by enthalpy and entropy, and hydrophobic forces and electrostatic attraction have a significant role in the reactions. Binding site I was found to be the primary binding site for β-carotene and astaxanthin. In addition, as shown by synchronous fluorescence spectroscopy and FT-IR, the two carotenoids may induce conformational and micro-environmental changes in HSA/BSA. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. Trypsin inhibitor complexes with human and bovine serum albumins: TEM and spectroscopic analysis.

    Science.gov (United States)

    Hebia, C; Bekale, L; Chanphai, P; Agbebavi, J; Tajmir-Riahi, H A

    2014-01-05

    We report the binding of trypsin inhibitor (TI) with human serum albumin (HSA) and bovine serum albumin (BSA) at physiological conditions, using FTIR, CD, UV-Visible spectroscopic methods and transmission electron microscopy (TEM). Structural analysis showed that trypsin inhibitor binds HSA and BSA via hydrophilic and hydrophobic contacts with overall binding constants of KTI-HSA=1.4 (±0.5)×10(4)M(-1) and KTI-BSA=1.1 (±0.4)×10(6)M(-1). Trypsin inhibitor complexation induces minor reduction of the protein α-helix and a major increase in β-sheet structure. TEM images show that trypsin inhibitor complex formation leads to the protein aggregation and fibrillation. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Intestinal absorption of dinitrophenyl-lysine and effect of immunization with dinitrophenylated bovine serum albumin

    International Nuclear Information System (INIS)

    Shimura, Fumio; Shimura, Junko; Shimazaki, Shigeki; Hosoya, Norimasa

    1983-01-01

    The intestinal absorption of dinitrophenyl-lysine (DNP-lys) was studied with a special interest on the role of the immune system in the absorption of small molecules which are recognized as nonself. [ 3 H]-DNP- lys was rapidly absorbed by ligated intestinal loops in situ via a saturable and unique route. When [ 3 H]-DNP-lys was preincubated with the immume serum obtained from rats immunized with dinitrophenylated bovine serum albumin (DNP-BSA), the [ 3 H]-DNP-lys absorption was depressed. The absorption of [ 3 H]-DNP-lys in DNP-BSA-immunized rats was depressed compared to the control. The results obtained suggest that the immune system play a role in avoiding the absorption of small molecules with antigenicity. (author)

  7. Spectroscopic and voltammetric characterizations of the interaction of two local anesthetics with bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Bi Shuyun, E-mail: sy_bi@sina.co [College of Chemistry, Changchun Normal University, Changchun 130032 (China); Yan Lili; Wang Binbin; Bian Jiangyu [College of Chemistry, Changchun Normal University, Changchun 130032 (China); Sun Yantao [College of Chemistry, Jilin Normal University, Sipin 136000 (China)

    2011-05-15

    The interactions of bovine serum albumin (BSA) with two local anesthetics, procaine hydrochloride (PCH) and tetracaine hydrochloride (TCH) were studied using spectroscopic methods such as fluorescence and ultraviolet visible (UV-vis), and electrochemical techniques including cyclic voltammetry (CV) and differential pulsed stripping voltammetry (DPSV). The results obtained from these techniques turned out that both PCH and TCH could bind to BSA. The binding constants (K{sub A}) and the number of binding sites (n) of the two drugs with BSA at different temperatures were determined, respectively. At 291 K, K{sub A} was found as 2.40x10{sup 4} and 1.42x10{sup 4} L mol{sup -1} and n was 1.03 and 0.99 for PCH-BSA and TCH-BSA, respectively. According to van't Hoff equation, the thermodynamic parameters, {Delta}G, {Delta}H and {Delta}S, were obtained, showing the involvement of hydrophobic and electrostatic force in these interactions. Based on the theory of the Foerster energy transference, the distance between the acceptor (PCH or TCH) and the donor (BSA) were determined as 2.32 and 3.62 nm for PCH and TCH, respectively. The effects of Fe{sup 3+}, Cu{sup 2+}, Mg{sup 2+}, Mn{sup 2+}, Zn{sup 2+} and Ca{sup 2+} on the binding of PCH or TCH to BSA were also evaluated. - Research highlights: Procaine or tetracaine hydrochloride quenching the fluorescence of Trp in BSA was a static quenching process. Synchronous fluorescence was applied to study the structural change of BSA. Binding constant, binding site and binding force were determined. Voltammetry techniques further characterized the nature of the interactions of the two drugs with BSA.

  8. Investigation of three flavonoids binding to bovine serum albumin using molecular fluorescence technique

    International Nuclear Information System (INIS)

    Bi Shuyun; Yan Lili; Pang Bo; Wang Yu

    2012-01-01

    The three flavonoids including naringenin, hesperetin and apigenin binding to bovine serum albumin (BSA) at pH 7.4 was studied by fluorescence quenching, synchronous fluorescence and UV-vis absorption spectroscopic techniques. The results obtained revealed that naringenin, hesperetin and apigenin strongly quenched the intrinsic fluorescence of BSA. The Stern-Volmer curves suggested that these quenching processes were all static quenching processes. At 291 K, the value and the order of the binding constant were K A n aringenin) =4.08x10 4 A(hesperetin) =5.40x10 4 ∼K A(apigenin) =5.32x10 4 L mol -1 . The main binding force between the flavonoid and BSA was hydrophobic and electrostatic force. According to the Foerster theory of non-radiation energy transfer, the binding distances (r 0 ) were obtained as 3.36, 3.47 and 3.30 nm for naringenin-BSA, hesperetin-BSA and apigenin-BSA, respectively. The effect of some common ions such as Fe 3+ , Cu 2+ , Mg 2+ , Mn 2+ , Zn 2+ and Ca 2+ on the binding was also studied in detail. The competition binding was also performed. The apparent binding constant (K' A ) obtained suggested that one flavonoid had an obvious effect on the binding of another flavonoid to protein when they coexisted in BSA solution. - Highlights: → Quenchings of BSA fluorescence by the flavonoids was all static quenchings. → Synchronous fluorescence was applied to study the structural change of BSA. → Binding constant, binding site and binding force were determined. → Competition binding experiments were performed. → One flavonoid had an obvious effect on the binding of another one to BSA.

  9. A radioimmunoassay for human parathyroid hormone utilizing a goat anto-bovine PTH serum

    International Nuclear Information System (INIS)

    Mallette, L.E.

    1981-01-01

    An antiserum (NG-1) against bovine PTH (hPTH) generated in the domestic goat was characterized for use in the radioimmunoassay of PTH in human serum. When a carboxyterminal fragment of bPTH is used as radioligand, this antiserum detects only an antigenic site in the central region of the hPTH molecule. The synthetic hormone fragment, hPTH-(44-68), will displace 93% of the tracer, after which the addition of intact hPTH causes no further displacement. The assay does not detect the synthetic aminoterminal 1-34 fragment of the bovine or human hormones, nor the carboxyterminal fragment of the human hormone, hPTH-(53-84). Standard curves with bPTH-(1-84) and partially purified hPTH are not parallel, so that hPTH is used as standard. Serum from subjects with uraemia or primary hyperparathyroidism gives dilution curves parallel to that with the hPTH standard. The assay with NG-1 has been applied to the diagnosis of primary and secondary hyperparathyroidism, used to monitor the disappearance of PTH after parathyroidectomy, and for measurement of PTH in selective venous samples. (author)

  10. Binding of the neuroleptic drug, gabapentin, to bovine serum albumin: Insights from experimental and computational studies

    Energy Technology Data Exchange (ETDEWEB)

    Jalali, Fahimeh, E-mail: fahimehjalali@yahoo.com [Department of Chemistry, Razi University, 67346 Kermanshah (Iran, Islamic Republic of); Dorraji, Parisa S. [Department of Chemistry, Razi University, 67346 Kermanshah (Iran, Islamic Republic of); Mahdiuni, Hamid [Department of Biology, Razi University, 67346 Kermanshah (Iran, Islamic Republic of)

    2014-04-15

    The interaction between antiepileptic drug, gabapentin (GP), and bovin serum albumin (BSA) was studied by spectroscopic and computational methods. The native fluorescence of BSA was quenched by GP. Stern–Volmer quenching constant was calculated at different temperatures which suggested a static mechanism. The association constant (K{sub a}) was calculated from fluorescence quenching studies, which increased with temperature rising. GP competed well with warfarine for hydrophobic subdomain IIA (Sudlow's site I) on the protein. Enthalpy and entropy changes during the interaction of GP with BSA were obtained using van't Hoff plot, which showed an entropy-driven process and involvement of hydrophobic forces (ΔH>0 and ΔS>0). Synchronous fluorescence measurements of BSA solution in the presence of GP showed a considerable blue shift when Δλ=15 nm, therefore, GP interacts with tyrosine-rich sites on BSA. Optimized docked model of BSA–GP mixture confirmed the experimental results. -- Highlights: • Interaction of gabapentin and bovine serum albumin (BSA) is investigated by spectroscopic techniques. • Gabapentin can quench the fluorescence of BSA through a static quenching procedure. • The binding of gabapentin to BSA is driven mainly by hydrophobic interactions. • Subdomain IIA (Sudlow's site I) of BSA is found to be the main binding site for gabapentin. • Molecular docking modeling confirmed the experimental results.

  11. Synthesis and characterization of imprinted sorbent for separation of gramine from bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Luliński, Piotr; Klejn, Dorota; Maciejewska, Dorota, E-mail: dmaciejewska@wum.edu.pl

    2016-08-01

    The aim of this study was to develop an efficient sorbent for separation of N,N-dimethyl-3-aminomethylindole (gramine) from bovine serum albumin. An imprinting technology was involved in the synthesis of polymers from nine different functional monomers in the presence of ethylene glycol dimethacrylate as a cross-linker. The analysis of binding capacities showed that the highest specificity towards gramine was achieved when 4-vinylbenzoic acid was used as the functional monomer in methanol to form the bulk imprinted polymer, MIP1 (imprinting factor equal to 21.3). The Scatchard analysis of MIP1 showed two classes of binding sites with the dissociation constants K{sub d} equal to 0.105 and 6.52 μmol L{sup −1}. The composition and morphology of polymers were defined by {sup 13}C CP/MAS NMR, BET and SEM-EDS analyses. The recognition mechanism of MIP1 was tested using the structurally related bioanalytes, and the dominant role of indole moiety and ethylamine side chain was revealed. A new MISPE protocol was optimized for separation of gramine. The total recoveries on MIP1 were equal to 94 ± 12 % from standard solutions and 85 ± 11 % from bovine serum albumin. - Highlights: • Indole alkaloid (gramine) imprinted polymer was synthesized. • Very high specifity of sorbent towards gramine was achieved. • Physico-chemical characteristics of novel material was presented. • Efficient MISPE protocol was proposed for separation of gramine from model sample.

  12. Synthesis and characterization of imprinted sorbent for separation of gramine from bovine serum albumin.

    Science.gov (United States)

    Luliński, Piotr; Klejn, Dorota; Maciejewska, Dorota

    2016-08-01

    The aim of this study was to develop an efficient sorbent for separation of N,N-dimethyl-3-aminomethylindole (gramine) from bovine serum albumin. An imprinting technology was involved in the synthesis of polymers from nine different functional monomers in the presence of ethylene glycol dimethacrylate as a cross-linker. The analysis of binding capacities showed that the highest specificity towards gramine was achieved when 4-vinylbenzoic acid was used as the functional monomer in methanol to form the bulk imprinted polymer, MIP1 (imprinting factor equal to 21.3). The Scatchard analysis of MIP1 showed two classes of binding sites with the dissociation constants Kd equal to 0.105 and 6.52μmolL(-1). The composition and morphology of polymers were defined by (13)C CP/MAS NMR, BET and SEM-EDS analyses. The recognition mechanism of MIP1 was tested using the structurally related bioanalytes, and the dominant role of indole moiety and ethylamine side chain was revealed. A new MISPE protocol was optimized for separation of gramine. The total recoveries on MIP1 were equal to 94±12 % from standard solutions and 85±11 % from bovine serum albumin. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Bovine serum albumin contained in culture medium used in artificial insemination is an important anaphylaxis risk factor.

    Science.gov (United States)

    Pagán, Juan A; Postigo, Idoia; Rodríguez-Pacheco, Jorge R; Peña, Maribel; Guisantes, Jorge A; Martínez, Jorge

    2008-11-01

    To analyze the cause of the anaphylactic reaction after a standard artificial insemination process in a patient diagnosed with asthma. Case report. Residencia Sanitaria Virgen de la Arrixaca (Murcia, Spain) and University of the Basque Country (Vitoria, Spain). A 30-year-old woman with a previous medical history compatible with respiratory allergy who suffered an anaphylactic reaction after an artificial insemination with spermatozoids in capable medium (Upgraded B2 INRA medium; Laboratories CCD, Paris, France). Cutaneous tests and specific IgE levels to inhalant allergens, grass and Olea pollens, and insemination medium were performed. Specific IgE levels to mammal epithelia and bovine serum albumin (BSA). Skin prick tests were positive for inhalant allergens such as mites, cat, dog, horse, and rabbit epithelia, grasses and Olea pollens, and the insemination medium. The beta-lactamic tests were negative. The determination of specific IgE demonstrated positive values to mammal epithelia and mammal serum albumins including BSA. We report a case of an anaphylactic reaction to the BSA included in the insemination culture medium induced by a subclinical sensitivity to serum albumins of mammal epithelia. A previous testing with the medium is recommended and specific testing might be needed in women who have a history of animal epithelium allergies.

  14. Pathogenic role of effector cells and immunoglobulins in cationic bovine serum albumin-induced membranous nephropathy.

    Science.gov (United States)

    Wu, Chia-Chao; Lu, Kuo-Cheng; Lin, Yuh-Feng; Chen, Jin-Shuen; Huang, Ching-Feng; Chen, Chun-Chi; Lin, Shih-Hua; Chu, Pauling; Sytwu, Huey-Kang

    2012-02-01

    Membranous nephropathy (MN) is an autoimmune-mediated glomerulonephritis. The roles of effector cells and immunoglobulins (Igs) in the mediation of glomerular injury in MN have not been fully elucidated. MN was induced by cationic bovine serum albumin (cBSA), and passive disease was induced by transferring effector cells or serum into severe combined immunodeficient (SCID) mice. MN could not be induced in SCID mice. Transfer of serum from MN mice, but not from normal control mice, to SCID mice induced granular immune complex deposits and pathologic proteinuria. Increased immunofluorescent staining for complement, oxidative stress, terminal deoxynucleotidyl transferase-mediated nick end-labeling assay-positive cells, and augmented phospho-NF-κB staining were evident in the kidneys of MN serum recipients. However, no histological or clinical manifestations were exhibited by SCID mice that received an adoptive transfer of splenocytes. Adaptive immunity was essential for the development of MN. Specific Igs and their subsequent response contribute to the development of renal injury in cBSA-induced MN.

  15. Evaluation of human and bovine serum albumin on oxidation characteristics by a photosensitization reaction under complete binding of talaporfin sodium.

    Science.gov (United States)

    Kurotsu, Mariko; Yajima, Masahiro; Takahashi, Mei; Ogawa, Emiyu; Arai, Tsunenori

    2015-09-01

    In order to investigate the therapeutic interaction of an extra-cellular photosensitization reaction, we evaluated the oxidation characteristics of human and bovine serum albumin by this reaction with talaporfin sodium under complete binding with albumin by spectroscopic analysis in a cell-free solution. The solution was composed of 20μg/ml talaporfin sodium and 2.1mg/ml human or bovine serum albumin. A 662nm laser light was used to irradiate the solution. Visible absorbance spectra of solutions were measured to obtain the oxidized and non-oxidized relative densities of albumin and talaporfin sodium before and after the photosensitization reaction. The defined oxidation path ratio of talaporfin sodium to albumin reflected the oxidation of the solution. Absorbance wavelengths at approximately 240 and 660nm were used to calculate normalized molecular densities of oxidized albumin and talaporfin sodium, respectively. The oxidation path ratio of talaporfin sodium to albumin when binding human serum albumin was approximately 1.8 times larger than that of bovine serum albumin during the photosensitization reaction from 1 to 50J/cm(2). We hypothesized that the oxidation path ratio results might have been caused by talaporfin sodium binding affinity or binding location difference between the two albumins, because the fluorescence lifetimes of talaporfin sodium bound to human and bovine serum albumin were 7.0 and 4.9ns, respectively. Therefore, the photodynamic therapeutic interaction might be stronger with human serum albumin than with bovine serum albumin in the case of extracellular photosensitization reaction. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Sensitization to bovine serum albumin as a possible cause of allergic reactions to vaccines.

    Science.gov (United States)

    de Silva, Rajiva; Dasanayake, W M D K; Wickramasinhe, G D; Karunatilake, Chandima; Weerasinghe, Nayani; Gunasekera, Peshala; Malavige, Gathsaurie Neelika

    2017-03-13

    Immediate type hypersensitivity to vaccines containing bovine/porcine excipients, such as the measles, mumps and rubella (MMR) vaccine is probably due to sensitization to bovine/porcine gelatin. Most patients with such reactions in Sri Lanka have cow's milk (CM) or beef allergy. We investigated whether those who had beef and CM allergy had a higher incidence of hypersensitivity reactions to vaccines and the possible trigger of such reactions. Twenty patients with immediate type hypersensitivity reactions to vaccines containing bovine/porcine excipients, controls with allergy to beef/pork (n=11) or CM (n=11), and 8 non atopic controls were recruited. Total serum IgE, specific IgE to beef, CM, casein, beta lactoglobulin, gelatin and bovine serum albumin (BSA) by Phadia ImmunoCap and IgE to porcine gelatin by Western blot were evaluated. 11/20, 5/20, 2/20, 2/20, 1/20 and 1/20 patients reported allergic reactions to measles containing, JE, rabies primary chick embryo, pentavalent, diphtheria and tetanus, and adult diphtheria and tetanus vaccines, respectively. Only one patient with allergy to vaccines had gelatin specific IgE, whereas IgE to BSA was seen in 73.3%, 90%, 66.6% and 0 of vaccine, beef or CM allergic and non-atopic controls, respectively. The mean IgE to BSA was higher in patients with allergy to vaccines, although not significant. Specific IgE to BSA was present in 54.7% of children with allergy to CM, of whom 11.8% had high levels (>17.5kUA/L). In contrast, 66.6% of these children did not have specific IgE to β-lactoglobulin, which is one of the major components of whey protein. Gelatin does not appear to play a major role in Sri Lankan children with allergy to vaccines. In contrast, due to the higher levels of BSA specific IgE, sensitization to BSA is possibly playing a role. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Iron and Ferritin Levels in the Serum and Milk of Bovine Leukemia Virus-Infected Dairy Cows

    OpenAIRE

    Schnell, Star A.; Ohtsuka, Hiromichi; Kakinuma, Seiichi; Yoshikawa, Yasunaga; Watanabe, Kiyotaka; Orino, Koichi

    2015-01-01

    Iron metabolism was examined in 15 bovine leukemia virus (BLV)-infected dairy cows (2.6–7.8 years old). BLV infection was detected by measuring serum antibody titer against BLV virus antigen (gp51). The anti-BLV antibody titers of the BLV-infected cows were significantly higher in serum than in milk; a single serum-positive animal lacked detectable anti-BLV antibodies in its milk. Iron and ferritin concentrations also were significantly higher in serum than in milk. Although most of the BLV-i...

  18. Development of a flow cytometric immunoassay for recombinant bovine somatotropin-induced antibodies in serum of dairy cows

    NARCIS (Netherlands)

    Smits, N.G.E.; Bremer, M.G.E.G.; Ludwig, S.K.J.; Nielen, M.W.F.

    2012-01-01

    Administration of recombinant bovine somatotropin (rbST) to enhance milk production in dairy cows is banned within the European Union. Therefore, methods for pinpointing rbST abuse are required. Due to the problematic detection of rbST itself in serum, methods are also focused on detecting changes

  19. Adsorption behavior of cation-exchange resin-mixed polyethersulfone-based fibrous adsorbents with bovine serum albumin

    NARCIS (Netherlands)

    Zhang, Y.; Zhang, Yuzhong; Borneman, Zandrie; Koops, G.H.; Wessling, Matthias

    2006-01-01

    The cation-exchange resin-mixed polyethersulfone (PES)-based fibrous adsorbents were developed to study their adsorption behavior with bovine serum albumin (BSA). A fibrous adsorbent with an open pore surface had much better adsorption behavior with a higher adsorbing rate. The adsorption capacity

  20. Lack of evidence for the presence of emerging HoBi-like viruses in North American fetal bovine serum lots.

    Science.gov (United States)

    The detection of HoBi-like virus in fetal bovine serum (FBS) labeled as United States of America (USA) origin, but packaged in Europe, raised concerns that HoBi-like virus may have entered the USA. In this study, 90 lots of FBS originating in North America (NA) were screened for pestivirus antigen ...

  1. Interaction of the flavonoid hesperidin with bovine serum albumin: A fluorescence quenching study

    Energy Technology Data Exchange (ETDEWEB)

    Wang Yanqing [Jiangsu Provincial Key Laboratory of Coastal Wetland Bioresources and Environmental Protection, Yancheng Normal College, Yancheng City, Jiangsu Province 224002 (China) and Institute of Applied Chemistry and Environmental Engineering, Yancheng Normal College, Yancheng City, Jiangsu Province 224002 (China)]. E-mail: wyqing76@126.com; Zhang Hongmei [Institute of Applied Chemistry and Environmental Engineering, Yancheng Normal College, Yancheng City, Jiangsu Province 224002 (China); Zhang Gencheng [Jiangsu Provincial Key Laboratory of Coastal Wetland Bioresources and Environmental Protection, Yancheng Normal College, Yancheng City, Jiangsu Province 224002 (China); Institute of Applied Chemistry and Environmental Engineering, Yancheng Normal College, Yancheng City, Jiangsu Province 224002 (China); Tao Weihua [Institute of Applied Chemistry and Environmental Engineering, Yancheng Normal College, Yancheng City, Jiangsu Province 224002 (China); Tang Shuhe [Institute of Applied Chemistry and Environmental Engineering, Yancheng Normal College, Yancheng City, Jiangsu Province 224002 (China)

    2007-09-15

    The interaction between the flavonoid hesperidin and bovine serum albumin (BSA) was investigated by fluorescence and UV/Vis absorption spectroscopy. The results revealed that hesperidin caused the fluorescence quenching of BSA through a static quenching procedure. The hydrophobic and electrostatic interactions play a major role in stabilizing the complex. The binding site number n, and apparent binding constant K {sub A}, corresponding thermodynamic parameters {delta}G {sup o}, {delta}H {sup o}, {delta}S {sup o} at different temperatures were calculated. The distance r between donor (BSA) and acceptor (hesperidin) was obtained according to fluorescence resonance energy transfer. The effect of Cu{sup 2+}, Zn{sup 2+}, Ni{sup 2+}, Co{sup 2+}, and Mn{sup 2+} on the binding constants between hesperidin and BSA were studied. The effect of hesperidin on the conformation of BSA was analyzed using synchronous fluorescence spectroscopy and UV/Vis absorption spectroscopy.

  2. The interaction between 4-aminoantipyrine and bovine serum albumin: Multiple spectroscopic and molecular docking investigations

    International Nuclear Information System (INIS)

    Teng Yue; Liu Rutao; Li Chao; Xia Qing; Zhang Pengjun

    2011-01-01

    4-Aminoantipyrine (AAP) is widely used in the pharmaceutical industry, in biochemical experiments and in environmental monitoring. AAP as an aromatic pollutant in the environment poses a great threat to human health. To evaluate the toxicity of AAP at the protein level, the effects of AAP on bovine serum albumin (BSA) were investigated by multiple spectroscopic techniques and molecular modeling. After the inner filter effect was eliminated, the experimental results showed that AAP effectively quenched the intrinsic fluorescence of BSA via static quenching. The number of binding sites, the binding constant, the thermodynamic parameters and binding subdomain were measured, and indicated that AAP could spontaneously bind with BSA on subdomain IIIA through electrostatic forces. Molecular docking results revealed that AAP interacted with the Glu 488 and Glu 502 residues of BSA. Furthermore, the conformation of BSA was demonstrably changed in the presence of AAP. The skeletal structure of BSA loosened, exposing internal hydrophobic aromatic ring amino acids and peptide strands to the solution.

  3. Gold nanoparticles synthesized by gamma radiation and stabilized by bovine serum albumin

    International Nuclear Information System (INIS)

    Leal, Jessica; Silva, Andressa A.; Geraldes, Adriana N.; Lugao, Ademar B.; Grasselli, Mariano

    2015-01-01

    Gold nanoparticles (AuNPs) are a new option for pharmaceutical and cosmetic industries due to their interesting chemical, electrical and catalytic properties. Research for cancer treatments have been developed using this promising radiotherapy agent. The challenge of gold nanoparticles is to keep them stable, due to metallic behavior. It is know that surface plasma resonance promotes agglomeration of metallic nanoparticles, but they are not stable. Stabilizers have been used to reduce agglomeration. The aim of this work is reduction of HAuCl 4 salt to AuNPs performed by gamma radiation 60 Co source and the stabilization of gold nanoparticles using bovine serum albumin (BSA) fraction V as stabilizer agent. AuNPs were characterized by UV-visible to verify the nanoparticles formation. Samples containing BSA and samples obtained by the conventional method (without stabilizer) were monitored for two weeks and analyzed. Results were compared. (author)

  4. Structural changes of ultrasonicated bovine serum albumin revealed by hydrogen-deuterium exchange and mass spectrometry.

    Science.gov (United States)

    Zhang, Qiuting; Tu, Zongcai; Wang, Hui; Huang, Xiaoqin; Sha, Xiaomei; Xiao, Hui

    2014-11-01

    The structural changes of bovine serum albumin (BSA) under high-intensity ultrasonication were investigated by fluorescence spectroscopy and mass spectrometry. Evidence for the ultrasonication-induced conformational changes of BSA was provided by the intensity changes and maximum-wavelength shift in fluorescence spectrometry. Matrix-assisted laser desorption-ionization time-of-flight mass spectroscopy (MALDI-TOF MS) revealed the increased intensity of the peak at the charge state +5 and a newly emerged peak at charge state +6, indicating that the protein became unfolded after ultrasonication. Prevalent unfolding of BSA after ultrasonication was revealed by hydrogen-deuterium exchange coupled with mass spectrometry (HDX-MS). Increased intensity and duration of ultrasonication further promoted the unfolding of the protein. The unfolding induced by ultrasonication goes through an intermediate state similar to that induced by a low concentration of denaturant.

  5. UV radiation sensitivity of bovine serum albumin bound to silver nanoparticles

    Directory of Open Access Journals (Sweden)

    Meena Pandey Pant

    2014-10-01

    Full Text Available We report here the UV radiation sensitivity of Bovine Serum Albumin (BSA bound silver nanoparticles (62 nm diameter to various power density between 468 mJ/cm2 to 1872 mJ/cm2 under physiological conditions. The functional properties associated with BSA such as esterase activity, free thiols and copper ion binding have been studied. Decrease in free thiols, with increase in copper ion binding and P- nitrophenyl acetate (PNPA turnover were observed in BSA bound silver nanoparticles (SNP in the presence of UV radiation. Intrinsic fluorescence intensity of BSA bound SNP was decreased with UV radiation. Circular Dichroism results indicated a decrease in alpha helical content of BSA bound SNP. The overall results suggest modifications in structure–function properties of BSA bound to SNP in the presence of UV radiation. The possible mechanisms of interaction between BSA and SNP have been explained in presence of UV radiation.

  6. Adsorption behavior of oxidized galactomannans onto amino terminated surfaces and their interaction with bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Sierakowski, M.-R; Silva, Maria R.V. da [Universidade Federal do Parana, Curitiba, PR (Brazil). Dept. de Quimica. Lab. de Biopolimeros]. E-mail: mrbiopol@quimica.ufpr.br; Freitas, R.A.; Moreira, Jose S.R. [Universidade Federal do Parana, Curitiba, PR (Brazil). Dept. de Bioquimica; Fujimoto, J.; Petri, D.F.S.; Cordeiro, Paulo R.D. [Sao Paulo Univ., SP (Brazil). Inst. de Quimica]. E-mail: dfsp@quim.iq.usp.br; Andrade, Fabiana D

    2001-07-01

    A galactomannan (CF) extracted from Cassia fastuosa seeds was purified and oxidized with (2,2,6,6- tetramethylpiperidine-1-oxyl) to form a uronic acid-containing polysaccharide (CFOX) with a degree of oxidation (DO) of 0.22. The chemical structures of CF and CFOX were characterized. The adsorption behavior of CF and CFOX onto amino-terminated surfaces was studied by means of ellipsometric measurements. The influence of p H and ionic strength on the adsorption was also investigated. At p H 4, there was a maximum in the adsorbed amount caused by strong electrostatic attraction between the substrate and the oxidized galactomannans. There was no ionic strength effect on the adsorption behavior. The immobilization of bovine serum albumin onto CF and CFOX was studied as a function of p H. At the isoelectric point a maximum in the adsorbed amount was found. (author)

  7. Adsorption behavior of oxidized galactomannans onto amino terminated surfaces and their interaction with bovine serum albumin

    International Nuclear Information System (INIS)

    Sierakowski, M.-R; Silva, Maria R.V. da; Freitas, R.A.; Moreira, Jose S.R.; Fujimoto, J.; Petri, D.F.S.; Cordeiro, Paulo R.D.; Andrade, Fabiana D.

    2001-01-01

    A galactomannan (CF) extracted from Cassia fastuosa seeds was purified and oxidized with (2,2,6,6- tetramethylpiperidine-1-oxyl) to form a uronic acid-containing polysaccharide (CFOX) with a degree of oxidation (DO) of 0.22. The chemical structures of CF and CFOX were characterized. The adsorption behavior of CF and CFOX onto amino-terminated surfaces was studied by means of ellipsometric measurements. The influence of p H and ionic strength on the adsorption was also investigated. At p H 4, there was a maximum in the adsorbed amount caused by strong electrostatic attraction between the substrate and the oxidized galactomannans. There was no ionic strength effect on the adsorption behavior. The immobilization of bovine serum albumin onto CF and CFOX was studied as a function of p H. At the isoelectric point a maximum in the adsorbed amount was found. (author)

  8. Formation and Properties of Multilayer Films Based on Polyethyleneimine and Bovine Serum Albumin

    Science.gov (United States)

    Kulikouskaya, V. I.; Lazouskaya, M. E.; Kraskouski, A. N.; Agabekov, V. E.

    2018-01-01

    (Polyethyleneimine/bovine serum albumin) n ((PEI/BSA) n) multilayer films ( n = 1-10) are produced via the layer-by-later deposition of polyelectrolytes. It is shown that thickness and morphology of the formed coatings can be controlled by varying the solution's ionic strength during alternating adsorption of the components. (PEI/BSA)10 multilayer systems that contain up to 0.6 mg of antiseptic miramistin per 1 cm2 of film were created. It is established that the kinetics of miramistin release from (PEI/BSA)10 films in phosphate buffers and physiological solutions obey the Korsmeyer-Peppas equation with a high degree of accuracy ( R 2 > 0.95).

  9. Performance study of ultrafiltration membrane with bovine serum albumin as feed solution

    International Nuclear Information System (INIS)

    Syahril Ahmad

    2009-01-01

    Bovine serum albumin solutions at different temperature, pH, flow rate and operation pressure have been used as feed solution for studying performance of ultrafiltration membrane. Polysulfone membranes used for this experiment were in form of hollow fibers that have Molecular Weight Cut Off (MWCO) 60 kDa. Observation was focused on flux parameter and rejection coefficient towards protein during the process. Result shows that temperature, pH of BSA feed solution, flow rate and operation pressure can affect the flux and rejection coefficient of membrane. High temperature feed solution tend to decrease the flux but increase rejection coefficient. Rejection coefficient of membrane will increase while flux decreasing at pH of feed solution near to protein isoelectric point. High pressure of feed solution will increase flux but decrease rejection of membrane. Rejection of membrane will decrease and flux will increase when the process operated in slow flow rate. (author)

  10. Experimental and theoretical study on the binding of 2-mercaptothiazoline to bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Teng, Yue, E-mail: tengyue@jiangnan.edu.cn; Wang, Xiang; Zou, Luyi; Huang, Ming; Du, Xianzheng

    2015-05-15

    2-Mercaptothiazoline (MTZ) is widely utilized as a brightening and stabilization agent, corrosion inhibitor and antifungal reagent. The residue of MTZ in the environment is potentially hazardous to human health. In this study, the binding mode of MTZ with bovine serum albumin (BSA) was investigated using spectroscopic and molecular docking methods under physiological conditions. MTZ could spontaneously bind with BSA through hydrogen bond and van der Waals interactions with one binding site. The site marker displacement experiments and the molecular docking revealed that MTZ bound into site II (subdomain IIIA) of BSA, which further resulted in some backbone structures and microenvironmental changes of BSA. This work is helpful for understanding the transportation, distribution and toxicity effects of MTZ in blood. - Highlights: • The mechanism was explored by multiple spectroscopic and molecular docking methods. • MTZ can spontaneously bind with BSA at subdomain IIIA (site II). • MTZ can lead to some conformational changes of BSA.

  11. FTIR study of secondary structure of bovine serum albumin and ovalbumin

    International Nuclear Information System (INIS)

    Abrosimova, K V; Shulenina, O V; Paston, S V

    2016-01-01

    Proteins structure is the critical factor for their functioning. Fourier transform infrared spectroscopy provides a possibility to obtain information about secondary structure of proteins in different states and also in a whole biological samples. Infrared spectra of egg white from the untreated and hard-boiled hen's egg, and also of chicken ovalbumin and bovine serum albumin in lyophilic form and in aqueous solution were studied. Lyophilization of investigated globular proteins is accompanied by the decrease of a-helix structures and the increase in amount of intermolecular β-sheets. Analysis of infrared spectrum of egg white allowed to make an estimation of OVA secondary structure and to observe α-to-β structural transformation as a result of the heat denaturation. (paper)

  12. Characterization of the binding of 2-mercaptobenzimidazole to bovine serum albumin.

    Science.gov (United States)

    Teng, Yue; Zou, Luyi; Huang, Ming; Zong, Wansong

    2015-04-01

    2-Mercaptobenzimidazole (MBI) is widely utilized as a corrosion inhibitor, copper-plating brightener and rubber accelerator. The residue of MBI in the environment is potentially harmful to human health. In this article, the interaction of MBI with bovine serum albumin (BSA) was explored using spectroscopic and molecular docking methods under physiological conditions. The positively charged MBI can spontaneously bind with the negatively charged BSA through electrostatic forces with one binding site. The site marker competition experiments and the molecular docking study revealed that MBI bound into site II (subdomain IIIA) of BSA, which further led to some secondary structure and microenvironmental changes of BSA. This work provides useful information on understanding the toxicological actions of MBI at the molecular level. Copyright © 2015 John Wiley & Sons, Ltd.

  13. [Synthesis of purpure-18-imide and study on its interaction with bovine serum albumins by fluorescence].

    Science.gov (United States)

    Wang, Jin-jun; Liu, Yong-ming; Li, Li

    2005-04-01

    Purpure-18-imide was synthesized based on the porphyrin's special affinity for cancer cells and the antitumor activity of pyrrolidine compound. The binding reaction between purpure-18-imide and bovine serum albumins (BSA) in aqueous was studied by fluorescence and UV-Vis absorption spectra. The research results indicated that the combination reaction of them was a single static quenching process. In aqueous, purpure-18-imide strongly bound BSA with molar ratio of 1:1. The binding constant K0 was 5.386 x 10(5) L x mol(-1). The shortest binding distance (r = 3.54 nm) and energy transfer efficiencies (E = 0.26) between donor (BSA) and acceptor (purpure-18-imide) were obtained by Forster's nonradiative energy transfer mechanism.

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

    Science.gov (United States)

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

    2013-01-01

    This study investigated the interaction between eupatorin and bovine serum albumin (BSA) using ultraviolet-visible (UV-vis) absorption, fluorescence, synchronous fluorescence, circular dichroism (CD) spectroscopies, and molecular modeling at pH 7.4. Results of UV-vis and fluorescence spectroscopies illustrated that BSA fluorescence was quenched by eupatorin via a static quenching mechanism. Thermodynamic parameters revealed that hydrophobic and electrostatic interactions played major roles in the interaction. Moreover, the efficiency of energy transfer, and the distance between BSA and acceptor eupatorin, were calculated. The effects of eupatorin on the BSA conformation were analyzed using UV-vis, CD, and synchronous fluorescence. Finally, the binding of eupatorin to BSA was modeled using the molecular docking method. PMID:23839090

  15. Fetal bovine serum adsorption onto hydroxyapatite sensor monitoring by quartz crystal microbalance with dissipation technique

    Energy Technology Data Exchange (ETDEWEB)

    Tagaya, Motohiro [Biomaterials Center, National Institute for Materials Science, Sengen 1-2-1, Tsukuba, Ibaraki 305-0047 (Japan); Graduate School of Science and Engineering, Tokyo Institute of Technology, O-okayama 2-12-1, Meguro-ku, Tokyo 152-8550 (Japan); Ikoma, Toshiyuki, E-mail: IKOMA.Toshiyuki@nims.go.jp [Biomaterials Center, National Institute for Materials Science, Sengen 1-2-1, Tsukuba, Ibaraki 305-0047 (Japan); Migita, Satoshi; Okuda, Mitsuhiro; Takemura, Taro; Hanagata, Nobutaka [Biomaterials Center, National Institute for Materials Science, Sengen 1-2-1, Tsukuba, Ibaraki 305-0047 (Japan); Yoshioka, Tomohiko; Tanaka, Junzo [Graduate School of Science and Engineering, Tokyo Institute of Technology, O-okayama 2-12-1, Meguro-ku, Tokyo 152-8550 (Japan)

    2010-10-15

    Adsorption behavior of fetal bovine serum (FBS) onto hydroxyapatite (HAp) was analyzed in situ in phosphate buffer saline (PBS), minimum essential mediums ({alpha}MEM or DMEM), and different carbonate-concentrated buffers by using a quartz crystal microbalance with dissipation (QCM-D) technique. Although the adsorption behaviors of FBS onto gold showed no dependence on the solvents, those onto HAp clearly depended on the adsorbed ions in the solvents (PBS or MEMs). The amount and viscoelasticity of FBS adsorbed onto HAp in PBS were approximately two times larger than those in MEMs. The adsorption amounts of carbonate ions on HAp in the carbonate buffers increased with increasing carbonate concentrations, which inhibited the adsorption of FBS and decreased the viscoelastic property of the adsorption layers.

  16. Probing Conformational Change of Bovine Serum Albumin–Dextran Conjugates under Controlled Dry Heating

    Energy Technology Data Exchange (ETDEWEB)

    Xia, Shuqin; Li, Yunqi; Zhao, Qin; Li, Ji; Xia, Qiuyang; Zhang, Xiaoming; Huang, Qingrong (Rutgers); (Chinese Aca. Sci.); (Jiangnan)

    2015-04-29

    The time-dependent conformational change of bovine serum album (BSA) during Maillard reaction with dextran under controlled dry heating has been studied by small-angle X-ray scattering, fluorescence spectroscopy, dynamic light scattering, and circular dichroism analysis. Through the research on the radii of gyration (Rg), intrinsic fluorescence, and secondary structure, conjugates with dextran coating were found to inhibit BSA aggregation and preserve the secondary structure of native BSA against long-time heat treatment during Maillard reaction. The results suggested that the hydrophilic dextran was conjugated to the compact protein surface and enclosed it and more dextran chains were attached to BSA with the increase of the heating time. The study presented here will be beneficial to the understanding of the conformational evolution of BSA molecules during the dry-heating Maillard reaction and to the control of the protein–polysaccharide conjugate structure.

  17. Investigation on the interaction of pyrene with bovine serum albumin using spectroscopic methods.

    Science.gov (United States)

    Xu, Chengbin; Gu, Jiali; Ma, Xiping; Dong, Tian; Meng, Xuelian

    2014-05-05

    This paper was designed to investigate the interaction of pyrene with bovine serum albumin (BSA) under physiological condition by spectroscopic methods. Spectroscopic analysis of the emission quenching revealed that the quenching mechanism of BSA by pyrene was static. The binding sites and constants of pyrene-BSA complex were observed to be 1.20 and 2.63×10(6) L mol(-1) at 298 K, respectively. The enthalpy change (ΔH) and entropy change (ΔS) revealed that van der Waals forces and hydrogen bonds stabilized the pyrene-BSA complex. Energy transfer from tryptophan to pyrene occurred by a FRET (fluorescence resonance energy transfer) mechanism, and the distance (r=2.72 nm) had been determined. The results of synchronous, three-dimensional fluorescence, and circular dichroism spectra showed that the pyrene induced conformational changes of BSA. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Dietary Compound Kaempferol Inhibits Airway Thickening Induced by Allergic Reaction in a Bovine Serum Albumin-Induced Model of Asthma.

    Science.gov (United States)

    Shin, Daekeun; Park, Sin-Hye; Choi, Yean-Jung; Kim, Yun-Ho; Antika, Lucia Dwi; Habibah, Nurina Umy; Kang, Min-Kyung; Kang, Young-Hee

    2015-12-16

    Asthma is characterized by aberrant airways including epithelial thickening, goblet cell hyperplasia, and smooth muscle hypertrophy within the airway wall. The current study examined whether kaempferol inhibited mast cell degranulation and prostaglandin (PG) release leading to the development of aberrant airways, using an in vitro model of dinitrophenylated bovine serum albumin (DNP-BSA)-sensitized rat basophilic leukemia (RBL-2H3) mast cells and an in vivo model of BSA-challenged asthmatic mice. Nontoxic kaempferol at 10-20 μM suppressed β-hexosaminidase release and cyclooxygenase 2 (COX2)-mediated production of prostaglandin D2 (PGD2) and prostaglandin F2α (PGF2α) in sensitized mast cells. Oral administration of ≤20 mg/kg kaempferol blocked bovine serum albumin (BSA) inhalation-induced epithelial cell excrescence and smooth muscle hypertrophy by attenuating the induction of COX2 and the formation of PGD2 and PGF2α, together with reducing the anti-α-smooth muscle actin (α-SMA) expression in mouse airways. Kaempferol deterred the antigen-induced mast cell activation of cytosolic phospholipase A2 (cPLA2) responsive to protein kinase Cμ (PKCμ) and extracellular signal-regulated kinase (ERK). Furthermore, the antigen-challenged activation of Syk-phospholipase Cγ (PLCγ) pathway was dampened in kaempferol-supplemented mast cells. These results demonstrated that kaempferol inhibited airway wall thickening through disturbing Syk-PLCγ signaling and PKCμ-ERK-cPLA2-COX2 signaling in antigen-exposed mast cells. Thus, kaempferol may be a potent anti-allergic compound targeting allergic asthma typical of airway hyperplasia and hypertrophy.

  19. Interaction of Palmitic Acid with Metoprolol Succinate at the Binding Sites of Bovine Serum Albumin

    Directory of Open Access Journals (Sweden)

    Mashiur Rahman

    2014-12-01

    Full Text Available Purpose: The aim of this study was to characterize the binding profile as well as to notify the interaction of palmitic acid with metoprolol succinate at its binding site on albumin. Methods: The binding of metoprolol succinate to bovine serum albumin (BSA was studied by equilibrium dialysis method (ED at 27°C and pH 7.4, in order to have an insight in the binding chemistry of the drug to BSA in presence and absence of palmitic acid. The study was carried out using ranitidine as site-1 and diazepam as site-2 specific probe. Results: Different analysis of binding of metoprolol succinate to bovine serum albumin suggested two sets of association constants: high affinity association constant (k1 = 11.0 x 105 M-1 with low capacity (n1 = 2 and low affinity association (k2 = 4.0×105 M-1 constant with high capacity (n2 = 8 at pH 7.4 and 27°C. During concurrent administration of palmitic acid and metoprolol succinate in presence or absence of ranitidine or diazepam, it was found that palmitic acid displaced metoprolol succinate from its binding site on BSA resulting reduced binding of metoprolol succinate to BSA. The increment in free fraction of metoprolol succinate was from 26.27% to 55.08% upon the addition of increased concentration of palmitic acid at a concentration of 0×10-5 M to 16×10-5 M. In presence of ranitidine and diazepam, palmitic acid further increases the free fraction of metoprolol succinate from 33.05% to 66.95% and 40.68% to 72.88%, respectively. Conclusion: This data provided the evidence of interaction at higher concentration of palmitic acid at the binding sites on BSA, which might change the pharmacokinetic properties of metoprolol succinate.

  20. Fetal Bovine Serum Meningkatkan Maturasi Inti Oosit Kelinci Setelah Dimaturasi Secara In Vitro

    Directory of Open Access Journals (Sweden)

    Ni Wayan Kurniani Karja

    2010-09-01

    Full Text Available This study was conducted to examine the meiotic competence or nuclear maturation of rabbit oocytesmatured in vitro. Oocytes were recovered by mincing the ovaries in modified phosphate buffer saline (m-PBS. Selected cumulus-oocyte complexes (COCs with compact cumulus cell mass and a dark, homogenousooplasm were cultured in maturation medium at 38°C in a humidified atmosphere of 5% CO2 in air, andthen stained for nuclear maturation. Three experiments were carried out. In Experiment 1, COCs werecultured in maturation medium for 18-20, 22-24, and 28-30 h. The proportion of oocytes at metaphase II(MII was similar (P>0.05 at 18-20 (69.2%, 22-24 (66.4%, and 28-30 (71.1 h of culture. In Experiment 2,COCs were cultured in either maturation medium containing 0.04% bovine serum albumine (BSA or 5%fetal bovine serum (FBS for 24 h. The proprotion of oocytes that reached MII were higher (P<0.05 in FBSgroup (79.2% than those of oocytes in BSA group (64%. In Experiment 3, based on the presence or absenceof follicles and corpora lutea, the ovarian pairs were classified into follicular or luteal stages. There was nodifference (P>0.05 among oocytes collected from ovaries in follicular (79.7% and luteal stages (78.7% inthe ability to achieve nuclear maturation. These results indicated that nuclear maturation of rabbitoocytes in vitro was completed at 18-20 h of maturation culture and was not affected by ovary’s reproductivestage. The presence of FBS in the maturation medium enhanced the ability of rabbit oocytes to achievenuclear maturation.

  1. Protections of bovine serum albumin protein from damage on functionalized graphene-based electrodes by flavonoids

    International Nuclear Information System (INIS)

    Sun, Bolu; Gou, Yuqiang; Xue, Zhiyuan; Zheng, Xiaoping; Ma, Yuling; Hu, Fangdi; Zhao, Wanghong

    2016-01-01

    A sensitive electrochemical sensor based on bovine serum albumin (BSA)/poly (diallyldimethylammonium chloride) (PDDA) functionalized graphene nanosheets (PDDA-G) composite film modified glassy carbon electrode (BSA/PDDA-G/GCE) had been developed to investigate the oxidative protein damage and protections of protein from damage by flavonoids. The performance of this sensor was remarkably improved due to excellent electrical conductivity, strong adsorptive ability, and large effective surface area of PDDA-G. The BSA/PDDA-G/GCE displayed the greatest degree of BSA oxidation damage at 40 min incubation time and in the pH 5.0 Fenton reagent system (12.5 mM FeSO 4 , 50 mM H 2 O 2 ). The antioxidant activities of four flavonoids had been compared by fabricated sensor based on the relative peak current ratio of SWV, because flavonoids prevented BSA damage caused by Fenton reagent and affected the BSA signal in a solution containing Co(bpy) 3 3+ . The sensor was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM). UV–vis spectrophotometry and FTIR were also used to investigate the generation of hydroxyl radical and BSA damage, respectively. On the basis of results from electrochemical methods, the order of the antioxidant activities of flavonoids is as follows: (+)-catechin > kaempferol > apigenin > naringenin. A novel, direct SWV analytical method for detection of BSA damage and assessment of the antioxidant activities of four flavonoids was developed and this electrochemical method provided a simple, inexpensive and rapid detection of BSA damage and evaluation of the antioxidant activities of samples. - Highlights: • Hydroxyl radicals were produced by Fenton reagents. • An electrochemical bovine serum albumin (BSA) damage sensor was successfully fabricated. • The proposed biosensor can assess the antioxidant capacity of four flavonoids. • The order of antioxidant activities of flavonoids

  2. Protections of bovine serum albumin protein from damage on functionalized graphene-based electrodes by flavonoids

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Bolu [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Gou, Yuqiang [Lanzhou Military Command Center for Disease Prevention and Control, Lanzhou 730000 (China); Xue, Zhiyuan; Zheng, Xiaoping; Ma, Yuling [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Hu, Fangdi, E-mail: hufd@lzu.edu.cn [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Zhao, Wanghong, E-mail: wanghongzhao@sina.com [Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou 51515 (China)

    2016-05-01

    A sensitive electrochemical sensor based on bovine serum albumin (BSA)/poly (diallyldimethylammonium chloride) (PDDA) functionalized graphene nanosheets (PDDA-G) composite film modified glassy carbon electrode (BSA/PDDA-G/GCE) had been developed to investigate the oxidative protein damage and protections of protein from damage by flavonoids. The performance of this sensor was remarkably improved due to excellent electrical conductivity, strong adsorptive ability, and large effective surface area of PDDA-G. The BSA/PDDA-G/GCE displayed the greatest degree of BSA oxidation damage at 40 min incubation time and in the pH 5.0 Fenton reagent system (12.5 mM FeSO{sub 4}, 50 mM H{sub 2}O{sub 2}). The antioxidant activities of four flavonoids had been compared by fabricated sensor based on the relative peak current ratio of SWV, because flavonoids prevented BSA damage caused by Fenton reagent and affected the BSA signal in a solution containing Co(bpy){sub 3}{sup 3+}. The sensor was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM). UV–vis spectrophotometry and FTIR were also used to investigate the generation of hydroxyl radical and BSA damage, respectively. On the basis of results from electrochemical methods, the order of the antioxidant activities of flavonoids is as follows: (+)-catechin > kaempferol > apigenin > naringenin. A novel, direct SWV analytical method for detection of BSA damage and assessment of the antioxidant activities of four flavonoids was developed and this electrochemical method provided a simple, inexpensive and rapid detection of BSA damage and evaluation of the antioxidant activities of samples. - Highlights: • Hydroxyl radicals were produced by Fenton reagents. • An electrochemical bovine serum albumin (BSA) damage sensor was successfully fabricated. • The proposed biosensor can assess the antioxidant capacity of four flavonoids. • The order of antioxidant

  3. Investigating the influence of effective parameters on molecular characteristics of bovine serum albumin nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Rohiwal, S.S.; Satvekar, R.K.; Tiwari, A.P.; Raut, A.V.; Kumbhar, S.G.; Pawar, S.H., E-mail: pawar_s_h@yahoo.com

    2015-04-15

    Graphical abstract: The physiochemical properties of nanoparticles provide the basic aspects about the conformational transitions which could have a strong bearing on the bioavailability for bioactive molecules such as peptides and hormones. - Highlights: • Synthesis and surface and structural properties of Bovine Serum Albumin nanoparticles (BSANPs). • Study of conformational transitions of BSANPs by spectroscopic techniques. • Studies on the effect of pH and protein concentration on formulation of BSANPs. - Abstract: The protein nanoparticles formulation is a challenging task as they are prone to undergo conformational transitions while processing which may affect bioavailability for bioactive compounds. Herein, a modified desolvation method is employed to prepare Bovine Serum Albumin nanoparticles, with controllable particle size ranging from 100 to 300 nm and low polydispersity index. The factors influencing the size and structure of BSA NPs viz. protein concentration, pH and the conditions for purification are well investigated. The structure of BSA NPs is altered due to processing, and may affect the effective binding ability with drugs and bioactive compounds. With that aims, investigations of molecular characteristics of BSA NPs are carried out in detail by using spectroscopic techniques. UV–visible absorption and Fourier Transform Infrared demonstrate the alteration in protein structure of BSA NPs whereas the FT-Raman spectroscopy investigates changes in the secondary and tertiary structures of the protein. The conformational changes of BSA NPs are observed by change in fluorescence intensity and emission maximum wavelength of tryptophan residue by fluorescence spectroscopy. The field emission scanning electron and atomic force microscopy micrographs confirm the size and semi-spherical morphology of the BSA NPs. The effect of concentration and pH on particle size distribution is studied by particle size analyzer.

  4. Interaction mode and nanoparticle formation of bovine serum albumin and anthocyanin in three buffer solutions

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Rui; Dong, Xueyan; Song, Lanlan; Jing, Hao, E-mail: hao.haojing@gmail.com

    2014-11-15

    Investigation of interaction mode of bovine serum albumin (BSA) and anthocyanin (ACN) in different solutions will help us understand the interaction mechanism and functional change of bioactive small molecule and biomacromolecule. This study investigated the binding mode, including binding constant, number of binding sites, binding force of BSA and ACN interaction in three buffer solutions of phosphate (PBS), sodium chloride (NaCl), and PBS-NaCl, using fluorescence spectroscopy and synchronous fluorescence spectroscopy. Formation and characteristics of BSA–ACN complex were also investigated using dynamic light scattering (DLS) and transmission electron microscopy (TEM). The results showed that ACN could interact with BSA at both tyrosine (Tyr) and tryptophan (Trp) residues through both hydrogen bonds and van der Waals force, and the same binding mode was seen in dH{sub 2}O and three buffer solutions. The value of binding constant K was decreased as the temperature increased from 298 K to 308 K, and the decreasing degree was in the order of dH{sub 2}O (9.0×10{sup 4})>NaCl (2.64×10{sup 4})/PBS (2.37×10{sup 4})>PBS-NaCl (0.88×10{sup 4}), which was inversely correlated with the ionic strength of the buffer solutions of PBS-NaCl>NaCl>PBS. It indicated that stability of BSA–ACN complex was affected most in dH{sub 2}O than in three buffer solutions. The BSA and ACN interaction led to formation of BSA–ACN nanoparticles. The sizes of BSA–ACN nanoparticles in dH{sub 2}O were smaller than that in three buffer solutions, which correlated with stronger binding force between BSA and ACN in dH{sub 2}O than in three buffer solutions at room temperature (25 °C, 298 K). - Highlights: • We report the influences of four solutions on the BSA–ACN interaction. • We report the relationship between BSA–ACN interaction and particle size of complex. • The stability of BSA–ACN complex was affected most in dH{sub 2}O than in buffer solutions.

  5. Interaction mode and nanoparticle formation of bovine serum albumin and anthocyanin in three buffer solutions

    International Nuclear Information System (INIS)

    Zhou, Rui; Dong, Xueyan; Song, Lanlan; Jing, Hao

    2014-01-01

    Investigation of interaction mode of bovine serum albumin (BSA) and anthocyanin (ACN) in different solutions will help us understand the interaction mechanism and functional change of bioactive small molecule and biomacromolecule. This study investigated the binding mode, including binding constant, number of binding sites, binding force of BSA and ACN interaction in three buffer solutions of phosphate (PBS), sodium chloride (NaCl), and PBS-NaCl, using fluorescence spectroscopy and synchronous fluorescence spectroscopy. Formation and characteristics of BSA–ACN complex were also investigated using dynamic light scattering (DLS) and transmission electron microscopy (TEM). The results showed that ACN could interact with BSA at both tyrosine (Tyr) and tryptophan (Trp) residues through both hydrogen bonds and van der Waals force, and the same binding mode was seen in dH 2 O and three buffer solutions. The value of binding constant K was decreased as the temperature increased from 298 K to 308 K, and the decreasing degree was in the order of dH 2 O (9.0×10 4 )>NaCl (2.64×10 4 )/PBS (2.37×10 4 )>PBS-NaCl (0.88×10 4 ), which was inversely correlated with the ionic strength of the buffer solutions of PBS-NaCl>NaCl>PBS. It indicated that stability of BSA–ACN complex was affected most in dH 2 O than in three buffer solutions. The BSA and ACN interaction led to formation of BSA–ACN nanoparticles. The sizes of BSA–ACN nanoparticles in dH 2 O were smaller than that in three buffer solutions, which correlated with stronger binding force between BSA and ACN in dH 2 O than in three buffer solutions at room temperature (25 °C, 298 K). - Highlights: • We report the influences of four solutions on the BSA–ACN interaction. • We report the relationship between BSA–ACN interaction and particle size of complex. • The stability of BSA–ACN complex was affected most in dH 2 O than in buffer solutions

  6. Human Umbilical Cord Blood Serum Has Higher Potential in Inducing Proliferation of Fibroblast than Fetal Bovine Serum

    Directory of Open Access Journals (Sweden)

    Ferry Sandra

    2017-09-01

    Full Text Available Background: Cytokines and growth factors were reported to play an important role in stimulating fibroblast proliferation. In vitro culture, fibroblast is mostly culture in medium containing fetal bovine serum (FBS.  Human umbilical cord blood (hUCB has been reported to have low immunogenic property and potential in wound healing, so therefore hUCB serum (hUCBS could be potential and were investigated in current study. Materials and Methods: Five hUCBs were collected from healthy volunteers with normal delivering procedure. hUCB was ex utero immediately collected from umbilical vein in vacutainers and processed. NIH3T3 cells were cultured in DMEM with 10% FBS or 5-20% hUCBS for 48 hours. Cells were then quantified using MTT assay. Protein concentration of FBS and hUCBS were quantified using Bradford assay. Results: NIH3T3 cells density grown in DMEM with 10% FBS was the lowest. NIH3T3 cells densities were increased along with the increment of hUCBS concentrations. MTT results showed that average number of NIH3T3 cells grown in DMEM with 10% FBS was 6,185±1,243. Meanwhile average numbers of NIH3T3 cells grown in DMEM with 5%, 10% and 20% hUCBS were 8,126±628, 9,685±313 and 12,200±304, respectively. Average numbers of NIH3T3 cells grown in DMEM with 5% hUCBS were significantly higher than the ones with 10% FBS (p=0.000. Bradford results showed that concentration of hUCBS was significantly higher than the one of FBS (p=0.000. Conclusion: hUCBS could induce higher proliferation rate of NIH3T3 cells than FBS. Hence hUCBS could be suggested as an alternate of FBS in inducing fibroblast. Keywords: NIH3T3, fibroblast, UCB, serum, FBS, proliferation

  7. Detection by radioimmunoassay and enzyme-linked immunosorbent assay of coronavirus antibodies in bovine serum and lacteal secretions.

    Science.gov (United States)

    Rodak, L; Babiuk, L A; Acres, S D

    1982-07-01

    The sensitivity of a radioimmunoassay (RIA), an enzyme-linked immunosorbent assay (ELISA), and a serum neutralization assay (SN) for detecting antibodies to bovine coronavirus in serum and colostrum were compared. Although there proved to be a good correlation among all three assays (r = 0.915 and 0.964 for RIA with SN and ELISA, respectively), RIA and ELISA proved to be at least 10 times more sensitive than neutralization tests. By using these techniques, it was possible to detect a time-dependent decrease in antibody levels in bovine colostrum after parturition. Using ELISA, we demonstrated that 12 of 12 herds in Saskatchewan, and 109 of 110 animals tested, and antibody to bovine coronavirus. There was no elevated antibody response in serum or lacteal secretions of cows vaccinated once or twice with a commercially available modified live rota-coronavirus vaccine. In addition to being more sensitive than SN, ELISA and RIA proved to have other advantages for measuring antibody levels to bovine coronavirus and therefore warrant wider use as tools in diagnostic virology.

  8. Effect of fetal bovine serum on mineralization in silk fibroin scaffolds.

    Science.gov (United States)

    Vetsch, Jolanda R; Paulsen, Samantha J; Müller, Ralph; Hofmann, Sandra

    2015-02-01

    Fetal bovine serum (FBS) is a common media supplement used in tissue engineering (TE) cultures. The chemical composition of FBS is known to be highly variable between different brands, types or batches and can have a significant impact on cell function. This study investigated the influence of four different FBS types in osteogenic or control medium on mineralization of acellular and cell-seeded silk fibroin (SF) scaffolds. In bone TE, mineralized tissue is considered as the final product of a successful cell culture. Calcium assays and micro-computed tomography scans revealed spontaneous mineralization on SF scaffolds with certain FBS types, even without cells present. In contrast, cell-mediated mineralization was found under osteogenic conditions only. Fourier transform infrared spectroscopy analysis demonstrated a similar ion composition of the mineralization present in scaffolds, whether cell-mediated or spontaneous. These results were confirmed by scanning electron microscopy. This study shows clear evidence for the influence of FBS type on mineralization on SF scaffolds. The suitability of FBS medium supplementation in TE studies is highly questionable with regard to reproducibility of studies and comparability of obtained results. For future TE studies, alternatives to conventional FBS such as defined FBS or serum-free media should be considered, as suggested decades ago. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  9. The binding of Curcuma longa extract with bovine serum albumin monitored via time-resolved fluorescence.

    Science.gov (United States)

    Lemos, M Adília; Hungerford, Graham

    2013-01-01

    Turmeric (Curcuma longa L.) is obtained from the rhizome of the Zingberaceae family and has a long history as an ingredient in cooking. It has been used as a dye and recently research has concentrated on its possible health benefits, specifically because of its antioxidant activity. The principal compound that is responsible for this activity is curcumin, which is present with the other curcuminoids; demethoxycurcumin and bisdemethoxycurcumin. Curcumin exhibits fluorescence and its photophysics are markedly affected by the polarity, hydrogen bonding and pH. This provides a means to examine its interaction with proteins, which is important if its potential health role is to be fully investigated. In this work, we monitor the binding kinetics using time-resolved fluorescence measurements, enabled by the use of low dead time electronics coupled with a high repetition rate excitation source and time-resolved emission spectra of the extracted curcuminoids upon interaction with bovine serum albumin. From these measurements the decay-associated spectra of the different lifetime components were obtained, which is consistent with reports of more than one binding site. Monitoring changes in these spectra with increasing temperature also allows for the denaturing of the serum albumin to be inferred. © 2013 The American Society of Photobiology.

  10. The Process of Separating Bovine Serum Albumin Using Hydroxyapatite and Active Babassu Coal (Orbignya martiana).

    Science.gov (United States)

    Ribeiro Alves, Márcia Regina; Zuñiga, Abraham Damian Giraldo; Sousa, Rita de Cássia Superbi; Zacchi Scolforo, Carmelita

    2016-01-01

    Bovine serum albumin is one of the major serum proteins; it plays an important role as a result of its functional and nutritional properties which have bioactive peptides. Adsorption method was used to separate protein, which involves hydroxyapatite, synthetic hydroxyapatite, and active babassu coal. Initially, characterization was carried out using the zeta potential of the adsorbents. Kinetic pseudo-first- and pseudo-second-order models were applied. For isotherms, equilibrium data studies were carried out using the Langmuir and Freundlich models, in addition to determining the efficiency of adsorptive process. The results of the zeta potential showed loads ranging from +6.9 to -42.8 mV. The kinetic data were better represented in the pseudo-second-order model with chemisorption characteristics. The adsorption capacity of the adsorbents decreased as pH increased, indicating that the electrostatic bonds and some functional groups of active babassu coal contributed to the reduction of adsorption, especially oxygen linked to carbon atoms. The value of pH 4.0 showed the best results of adsorption, being obtained as the maximum adsorption capacity (q m ) and yield (%) (where q m = 87.95 mg g(-1) and 74.2%; 68.26 mg g(-1) and 68.6%; and 36.18 mg g(-1), 37.4%) of hydroxyapatite, synthetic hydroxyapatite, and active babassu coal, respectively.

  11. The Process of Separating Bovine Serum Albumin Using Hydroxyapatite and Active Babassu Coal (Orbignya martiana

    Directory of Open Access Journals (Sweden)

    Márcia Regina Ribeiro Alves

    2016-01-01

    Full Text Available Bovine serum albumin is one of the major serum proteins; it plays an important role as a result of its functional and nutritional properties which have bioactive peptides. Adsorption method was used to separate protein, which involves hydroxyapatite, synthetic hydroxyapatite, and active babassu coal. Initially, characterization was carried out using the zeta potential of the adsorbents. Kinetic pseudo-first- and pseudo-second-order models were applied. For isotherms, equilibrium data studies were carried out using the Langmuir and Freundlich models, in addition to determining the efficiency of adsorptive process. The results of the zeta potential showed loads ranging from +6.9 to −42.8 mV. The kinetic data were better represented in the pseudo-second-order model with chemisorption characteristics. The adsorption capacity of the adsorbents decreased as pH increased, indicating that the electrostatic bonds and some functional groups of active babassu coal contributed to the reduction of adsorption, especially oxygen linked to carbon atoms. The value of pH 4.0 showed the best results of adsorption, being obtained as the maximum adsorption capacity (qm and yield (% (where qm=87.95 mg g−1 and 74.2%; 68.26 mg g−1 and 68.6%; and 36.18 mg g−1, 37.4% of hydroxyapatite, synthetic hydroxyapatite, and active babassu coal, respectively.

  12. Multi-spectroscopic investigation of the binding interaction of fosfomycin with bovine serum albumin

    Directory of Open Access Journals (Sweden)

    Manjunath D. Meti

    2015-08-01

    Full Text Available The interaction between fosfomycin (FOS and bovine serum albumin (BSA has been investigated effectively by multi-spectroscopic techniques under physiological pH 7.4. FOS quenched the intrinsic fluorescence of BSA via static quenching. The number of binding sites n and observed binding constant KA were measured by the fluorescence quenching method. The thermodynamic parameters ΔG0, ΔH0 and ΔS0 were calculated at different temperatures according to the van’t Hoff equation. The site of binding of FOS in the protein was proposed to be Sudlow’s site I based on displacement experiments using site markers viz. warfarin, ibuprofen and digitoxin. The distance r between the donor (BSA and acceptor (FOS molecules was obtained according to the Förster theory. The effect of FOS on the conformation of BSA was analyzed using synchronous fluorescence spectra (SFS, circular dichroism (CD and 3D fluorescence spectra. A molecular modeling study further confirmed the binding mode obtained by the experimental studies. Keywords: Fosfomycin, Serum albumin, Spectroscopic methods, Synchronous fluorescence, 3D spectra

  13. The Process of Separating Bovine Serum Albumin Using Hydroxyapatite and Active Babassu Coal (Orbignya martiana)

    Science.gov (United States)

    Zuñiga, Abraham Damian Giraldo; Sousa, Rita de Cássia Superbi; Zacchi Scolforo, Carmelita

    2016-01-01

    Bovine serum albumin is one of the major serum proteins; it plays an important role as a result of its functional and nutritional properties which have bioactive peptides. Adsorption method was used to separate protein, which involves hydroxyapatite, synthetic hydroxyapatite, and active babassu coal. Initially, characterization was carried out using the zeta potential of the adsorbents. Kinetic pseudo-first- and pseudo-second-order models were applied. For isotherms, equilibrium data studies were carried out using the Langmuir and Freundlich models, in addition to determining the efficiency of adsorptive process. The results of the zeta potential showed loads ranging from +6.9 to −42.8 mV. The kinetic data were better represented in the pseudo-second-order model with chemisorption characteristics. The adsorption capacity of the adsorbents decreased as pH increased, indicating that the electrostatic bonds and some functional groups of active babassu coal contributed to the reduction of adsorption, especially oxygen linked to carbon atoms. The value of pH 4.0 showed the best results of adsorption, being obtained as the maximum adsorption capacity (q m) and yield (%) (where q m = 87.95 mg g−1 and 74.2%; 68.26 mg g−1 and 68.6%; and 36.18 mg g−1, 37.4%) of hydroxyapatite, synthetic hydroxyapatite, and active babassu coal, respectively. PMID:27376149

  14. Biomimetic synthesis of calcium carbonate with different morphologies and polymorphs in the presence of bovine serum albumin and soluble starch.

    Science.gov (United States)

    Liu, Yuxi; Chen, Yuping; Huang, Xuechen; Wu, Gang

    2017-10-01

    Calcium carbonate has been synthesized by the reaction of Na 2 CO 3 and CaCl 2 in the presence of bovine serum albumin (BSA) and soluble starch. Effects of various bovine serum albumin (BSA) and soluble starch on the polymorph and morphology of CaCO 3 crystals were investigated. Crystallization of vaterite is favored in the presence of BSA and soluble starch, respectively, while calcite is favored in the presence of a mixture of BSA and soluble starch. The morphologies of CaCO 3 particles in the presence of mixture of BSA and soluble starch are mainly rod-like, suggesting that the BSA, soluble and their assemblies play key roles in stabilizing and directing the CaCO 3 crystal growth. Copyright © 2017. Published by Elsevier B.V.

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

    Science.gov (United States)

    Kasálková, Nikola Slepičková; Slepička, Petr; Kolská, Zdeňka; Hodačová, Petra; Kučková, Štěpánka; Švorčík, Václav

    2014-04-01

    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.

  16. Binding of several anti-tumor drugs to bovine serum albumin: Fluorescence study

    Energy Technology Data Exchange (ETDEWEB)

    Bi Shuyun [College of Chemistry, Changchun Normal University, Changchun 130032 (China)], E-mail: sy_bi@sina.com; Sun Yantao [College of Chemistry, Jilin University, Changchun 130023 (China); College of Chemistry, Jilin Normal University, Siping 136000 (China); Qiao Chunyu; Zhang Hanqi [College of Chemistry, Jilin University, Changchun 130023 (China); Liu Chunming [College of Chemistry, Changchun Normal University, Changchun 130032 (China)

    2009-05-15

    The interactions of mitomycin C (MMC), fluorouracil (FU), mercaptopurine (MP) and doxorubicin hydrochloride (DXR) with bovine serum albumin (BSA) were studied by spectroscopic method. Quenching of fluorescence of serum albumin by these drugs was found to be a static quenching process. The binding constants (K{sub A}) were 9.66x10{sup 3}, 2.08x10{sup 3}, 8.20x10{sup 2} and 7.50x10{sup 3} L mol{sup -1} for MMC-, FU-, MP- and DXR-BSA, respectively, at pH 7.4 Britton-Robinson buffer at 28 deg. C. The thermodynamic functions such as enthalpy change ({delta}H), entropy change ({delta}S) and Gibbs free-energy change ({delta}G) for the reactions were also calculated according to the thermodynamic equations. The main forces in the interactions of these drugs with BSA were evaluated. It was found that the interactions of MMC and FU with BSA were exothermic processes and those of MP and DXR with BSA were endothermic. In addition, the binding sites on BSA for the four drugs were probed by the changes of binding properties of these drugs with BSA in the presence of two important site markers such as ibuprofen and indomethacin. Based on the Foester theory of non-radiation energy transfer, the binding distances between the drugs and tryptophane were calculated and they were 3.00, 1.14, 2.85, and 2.79 nm for MMC, FU, MP and DXR, respectively.

  17. The influence of salt type on the retention of bovine serum albumin in ion-exchange chromatography

    DEFF Research Database (Denmark)

    Al-Jibbouri, Sattar

    2007-01-01

    In this paper, an analysis of the influence of the salt types, NaCl, NaCH"3COO, Na"2SO"4 and Na"3C"6H"5O"7, on the isocratic retention behaviour of bovine serum albumin (BSA) on two anion-exchangers media (Source 30Q and TSK Gel Super Q 5 PW) has been presented. The retention data demonstrated...

  18. Interactions of hemin with bovine serum albumin and human hemoglobin: A fluorescence quenching study

    Science.gov (United States)

    Makarska-Bialokoz, Magdalena

    2018-03-01

    The binding interactions between hemin (Hmi) and bovine serum albumin (BSA) or human hemoglobin (HHb), respectively, have been examined in aqueous solution at pH = 7.4, applying UV-vis absorption, as well as steady-state, synchronous and three-dimensional fluorescence spectra techniques. Representative results received for both BSA and HHb intrinsic fluorescence proceeding from the interactions with hemin suggest the formation of stacking non-covalent and non-fluorescent complexes in both the Hmi-BSA and Hmi-HHb systems, with highly possible concurrent formation of a coordinate bond between a group on the protein surface and the metal in Hmi molecule. All the values of calculated parameters, the binding, fluorescence quenching and bimolecular quenching rate constants point to the involvement of static quenching in both the systems studied. The blue shift in the synchronous fluorescence spectra imply the participation of both tryptophan and tyrosine residues in quenching of BSA and HHb intrinsic fluorescence. Depicted outcomes suggest that hemin is supposedly able to influence the physiological functions of BSA and HHb, the most important blood proteins, particularly in case of its overuse.

  19. Formulation for Oral Delivery of Lactoferrin Based on Bovine Serum Albumin and Tannic Acid Multilayer Microcapsules

    Science.gov (United States)

    Kilic, Ece; Novoselova, Marina V.; Lim, Su Hui; Pyataev, Nikolay A.; Pinyaev, Sergey I.; Kulikov, Oleg A.; Sindeeva, Olga A.; Mayorova, Oksana A.; Murney, Regan; Antipina, Maria N.; Haigh, Brendan; Sukhorukov, Gleb B.; Kiryukhin, Maxim V.

    2017-03-01

    Lactoferrin (Lf) has considerable potential as a functional ingredient in food, cosmetic and pharmaceutical applications. However, the bioavailability of Lf is limited as it is susceptible to digestive enzymes in gastrointestinal tract. The shells comprising alternate layers of bovine serum albumin (BSA) and tannic acid (TA) were tested as Lf encapsulation system for oral administration. Lf absorption by freshly prepared porous 3 μm CaCO3 particles followed by Layer-by-Layer assembly of the BSA-TA shells and dissolution of the CaCO3 cores was suggested as the most efficient and harmless Lf loading method. The microcapsules showed high stability in gastric conditions and effectively protected encapsulated proteins from digestion. Protective efficiency was found to be 76 ± 6% and 85 ± 2%, for (BSA-TA)4 and (BSA-TA)8 shells, respectively. The transit of Lf along the gastrointestinal tract (GIT) of mice was followed in vivo and ex vivo using NIR luminescence. We have demonstrated that microcapsules released Lf in small intestine allowing 6.5 times higher concentration than in control group dosed with the same amount of free Lf. Significant amounts of Lf released from microcapsules were then absorbed into bloodstream and accumulated in liver. Suggested encapsulation system has a great potential for functional foods providing lactoferrin.

  20. Electrostatic compared with hydrophobic interactions between bovine serum amine oxidase and its substrates.

    Science.gov (United States)

    Di Paolo, Maria Luisa; Stevanato, Roberto; Corazza, Alessandra; Vianello, Fabio; Lunelli, Lorenzo; Scarpa, Marina; Rigo, Adelio

    2003-01-01

    A steady-state kinetic study of bovine serum amine oxidase activity was performed, over a wide range of pH values (5.4-10.2) and ionic strength (10-200 mM), using various (physiological and analogue) substrates as specific probes of the active-site binding region. Relatively small changes in k (cat) values (approx. one order of magnitude) accompanied by marked changes in K(m) and k(cat)/K(m) values (approx. six orders of magnitude) were observed. This behaviour was correlated with the presence of positively charged groups or apolar chains in the substrates. In particular, it was found that the docking of the physiological polyamines, i.e. spermidine and spermine, appears to be modulated by three amino acid residues of the active site, which we have named L(-)H(+), G(-)H(+) and IH(+), characterized by p K (a) values of 6.2+/-0.2 [Di Paolo, Scarpa, Corazza, Stevanato and Rigo (2002) Biophys. J. 83, 2231-2239], 8.2+/-0.3 and 7.8+/-0.4 respectively. The electrostatic interaction between the protonated substrates and the enzyme containing the residues L(-)H(+), G(-)H(+) and IH(+) in the deprotonated form, the on/off role of the IH(+) residue and the role of hydrophobic interactions with substrates characterized by apolar chains are discussed. PMID:12529179

  1. Bovine serum albumin nanoparticles loaded with Photosens photosensitizer for effective photodynamic therapy

    Science.gov (United States)

    Khanadeev, Vitaly; Khlebtsov, Boris; Packirisamy, Gopinath; Khlebtsov, Nikolai

    2017-03-01

    Polymeric nanoparticles (NPs) are widely used for drug delivery applications due to high biodegradability, low toxicity and high loading capacity. The focus of this study is the development of photosensitizer Photosens (PS) loaded albumin NPs for efficient photodynamic therapy (PDT). To fabricate PS-loaded bovine serum albumin nanoparticles (BSA-PS NPs), we used a coacervation method with glutaraldehyde followed by passive loading of PS. Successful loading of PS was confirmed by appearance of characteristic peak in absorption spectrum which allows to determine the PS loading in BSA NPs. The synthesized BSA-PS NPs demonstrated low toxicity to HeLa cells at therapeutic concentrations of loaded PS. Compared to free PS solution, the synthesized BSA-PS NPs generated the singlet oxygen more effectively under laser irradiation at 660 nm. In addition, due to presence of various chemical groups on the surface of BSA-PS NPs, they are capable to adsorb on cell surface and accumulate in cells due to cellular uptake mechanisms. Owing to combination of PD and cell uptake advantages, BSA-PS NPs demonstrated higher efficacy of photodynamic damage to cancer cells as compared to free PS at equivalent concentrations. These results suggest that non-targeted BSA-PS NPs with high PD activity and low-fabrication costs of are promising candidates for transfer to PD clinic treatments.

  2. Functionalized polypropylene non-woven fabric membrane with bovine serum albumin and its hemocompatibility enhancement.

    Science.gov (United States)

    Zhang, Chang; Jin, Jing; Zhao, Jie; Jiang, Wei; Yin, Jinghua

    2013-02-01

    Bovine serum albumin (BSA) was successfully immobilized onto polypropylene non-woven fabric (PP(NWF)) membranes using poly(acrylic acid) (PAA) as a spacer. Firstly, O(2) plasma treatment and UV-irradiated technique were combined to graft PAA onto the membranes. BSA was then immobilized onto the PAA grafted surface through the coupling of amino groups of BSA to the carboxyl groups of PAA. The immobilization of PAA and BSA onto the membrane was confirmed by attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), and water contact angle measurement. The water contact angle measurement results revealed that the membrane hydrophilicity improved after modification with PAA and BSA. After BSA immobilization, the amount of protein adsorption and the number of platelet adhesion on the modified membrane significantly decreased, which indicated that hemocompatibility had been considerably improved compared with neat and PAA grafted PP(NWF). The whole blood clotting time measurement showed that the anticoagulant property of the modified membrane was also significantly enhanced. Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.

  3. Properties and stability of blueberry anthocyanin--bovine serum albumin nanoparticles.

    Science.gov (United States)

    Chen, Jian; Tao, Xiaoyun; Zhang, Meng; Sun, Aidong; Zhao, Liyi

    2014-07-01

    Since they would be easily decomposed under alkaline conditions, anthocyanins are likely to have poor oxidation stability. However, encapsulated with protein molecules, anthocyanins could be protected owing to the slowing down of the oxidation process. In this study, the characteristics of nanoparticles, formed by the interactions of anthocyanins with bovine serum albumin (BSA), and their impact on the oxidation stability of anthocyanins were investigated. Both BSA and anthocyanin-bound BSA could form self-assembled nanoparticles in phosphate buffer (pH 7.4), and the particle size of anthocyanin-bound BSA (20-25 nm) was smaller than that of BSA (35-40 nm). The ratio of BSA to anthocyanin was 1:10. The radical scavenging rates of BSA-bound anthocyanin were lower than those of the unbound anthocyanin. No significant difference was seen in the stability between the unbound and BSA-bound anthocyanin in the simulated gastric system, whereas a difference was seen in the simulated intestinal system. The amount of unbound anthocyanin decreased by 70% after 6 h, while BSA-bound anthocyanin was almost unchanged. BSA exhibited a remarkable effect on the oxidation stability of anthocyanins. BSA nanocarriers could improve the stability of anthocyanin under neutral conditions, which has great potential for applications. © 2013 Society of Chemical Industry.

  4. Spectroscopic studies on the interaction between chalcone and bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Naik, Keerti M.; Nandibewoor, Sharanappa T., E-mail: stnandibewoor@yahoo.com

    2013-11-15

    The interaction between chalcone and bovine serum albumin (BSA) has been studied by spectroscopic techniques under physiological condition. By the analysis of fluorescence spectrum and fluorescence intensity, it was observed that the chalcone has a strong ability to quench the intrinsic fluorescence with BSA through a static quenching procedure and non-radiation energy transfer were the main reasons for the fluorescence quenching. The association constants of chalcone with BSA were determined at different temperatures based on fluorescence quenching results. The positive entropy change and enthalpy change indicated that the interaction of chalcone and BSA was driven mainly by hydrophobic forces. The process of binding was a spontaneous process in which Gibbs free energy change was negative. The distance, r, between donor (BSA) and acceptor (chalcone) was obtained according to the Forster's theory of non-radiation energy transfer. The UV–vis, CD, FT-IR, synchronous and 3-D spectral results revealed the changes in the secondary structure of BSA upon interaction with chalcone. The effects of some common metal ions on binding of BSA–chalcone complex were also investigated. -- Highlights: • We explored the interaction between chalcone and BSA by fluorescence spectroscopy. • The fluorescence quenching mechanism was static quenching. • The binding constants and thermodynamic parameters were calculated. • The interaction is driven mainly by hydrophobic force. • The binding of chalcone to BSA induced changes in the secondary structure of BSA.

  5. Investigation on the interaction between bovine serum albumin and 2,2-diphenyl-1-picrylhydrazyl

    Energy Technology Data Exchange (ETDEWEB)

    Li, Xiangrong [Department of Chemistry, School of Basic Medicine, Xinxiang Medical University, Xinxiang, Henan 453003 (China); Chen, Dejun; Wang, Gongke [School of Chemistry and Chemical Engineering, Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, Henan Normal University, 46 Jian-she Road, Mu Ye District, Xinxiang, Henan 453007 (China); Lu, Yan, E-mail: 1842457577@qq.com [School of Chemistry and Chemical Engineering, Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, Henan Normal University, 46 Jian-she Road, Mu Ye District, Xinxiang, Henan 453007 (China)

    2014-12-15

    Albumin represents a very abundant and important circulating antioxidant in plasma. In this paper, the ability of bovine serum albumin (BSA) to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical has been investigated using UV–vis absorption spectra. The result shows that the antioxidant activity of BSA against DPPH radical is similar to glutathione and the value of IC{sub 50} is 5.153×10{sup −5} mol L{sup −1}. The interaction between BSA and DPPH has been investigated without or with the eight popular antioxidants (L-ascorbic acid, α-tocopherol, glutathione, melatonin, (+)-catechin hydrate, procyanidine B3, β-carotene and astaxanthin) by means of fluorescence spectroscopy and circular dichroism (CD) spectroscopy. The fluorescence experiments show that DPPH quenches the fluorescence intensity of BSA through a static mechanism. The quenching process of DPPH with BSA is easily affected by the eight antioxidants, however, they cannot change the quenching mechanism of DPPH with BSA. Additionally, as shown by synchronous fluorescence spectroscopy and CD, DPPH may induce conformational and microenvironmental changes of BSA. - Highlights: • The antioxidant activity of BSA against DPPH is similar to glutathione. • DPPH can quench the fluorescence of BSA through a static quenching. • One molecule of DPPH radical reduced by one molecule of BSA. • The eight antioxidants cannot change the quenching mechanism of DPPH with BSA. • The binding parameters are decreased by the introduction of the eight antioxidants.

  6. Urea-induced binding between diclofenac sodium and bovine serum albumin: a spectroscopic insight.

    Science.gov (United States)

    Dohare, Neeraj; Khan, Abbul Bashar; Athar, Fareeda; Thakur, Sonu Chand; Patel, Rajan

    2016-06-01

    We investigated the interaction of diclofenac sodium (Dic.Na) with bovine serum albumin (BSA) in the absence and presence of urea using different spectroscopic techniques. A fluorescence quenching study revealed that the Stern-Volmer quenching constant decreases in the presence of urea, decreasing further at higher urea concentrations. The binding constant and number of binding sites were also evaluated for the BSA-Dic.Na interaction system in the absence and presence of urea using a modified Stern-Volmer equation. The binding constant is greater at high urea concentrations, as shown by the fluorescence results. In addition, for the BSA-Dic.Na interaction system, a static quenching mechanism was observed, which was further confirmed using time-resolved fluorescence spectroscopy. UV-vis spectroscopy provided information about the formation of a complex between BSA and Dic.Na. Circular dichroism was carried out to explain the conformational changes in BSA induced by Dic.Na in the absence and presence of urea. The presence of urea reduced the α-helical content of BSA as the Dic.Na concentration varied. The distance r between the donor (BSA) and acceptor (Dic.Na) was also obtained in the absence and presence of urea, using fluorescence resonance energy transfer. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  7. Analysis of the hydration water around bovine serum albumin using terahertz coherent synchrotron radiation.

    Science.gov (United States)

    Bye, Jordan W; Meliga, Stefano; Ferachou, Denis; Cinque, Gianfelice; Zeitler, J Axel; Falconer, Robert J

    2014-01-09

    Terahertz spectroscopy was used to study the absorption of bovine serum albumin (BSA) in water. The Diamond Light Source operating in a low alpha mode generated coherent synchrotron radiation that covered a useable spectral bandwidth of 0.3-3.3 THz (10-110 cm(-1)). As the BSA concentration was raised, there was a nonlinear change in absorption inconsistent with Beer's law. At low BSA concentrations (0-1 mM), the absorption remained constant or rose slightly. Above a concentration of 1 mM BSA, a steady decrease in absorption was observed, which was followed by a plateau that started at 2.5 mM. Using a overlapping hydration layer model, the hydration layer was estimated to extend 15 Å from the protein. Calculation of the corrected absorption coefficient (αcorr) for the water around BSA by subtracting the excluded volume of the protein provides an alternative approach to studying the hydration layer that provides evidence for complexity in the population of water around BSA.

  8. Bovine serum albumin interacts with silver nanoparticles with a "side-on" or "end on" conformation.

    Science.gov (United States)

    Dasgupta, Nandita; Ranjan, Shivendu; Patra, Dhabaleswar; Srivastava, Priyanka; Kumar, Ashutosh; Ramalingam, Chidambaram

    2016-06-25

    As the nanoparticles (NPs) enter into the biological interface, they have to encounter immediate and first exposure to many proteins of different concentrations. The physicochemical interaction of NPs and proteins is greatly influenced not only by the number and type of proteins; but also the surface chemistry of NPs. To analyze the effects of NPs on proteins, the interaction between bovine serum albumin (BSA) and silver nanoparticles (AgNPs) at different concentrations were investigated. The interaction, BSA conformations, kinetics and adsorption were analyzed by UV-Visible spectrophotometer, dynamic light scattering (DLS), FT-IR spectroscopy and fluorescence quenching. DLS, FTIR and UV-visible spectrophotometric analysis confirms the interaction with minor alterations in size of the protein. Fluorescence quenching analysis confirms the side-on or end-on interaction of 1.5 molecules of BSA to AgNP. Further, pseudo-second order kinetics was determined with equilibrium contact-time of 30 min. The data of the present study determines the detailed evaluation of BSA adsorption on AgNP along with mechanism, kinetics and isotherm of the adsorption. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  9. Characterization of zigzag patterns on the surface of bovine serum albumin films

    Directory of Open Access Journals (Sweden)

    D. M. Glibitskiy

    2017-09-01

    Full Text Available Analysis of the patterns formed during the drying of biological fluids is employed for research and diagnosis in medicine and agriculture. Saline solutions of native proteins and DNA are characterized by zigzag patterns, which could be quantitatively described using the specific length of zigzags Lspecific. The aim of this study was to analyze a wide number of characteristics in order to identify those most sensitive to the effects of various physical and chemical factors on the protein structure. We examined the films prepared from salt solutions of bovine serum albumin (BSA under varying conditions, viz.: a proportional change in the concentration of the components, substitution of NaCl with NaF and NaBr, temperature treatment, gamma irradiation, and addition of trivalent iron and biologically active flavin mononucleotide. The results revealed that the distribution of zigzag segment lengths was approximately lognormal, and the distribution of angles between the segments was approximately logistic. Two parameters appeared to be the most informative, viz. the average length of the segments, mainly depending on Cl– concentration, and the number of segments, depending on: Cl– concentration, concentration of the non-aggregated (but not necessarily unfragmented protein, and the excess concentration of ions and low molecular weight compounds.

  10. Effect of cationic grafted copolymer structure on the encapsulation of bovine serum albumin

    International Nuclear Information System (INIS)

    Flynn, Nicholas; Topal, Ç. Özge; Hikkaduwa Koralege, Rangika S.; Hartson, Steve; Ranjan, Ashish; Liu, Jing; Pope, Carey; Ramsey, Joshua D.

    2016-01-01

    The aim of the present study was to evaluate a library of poly-L-lysine (PLL)-graft (g)-polyethylene glycol (PEG) copolymers for the ability to encapsulate effectively a model protein, bovine serum albumin (BSA), and to characterize the stability and protein function of the resulting nanoparticle. A library of nine grafted copolymers was produced by varying PLL molecular weight and PEG grafting ratio. Electrostatic self-assembly of the protein and the grafted copolymer drove encapsulation. The formation of protein/polymer nanoparticles with a core/shell structure was confirmed using PAGE, dynamic light scattering, and electron microscopy. Encapsulation of the BSA into nanoparticles was strongly dependent on the copolymer-to-protein mass ratio, PEG grafting ratio, and PLL molecular weight. A copolymer-to-protein mass ratio of 7:1 and higher was generally required for high levels of encapsulation, and under these conditions, no loss of protein activity was observed. Copolymer characteristics also influenced nanoparticle resistance to polyanions and protease degradation. The results indicate that a copolymer of 15–30 kDa PLL, with a PEG grafting ratio of 10:1, is most promising for protein delivery. - Highlights: • A 4–70 kDa range of PLL-g-PEG copolymers was able to encapsulate BSA into NPs. • Encapsulation of BSA by PLL-g-PEG not only retained but increased esterolytic activity. • NPs were stable against protease degradation and polyanion dissociation.

  11. Solution combustion synthesis of calcium phosphate particles for controlled release of bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Junfeng, E-mail: daidai02304@163.com [School of Chemistry and Materials Engineering, Changshu Institute of Technology, Changshu (China); Jiangsu Laboratory of Advanced Functional Materials, Changshu Institute of Technology, Changshu (China); Zhao, Junjie; Qian, Yu; Zhang, Xiali; Zhou, Feifei; Zhang, Hong [School of Chemistry and Materials Engineering, Changshu Institute of Technology, Changshu (China); Lu, Hongbin [National Laboratory of Solid State Microstructures, College of Engineering and Applied Sciences, Nanjing University, Nanjing (China); Chen, JianHua; Wang, XuHong [School of Chemistry and Materials Engineering, Changshu Institute of Technology, Changshu (China); Jiangsu Laboratory of Advanced Functional Materials, Changshu Institute of Technology, Changshu (China); Yu, Wencong [School of Chemistry and Materials Engineering, Changshu Institute of Technology, Changshu (China)

    2015-05-01

    Four different phase compositions of calcium phosphate (CaP) particles were prepared via a solution combustion method. X-ray diffraction (XRD) and Rietveld analysis results revealed that the variations in the nominal Ca/P (molar) ratios were found to provide a favorable control in the different proportions of CaP materials. Bovine serum albumin (BSA) was used as a model protein to study the loading and release behavior. The release profile indicated that the BSA release rates depended on the phase compositions of the CaP particles, and showed an order of TCP-BSA > BCP-1-BSA > BCP-2-BSA > HA-BSA. The results suggested that the BSA protein release rate can be controlled by varying the phase compositions of CaP carriers. Moreover, the release process involved two stages: firstly surface diffusion via ion exchange and secondly intraparticle diffusion. - Highlights: • Solution combustion method was an efficient way to produced CaP powders. • Ca/P (molar) ratios provided a favorable control in the different proportions of phase composition. • BSA release rate varied depending on the phase composition of the CaP particles. • Two kinetic models were chosen to simulate the release kinetics of the drugs from CaP carriers.

  12. Spectroscopic studies on the binding of barbital to bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Ding Fei, E-mail: caudf@163.co [Department of Chemistry, China Agricultural University, Beijing 100193 (China); Pan Hong; Li Zhiyuan; Liu Feng [Department of Chemistry, China Agricultural University, Beijing 100193 (China); Sun Ying, E-mail: sunying@cau.edu.c [Department of Chemistry, China Agricultural University, Beijing 100193 (China)

    2009-06-15

    In this paper, the interaction between barbital and bovine serum albumin (BSA) was investigated by the method of fluorescence spectroscopy under simulative physiological conditions. Fluorescence data revealed that the fluorescence quenching of BSA by barbital was the result of the formation of BSA-barbital complex, and the effective quenching constants (K{sub a}) were 1.468x10{sup 4}, 1.445x10{sup 4} and 1.403x10{sup 4} M{sup -1} at 297, 303 and 310 K, respectively. The thermodynamic parameters enthalpy change (DELTAH) and entropy change (DELTAS) for the reaction were calculated to be -2.679 kJ mol{sup -1} and 70.76 J mol{sup -1} K{sup -1}, respectively, according to the van't Hoff equation. The results indicated that hydrophobic and electrostatic interactions were the dominant intermolecular force in stabilizing the complex. The results of synchronous fluorescence spectra showed that binding of barbital with BSA can induce conformational changes in BSA. In addition, the effects of Cu{sup 2+} and Zn{sup 2+} on the constants of BSA-barbital complex were also discussed.

  13. Effect of cationic grafted copolymer structure on the encapsulation of bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Flynn, Nicholas [School of Chemical Engineering, Oklahoma State University, Stillwater, OK 74078 (United States); Topal, Ç. Özge [School of Mechanical and Aerospace Engineering, Oklahoma State University, Stillwater, OK 74078 (United States); Hikkaduwa Koralege, Rangika S. [School of Chemical Engineering, Oklahoma State University, Stillwater, OK 74078 (United States); Hartson, Steve [Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK 74078 (United States); Ranjan, Ashish; Liu, Jing; Pope, Carey [Department of Physiological Sciences, Oklahoma State University, Stillwater, OK 74078 (United States); Ramsey, Joshua D., E-mail: josh.ramsey@okstate.edu [School of Chemical Engineering, Oklahoma State University, Stillwater, OK 74078 (United States)

    2016-05-01

    The aim of the present study was to evaluate a library of poly-L-lysine (PLL)-graft (g)-polyethylene glycol (PEG) copolymers for the ability to encapsulate effectively a model protein, bovine serum albumin (BSA), and to characterize the stability and protein function of the resulting nanoparticle. A library of nine grafted copolymers was produced by varying PLL molecular weight and PEG grafting ratio. Electrostatic self-assembly of the protein and the grafted copolymer drove encapsulation. The formation of protein/polymer nanoparticles with a core/shell structure was confirmed using PAGE, dynamic light scattering, and electron microscopy. Encapsulation of the BSA into nanoparticles was strongly dependent on the copolymer-to-protein mass ratio, PEG grafting ratio, and PLL molecular weight. A copolymer-to-protein mass ratio of 7:1 and higher was generally required for high levels of encapsulation, and under these conditions, no loss of protein activity was observed. Copolymer characteristics also influenced nanoparticle resistance to polyanions and protease degradation. The results indicate that a copolymer of 15–30 kDa PLL, with a PEG grafting ratio of 10:1, is most promising for protein delivery. - Highlights: • A 4–70 kDa range of PLL-g-PEG copolymers was able to encapsulate BSA into NPs. • Encapsulation of BSA by PLL-g-PEG not only retained but increased esterolytic activity. • NPs were stable against protease degradation and polyanion dissociation.

  14. Conformational Change-Induced Fluorescence of Bovine Serum Albumin-Gold Complexes.

    Science.gov (United States)

    Dixon, Jacob M; Egusa, Shunji

    2018-02-14

    We report new findings on the red fluorescent (λ em = 640 nm) bovine serum albumin (BSA)-gold (Au) compound initially described by Xie et al. (J. Am. Chem. Soc. 2009, 131, 888-889) as Au 25 nanoclusters. The BSA-Au compounds were further reducible to yield nanoparticles, suggesting that these compounds were BSA-cationic Au complexes. We examined the correlations between BSA conformations (pH-induced as well as denatured) and the resulting fluorescence of BSA-Au complexes, to understand the possible cationic Au binding sites. The red fluorescence of the BSA-Au complex was associated with a particular isoform of BSA, the aged form (pH > 10) of the five pH-dependent BSA conformations, while the other conformations, expanded (pH < 2.7), fast (2.7 < pH < 4.3), normal (4.3 < pH < 8), and basic (8 < pH < 10) did not result in red fluorescence. There could be internal energy transfer mechanisms to produce red fluorescence, deduced from excitation-emission map measurements. The ensemble minimum number of Au(III) per BSA to yield red fluorescence was <7. We illustrate the presence of multiple specific Au binding sites in BSA, and present an interpretation of the fluorescence of the BSA-Au complex, alternative to a single-site nucleation of a neutral Au 25 nanocluster.

  15. Interaction and sonodynamic damage activity of acridine red (AD-R) to bovine serum albumin (BSA)

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Dandan; Xie, Jinhui; Wu, Qiong; Fan, Ping; Wang, Jun, E-mail: wangjun888tg@126.com

    2015-04-15

    The sonodynamic therapy (SDT) has become an attractive antitumor treatment method in recent years, but the selection of sonosensitizer, mechanism of damage biomolecule and kind of reactive oxygen species (ROS) generated during sonodynamic process have not been investigated in detail. In this paper, the acridine red (AD-R), as a sonosensitizer, combining with ultrasonic irradiation to damage bovine serum albumin (BSA) was investigated. At first, the interaction of AD-R to BSA molecules in aqueous solution was studied by fluorescence spectroscopy. As judged from the experimental results, the quenching mechanism of BSA fluorescence belongs to a static process. Synchronous fluorescence spectra demonstrate that the binding and damage sites to BSA molecules are mainly on the tryptophan residues. The generation and kind of generated ROS were also estimated by the method of oxidation and extraction photometry. This paper may offer some valuable references for the study of the sonodynamic activity and application of AD-R in SDT for tumor treatment. - Highlights: ●Acridine red (AD-R) is used to study interaction with BSA. ●Spectroscopy is used to study sonodynamic damage activity of AD-R to BSA. ●Generation of ROS caused by AD-R under ultrasonic irradiation was determined.

  16. Extraction and stability of bovine serum albumin (BSA) using cholinium-based Good's buffers ionic liquids.

    Science.gov (United States)

    Taha, Mohamed; Quental, Maria V; Correia, Isabel; Freire, Mara G; Coutinho, João A P

    2015-07-01

    Good's buffers ionic liquids (GB-ILs), composed of cholinium-based cations and Good's buffers anions, display self-buffering characteristics in the biological pH range, and their polarity and hydrophobicity can be easily tuned by a proper manipulation of their ions chemical structures. In this work, the extraction ability for bovine serum albumin (BSA) of aqueous biphasic systems (ABS) formed by polypropylene glycol 400 (PPG 400) and several GB-ILs was evaluated. ABS formed by PPG 400 and cholinium chloride ([Ch]Cl), GBs, and sucrose were also investigated for comparison purposes. It is shown that BSA preferentially migrates for the GB-IL-rich phase, with extraction efficiencies of 100%, achieved in a single-step. Dynamic light scattering, and circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopies were employed to evaluate the effect of the investigated cholinium-based GB-ILs on the BSA stability, and compared with results obtained for the respective GBs precursors, [Ch]Cl and sucrose, a well-known protein stabilizer. Molecular docking studies were also carried out to investigate on the binding sites of GB-IL ions to BSA. The experimental results confirm that BSA has a higher stability in GB-ILs than in any of the other compounds investigated.

  17. Spectroscopic study on the interaction between mononaphthalimide spermidine (MINS) and bovine serum albumin (BSA).

    Science.gov (United States)

    Tian, Zhiyong; Zang, Fenglei; Luo, Wen; Zhao, Zhonghua; Wang, Yueqiao; Xu, Xuejun; Wang, Chaojie

    2015-01-01

    The interaction mononaphthalimide spermidine (MINS, 1) and bovine serum albumin (BSA) was studied by UV/vis absorption, fluorescence and circular dichroism spectra (CD) under physiological conditions (pH=7.4). The observed spectral quenching of BSA by compound 1 indicated compound 1 could bind to BSA. Further fluorescent tests revealed that the quenching mechanism of BSA by compound 1 was overall static. Meanwhile, the obtained binding constant and thermodynamic parameters on compound-BSA interaction showed that the type of interaction force of compound 1 and BSA was mainly hydrophobic. The analysis of synchronous, three-dimensional fluorescence and CD showed that compound 1 had weak influence on the conformational changes in BSA. Molecular docking simulation was performed and docking model in silico suggested that the configuration of compound 1 was localized in enzymatic drug site II in BSA. Furthermore, naphthalimide moiety of compound 1 greatly contributed to the hydrophobic interaction between compound 1 and BSA protein, as confirmed by experimental data. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Doxorubicin hinders DNA condensation promoted by the protein bovine serum albumin (BSA).

    Science.gov (United States)

    Lima, C H M; de Paula, H M C; da Silva, L H M; Rocha, M S

    2017-12-01

    In this work, we have studied the interaction between the anticancer drug doxorubicin (doxo) and condensed DNA, using optical tweezers. To perform this task, we use the protein bovine serum albumin (BSA) in the working buffer to mimic two key conditions present in the real intracellular environment: the condensed state of the DNA and the abundant presence of charged macromolecules in the surrounding medium. In particular, we have found that, when doxo is previously intercalated in disperse DNA, the drug hinders the DNA condensation process upon the addition of BSA in the buffer. On the other hand, when bare DNA is firstly condensed by BSA, doxo is capable to intercalate and to unfold the DNA condensates at relatively high concentrations. In addition, a specific interaction between BSA and doxo was verified, which significantly changes the chemical equilibrium of the DNA-doxo interaction. Finally, the presence of BSA in the buffer stabilizes the double-helix structure of the DNA-doxo complexes, preventing partial DNA denaturation induced by the stretching forces. © 2017 Wiley Periodicals, Inc.

  19. Interaction of different thiol-capped CdTe quantum dots with bovine serum albumin

    International Nuclear Information System (INIS)

    Wang Qisui; Zhang Xiaolei; Zhou Xiaolan; Fang Tingting; Liu Pengfei; Liu Peng; Min Xinmin; Li, Xi

    2012-01-01

    Due to their unique optical properties, quantum dots (QDs) are rapidly revolutionizing many areas of medicine and biology. Despite the remarkable speed of development of nanoscience, relatively little is known about the interaction of nanoscale objects with organism. In this work, interaction of CdTe QDs coated with mercaptopropanoic acid (MPA), L-cysteine (L-cys), and glutathione (GSH) with bovine serum albumin (BSA) was investigated. Fluorescence (FL), UV–vis absorption, and circular dichroism (CD) spectra methods were used. The Stern-Volmer quenching constant (K sv ) at different temperatures, corresponding thermodynamic parameters (ΔH, ΔG and ΔS), and information of the structural features of BSA were gained. We found that QDs can effectively quench the FL of BSA in a ligand-dependent manner, electrostatic interactions play a major role in the binding reaction, and the nature of quenching is static, resulting in forming QDs-BSA complexes. The CD spectra showed that the secondary and tertiary structure of BSA was changed. This study contributes to a better understanding of the ligand effects on QDs-proteins interactions, which is a critical issue for the applications in vivo. - Highlights: ► The interaction between three thiol-capped QDs and BSA by UV–vis, FL, and CD spectra. ► The bio-effect of CdTe QDs on BSA was a ligand-dependent manner. ► The thermodynamic parameters and the structural features of BSA were gained.

  20. Facile synthesis of bovine serum albumin conjugated low-dimensional ZnS nanocrystals.

    Science.gov (United States)

    Mullamuri, Bhanu; Bhagavathula, Subrahmanya Diwakar; Kasturi, Krishna C S B; Reddy, Venu

    2017-08-01

    We present a facile synthesis of bovine serum albumin (BSA) conjugated low-dimensional ZnS nanocrystals. The experimental parameters such as effects of BSA concentration and precursor vol ratios of Zn:S on the formation of ZnS nanoparticles in BSA matrix were investigated. The ZnS crystalline sizes of 1.9, 1.8 and 1.6nm were obtained by using the BSA concentrations of 1×10 -4 , 5×10 -4 and 10×10 -4 g/mL, respectively, with a fixed Zn:S vol ratio of 1:1. The ZnS samples prepared from 1:10 and 10:1 vol ratios of Zn:S at BSA concentration of 5×10 -4 g/mL shows the crystalline sizes of ZnS are 2.1 and 1.5nm, respectively. FT-IR analysis suggests that the prepared ZnS nanoparticles might be conjugated through the interactions of hydroxyl and amine groups present in BSA. We evaluate the cytotoxicity of the prepared ZnS nanoparticles, the THP-1 cells showed a good viability (>88%) for all the prepared ZnS samples. The plausible mechanism for the formation of ZnS-BSA composite has also been discussed. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Development of etoposide-loaded bovine serum albumin nanosuspensions for parenteral delivery.

    Science.gov (United States)

    Wang, Zhonglan; Li, Zhongwen; Zhang, Dong; Miao, Lei; Huang, Guihua

    2015-01-01

    Nanosuspensions emerge as a promising strategy for delivery of poorly water-soluble drugs. Albumin is a versatile protein carrier for drug delivery and targeting. The purpose of this study was to develop a formulation of etoposide-loaded bovine serum albumin (BSA) nanosuspensions, to study in vitro characterization, and to estimate the in vivo safety and tissue distribution of etoposide-loaded BSA nanosuspensions for parenteral delivery. Etoposide-loaded BSA nanosuspensions were prepared by high-pressure homogenization-solvent precipitation method. The particle size, zeta potential, drug entrapment efficiency, and drug loading of the lyophilized formulation were 182.3 nm, -22.18 mV, 86.44%, and 8.49% respectively. In vitro release files of the formulation presented sustained release properties. Preliminary safety study was conducted to evaluate the delivery system, and results indicated that myelosuppression effect of the etoposide-loaded BSA nanosuspensions group was significantly lower than the Injection® group. Furthermore, results of tissue distribution studies showed that the concentration and AUC of etoposide were increased significantly in lung, liver, spleen while reduced in heart, kidney compared with the etoposide injection® group after i.v. administration of etoposide-loaded BSA nanosuspensions. The formulation played a role in targeting delivery to lung, reduce toxicity, and side effects of etoposide. In conclusion, etoposide-loaded BSA nanosuspensions were promising for parenteral delivery of etoposide.

  2. Improved activity of immobilized horseradish peroxidase on gold nanoparticles in the presence of bovine serum albumin

    International Nuclear Information System (INIS)

    Ni, Yuyang; Li, Jun; Huang, Zhenzhen; He, Ke; Zhuang, Jiaqi; Yang, Wensheng

    2013-01-01

    The using of macromolecular additives is known to be a simple and effective way to improve the activity of immobilized enzymes on solid support, yet the mechanism has not been well understood. Taking horseradish peroxidase (HRP) as an example, only 30 % of its catalytic activity was kept after being immobilized on the surface of 25-nm Au nanoparticles, mainly attributed to the conformational change of the heme-containing active site. The catalytic activity of HRP was significantly improved to 80 % when a certain amount of bovine serum albumin (BSA) was added at the initial stage of the immobilization. Systematic spectral investigation indicated that the addition of BSA inhibited the tertiary structure change around the active site, which was a prerequisite for improved activity of the immobilized HRP. Steady-state kinetic analyses revealed that the introduction of BSA could effectively improve the turnover rate of substrate to product in spite of slight reduced affinity to substrates, which also contributed to the improved catalytic activity

  3. Bovine serum albumin adsorption to iron-oxide coated sands can change microsphere deposition mechanisms.

    Science.gov (United States)

    Flynn, Raymond M; Yang, Xinyao; Hofmann, Thilo; von der Kammer, Frank

    2012-03-06

    Particulate colloids often occur together with proteins in sewage-impacted water. Using Bovine Serum Albumin (BSA) as a surrogate for protein in sewage, column experiments investigating the capacity of iron-oxide coated sands to remove latex microspheres from water revealed that microsphere attenuation mechanisms depended on antecedent BSA coverage. Dual pulse experiment (DPE) results suggested that where all BSA was adsorbed, subsequent multiple pore volume microsphere breakthrough curves reflected progressively reduced colloid deposition rates with increasing adsorbed BSA content. Modeling colloid responses suggested adsorption of 1 μg BSA generated the same response as blockage by between 7.1 × 10(8) and 2.3 × 10(9) deposited microspheres. By contrast, microsphere responses in DPEs where BSA coverage of the deposition sites approached/reached saturation revealed the coated sand maintained a finite capacity to attenuate microspheres, even when incapable of further BSA adsorption. Subsequent microsphere breakthrough curves demonstrated the matrix's colloid attenuation capacity progressively increased with continued microsphere deposition. Experimental findings suggested BSA adsorption on the sand surface approaching/reaching saturation generated attractive deposition sites for colloids, which became progressively more attractive with further colloid deposition (filter ripening). Results demonstrate that adsorption of a single type of protein may either enhance or inhibit colloid mobility in saturated porous media.

  4. Adsorption of bovine serum albumin on amorphous carbon surfaces studied with dip pen nanolithography

    Science.gov (United States)

    Yadav, Pradeep K.; McKavanagh, Fiona; Maguire, Paul D.; Lemoine, Patrick

    2011-10-01

    This article reports the use of dip pen nanolithography (DPN) for the study of adsorption of bovine serum albumin (BSA) proteins on amorphous carbon surfaces; tetrahedral amorphous carbon (t-aC) and silicon doped hydrogenated amorphous carbon (a-C:H:Si). Contact angle study shows that the BSA proteins reduce the contact angle on both carbon materials. We also noticed that the drop volume dependence is consistent with a negative line tension, i.e. due to an attractive protein/surface interaction. The DPN technique was used to write short-spaced (100 nm) BSA line patterns on both samples. We found a line merging effect, stronger in the case of the a-C:H:Si material. We discuss possible contributions from tip blunting, scratching, cross-talk between lever torsion and bending and nano-shaving of the patterns. We conclude that the observed effect is caused in large measure by the diffusion of BSA proteins on the amorphous carbon surfaces. This interpretation of the result is consistent with the contact angle data and AFM force curve analysis indicating larger tip/surface adhesion and spreading for the a-C:H:Si material. We conclude by discussing the advantages and limitations of DPN lithography to study biomolecular adsorption in nanoscale wetting environments.

  5. Photophysical studies of PET based acridinedione dyes with globular protein: Bovine serum albumin

    International Nuclear Information System (INIS)

    Rajendran, Kumaran; Perumal, Ramamurthy

    2010-01-01

    Interaction of acridinedione dyes with model transport proteins, bovine serum albumin (BSA) in aqueous solution were investigated by fluorescence spectral studies. A fluorescence enhancement was observed on the addition of BSA to photoinduced electron transfer (PET) based acridinedione dyes, which posses C 6 H 4 (p-OCH 3 ) in the 9th position of the basic acridinedione ring. On the contrary, the addition of BSA to non-PET based acridinedione dyes with methyl or phenyl substitution in the 9th position does not result in any fluorescence enhancement. The enhancement in the fluorescence intensity is attributed to the suppression of PET process through space between -OCH 3 group and the acridinedione moiety is elucidated by steady state fluorescence measurements. The fluorescence anisotropy value (r) of 0.40 reveals that the motion of the dye molecule is highly constrained and is largely confined to the rigid microenvironment of the protein molecule. The binding constant (K) was found to be in the order of 6.0x10 3 [M] -1 , which implies the existence of hydrophobic interaction between the PET based dye and BSA. Time resolved fluorescence lifetime measurements reveal that the PET based acridinedione dye preferably binds in the hydrophobic interior of BSA.

  6. Kinetics of adsorption of bovine serum albumin on magnetic carboxymethyl chitosan nanoparticles.

    Science.gov (United States)

    Wang, Zhouli; Yue, Tianli; Yuan, Yahong; Cai, Rui; Niu, Chen; Guo, Caixia

    2013-07-01

    The magnetic carboxymethyl chitosan nanoparticles (MNPs-CMC) were developed as effective magnetic affinity adsorbent for Bovine serum albumin and the adsorption reactions were investigated. The obtained experimental data were compared with the adsorption kinetics models and equilibrium isotherms. The experimental kinetic data were modeled using Pseudo-first order, Pseudo-second order, Bangham's equation, Intra-particle diffusion model and Elovich equations. It was found that the adsorption reactions followed the Pseudo-second order kinetics equation. The experimental isotherm data were analyzed using Langmuir, Freundlich, Dubinin-Radushkevich and Temkin equations. By comparing the correlation coefficients determined for each linear transformation of isotherm analysis, it was found that the Langmuir equation was the best fit equilibrium model for the adsorption of BSA. Error functions have been used to determine the alternative single component parameters by nonlinear regression due to the inherent bias in using the correlation coefficient resulting from linearization. It showed that the Langmuir equation resulted in the lowest values for the error function and thus fitted the data better than the other isotherm. Various thermodynamic parameters such as enthalpy (ΔH°), entropy (ΔS°) and Gibbs free energy (ΔG°) were evaluated. MNPs-CMC nanoaprticles were shown to be a promising material for adsorption of BSA from aqueous solutions. Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

  7. Silkworm (Bombyx mori) hemolymph unable to substitute fetal bovine serum in insect cell culture

    Science.gov (United States)

    Suparto, Irma H.; Khalam, Chandra Nur; Praira, Willy; Sajuthi, Dondin

    2014-03-01

    Fetal Bovine Serum (FBS) in animal cell culture media is an important source of nutrients for cell growth. However, the harvest and collection of FBS cause bioethical concerns. Efforts to reduce and preferably replace FBS with synthetic or other natural alternatives are continually being explored. Hemolymph silkworm (Bombyx mori) contains many nutrients needed for the process of metamorphosis. Therefore, there is possibility as an alternative nutritional supplement for cell culture to reduce the use of FBS. The objective of this study was to evaluate the macrocomponent of hemolymph and the possibility as medium supplement for Spodoptera fugiperda (Sf9) cell culture. Proximate analyses showed that hemolymph contains 89.76% of water, 2.52 mg/mL carbohydrate, 2.35% fat and 55.61 mg/mL protein. Further protein analysis, it consists of 15 fractions containing molecular weight of 22 - 152 kDa. The use of hemolymph as FBS substitution in Sf9 cell culture with various concentrations was unable to maintain and support cell growth. Further research still needed by prior adaptation of the tissue culture to minimal nutrition media before introduction of the hemolymph as supplement.

  8. Tuning the solution organization of cationic polymers through interactions with bovine serum albumin.

    Science.gov (United States)

    Papagiannopoulos, Aristeidis; Vlassi, Eleni; Pispas, Stergios; Jafta, Charl J

    2017-07-19

    The interactions of bovine serum albumin (BSA) with aggregates of cationic polymers, i.e. quaternized poly(chloromethyl styrene) chains (QIm-PCMS), in aqueous solutions are investigated using small angle neutron scattering on length scales relevant to the size of BSA. The arrangement of the macromolecular chains within their aggregates is consistent with a blob description of overlapping chains that contain hydrophobic domains. The local conformations depend on the salt content as in typical linear polyelectrolytes. Although the hydrophobic content of the cationic polymers does not cause measurable local morphology differences, the interactions with BSA are enhanced in the case of the not fully quaternized polymer. The secondary structure of BSA is critically compromised by the interaction with the quaternized polymers as the signature of the alpha helix conformation is lost. The complexation with BSA and the resulting enhancement of interchain associations on higher length scales are verified using dynamic light scattering experiments. This study demonstrates the ability to tune the polyelectrolyte/protein interactions and polyelectrolyte chain-chain associations by modifying the hydrophobic content of the polyelectrolytes.

  9. Binding of carbendazim to bovine serum albumin: Insights from experimental and molecular modeling studies

    Science.gov (United States)

    Li, Jinhua; Zhang, Yulei; Hu, Lin; Kong, Yaling; Jin, Changqing; Xi, Zengzhe

    2017-07-01

    Carbendazim (CBZ) is a widely used benzimidazole fungicide in agriculture to control a wide range of fruit and vegetable pathogens, which may lead to potential health hazards. To evaluate the potential toxicity of CBZ, the binding mechanism of bovine serum albumin (BSA) with CBZ was investigated by the fluorescence quenching technology, UV absorbance spectra, circular dichroism (CD), and molecular modeling. The fluorescence titration and UV absorbance spectra revealed that the fluorescence quenching mechanism of BSA by CBZ was a combined quenching process. In addition, the studies of CD spectra suggested that the binding of CBZ to BSA changed the secondary structure of protein. Furthermore, the thermodynamic functions of enthalpy change (ΔH0) and entropy change (ΔS0) for the reaction were calculated to be 24.87 kJ mol-1 and 162.95 J mol-1 K-1 according to Van't Hoff equation. These data suggested that hydrophobic interaction play a major role in the binding of CBZ to BSA, which was in good agreement with the result of molecular modeling study.

  10. Impact of condensed tannin size as individual and mixed polymers on bovine serum albumin precipitation.

    Science.gov (United States)

    Harbertson, James F; Kilmister, Rachel L; Kelm, Mark A; Downey, Mark O

    2014-10-01

    Condensed tannins composed of epicatechin from monomer to octamer were isolated from cacao (Theobroma cacao, L.) seeds and added to bovine serum albumin (BSA) individually and combined as mixtures. When added to excess BSA the amount of tannin precipitated increased with tannin size. The amount of tannin required to precipitate BSA varied among the polymers with the trimer requiring the most to precipitate BSA (1000 μg) and octamer the least (50 μg). The efficacy of condensed tannins for protein precipitation increased with increased degree of polymerisation (or size) from trimers to octamers (monomers and dimers did not precipitate BSA), while mixtures of two sizes primarily had an additive effect. This study demonstrates that astringent perception is likely to increase with increasing polymer size. Further research to expand our understanding of astringent perception and its correlation with protein precipitation would benefit from sensory analysis of condensed tannins across a range of polymer sizes. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. Investigation of proton pump inhibitors binding with bovine serum albumin and their relationship to molecular structure

    International Nuclear Information System (INIS)

    Zhang Yuping; Shi Shuyun; Peng Mijun

    2012-01-01

    The interactions of three proton pump inhibitors (PPIs), omeprazole, pantoprazole and ilaprazole with bovine serum albumin (BSA) have been investigated by fluorescence, synchronous fluorescence, ultraviolet–visible (UV–vis) and circular dichroism (CD). Various binding parameters have been calculated at various temperatures. The results indicated that omeprazole, pantoprazole and ilaprazole had a strong ability to quench the intrinsic fluorescence of BSA with static quenching mechanism, and the binding affinities were significantly affected by different substituents and polarities as the order ilaprazole>pantoprazole>omeprazole. The site marker competitive experiments indicated that the binding of omeprazole, pantoprazole and ilaprazole to BSA primarily took place in subdomain IIA. The results of thermodynamic parameters ΔG, ΔH and ΔS indicated that electrostatic interaction played a major role for PPIs–BSA association. The distance r between PPIs and BSA was evaluated according to the theory of Förster's energy transfer. The quantitative analysis of synchronous fluorescence and CD spectra showed the change in secondary structure of the BSA upon interaction with PPIs by a reduction of α-helix. All the above results many have relevant insight into the PPIs' availability and distribution. - Highlights: ► The interactions of three PPIs with BSA have been investigated. ► The fluorescence quenching mechanism is static quenching. ► Binding affinities were greatly affected by the substituents and polarities. ► The binding of three PPIs to BSA primarily took place in subdomain IIA.

  12. Investigation on the interactions of clenbuterol to bovine serum albumin and lysozyme by molecular fluorescence technique.

    Science.gov (United States)

    Bi, Shuyun; Pang, Bo; Wang, Tianjiao; Zhao, Tingting; Yu, Wang

    2014-01-01

    Clenbuterol interacting with bovine serum albumin (BSA) or lysozyme (LYS) in physiological buffer (pH 7.4) was investigated by the fluorescence spectroscopy and UV-vis absorption spectroscopy. The results indicated that clenbuterol quenched the intrinsic fluorescence of BSA and LYS via a static quenching procedure. The binding constants of clenbuterol with BSA and LYS were 1.16×10(3) and 1.49×10(3) L mol(-1) at 291 K. The values of ΔH and ΔS implied that hydrophobic and electrostatic interaction played a major role in stabilizing the complex (clenbuterol-BSA or clenbuterol-LYS). In the presence of Fe2+, Fe3+, Cu2+, Mg2+, Ca2+, or Zn2+, the binding constants of clenbuterol to BSA or LYS had no significant differences. The distances between the donor (BSA or LYS) and acceptor (clenbuterol) were 2.61 and 2.19 nm for clenbuterol-BSA and clenbuterol-LYS respectively. Furthermore, synchronous fluorescence spectrometry was used to analyze the conformational changes of BSA and LYS. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. The investigation of the interaction between piracetam and bovine serum albumin by spectroscopic methods

    Science.gov (United States)

    Guo, Xingjia; Han, Xiaowei; Tong, Jian; Guo, Chuang; Yang, Wenfeng; Zhu, Jifen; Fu, Bing

    2010-03-01

    The interaction between piracetam (OPA) with bovine serum albumin (BSA) has been thoroughly studied by fluorescence quenching technique in combination with UV-vis absorption, Fourier transform infrared (FT-IR), and circular dichroism (CD) spectroscopies under the simulative physiological conditions. The quenching of BSA fluorescence by OPA was found to be a static quenching process. The binding constants ( K a) are 3.014, 2.926, and 2.503 × 10 3 M -1 at 292, 298, and 309 K, respectively. According to the van't Hoff equation, the thermodynamic functions standard enthalpy (Δ H) and standard entropy (Δ S) for the reaction were calculated to be -74.560 kJ mol -1 and -159.380 J mol -1 K -1, which indicated that OPA binds to BSA mainly by hydrogen bonds and van der Waals interactions. The binding distance between BSA and OPA was calculated to be 4.10 nm according to the theory of FÖrster's non-radiation energy transfer. The displacement experiments confirmed that OPA could bind to the site I of BSA. Furthermore, the effects of pH and some common ions on the binding constant were also examined. And the alterations of protein secondary structure in the presence of OPA were observed by the CD and FT-IR spectra.

  14. The wettability of a cellulose acetate membrane in the presence of bovine serum albumin

    Science.gov (United States)

    Białopiotrowicz, Tomasz; Jańczuk, Bronisław

    2002-11-01

    The measurements of the contact angle for water (W), glycerol (G), formamide (F), ethylene glycol (E) and diiodomethane (D) on a bare cellulose acetate membrane and covered by adsorptive bovine serum albumin (BSA) films were made. The adsorption was performed from solutions in concentration range 0-100 mg/ml. An influence of the membrane porosity on an apparent contact angle was discussed and Cassie and Baxter equation was used for that purpose. It was suggested that some liquids could penetrate in to membrane pores reducing its apparent porosity. To explain such behaviour, the spreading coefficient and the work of adhesion was calculated for the studied liquids. Components and parameters of the surface free energy of a bare cellulose acetate membrane and covered by an adsorptive BSA film were determined for W-G-D, W-F-D and G-F-D three-liquid systems and they were similar for these systems. However, for the hydrated BSA layer those components and parameters for the systems W-G-D, W-F-D were different than those for the system G-F-D. It was stated that after BSA adsorption on that membrane percentage of empty pores decreased, reducing their number almost to 0, at the highest BSA concentrations.

  15. Characterization of chemically modified chitosan microspheres as adsorbents using standard Proteins (bovine serum albumin and lysozyme

    Directory of Open Access Journals (Sweden)

    M. A. Torres

    2007-09-01

    Full Text Available Chitosan microspheres with a mean size of 140 ± 119 µm were produced by the spray and coagulation methods. The microspheres were chemically modified using the following routes: a crosslinking with glutaraldehyde b crosslinking with epychlorohydrin and c acetylation. For investigation of their ability as adsorbents, the following standard proteins were chosen as adsorbates: bovine serum albumin - BSA (pI = 4.8 and MW = 66 kDa and lysozyme (pI = 11 and MW = 14 kDa. The adsorption experiments were performed using a static method. The adsorption media and equilibrium concentration of adsorbates were varied in the ranges of pH 4-11 and 0.07-0.70 mg.ml-1, respectively. The maximum adsorption capacities (q m and the constant of the Langmuir model (Ks were shown to be dependent on charge interactions and on the kind of treatment performed on chitosan microspheres. The satisfactory fit of a kinetic model to the experimental data shows that the step that controls the adsorption kinetics is probably the initial adsorbate transport.

  16. The investigation of the interaction between edaravone and bovine serum albumin by spectroscopic approaches

    Energy Technology Data Exchange (ETDEWEB)

    Yu Xianyong; Yang Ying; Liu Ronghua; Huang Haowen; Chen Jian; Ji Danhong [Key Laboratory of Theoretical Chemistry and Molecular Simulation of Ministry of Education, Hunan Province College Key Laboratory of QSAR/QSPR, School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan 411201 (China); Li Xiaofang, E-mail: fine_chem@163.co [Key Laboratory of Theoretical Chemistry and Molecular Simulation of Ministry of Education, Hunan Province College Key Laboratory of QSAR/QSPR, School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan 411201 (China); Yang Fengxian [Key Laboratory of Theoretical Chemistry and Molecular Simulation of Ministry of Education, Hunan Province College Key Laboratory of QSAR/QSPR, School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan 411201 (China); Yi Pinggui, E-mail: pgyi@hnust.c [Key Laboratory of Theoretical Chemistry and Molecular Simulation of Ministry of Education, Hunan Province College Key Laboratory of QSAR/QSPR, School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan 411201 (China)

    2011-07-15

    The fluorescence and ultraviolet spectroscopies were explored to study the interaction between edaravone (EDA) and bovine serum albumin (BSA) under imitated physiological condition. The experimental results show that the fluorescence quenching mechanism between EDA and BSA is a combined quenching (dynamic and static quenching). The binding constants, binding sites, and the corresponding thermodynamic parameters ({Delta}G, {Delta}H, and {Delta}S) of the interaction system were calculated at different temperatures. According to Foerster non-radiation energy transfer theory, the binding distance between EDA and BSA was calculated to be 3.10 nm. The effect of EDA on the conformation of BSA was analyzed using synchronous fluorescence spectroscopy. In addition, the effects of some common metal ions Mg{sup 2+}, Ca{sup 2+}, Cu{sup 2+}, and Ni{sup 2+} on the binding constant between EDA and BSA were examined. - Highlights: {yields} We explored the interaction of BSA and EDA using spectroscopic methods. {yields} The fluorescence quenching mechanism is combined quenching. {yields} Hydrophobic interaction force plays a major role in stabilizing the complex. {yields} The binding constants, binding sites, and thermodynamic parameters were calculated. {yields} EDA affects the conformation of tryptophan residue's microregion.

  17. Study on the interaction between promethazine hydrochloride and bovine serum albumin by fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    He Lingling [Department of Chemistry, Northeastern University, Shenyang 110004 (China); College of Applied Chemistry, Shenyang University of Chemical Technology, Shenyang 110142 (China); Wang Xin [School of Pharmaceutical Sciences, Liaoning University, Shenyang 110036 (China); Liu Bin [Department of Chemistry, Northeastern University, Shenyang 110004 (China); School of Pharmaceutical Sciences, Liaoning University, Shenyang 110036 (China); Wang Jun [School of Pharmaceutical Sciences, Liaoning University, Shenyang 110036 (China); Sun Yaguang; Gao Enjun [College of Applied Chemistry, Shenyang University of Chemical Technology, Shenyang 110142 (China); Xu Shukun, E-mail: xushukun46@126.co [Department of Chemistry, Northeastern University, Shenyang 110004 (China)

    2011-02-15

    The interaction between promethazine hydrochloride (PMT) and bovine serum albumin (BSA) in vitro was investigated by means of fluorescence spectroscopy and absorption spectroscopy. The fluorescence of BSA was quenched remarkably by PMT and the quenching mechanism was considered as static quenching by forming a complex. The association constants K{sub a} and the number of binding sites n were calculated at different temperatures. The BSA-PMT binding distance was determined to be less than 8 nm, suggesting that energy transfer from BSA to PMT may occur. The thermodynamic parameters of the interaction between PMT and BSA were measured according to the van't Hoff equation. The enthalpy change ({Delta}H) and entropy change ({Delta}S) were calculated to be -23.62 kJ mol{sup -1} and -0.10 J mol{sup -1} K{sup -1}, respectively, which indicated that the interaction of PMT with BSA was driven mainly by van der Waals forces and hydrogen bonds. The binding process was a spontaneous process in which Gibbs free energy change ({Delta}G) was negative. In addition, the results of synchronous fluorescence spectra and three-dimensional fluorescence spectra showed that binding of PMT with BSA can induce conformational changes in BSA.

  18. Multi-spectroscopic characterization of bovine serum albumin upon interaction with atomoxetine

    Directory of Open Access Journals (Sweden)

    Arunkumar T. Buddanavar

    2017-06-01

    Full Text Available The quenching interaction of atomoxetine (ATX with bovine serum albumin (BSA was studied in vitro under optimal physiological condition (pH=7.4 by multi-spectroscopic techniques. The mechanism of ATX-BSA system was a dynamic quenching process and was confirmed by the fluorescence spectra and lifetime measurements. The number of binding sites, binding constants and other binding characteristics were computed. Thermodynamic parameters ∆H° and ∆S° indicated that intermolecular hydrophobic forces predominantly stabilized the drug-protein system. The average binding distance between BSA and ATX was studied by Försters theory. UV-absorption, Fourier transform infrared spectroscopy (FT-IR, circular dichroism (CD, synchronous spectra and three-dimensional (3D fluorescence spectral results revealed the changes in micro-environment of secondary structure of protein upon the interaction with ATX. Displacement of site probes and the effects of some common metal ions on the binding of ATX with BSA interaction were also studied.

  19. Structural Relationship and Binding Mechanisms of Five Flavonoids with Bovine Serum Albumin

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    Ping Li

    2010-12-01

    Full Text Available Flavonoids are structurally diverse and the most ubiquitous groups of dietary polyphenols distributed in various fruits and vegetables. In this study, the interaction between five flavonoids, namely formononetin-7-O-β-D-glucoside, calycosin- 7-O-β-D-glucoside, calycosin, rutin, and quercetin, and bovine serum albumin (BSA was investigated by fluorescence and UV-vis absorbance spectroscopy. In the discussion, it was proved that the fluorescence quenching of BSA by flavonoids was a result of the formation of a flavonoid-BSA complex. Fluorescence quenching constants were determined using the Stern-Volmer and Lineweaver-Burk equations to provide a measure of the binding affinity between the flavonoids and BSA. The binding constants ranked in the order quercetin > rutin > calycosin > calycosin-7-O-β-D-glucoside ≈ formononetin-7-O-β-D-glucoside. The results of thermodynamic parameters ΔG, ΔH, and ΔS at different temperatures indicated that the hydrophobic interaction played a major role in flavonoid-BSA association. The distance r between BSA and acceptor flavonoids was also obtained according to Förster’s theory of non-radiative energy transfer.

  20. Bovine Serum Albumin-Glutaraldehyde Sealed Fish-Mouth Closure of the Pancreatic Remnant during Distal Pancreatectomy

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    Fritz Klein

    2017-01-01

    Full Text Available Introduction. Postoperative pancreatic fistula formation remains the major complication after distal pancreatectomy. At our institution, we have recently developed a novel bovine serum albumin-glutaraldehyde sealed hand sutured fish-mouth closure technique of the pancreatic remnant during distal pancreatectomy. The aim of this study was to analyze the impact of this approach with regard to technical feasibility and overall postoperative outcome. Patients and Methods. 32 patients who underwent a bovine serum albumin-glutaraldehyde sealed hand sutured fish-mouth closure of the pancreatic remnant during distal pancreatectomy between 2012 and 2014 at our institution were analyzed for clinically relevant postoperative pancreatic fistula formation (Grades B and C according to ISGPF definition and overall postoperative morbidity. Results. Three out of 32 patients (9.4% developed Grade B pancreatic fistula, which could be treated conservatively. No Grade C pancreatic fistulas were observed. Postpancreatectomy hemorrhage occurred in 1 patient (3.1%. Overall postoperative complications > Clavien II were observed in 5 patients (15.6%. There was no postoperative mortality. Conclusion. The performance of a bovine serum albumin-glutaraldehyde sealed hand sutured fish-mouth closure of the pancreatic remnant was shown to be technically feasible and may lead to a significant decrease of postoperative pancreatic fistula formation after distal pancreatectomy.

  1. Plating Efficiency for Primary Hamster Embryo Cells as an Index of Efficacy of Fetal Bovine Serum for Cell Culture

    Science.gov (United States)

    Goodheart, C. R.; Casto, B. C.; Zwiers, A.; Regnier, P. R.

    1973-01-01

    Attachment and growth of mammalian cells plated at low cell density require optimum conditions for the cells to form colonies. Reliability, reproducibility, and validity of the plating efficiency test for evaluating cell culture sera were determined by measuring the plating efficiency of 37 lots of fetal bovine serum obtained from 8 suppliers (5 lots from each of 7, 2 lots from 1 supplier), by using hamster embryo fibroblasts plated at low cell density. The test revealed considerable variation between lots of serum and between suppliers. The five lots from some suppliers had consistently high plating efficiencies, whereas one or more lots from other suppliers had quite low efficiencies. The results were reproducible in repeated tests, and control experiments indicated that the test measured the efficiency of the test serum independently of the efficiency of the serum used for the primary outgrowth of the hamster embryo cells. PMID:4584591

  2. Interaction of Flavonoids from Woodwardia unigemmata with Bovine Serum Albumin (BSA): Application of Spectroscopic Techniques and Molecular Modeling Methods.

    Science.gov (United States)

    Ma, Rui; Pan, Hong; Shen, Tao; Li, Peng; Chen, Yanan; Li, Zhenyu; Di, Xiaxia; Wang, Shuqi

    2017-08-09

    Phytochemical investigation on the methanol extract of Woodwardia unigemmata resulted in the isolation of seven flavonoids, including one new flavonol acylglycoside ( 1 ). The structures of these compounds were elucidated on the basis of extensive spectroscopic analysis and comparison of literature data. The multidrug resistance (MDR) reversing activity was evaluated for the isolated compounds using doxorubicin-resistant K562/A02 cells model. Compound 6 showed comparable MDR reversing effect to verapamil. Furthermore, the interaction between compounds and bovine serum albumin (BSA) was investigated by spectroscopic methods, including steady-state fluorescence, synchronous fluorescence, circular dichroism (CD) spectroscopies, and molecular docking approach. The experimental results indicated that the seven flavonoids bind to BSA by static quenching mechanisms. The negative ΔH and ΔS values indicated that van der Waals interactions and hydrogen bonds contributed in the binding of compounds 2 - 6 to BSA. In the case of compounds 1 and 7 systems, the hydrophobic interactions play a major role. The binding of compounds to BSA causes slight changes in the secondary structure of BSA. There are two binding sites of compound 6 on BSA and site I is the main site according to the molecular docking studies and the site marker competitive binding assay.

  3. Attenuation of Colitis by Serum-Derived Bovine Immunoglobulin/Protein Isolate in a Defined Microbiota Mouse Model.

    Science.gov (United States)

    Henderson, Abigail L; Brand, Meghan Wymore; Darling, Ross J; Maas, Kenneth J; Detzel, Christopher J; Hostetter, Jesse; Wannemuehler, Michael J; Weaver, Eric M

    2015-11-01

    The pathogenesis of inflammatory bowel disease (IBD) is complex and multifaceted including genetic predisposition, environmental components, microbial dysbiosis, and inappropriate immune activation to microbial components. Pathogenic bacterial provocateurs like adherent and invasive E. coli have been reported to increase susceptibility to Crohn's disease. Serum-derived bovine immunoglobulin/protein isolate (SBI) is comprised primarily of immunoglobulins (Igs) that bind to conserved microbial components and neutralize exotoxins. To demonstrate that oral administration of SBI may modulate mucosal inflammation following colonization with E. coli, LF82, and exposure to dextran sodium sulfate (DSS). Defined microbiota mice harboring the altered Schaedler flora (ASF) were administered SBI or hydrolyzed collagen twice daily starting 7 days prior to challenge with E. coli LF82 and continuing for the remainder of the experiment. Mice were treated with DSS for 7 days and then evaluated for evidence of local and peripheral inflammation. Igs within SBI bound multiple antigens from all eight members of the ASF and E. coli LF82 by western blot analysis. Multiple parameters of LF82/DSS-induced colitis were reduced following administration of SBI, including histological lesion scores, secretion of cytokines and chemokines from cecal biopsies, intestinal fatty acid binding protein (I-FABP) and serum amyloid A from plasma. Oral administration of SBI attenuated clinical signs of LF82/DSS-induced colitis in mice. The data are consistent with the hypothesis that SBI immunoglobulin binding of bacterial antigens in the intestinal lumen may inhibit the inflammatory cascades that contribute to IBD, thus attenuating DSS-induced colitis.

  4. Importance of exosome depletion protocols to eliminate functional and RNA-containing extracellular vesicles from fetal bovine serum

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    Ganesh Vilas Shelke

    2014-09-01

    Full Text Available Extracellular vesicles (EVs, including the nano-sized exosomes, have the capacity to transfer multiple functional molecules between cells. In cell culture experiments, fetal bovine serum (FBS is often used to supplement cell culture medium as a nutrient, but it is important to know that the FBS also contain significant quantities of EVs. The aim of the current study was to determine whether the FBS EVs can influence cultured cell phenotype, and secondly to determine the efficiency of FBS-EV elimination protocols. Firstly, FBS that had not been depleted of EVs induced a migratory phenotype in a lung cancer epithelial cell line (A549 cells, an effect that could be mimicked by isolated FBS EVs alone. FBS-derived EVs also contained RNA, which was protected from consecutive proteinase K and RNase A treatment. Comparison of common isolation protocols suggested that an 18-hour centrifugation period eliminates approximately 95% of RNA-containing FBS EVs, whereas a 1.5-hour protocol is insufficient. In conclusion, this study shows that FBS EVs substantially influence cultured cell behaviour, but also that they can be virtually removed by an 18-hour ultracentrifugation protocol.

  5. Effects of Oxidized Glutathione, Bovine Serum Albumin, Cysteine and Lycopene on the Quality of Frozen-Thawed Ram Semen

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    O. Uysal

    2007-01-01

    Full Text Available Free radicals are known to be involved in lipid peroxidation as well as DNA and sperm membrane damages that may lead to decreased sperm motility or cell death. The balance between free radical production and their detoxification may be an important factor in sperm survival and function before, during and after cryopreservation. The aim of this study was to determine the effects of the addition of the antioxidants of oxidized glutathione (GSSG, bovine serum albumin (BSA, cysteine and lycopene to freezing media on the post-thawing sperm characteristics, including motility, morphology, acrosome integrity, viability and membrane integrity. A total number of 42 ejaculates were collected using the artifi cial vagina from 4 Akkaraman rams and 10 replicates of the ejaculates were diluted with a Tris-based extender containing additives and no additives as control. GSSG (5 mM, BSA (20 mg/ml, cysteine (10 mM and lycopene (800 μg showed more positive effects than other concentrations of the supplements and controls in protecting sperm characteristics after the freezing-thawing process (P < 0.001. Many aspects of sperm protection, e.g. sperm motility, viability and membrane stabilisation of the sperm cells during relative cryopreservation, are the key factors in determining the preservation of sperm function. The results of this study provide a new approach to the cryopreservation of sperm from rams and related breeds, and thereby contribute to the improvement of these breeds for the world sheep industry.

  6. Effect of gamma-irradiation of bovine serum albumin solution on the formation of zigzag film textures

    Science.gov (United States)

    Glibitskiy, Dmitriy M.; Gorobchenko, Olga A.; Nikolov, Oleg T.; Cheipesh, Tatiana A.; Roshal, Alexander D.; Zibarov, Artem M.; Shestopalova, Anna V.; Semenov, Mikhail A.; Glibitskiy, Gennadiy M.

    2018-03-01

    Formation of patterns on the surface of dried films of saline biopolymer solutions is influenced by many factors, including particle size and structure. Proteins may be modified under the influence of ionizing radiation. By irradiating protein solutions with gamma rays, it is possible to affect the formation of zigzag (Z) structures on the film surface. In our study, the films were obtained by desiccation of bovine serum albumin (BSA) solutions, which were irradiated by a 60Co gamma-source at doses ranging from 1 Gy to 12 kGy. The analysis of the resulting textures on the surface of the films was carried out by calculating the specific length of Z-structures. The results are compared against the absorption and fluorescence spectroscopy and dynamic light scattering (DLS) data. Gamma-irradiation of BSA solutions in the 1-200 Gy range practically does not influence the amount of Z-structures on the film surface. The decrease in fluorescence intensity and increase in absorbance intensity point to the destruction of BSA structure at 2 and 12 kGy, and DLS shows a more than 160% increase in particle size as a result of BSA aggregation at 2 kGy. This prevents the formation of Z-structures, which is reflected in the decrease of their specific length.

  7. Spectroscopic study on the sonodynamic and sonocatalytic damage of anthraquinone derivants to bovine serum albumin under ultrasonic irradiation

    International Nuclear Information System (INIS)

    Wang Zhiqiu; Gao Jingqun; Wang Jun; Li Ying; Li Kai; Kang Pingli; Zhang Xiangdong

    2012-01-01

    In this work, three anthraquinone derivants (Alizarin: 1,2-dihydroxy-9, 10-anthraquinone, Alizarin–DA: 1,2-dihydroxy-9, 10-anthraquinone-3-aminomethyl-N, N-diacetic acid and Alizarin–DA–Fe: 1,2-dihydroxy-9, 10-anthraquinone-3-aminomethyl-N, N-diacetate-Ferrous(III)) were used to study the sonodynamic and sonocatalytic damage of bovine serum albumin (BSA) molecules according to the hyperchromic effect of UV–vis spectra and quenching effect of intrinsic fluorescence. Meanwhile, some influencing factors such as ultrasonic irradiation time, anthraquinone derivants concentration and ionic strength on the damage of BSA molecules were also considered. The results show that the synergetic effect of anthraquinone derivants and ultrasonic irradiation can efficiently damage the BSA molecules. Finally, some special radical scavengers were used to determine the kind of generated reactive oxygen species (ROS) in the presence of three anthraquinone derivants under ultrasonic irradiation. The results show that the ROS, at least, including singlet oxygen ( 1 O 2 ) and hydroxyl radicals (OH) are generated during the sonodynamic and sonocatalytic processes. It is wished that this paper could offer some valuable references for the application of anthraquinone derivants in sonodynamic therapy (SDT) and sonocatalytic therapy (SCT) for tumor treatment. - Highlights: ► Anthraquinone derivants were used to study the sonodynamic and sonocatalytic damage to BSA. ► The generations of ROS during sonodynamic and sonocatalytic process were estimated. ► Some quenchers were used to determine the kind of the ROS.

  8. Molecular Structure-Affinity Relationship of Flavonoids in Lotus Leaf (Nelumbo nucifera Gaertn.) on Binding to Human Serum Albumin and Bovine Serum Albumin by Spectroscopic Method.

    Science.gov (United States)

    Tang, Xiaosheng; Tang, Ping; Liu, Liangliang

    2017-06-23

    Lotus leaf has gained growing popularity as an ingredient in herbal formulations due to its various activities. As main functional components of lotus leaf, the difference in structure of flavonoids affected their binding properties and activities. In this paper, the existence of 11 flavonoids in lotus leaf extract was confirmed by High Performance Liquid Chromatography (HPLC) analysis and 11 flavonoids showed various contents in lotus leaf. The interactions between lotus leaf extract and two kinds of serum albumins (human serum albumin (HSA) and bovine serum albumin (BSA)) were investigated by spectroscopic methods. Based on the fluorescence quenching, the interactions between these flavonoids and serum albumins were further checked in detail. The relationship between the molecular properties of flavonoids and their affinities for serum albumins were analyzed and compared. The hydroxylation on 3 and 3' position increased the affinities for serum albumins. Moreover, both of the methylation on 3' position of quercetin and the C₂=C₃ double bond of apigenin and quercetin decreased the affinities for HSA and BSA. The glycosylation lowered the affinities for HSA and BSA depending on the type of sugar moiety. It revealed that the hydrogen bond force played an important role in binding flavonoids to HSA and BSA.

  9. Spectroscopic investigation of the interaction of water-soluble azocalix[4]arenes with bovine serum albumin.

    Science.gov (United States)

    Fan, Ping; Wan, Lu; Shang, Yunshan; Wang, Jun; Liu, Yulong; Sun, Xiaoyu; Chen, Chen

    2015-02-01

    In this work, three hydrosoluble azocalix[4]arene derivatives, 5-(o-methylphenylazo)-25,26,27-tris(carboxymethoxy)-28-hydroxycalix[4]arene (o-MAC-Calix), 5-(m-methylphenylazo)-25,26,27-tris(carboxymethoxy)-28-hydroxycalix[4]arene (m-MAC-Calix) and 5-(p-methylphenylazo)-25,26,27-tris(carboxymethoxy)-28-hydroxycalix[4]arene (p-MAC-Calix) were synthesized. Their structures were characterized by infrared spectrum (IR), nuclear magnetic resonance spectrum (1H NMR and 13C NMR) and mass spectrum (MS). The interactions between these compounds and bovine serum albumin (BSA) were studied by fluorescence spectroscopy, UV-vis spectrophotometry and circular dichroic spectroscopy. According to experimental results, three azocalix[4]arene derivatives can efficiently bind to BSA molecules and the o-MAC-Calix displays more efficient interactions with BSA molecules than m-MAC-Calix and p-MAC-Calix. Molecular docking showed that the o-MAC-Calix was embedded in the hydrophobic cavity of helical structure of BSA molecular and the tryptophan (Trp) residue of BSA molecular had strong interaction with o-MAC-Calix. The fluorescence quenching of BSA caused by azocalix[4]arene derivatives is attributed to the static quenching process. In addition, the synchronous fluorescence spectroscopy indicates that these azocalix[4]arene derivatives are more accessible to Trp residues of BSA molecules than the tyrosine (Tyr) residues. The circular dichroic spectroscopy further verified the binding of azocalix[4]arene derivatives and BSA. Copyright © 2014 Elsevier Inc. All rights reserved.

  10. Spectroscopic Study on the Interaction between Naphthalimide-Polyamine Conjugates and Bovine Serum Albumin (BSA

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    Zhi-Yong Tian

    2015-09-01

    Full Text Available The effect of a naphthalimide pharmacophore coupled with diverse substituents on the interaction between naphthalimide-polyamine conjugates 1–4 and bovine serum albumin (BSA was studied by UV absorption, fluorescence and circular dichroism (CD spectroscopy under physiological conditions (pH = 7.4. The observed spectral quenching of BSA by the compounds indicated that they could bind to BSA. Furthermore, caloric fluorescent tests revealed that the quenching mechanisms of compounds 1–3 were basically static type, but that of compound 4 was closer to a classical type. The Ksv values at room temperature for compound-BSA complexes-1-BSA, 2-BSA, 3-BSA and 4-BSA were 1.438 × 104, 3.190 × 104, 5.700 × 104 and 4.745 × 105, respectively, compared with the value of MINS, 2.863 × 104 at Ex = 280 nm. The obtained quenching constant, binding constant and thermodynamic parameter suggested that the binding between compounds 1–4 with BSA protein, significantly affected by the substituted groups on the naphthalene backbone, was formed by hydrogen bonds, and other principle forces mainly consisting of charged and hydrophobic interactions. Based on results from the analysis of synchronous three-dimensional fluorescence and CD spectra, we can conclude that the interaction between compounds 1–4 and BSA protein has little impact on the BSA conformation. Calculated results obtained from in silico molecular simulation showed that compound 1 did not prefer either enzymatic drug sites I or II over the other. However, DSII in BSA was more beneficial than DSI for the binding between compounds 2–4 and BSA protein. The binding between compounds 1–3 and BSA was hydrophobic in nature, compared with the electrostatic interaction between compound 4 and BSA.

  11. Induction of axial chirality in divanillin by interaction with bovine serum albumin.

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    Diego Venturini

    Full Text Available Vanillin is a plant secondary metabolite and has numerous beneficial health applications. Divanillin is the homodimer of vanillin and used as a taste enhancer compound and also a promissory anticancer drug. Here, divanillin was synthesized and studied in the context of its interaction with bovine serum albumin (BSA. We found that divanillin acquires axial chirality when complexed with BSA. This chiroptical property was demonstrated by a strong induced circular dichroism (ICD signal. In agreement with this finding, the association constant between BSA and divanillin (3.3 x 105 mol-1L was higher compared to its precursor vanillin (7.3 x 104 mol-1L. The ICD signal was used for evaluation of the association constant, demonstration of the reversibility of the interaction and determination of the binding site, revealing that divanillin has preference for Sudlow's site I in BSA. This property was confirmed by displacement of the fluorescent markers warfarin (site I and dansyl-L-proline (site II. Molecular docking simulation confirmed the higher affinity of divanillin to site I. The highest scored conformation obtained by docking (dihedral angle 242° was used for calculation of the circular dichroism spectrum of divanillin using Time-Dependent Density Functional Theory (TDDFT. The theoretical spectrum showed good similarity with the experimental ICD. In summary, we have demonstrated that by interacting with the chiral cavities in BSA, divanillin became a atropos biphenyl, i.e., the free rotation around the single bound that links the aromatic rings was impeded. This phenomenon can be explained considering the interactions of divanillin with amino acid residues in the binding site of the protein. This chiroptical property can be very useful for studying the effects of divanillin in biological systems. Considering the potential pharmacological application of divanillin, these findings will be helpful for researchers interested in the pharmacological

  12. Protections of bovine serum albumin protein from damage on functionalized graphene-based electrodes by flavonoids.

    Science.gov (United States)

    Sun, Bolu; Gou, Yuqiang; Xue, Zhiyuan; Zheng, Xiaoping; Ma, Yuling; Hu, Fangdi; Zhao, Wanghong

    2016-05-01

    A sensitive electrochemical sensor based on bovine serum albumin (BSA)/poly (diallyldimethylammonium chloride) (PDDA) functionalized graphene nanosheets (PDDA-G) composite film modified glassy carbon electrode (BSA/PDDA-G/GCE) had been developed to investigate the oxidative protein damage and protections of protein from damage by flavonoids. The performance of this sensor was remarkably improved due to excellent electrical conductivity, strong adsorptive ability, and large effective surface area of PDDA-G. The BSA/PDDA-G/GCE displayed the greatest degree of BSA oxidation damage at 40 min incubation time and in the pH 5.0 Fenton reagent system (12.5 mM FeSO4, 50 mM H2O2). The antioxidant activities of four flavonoids had been compared by fabricated sensor based on the relative peak current ratio of SWV, because flavonoids prevented BSA damage caused by Fenton reagent and affected the BSA signal in a solution containing Co(bpy)3(3+). The sensor was characterized by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM). UV-vis spectrophotometry and FTIR were also used to investigate the generation of hydroxyl radical and BSA damage, respectively. On the basis of results from electrochemical methods, the order of the antioxidant activities of flavonoids is as follows: (+)-catechin>kaempferol>apigenin>naringenin. A novel, direct SWV analytical method for detection of BSA damage and assessment of the antioxidant activities of four flavonoids was developed and this electrochemical method provided a simple, inexpensive and rapid detection of BSA damage and evaluation of the antioxidant activities of samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Serum-derived bovine immunoglobulin/protein isolate in the alleviation of chemotherapy-induced mucositis.

    Science.gov (United States)

    Bateman, Emma; Weaver, Eric; Klein, Gerald; Wignall, Anthony; Wozniak, Belinda; Plews, Erin; Mayo, Bronwen; White, Imogen; Keefe, Dorothy

    2016-01-01

    Gastrointestinal (GI) mucositis caused by chemotherapy is associated with diarrhoea and intestinal barrier disruption caused by apoptosis, immune dysfunction and microbiome alterations. Serum-derived bovine immunoglobulin/protein isolate (SBI) has been shown to manage HIV-associated enteropathy and irritable bowel syndrome with diarrhoea (IBS-D). We investigated in a rat model whether SBI was effective in alleviating symptoms of irinotecan-induced GI mucositis. Animals were gavaged with 250 or 500 mg/kg of SBI twice daily for 4 days, before intraperitoneal administration of 200 mg/kg irinotecan. Twice daily gavaging of SBI continued for 6 days post-irinotecan. Animals were monitored for bodyweight changes and incidence of diarrhoea and clinical symptoms of stress. Tissues and blood samples were collected at necropsy 6 h, and 2, 4 and 6 days post-irinotecan. H&E-stained colon and jejunum were analysed for histological damage. The overall incidence, severity and duration of diarrhoea, and clinical symptoms of mucositis were decreased in irinotecan-treated animals that had received SBI. Animals receiving 500 mg/kg SBI also tended to lose less bodyweight than animals treated only with irinotecan (P > 0.10). SBI-gavaged animals had less pronounced irinotecan-induced changes in neutrophil (P = 0.04959) and lymphocyte (P = 0.0035) levels, and lower tissue damage scores than those receiving irinotecan alone (P < 0.0001). Twice daily oral gavage of SBI was well-tolerated and reduced the incidence, severity and duration of irinotecan-induced mucositis. SBI was associated with less pronounced changes in inflammatory cell levels and tissue damage to colon and jejunum. Ongoing experiments aim to investigate the mechanisms of SBI-associated gastrointestinal protection.

  14. Sodium alginate microneedle arrays mediate the transdermal delivery of bovine serum albumin.

    Directory of Open Access Journals (Sweden)

    Yusuf K Demir

    Full Text Available BACKGROUND: The "poke and release" strategy for the delivery of macromolecules using polymeric microneedle (MN is of great importance because it eliminates microneedle reuse, the risks of biohazardous sharps and cross contamination, and it requires no special disposal mechanism. The main objective of this study was the determination of the stability and delivery of bovine serum albumin (BSA that was transported across human skin via sodium alginate (SA microneedle arrays (MNs and SA needle free patches using two different analytical methods. METHODOLOGY AND FINDINGS: The capability of two analytical methods, the bicinchoninic acid (BCA assay and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE, to precisely detect and quantify BSA within different types of polymeric MNs was assessed. The ex vivo protein release of BSA across dermatomed human abdominal skin from 10 w/w SA MNs was compared to that from needle-free patches using Franz diffusion cells. The developed applicator was mechanically characterized using a Texture Analyzer. The patch mold and its components were fabricated using a rapid prototyping machine. CONCLUSIONS/SIGNIFICANCE: The BCA method was able to precisely detect BSA that had been loaded into SA MNs. However, the use of SDS-PAGE as the analytical method resulted in significantly different amounts of BSA recovered from differently conditioned polymeric MNs. The permeation of BSA across dermatomed human abdominal skin by SA MNs, which were composed of 100 pyramidal needles, increased by approximately 15.4 fold compared to the permeation obtained with SA needle-free patches. The ease of use of the applicator during the release studies was also demonstrated, as was its mechanical characterization.

  15. Effect of high hydrostatic pressure on the enzymatic hydrolysis of bovine serum albumin.

    Science.gov (United States)

    De Maria, Serena; Ferrari, Giovanna; Maresca, Paola

    2017-08-01

    The extent of enzymatic proteolysis mainly depends on accessibility of the peptide bonds, which stabilize the protein structure. The high hydrostatic pressure (HHP) process is able to induce, at certain operating conditions, protein displacement, thus suggesting that this technology can be used to modify protein resistance to the enzymatic attack. This work aims at investigating the mechanism of enzymatic hydrolysis assisted by HHP performed under different processing conditions (pressure level, treatment time). Bovine serum albumin was selected for the experiments, solubilized in sodium phosphate buffer (25 mg mL -1 , pH 7.5) with α-chymotrypsin or trypsin (E/S ratio = 1/10) and HPP treatment (100-500 MPa, 15-25 min). HHP treatment enhanced the extent of the hydrolysis reaction of globular proteins, being more effective than conventional hydrolysis. At HHP treatment conditions maximizing the protein unfolding, the hydrolysis degree of proteins was increased as a consequence of the increased exposure of peptide bonds to the attack of proteolytic enzymes. The maximum hydrolysis degree (10% and 7% respectively for the samples hydrolyzed with α-chymotrypsin and trypsin) was observed for the samples processed at 400 MPa for 25 min. At pressure levels higher than 400 MPa the formation of aggregates was likely to occur; thus the degree of hydrolysis decreased. Protein unfolding represents the key factor controlling the efficiency of HHP-assisted hydrolysis treatments. The peptide produced under high pressure showed lower dimensions and a different structure with respect to those of the hydrolysates obtained when the hydrolysis was carried out at atmospheric pressure, thus opening new frontiers of application in food science and nutrition. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  16. Selective inhibition of aggregation/fibrillation of bovine serum albumin by osmolytes: Mechanistic and energetics insights.

    Directory of Open Access Journals (Sweden)

    Moumita Dasgupta

    Full Text Available Bovine serum albumin (BSA is an important transport protein of the blood and its aggregation/fibrillation would adversely affect its transport ability leading to metabolic disorder. Therefore, understanding the mechanism of fibrillation/aggregation of BSA and design of suitable inhibitor molecules for stabilizing its native conformation, are of utmost importance. The qualitative and quantitative aspects of the effect of osmolytes (proline, hydroxyproline, glycine betaine, sarcosine and sorbitol on heat induced aggregation/fibrillation of BSA at physiological pH (pH 7.4 have been studied employing a combination of fluorescence spectroscopy, Rayleigh scattering, isothermal titration calorimetry (ITC, dynamic light scattering (DLS and transmission electron microscopy (TEM. Formation of fibrils by BSA under the given conditions was confirmed from increase in fluorescence emission intensities of Thioflavin T over a time period of 600 minutes and TEM images. Absence of change in fluorescence emission intensities of 8-Anilinonaphthalene-1-sulfonic acid (ANS in presence of native and aggregated BSA signify the absence of any amorphous aggregates. ITC results have provided important insights on the energetics of interaction of these osmolytes with different stages of the fibrillar aggregates of BSA, thereby suggesting the possible modes/mechanism of inhibition of BSA fibrillation by these osmolytes. The heats of interaction of the osmolytes with different stages of fibrillation of BSA do not follow a trend, suggesting that the interactions of stages of BSA aggregates are osmolyte specific. Among the osmolytes used here, we found glycine betaine to be supporting and promoting the aggregation process while hydroxyproline to be maximally efficient in suppressing the fibrillation process of BSA, followed by sorbitol, sarcosine and proline in the following order of their decreasing potency: Hydroxyproline> Sorbitol> Sarcosine> Proline> Glycine betaine.

  17. Spectroscopic Study on the Interaction between Naphthalimide-Polyamine Conjugates and Bovine Serum Albumin (BSA).

    Science.gov (United States)

    Tian, Zhi-Yong; Song, Li-Na; Zhao, Yuan; Zang, Feng-Lei; Zhao, Zhong-Hua; Chen, Nan-Hao; Xu, Xue-Jun; Wang, Chao-Jie

    2015-09-11

    The effect of a naphthalimide pharmacophore coupled with diverse substituents on the interaction between naphthalimide-polyamine conjugates 1-4 and bovine serum albumin (BSA) was studied by UV absorption, fluorescence and circular dichroism (CD) spectroscopy under physiological conditions (pH = 7.4). The observed spectral quenching of BSA by the compounds indicated that they could bind to BSA. Furthermore, caloric fluorescent tests revealed that the quenching mechanisms of compounds 1-3 were basically static type, but that of compound 4 was closer to a classical type. The Ksv values at room temperature for compound-BSA complexes-1-BSA, 2-BSA, 3-BSA and 4-BSA were 1.438 × 10⁴, 3.190 × 10⁴, 5.700 × 10⁴ and 4.745 × 10⁵, respectively, compared with the value of MINS, 2.863 × 10⁴ at Ex = 280 nm. The obtained quenching constant, binding constant and thermodynamic parameter suggested that the binding between compounds 1-4 with BSA protein, significantly affected by the substituted groups on the naphthalene backbone, was formed by hydrogen bonds, and other principle forces mainly consisting of charged and hydrophobic interactions. Based on results from the analysis of synchronous three-dimensional fluorescence and CD spectra, we can conclude that the interaction between compounds 1-4 and BSA protein has little impact on the BSA conformation. Calculated results obtained from in silico molecular simulation showed that compound 1 did not prefer either enzymatic drug sites I or II over the other. However, DSII in BSA was more beneficial than DSI for the binding between compounds 2-4 and BSA protein. The binding between compounds 1-3 and BSA was hydrophobic in nature, compared with the electrostatic interaction between compound 4 and BSA.

  18. Characterization and antioxidant activity of bovine serum albumin and sulforaphane complex in different solvent systems

    Energy Technology Data Exchange (ETDEWEB)

    Dong, Xueyan; Zhou, Rui; Jing, Hao, E-mail: h200521@cau.edu.cn

    2014-02-15

    Modes and influencing factors of bovine serum albumin (BSA) and sulforaphane (SFN) interaction will help us understand the interaction mechanisms and functional changes of bioactive small molecule and biomacromolecule. This study investigated interaction mechanisms of BSA and SFN and associated antioxidant activity in three solvent systems of deionized water (dH{sub 2}O), dimethyl sulfoxide (DMSO) and ethanol (EtOH), using Fourier transform infrared spectroscopy (FT-IR), fluorescence spectroscopy, synchronous fluorescence spectroscopy, DPPH and ABTS radical scavenging assays. The results revealed that SFN had ability to quench BSA's fluorescence in static modes, and to interact with BSA at both tyrosine (Tyr) and tryptophan (Trp) residues, while the Trp residues were highly sensitive, which was demonstrated by fluorescence at 340 nm. Hydrophobic forces, hydrogen bonds and van der Waals interactions were all involved in BSA and SFN interaction, which were not significantly changed by three solvents. The binding constant values and binding site numbers were in a descending order of dH{sub 2}O>DMSO>EtOH. The values of free energy change were in a descending order of dH{sub 2}O>DMSO>EtOH, which indicated that the binding forces were in a descending order of dH{sub 2}O>DMSO>EtOH. There was no significant difference in antioxidant activity between SFN and BSA–SFN. Moreover, three solvents had not significant influence on antioxidant activity of SFN and BSA–SFN. -- Highlights: • We report interaction mechanisms of BSA and sulforaphane in three solvent systems. • We report antioxidant activity of BSA–sulforaphane complex in three solvent systems. • Decreasing the solvent polarity will decrease the binding of BSA and sulforaphane. • Three solvents had not influence on antioxidant activity of BSA–sulforaphane.

  19. Studies of the interaction of CS@ZnS:Mn with bovine serum albumin under illumination

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Li, E-mail: 2476625723@qq.com [Institute of Agricultural Quality Standards and Testing Technology Research, Hubei Academy of Agricultural Science, Wuhan 430064 (China); Xiao, Ling [School of Resource and Environmental Science, Hubei Biomass-Resource Chemistry and Environmental Biotechnology Key Laboratory, Wuhan University, Wuhan 430072 (China)

    2015-09-15

    Highlights: • The interaction and illumination damages of CS@ZnS:Mn D-dots to BSA were studied. • The quenching mechanism of CS@ZnS:Mn D-dots with BSA belongs to dynamic quenching. • The hydrophobic interaction plays a major role; the binding processes are spontaneous. • The FL enhancement of CS@ZnS:Mn D-dots by BSA under UV illumination was observed. • The probable mechanism is mainly a photo-induced free radical procedure. - Abstract: In this study, chitosan coated Mn-doped ZnS quantum dots (CS@ZnS:Mn D-dots) were obtained in aqueous media under ambient pressure. The interaction and illumination damages of CS@ZnS:Mn D-dots with bovine serum albumin (BSA) were studied by means of ultraviolet–visible (UV–vis) and fluorescence (FL) spectra. It was found that the FL of BSA was quenched by CS@ZnS:Mn D-dots. The dominating quenching mechanism of CS@ZnS:Mn D-dots with BSA belongs to dynamic quenching. Hydrophobic interaction plays a major role in the CS@ZnS:Mn–BSA interaction; binding processes are spontaneous. Influencing factors such as illumination time and CS@ZnS:Mn D-dots concentrations were considered. The FL quenching effect of BSA by CS@ZnS:Mn D-dots is enhanced with the increase of illumination time and CS@ZnS:Mn D-dots concentration. The FL enhancement of CS@ZnS:Mn D-dots by BSA under UV illumination was also observed. It was proved that, the interaction of CS@ZnS:Mn D-dots with BSA under UV illumination is mainly a result of a photo-induced free radical procedure. CS@ZnS:Mn D-dots may be used as photosensitizers in photodynamic therapy.

  20. Thermodynamic Study of the Interaction of Bovine Serum Albumin and Amino Acids with Cellulose Nanocrystals.

    Science.gov (United States)

    Lombardo, Salvatore; Eyley, Samuel; Schütz, Christina; van Gorp, Hans; Rosenfeldt, Sabine; Van den Mooter, Guy; Thielemans, Wim

    2017-06-06

    The interaction of bovine serum albumin (BSA) with sulfated, carboxylated, and pyridinium-grafted cellulose nanocrystals (CNCs) was studied as a function of the degree of substitution by determining the adsorption isotherm and by directly measuring the thermodynamics of interaction. The adsorption of BSA onto positively charged pyridinium-grafted cellulose nanocrystals followed Langmuirian adsorption with the maximum amount of adsorbed protein increasing linearly with increasing degree of substitution. The binding mechanism between the positively charged pyridinum-grafted cellulose nanocrystals and BSA was found to be endothermic and based on charge neutralization. A positive entropy of adsorption associated with an increase of the degree of disorder upon addition of BSA compensated for the unfavorable endothermic enthalpy and enabled formation of pyridinium-g-CNC-BSA complexes. The endothermic enthalpy of adsorption was further found to decrease as a function of increasing degree of substitution. Negatively charged cellulose nanocrystals bearing sulfate and/or carboxylic functionalities were found to not interact significantly with the BSA protein. To investigate in more detail the role of single amino acids in the adsorption of proteins onto cellulose nanocrystals, we also studied the interaction of different types of amino acids with CNCs, i.e., charged (lysine, aspartic acid), aromatic (tryptophan, tyrosine), and polar (serine) amino acids. We found that none of the single amino acids bound with CNCs irrespective of surface charge and that therefore the binding of proteins with CNCs appears to require larger amino acid sequences that induce a greater entropic contribution to stabilize binding. Single amino acids are thus not adsorbed onto cellulose nanocrystals.

  1. Platelet-rich plasma can replace fetal bovine serum in human meniscus cell cultures.

    Science.gov (United States)

    Gonzales, Veronica K; de Mulder, Eric L W; de Boer, Trix; Hannink, Gerjon; van Tienen, Tony G; van Heerde, Waander L; Buma, Pieter

    2013-11-01

    Concerns over fetal bovine serum (FBS) limit the clinical application of cultured tissue-engineered constructs. Therefore, we investigated if platelet-rich plasma (PRP) can fully replace FBS for meniscus tissue engineering purposes. Human PRP and platelet-poor plasma (PPP) were isolated from three healthy adult donors. Human meniscal fibrochondrocytes (MFCs) were isolated from resected tissue after a partial meniscectomy on a young patient. Passage-4 MFCs were cultured in monolayer for 24 h, and 3 and 7 days. Six different culture media were used containing different amounts of either PRP or PPP and compared to a medium containing 10% FBS. dsDNA was quantified, and gene expression levels of collagen types I and II and aggrecan were measured at different time points with quantitative polymerase chain reaction in the cultured MFCs. After 7 days, the dsDNA quantity was significantly higher in MFCs cultured in 10% and 20% PRP compared to the other PRP and PPP conditions, but equal to 10% FBS. Collagen type I expression was lower in MFCs cultured with medium containing 5% PRP, 10% and 20% PPP compared to FBS. When medium with 10% PRP or 20% PRP was used, expressions were not significantly different from medium containing 10% FBS. Collagen type II expression was absent in all medium conditions. Aggrecan expression did not show differences between the different media used. However, after 7 days a higher aggrecan expression was measured in most culture conditions, except for 5% PRP, which was similar compared to FBS. Statistical significance was found between donors at various time points in DNA quantification and gene expression, but the same donors were not statistically different in all conditions. At 7 days cell cultured with 10% PRP and 20% PRP showed a higher density, with large areas of clusters, compared to other conditions. In an MFC culture medium, FBS can be replaced by 10% PRP or 20% PRP without altering proliferation and gene expression of human MFCs.

  2. Natural monomeric form of fetal bovine serum acetylcholinesterase lacks the C-terminal tetramerization domain.

    Science.gov (United States)

    Saxena, Ashima; Hur, Regina S; Luo, Chunyuan; Doctor, Bhupendra P

    2003-12-30

    Acetylcholinesterase isolated from fetal bovine serum (FBS AChE) was previously characterized as a globular tetrameric form. Analysis of purified preparations of FBS AChE by gel permeation chromatography revealed the presence of a stable, catalytically active, monomeric form of this enzyme. The two forms could be distinguished from each other based on their molecular weight, hydrodynamic properties, kinetic properties, thermal stability, and the type of glycans they carry. No differences between the two forms were observed for the binding of classical inhibitors such as edrophonium and propidium or inhibitors that are current or potential drugs for the treatment of Alzheimer's disease such as (-) huperzine A and E2020; tacrine inhibited the monomeric form 2-3-fold more potently than the tetrameric form. Sequencing of peptides obtained from an in-gel tryptic digest of the monomer and tetramer by tandem mass spectrometry indicated that the tetramer consists of 583 amino acid residues corresponding to the mature form of the enzyme, whereas the monomer consists of 543-547 amino acid residues. The subunit molecular weight of the protein component of the monomer (major species) was determined to be 59 414 Da and that of the tetramer as 64 239 Da. The N-terminal of the monomer and the tetramer was Glu, suggesting that the monomer is not a result of truncation at the N-terminal. The only differences detected were at the C-terminus. The tetramer yielded the expected C-terminus, CSDL, whereas the C-terminus of the monomer yielded a mixture of peptides, of which LLSATDTLD was the most abundant. These results suggest that monomeric FBS AChE is trimmed at the C-terminus, and the results are consistent with the involvement of C-terminal amino acids in the assembly of monomers into tetramers.

  3. Characterizing the interaction between oridonin and bovine serum albumin by a hybrid spectroscopic approach

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Zhen [Department of Chemistry, Shantou University, Shantou 515063 (China); Chen, Junhui, E-mail: chenjupush@126.com [Interventional Oncology and Minimally Invasive Therapies Department, Peking University Shenzhen Hospital, Shenzhen 518036 (China); Wang, Shaobin [The Fourth People' s Hospital of Shenzhen, Shenzhen 518033 (China); Chen, Zhanguang, E-mail: kqlu@stu.edu.cn [Department of Chemistry, Shantou University, Shantou 515063 (China)

    2013-02-15

    Oridonin is an effective anticancer drug which has high potency and low systemic toxicity. In this study, the interaction between oridonin and bovine serum albumin (BSA) was investigated by several spectroscopic approaches for the first time. The binding characteristics of oridonin and BSA were determined by fluorescence emission spectra and resonance light scattering spectra. It is showed that the oridonin quenches the fluorescence of BSA and the static quenching constant K{sub SV} is 1.30 Multiplication-Sign 10{sup 4} L mol{sup -1} at 298 K. Moreover, oridonin and BSA form a 1:1 complex with a binding constant of 0.62 Multiplication-Sign 10{sup 4} L mol{sup -1}. On the other hand, the thermodynamic parameters indicate that the binding process was a spontaneous molecular interaction procedure, in which hydrophobic forces played a major role. The structure analysis indicates that oridonin binding results in an increased hydrophobicity around the tryptophan residues of BSA. Additionally, as shown by the UV-vis absorption, synchronous fluorescence and three-dimensional fluorescence results, oridonin could lead to conformational and some microenvironmental changes of BSA. The work provides accurate and full basic data for clarifying the binding mechanism of oridonin with BSA in vitro and is helpful for understanding its effect on protein function during its transportation and distribution in blood. - Highlights: Black-Right-Pointing-Pointer Interaction between oridonin and BSA was evaluated by multi-spectroscopic methods. Black-Right-Pointing-Pointer Binding constant, number of binding sites and thermodynamic parameters were calculated. Black-Right-Pointing-Pointer Oridonin binds to Subdomain II site in BSA and form a 1:1 complex with it. Black-Right-Pointing-Pointer Oridonin-BSA complex is stabilized mainly by hydrophobic force. Black-Right-Pointing-Pointer Oridonin binding induces conformational and microenvironmental changes in BSA.

  4. Study on the interaction between amphiphilic drug and bovine serum albumin: A thermodynamic and spectroscopic description

    Energy Technology Data Exchange (ETDEWEB)

    Rub, Malik Abdul, E-mail: malikrub@gmail.com [Chemistry Department, Faculty of Science, King Abdulaziz University, Jeddah-21589 (Saudi Arabia); Khan, Javed Masood [Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh 202002 (India); Asiri, Abdullah M. [Chemistry Department, Faculty of Science, King Abdulaziz University, Jeddah-21589 (Saudi Arabia); Center of Excellence for Advanced Materials Research, King Abdulaziz University, Jeddah-21589 (Saudi Arabia); Khan, Rizwan Hasan, E-mail: rizwanhkhan1@gmail.com [Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh 202002 (India); Kabir-ud-Din [Department of Applied Chemistry, Aligarh Muslim University, Aligarh-202002 (India)

    2014-11-15

    Herein we report the interaction of amphiphilic drug clomipramine hydrochloride (CLP—a tricyclic antidepressant) with bovine serum albumin (BSA) studied by fluorescence, UV–vis, and circular dichroism (CD) spectroscopic techniques. Clomipramine hydrochloride is used to treat a variety of mental health problems. The quenching rate constant (k{sub q}) values, calculated according to the fluorescence data, decrease with increase in temperature indicating the static quenching procedure for the CLP–BSA interaction. The association binding constants (K{sub A}), evaluated at different conditions, and the thermodynamic parameters (free energy, enthalpy and entropy changes) indicate that the hydrophobic forces play a major role in the binding interaction of drug. The interaction of BSA with CLP was further confirmed by UV absorption spectra. Blue shift of position was detected due to the complex formation between the BSA–CLP. The molecular distance, r{sub 0}, between donor (BSA) and acceptor (CLP) was estimated by fluorescence resonance energy transfer (FRET) whose value (4.47 nm) suggests high probability of static quenching interaction. The CD results prove the conformational changes in the BSA on binding with the drug. Thus, the results supply qualitative and quantitative understanding of the binding of BSA to CLP, which is important in understanding their effect as therapeutic agents. - Highlights: • BSA can be considered as a good carrier for transportation of CLP in vivo. • The fluorescence results indicated the presence of static quenching mechanism in the binding process. • CD spectra showed the change in molecular conformation of BSA in the presence of CLP. • The results have applicability in model drug delivery.

  5. The digestibility of gamma-irradiated bovine serum albumin solution with thermolysin

    International Nuclear Information System (INIS)

    Hattori, Yukihiko

    1981-01-01

    Samples of bovine serum albumin (BSA) (fatty acid free, fraction V, purchased from Miles Laboratories Inc.) solution (in triply distilled water) were irradiated with 60 Co γ-rays in Pyrex tubes in the presence of air at room temperature. Taking the practical conditions of food irradiation into consideration, the concentration of BSA solution ranged from 0.6 to 9.6%, and the dose ranged from 5 to 15 kGy. Irradiated BSA solutions were subjected to the assay of proteolytic digestibility with thermolysin. The result indicates that all irradiated samples had larger digestibility with thermolysin than unirradiated control. Within the dose range of the experiment, the largest digestibility values for the irradiated samples of 0.6, 1.2, 2.4, 4.8 and 9.6% were 2.7, 2.5, 2.5, 2.3, and 1.9 times as large as that of unirradiated control, respectively. In the case of 0.6% solution the digestibility showed its maximum at 5 kGy, and at the higher doses (10 and 15 kGy) it decreased linearly. The irradiated BSA solutions of 1.2% or higher concentrations did not reveal such decrease in the digestibility. The results of analyses of amino acid composition and ultracentrifugal sedimentation pattern suggest that the decrease in the digestibility in the case of 0.6% solution can be attributed to the modification of amino acid residues, crosslinking and aggregation of BSA molecules. Furthermore, it is mentioned that the digestibility with thermolysin will be a possible measure for the denaturation of water containing protein irradiated with practical dose for the food irradiation. (J.P.N.)

  6. Templated ultrathin polyelectrolyte microreservoir for delivery of bovine serum albumin: fabrication and performance evaluation.

    Science.gov (United States)

    Gupta, Girish K; Jain, Vikas; Mishra, Prabhat Ranjan

    2011-03-01

    The aim of the study was to develop ultrathin polyelectrolyte microreservoir (UPM) using two combinations of synthetic/synthetic (S/s; poly(allylamine hydrochloride) (PAH)/sodium poly(styrenesulfonate)) and synthetic/natural (S/n; PAH/sodium alginate) polyelectrolytes over spherical porous CaCO(3) core particles (CP) followed by core removal and to evaluate its biocompatibility and integrity of loaded model protein bovine serum albumin (BSA). A novel process for synthesis of CP was developed to obtain maximum yield of monodisperse vaterite (spherical) polymorph. The prepared UPM was characterized for surface morphology, layer-by-layer growth, pay load efficiency, integrity of BSA, as well as viability and cell adhesion using murine J 774 macrophages (Φ). In vitro release profile revealed that both S/s and S/n UPM were able to provide sufficient diffusion barrier to release protein at physiological pH. It has been observed that S/n UPM are fully biocompatible due to obvious reason of using natural polymer. In a separate experiment, the S/s UPM surface was modified with pluronic F-68 to tune biocompatibility which provides evidences for safety and tolerability of the S/s UPM as well. In nutshell, the proposed system could successfully be used for the delivery of proteins, and moreover, the system can be tailored to impart desired properties at any stage of layering especially in terms of drug release and to retain the integrity of proteins. © 2011 American Association of Pharmaceutical Scientists

  7. Disulfide-bond scrambling promotes amorphous aggregates in lysozyme and bovine serum albumin.

    Science.gov (United States)

    Yang, Mu; Dutta, Colina; Tiwari, Ashutosh

    2015-03-12

    Disulfide bonds are naturally formed in more than 50% of amyloidogenic proteins, but the exact role of disulfide bonds in protein aggregation is still not well-understood. The intracellular reducing agents and/or improper use of antioxidants in extracellular environment can break proteins disulfide bonds, making them unstable and prone to misfolding and aggregation. In this study, we report the effect of disulfide-reducing agent dithiothreitol (DTT) on hen egg white lysozyme (lysozyme) and bovine serum albumin (BSA) aggregation at pH 7.2 and 37 °C. BSA and lysozyme proteins treated with disulfide-reducing agents form very distinct amorphous aggregates as observed by scanning electron microscope. However, proteins with intact disulfide bonds were stable and did not aggregate over time. BSA and lysozyme aggregates show unique but measurable differences in 8-anilino-1-naphthalenesulfonic acid (ANS) and 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid (bis-ANS) fluorescence, suggesting a loose and flexible aggregate structure for lysozyme but a more compact aggregate structure for BSA. Scrambled disulfide-bonded protein aggregates were observed by nonreducing sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for both proteins. Similar amorphous aggregates were also generated using a nonthiol-based reducing agent, tris(2-carboxyethyl)phosphine (TCEP), at pH 7.2 and 37 °C. In summary, formation of distinct amorphous aggregates by disulfide-reduced BSA and lysozyme suggests an alternate pathway for protein aggregation that may be relevant to several proteins.

  8. Effects of urea, metal ions and surfactants on the binding of baicalein with bovine serum albumin

    Directory of Open Access Journals (Sweden)

    Atanu Singha Roy

    2016-08-01

    Full Text Available The interaction of baicalein with bovine serum albumin (BSA was investigated with the help of spectroscopic and molecular docking studies. The binding affinity of baicalein towards BSA was estimated to be in order of 105 M−1 from fluorescence quenching studies. Negative ΔH° (−5.66±0.14 kJ/mol and positive (ΔS° (+79.96±0.65 J/mol K indicate the presence of electrostatic interactions along with the hydrophobic forces that result in a positive ΔS°. The hydrophobic association of baicalein with BSA diminishes in the presence of sodium dodecyl sulfate (SDS due to probable hydrophobic association of baicalein with SDS, resulting in a negative ΔS° (−40.65±0.87 J/mol K. Matrix-assisted laser desorption ionization/time of flight (MALDI--TOF experiments indicate a 1:1 complexation between baicalein and BSA. The unfolding and refolding phenomena of BSA were investigated in the absence and presence of baicalein using steady-state and fluorescence lifetime measurements. It was observed that the presence of urea ruptured the non-covalent interaction between baicalein and BSA. The presence of metal ions (Ag+, Mg2+, Ni2+, Mn2+, Co2+and Zn2+ increased the binding affinity of ligand towards BSA. The changes in conformational aspects of BSA after ligand binding were also investigated using circular dichroism (CD and Fourier transform infrared (FT-IR spectroscopic techniques. Site selectivity studies following molecular docking analyses indicated the binding of baicalein to site 1 (subdomain IIA of BSA.

  9. Characterisation of the de-agglomeration effects of bovine serum albumin on nanoparticles in aqueous suspension.

    Science.gov (United States)

    Tantra, Ratna; Tompkins, Jordan; Quincey, Paul

    2010-01-01

    This paper describes the use of nanoparticle characterisation tools to evaluate the interaction between bovine serum albumin (BSA) and dispersed nanoparticles in aqueous media. Dynamic light scattering, zeta-potential measurements and scanning electron microscopy were used to probe the state of zinc oxide (ZnO) and titanium dioxide (TiO(2)) nanoparticles in the presence of various concentrations of BSA, throughout a three-day period. BSA was shown to adhere to ZnO but not to TiO(2). The adsorption of BSA led to subsequent de-agglomeration of the sub-micron ZnO clusters into smaller fragments, even breaking them up into individual isolated nanoparticles. We propose that certain factors, such as adsorption kinetics of BSA on to the surface of ZnO, as well as the initial agglomerated state of the ZnO, prior to BSA addition, are responsible for promoting the de-agglomeration process. Hence, in the case of TiO(2) we see no de-agglomeration because: (a) the nanoparticles are more highly agglomerated to begin with and (b) BSA does not adsorb effectively on the surface of the nanoparticles. The zeta-potential results show that, for either ZnO or TiO(2), the presence of BSA resulted in enhanced stability. In the case of ZnO, the enhanced stability is limited to BSA concentrations below 0.5 wt.%. Steric and electrostatic repulsion are thought to be responsible for improved stability of the dispersion.

  10. Spectroscopic analysis on the binding interaction of biologically active pyrimidine derivative with bovine serum albumin

    Directory of Open Access Journals (Sweden)

    Vishwas D. Suryawanshi

    2016-02-01

    Full Text Available A biologically active antibacterial reagent, 2–amino-6-hydroxy–4–(4-N, N-dimethylaminophenyl-pyrimidine-5-carbonitrile (AHDMAPPC, was synthesized. It was employed to investigate the binding interaction with the bovine serum albumin (BSA in detail using different spectroscopic methods. It exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus which are common food poisoning bacteria. The experimental results showed that the fluorescence quenching of model carrier protein BSA by AHDMAPPC was due to static quenching. The site binding constants and number of binding sites (n≈1 were determined at three different temperatures based on fluorescence quenching results. The thermodynamic parameters, enthalpy change (ΔH, free energy (ΔG and entropy change (ΔS for the reaction were calculated to be 15.15 kJ/mol, –36.11 kJ/mol and 51.26 J/mol K according to van't Hoff equation, respectively. The results indicated that the reaction was an endothermic and spontaneous process, and hydrophobic interactions played a major role in the binding between drug and BSA. The distance between donor and acceptor is 2.79 nm according to Förster's theory. The alterations of the BSA secondary structure in the presence of AHDMAPPC were confirmed by UV–visible, synchronous fluorescence, circular dichroism (CD and three-dimensional fluorescence spectra. All these results indicated that AHDMAPPC can bind to BSA and be effectively transported and eliminated in the body. It can be a useful guideline for further drug design.

  11. Binding interaction of phosphorus heterocycles with bovine serum albumin: A biochemical study

    Directory of Open Access Journals (Sweden)

    Swarup Roy

    2017-02-01

    Full Text Available Interaction between bovine serum albumin (BSA and phosphorus heterocycles (PHs was studied using multi-spectroscopic techniques. The results indicated the high binding affinity of PHs to BSA as it quenches the intrinsic fluorescence of BSA. The experimental data suggested the fluorescence quenching mechanism between PHs and BSA as a dynamic quenching. From the UV–vis studies, the apparent association constant (Kapp was found to be 9.25×102, 1.27×104 and 9.01×102 L/mol for the interaction of BSA with PH-1, PH-2 and PH-3 respectively. According to the Förster's non-radiation energy transfer (FRET theory, the binding distances between BSA and PHs were calculated. The binding distances (r of PH-1, PH-2 and PH-3 were found to be 2.86, 3.03, and 5.12 nm, respectively, indicating energy transfer occurs between BSA and PHs. The binding constants of the PHs obtained from the fluorescence quenching data were found to be decreased with increase of temperature. The negative values of the thermodynamic parameters ΔH, ΔS and ΔG at different temperatures revealed that the binding process is spontaneous; hydrogen bonds and van der Waals interaction were the main force to stabilize the complex. The microenvironment of the protein-binding site was studied by synchronous fluorescence and circular dichroism (CD techniques and data indicated that the conformation of BSA changed in the presence of PHs. Finally, we studied the BSA-PHs docking using Autodock and results suggest that PHs is located in the cleft between the domains of BSA.

  12. Interaction of bovine serum albumin with a psychotropic drug alprazolam: Physicochemical, photophysical and molecular docking studies

    Energy Technology Data Exchange (ETDEWEB)

    Sarkar, Moumita; Paul, Shiv Shankar; Mukherjea, Kalyan K., E-mail: k_mukherjea@yahoo.com

    2013-10-15

    The interaction between alprazolam (Alp) and bovine serum albumin (BSA) has been investigated under physiological conditions by UV–vis, steady state as well as time-resolved fluorescence, circular dichroism (CD) spectroscopic and molecular docking studies. The binding constant K of Alp to BSA was found to be 1.8×10{sup 5} L mol{sup −1} from absorption data. Fluorometric studies suggested the formation of the Alp–BSA complex, while time-resolved fluorescence studies showed that the binding of Alp by BSA was mainly static and the effective rate constant is found to be 2.33×10{sup 13} L mol{sup −1} s{sup −1}. According to the modified Stern–Volmer equation, the Stern–Volmer quenching constants (K{sub SV}) between Alp and BSA at four different temperatures 295, 303, 308, 313 K were obtained to be 1.19×10{sup 5}, 1.05×10{sup 5}, 0.99×10{sup 5} and 0.90×10{sup 5} L mol{sup −1}, respectively. The change in enthalpy (ΔH) and entropy (ΔS) were calculated to be −11.66 and 57.64 J mol{sup −1} K{sup −1}, respectively, indicating that the interaction was hydrophobic in nature. Site marker competitive experiments suggested that the binding of Alp to BSA primarily took place in sub-domain IIA, whereas the binding distance (r) between Alp and the tryptophan residue of BSA was obtained to be 1.87 nm by Förster's theory of non-radiative energy transfer. The conformational studies by CD spectroscopy showed that the presence of Alp decreased the α-helical content of BSA and induced the unfolding of the polypeptide of the protein. The change in conformation was also supported by excitation–emission matrix spectroscopy (EEMS) studies. The molecular docking experiment supports the above results and effectively proves the binding of Alp to BSA. -- Highlights: • Alprazolam: a benzodiazepine drug with anxiolytic and anticonvulsant properties. • Alprazolam induces conformational change on the native as well as urea denatured BSA. • Alprazolam may

  13. Investigation of the interaction of naringin palmitate with bovine serum albumin: spectroscopic analysis and molecular docking.

    Directory of Open Access Journals (Sweden)

    Xia Zhang

    Full Text Available BACKGROUND: Bovine serum albumin (BSA contains high affinity binding sites for several endogenous and exogenous compounds and has been used to replace human serum albumin (HSA, as these two compounds share a similar structure. Naringin palmitate is a modified product of naringin that is produced by an acylation reaction with palmitic acid, which is considered to be an effective substance for enhancing naringin lipophilicity. In this study, the interaction of naringin palmitate with BSA was characterised by spectroscopic and molecular docking techniques. METHODOLOGY/PRINCIPAL FINDINGS: The goal of this study was to investigate the interactions between naringin palmitate and BSA under physiological conditions, and differences in naringin and naringin palmitate affinities for BSA were further compared and analysed. The formation of naringin palmitate-BSA was revealed by fluorescence quenching, and the Stern-Volmer quenching constant (KSV was found to decrease with increasing temperature, suggesting that a static quenching mechanism was involved. The changes in enthalpy (ΔH and entropy (ΔS for the interaction were detected at -4.11 ± 0.18 kJ·mol(-1 and -76.59 ± 0.32 J·mol(-1·K(-1, respectively, which indicated that the naringin palmitate-BSA interaction occurred mainly through van der Waals forces and hydrogen bond formation. The negative free energy change (ΔG values of naringin palmitate at different temperatures suggested a spontaneous interaction. Circular dichroism studies revealed that the α-helical content of BSA decreased after interacting with naringin palmitate. Displacement studies suggested that naringin palmitate was partially bound to site I (subdomain IIA of the BSA, which was also substantiated by the molecular docking studies. CONCLUSIONS/SIGNIFICANCE: In conclusion, naringin palmitate was transported by BSA and was easily removed afterwards. As a consequence, an extension of naringin applications for use in food, cosmetic

  14. Binding thermodynamics of Diclofenac and Naproxen with human and bovine serum albumins: A calorimetric and spectroscopic study

    International Nuclear Information System (INIS)

    Bou-Abdallah, Fadi; Sprague, Samuel E.; Smith, Britannia M.; Giffune, Thomas R.

    2016-01-01

    Highlights: • The binding affinity of Diclofenac and Naproxen to BSA and HSA is on the order of 10 4 –10 6 M −1 . • Two Diclofenac molecules bind per BSA or HSA but only 0.75 and 3 Naproxen molecules bind to BSA and HSA, respectively. • Drugs binding to BSA is only enthalpically favored and both enthalpically and entropically favored for HSA. • Fluorescence quenching data suggest dynamic collisions and the formation of ground-state protein-drug complexes. • DSC data show multiple sequential unfolding events and strong drug stabilization effects. - Abstract: Serum albumins are ubiquitous proteins able to bind a variety of exogenous and endogenous ligands including hydrophobic pharmaceuticals. Most drugs bind to two very active binding regions located within sub-domains IIA and IIIA of the protein, also known as Sudlow’s sites. The drug binding mode of serum albumin provides important pharmacological information and influences drug solubility, efficacy, biological distribution, and excretion. Here, the binding thermodynamics of Diclofenac and Naproxen, two non-steroidal anti-inflammatory drugs (NSAIDs) to bovine and human serum albumins (BSA and HSA, respectively) were studied by isothermal titration calorimetry (ITC), fluorescence spectroscopy and differential scanning calorimetry (DSC). The ITC data show that the binding affinity (K) of Diclofenac to BSA and HSA is on the order of 10 4 M −1 with a binding stoichiometry (n) of 2 drug molecules per protein. Naproxen binding to the two proteins exhibits a different profile with K and n values on the order of 10 6 M −1 and 0.75 for BSA, and 10 5 M −1 and 3 for HSA, respectively. The binding of the two drugs to HSA is found to be both enthalpically and entropically favored suggesting the formation of hydrogen bonds and van der Waals hydrophobic effects. Binding of the two drugs to BSA is only enthalpically favored with an unfavorable entropy term. Significant enthalpy–entropy compensation

  15. Protein hydroperoxides and carbonyl groups generated by porphyrin-induced photo-oxidation of bovine serum albumin

    DEFF Research Database (Denmark)

    Silvester, J A; Timmins, G S; Davies, Michael Jonathan

    1998-01-01

    Porphyrin-sensitized photo-oxidation of bovine serum albumin results in oxidation at specific sites to produce protein radical species: at the Cys-34 residue (to give a thiyl radical) and at one or both tryptophan residues (Trp-134 and Trp-214) to give tertiary carbon-centered radicals and cause...... disruption of the indole ring system. This study shows that these photo-oxidation processes also consume oxygen and give rise to hydrogen peroxide, protein hydroperoxides, and carbonyl functions. The yield of hydrogen peroxide, protein hydroperoxides, and carbonyl functions is shown to be dependent...

  16. Thermal-Induced Denaturation and Aggregation Behavior of Lysozyme and Bovine Serum Albumin: a Thermodynamic and Structural Study

    Science.gov (United States)

    Perez, Aline Sanches; Oliveira, Cristiano Luis Pinto

    2017-10-01

    Solution studies permit a direct investigation of the particles on a well-defined environment. Fluorescence, circular dichroism, scattering, and calorimetry provide, individually, very important information among the protein structure, overall shape, and thermodynamic equilibrium. In this work, a combination of these techniques is presented for the study of denaturation induced by temperature of two well-known proteins, Henn Egg lysozyme and bovine serum albumin. A detailed thermodynamic and structural investigation is shown for these proteins, providing interesting information on the thermal-induced changes in the protein structure and aggregation behavior.

  17. New therapeutic option for irritable bowel syndrome: serum-derived bovine immunoglobulin.

    Science.gov (United States)

    Good, Larry; Rosario, Roxanne; Panas, Raymond

    2015-03-21

    Oral prescription medical foods have long been used in hospital settings but are also appropriate therapies for gastrointestinal disorders in outpatient medical practice. Oral serum-derived bovine immunoglobulin/protein isolate (SBI) has been shown in clinical studies to reduce loose stools and improve stool consistency as well as other symptoms (i.e., abdominal pain, bloating, and urgency) in patients with irritable bowel syndrome with diarrhea (IBS-D) and human immunodeficiency virus-associated enteropathy. This case series reports the outcomes of 14 IBS patients who received SBI as an addition to standard of care at an individual physician's clinical practice. The patients: 2 IBS with constipation (IBS-C), 7 IBS-D, 2 mixed diarrhea and constipation IBS (IBS-M) and 3 undefined IBS (IBS-U; also described by some physicians as IBS-Bloating), ranged in age from 22-87 years. SBI (5 g or 10 g daily dose) was added to the patient's current standard care and followed for several weeks to determine if symptoms were improved with the addition of SBI. Overall, 12 of the 14 patients indicated some level of improvement through direct questioning of the patients regarding changes from the prior visit. One IBS-Bloating patient had a resolution of symptoms and two patients (1 IBS-Bloating and 1 IBS-C) discontinued therapy because of insufficient relief. The 12 patients who continued on therapy reported an overall improvement in symptoms with better stool consistency, decreased frequency as well as reductions in abdominal pain, bloating, distention, and incontinence. In most cases, therapeutic effects of SBI were seen within the first four weeks of therapy with continued improvements at subsequent visits. SBI has a multifaceted mechanism of action and may help to manage IBS by providing a distinct protein source required to normalize bowel function, gastrointestinal microbiota, and nutritionally enhance tight junction protein expression between intestinal epithelial cells. SBI

  18. Size-dependent interaction of silica nanoparticles with lysozyme and bovine serum albumin proteins

    Science.gov (United States)

    Yadav, Indresh; Aswal, Vinod K.; Kohlbrecher, Joachim

    2016-05-01

    The interaction of three different sized (diameter 10, 18, and 28 nm) anionic silica nanoparticles with two model proteins—cationic lysozyme [molecular weight (MW) 14.7 kDa)] and anionic bovine serum albumin (BSA) (MW 66.4 kDa) has been studied by UV-vis spectroscopy, dynamic light scattering (DLS), and small-angle neutron scattering (SANS). The adsorption behavior of proteins on the nanoparticles, measured by UV-vis spectroscopy, is found to be very different for lysozyme and BSA. Lysozyme adsorbs strongly on the nanoparticles and shows exponential behavior as a function of lysozyme concentration irrespective of the nanoparticle size. The total amount of adsorbed lysozyme, as governed by the surface-to-volume ratio, increases on lowering the size of the nanoparticles for a fixed volume fraction of the nanoparticles. On the other hand, BSA does not show any adsorption for all the different sizes of the nanoparticles. Despite having different interactions, both proteins induce similar phase behavior where the nanoparticle-protein system transforms from one phase (clear) to two phase (turbid) as a function of protein concentration. The phase behavior is modified towards the lower concentrations for both proteins with increasing the nanoparticle size. DLS suggests that the phase behavior arises as a result of the nanoparticles' aggregation on the addition of proteins. The size-dependent modifications in the interaction potential, responsible for the phase behavior, have been determined by SANS data as modeled using the two-Yukawa potential accounting for the repulsive and attractive interactions in the systems. The protein-induced interaction between the nanoparticles is found to be short-range attraction for lysozyme and long-range attraction for BSA. The magnitude of attractive interaction irrespective of protein type is enhanced with increase in the size of the nanoparticles. The total (attractive+repulsive) potential leading to two-phase formation is found to be

  19. Circulating MicroRNAs in Serum from Cattle Challenged with Bovine Viral Diarrhea Virus‡

    Directory of Open Access Journals (Sweden)

    Tasia M. Taxis

    2017-06-01

    Full Text Available Bovine viral diarrhea virus (BVDV is an RNA virus that is often associated with respiratory disease in cattle. MicroRNAs have been proposed as indicators of exposure to respiratory pathogens. The objective of this study was to identify microRNAs in cattle that had been challenged with a non-cytopathic field strain of BVDV. Five colostrum deprived neonate Holstein calves were inoculated with BVDV (challenged and 4 were mock challenged (control. Serum from all calves was collected at four different times: prior to challenge (day 0 and at 4, 9, and 16 days post-challenge. RNA was extracted from sera, and expression, via read counts, of small non-coding RNAs were obtained using next-generation sequencing. A total of 905,861 sequences identified 427 microRNAs. Sixty-two microRNAs had >1,000 total reads across all samples. Bta-miR-339a, bta-miR-185, bta-miR-486, Bta-miR-92a, bta-miR-30e-5p, bta-let-7c, and bta-miR-2284x were significantly different (P < 0.05 across time regardless of challenge status. Bta-miR-423-5p (P = 0.008 and bta-miR-151-3p (P = 0.005 were significantly different between challenged and control animals across time. In challenged animals, bta-miR-423-5p peaked in number of reads by day 4 and steadily declined from day 4 to day 16. In control animals, bta-miR-423-5p declined from day 0 to day 9 and increased in number by day 16. By day 16, both challenged and control animals had similar levels of bta-miR-423-5p, and these levels were similar to day 0 levels. Bta-miR-151-3p peaked at day 9 in challenged animals, while control animals decreased across time. By day 16, the number of reads of bta-miR-151-3p were similar between challenged and control animals. The level in challenged animals had returned to day 0 levels by day 16, whereas the levels for control animals was significantly lower (P = 0.006 than day 0. Further studies are needed to establish if bta-miR-423-5p or bta-miR-151-3p could be used as a biomarker for exposure to

  20. Spectrofluorimetric study on the interaction between antimicrobial drug sulfamethazine and bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Dawoud Bani-Yaseen, Abdulilah, E-mail: aayaseen@yahoo.co [Department of Chemistry, Faculty of Science, Taibah University, Al-MAdinah Al-Munawarah P.O. Box 30002 (Saudi Arabia)

    2011-05-15

    The interaction between the antimicrobial drug sulfamethazine (STM) and bovine serum albumin (BSA) has been studied using steady state and synchronous fluorescence spectroscopy. Fluorescence emission data revealed that BSA (2x10{sup -6} M) fluorescence was statically quenched by STM at various concentrations, which implies that STM-BSA complex has been formed. The fluorescence emission data was analyzed via applying the Stern-Volmer analysis in combination with thermodynamic investigation, where obtained results revealed that quenching is static with quenching constants of 2.371, 1.658, and 0.916x10{sup 5} M{sup -1} at 298, 304, and 310 K, respectively. Binding constants and number of binding sites at different temperatures were also determined by applying the Scatchard method, which in turn were used to construct the van't Hoff plot in order to estimate the enthalpy ({Delta}H) and entropy changes ({Delta}S) for the complexation process. An average of 1.00{+-}0.17 was estimated for the number of sites of BSA, which indicated that STM binds to BSA with stoichiometric ratio of 1:1. The values that were estimated from the van't Hoff plot for {Delta}H and ({Delta}S) were -36.8 kJ mol{sup -1} and -14.9 J mol{sup -1} K{sup -1}, respectively, which indicate that the STM-BSA complex is stabilized with hydrogen bonds and van der Waals interactions. Synchronous fluorescence data was obtained at {Delta}{lambda} of 15 and 60 nm, where obtained results confirmed that STM binds to BSA at the tryptophan residue (Trp. 213). In addition, the distance between STM and the Trp. 213 was estimated via employing the Foerster's non-radiative energy-transfer theory, and was found to be 2.73 nm, which in turn indicated that STM can bind to BSA with high probability. - Research highlights: {yields} Fluorescence data was used to demonstrate that BSA fluorescence can be quenched by STM. {yields} STM can bind to BSA with stoichiometric ratio of 1:1. {yields} STM-BSA complex is

  1. Unexpected effects of absorbed normal rabbit serum and bovine serum albumin on survival of Haemophilus influenzae type b in the infant rat.

    Science.gov (United States)

    Loeb, M R

    1988-01-01

    In the course of using the infant rat model to determine the ability of various rabbit antisera to protect against challenge by Haemophilus influenzae type b we made two unexpected observations. In these experiments 4-day-old rats were inoculated s.c. on the dorsum with either rabbit serum or physiological buffers (sham serum) and then were challenged the next day with H. influenzae type b injected i.p. Bacteremia, as a marker for disease, was measured 24 h later on day 6. We observed the following. (i) Pre-immune, i.e., normal rabbit serum, containing minimal levels of antibodies to outer membrane proteins and depleted of antibodies to capsule and lipopolysaccharide, nevertheless significantly (P less than 0.01) protected the rats from challenge with H. influenzae type b when compared to a sham inoculation of buffer; (ii) In the absence of a serum inoculation on day 4 (a buffer was used as a sham serum inoculation), the levels of bacteremia obtained after inoculation with bacteria on day 5 depended upon the composition of the buffer in which the H. influenzae inoculum was suspended. Use of phosphate buffered saline (PBS) resulted in higher levels of bacteremia than PBS containing 0.5% bovine serum albumin (PBS-BSA) (P less than 0.001), i.e. the BSA apparently acted to protect the rats from H. influenzae infection. In fact the use of PBS-BSA as an inoculum buffer masked the protective effect noted above of the absorbed normal rabbit serum.

  2. The effect of glycation on bovine serum albumin conformation and ligand binding properties with regard to gliclazide

    Science.gov (United States)

    Żurawska-Płaksej, Ewa; Rorbach-Dolata, Anna; Wiglusz, Katarzyna; Piwowar, Agnieszka

    2018-01-01

    Albumin, the major serum protein, plays a variety of functions, including binding and transporting endogenous and exogenous ligands. Its molecular structure is sensitive to different environmental modifiers, among which glucose is one of the most significant. In vivo albumin glycation occurs under physiological conditions, but it is increased in diabetes. Since bovine serum albumin (BSA) may serve as a model protein in in vitro experiments, we aimed to investigate the impact of glucose-mediated BSA glycation on the binding capacity towards gliclazide, as well as the ability of this drug to prevent glycation of the BSA molecule. To reflect normo- and hyperglycemia, the conditions of the glycation process were established. Structural changes of albumin after interaction with gliclazide (0-14 μM) were determined using fluorescence quenching and circular dichroism spectroscopy. Moreover, thermodynamic parameters as well as energy transfer parameters were determined. Calculated Stern-Volmer quenching constants, as well as binding constants for the BSA-gliclazide complex, were lower for the glycated form of albumin than for the unmodified protein. The largest, over 2-fold, decrease in values of binding parameters was observed for the sample with 30 mM of glucose, reflecting the poorly controlled diabetic state, which indicates that the degree of glycation had a critical influence on binding with gliclazide. In contrast to significant changes in the tertiary structure of BSA upon binding with gliclazide, only slight changes in the secondary structure were observed, which was reflected by about a 3% decrease of the α-helix content of glycated BSA (regardless of glucose concentration) in comparison to unmodified BSA. The presence of gliclazide during glycation did not affect its progress. The results of this study indicate that glycation significantly changed the binding ability of BSA towards gliclazide and the scale of these changes depended on glucose concentration. It

  3. Spectroscopic study on the sonodynamic and sonocatalytic damage of anthraquinone derivants to bovine serum albumin under ultrasonic irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Wang Zhiqiu; Gao Jingqun [College of Chemistry, Liaoning University, Shenyang 110036 (China); Wang Jun, E-mail: wangjun890@126.com [College of Chemistry, Liaoning University, Shenyang 110036 (China); Li Ying; Li Kai; Kang Pingli; Zhang Xiangdong [College of Chemistry, Liaoning University, Shenyang 110036 (China)

    2012-03-15

    In this work, three anthraquinone derivants (Alizarin: 1,2-dihydroxy-9, 10-anthraquinone, Alizarin-DA: 1,2-dihydroxy-9, 10-anthraquinone-3-aminomethyl-N, N-diacetic acid and Alizarin-DA-Fe: 1,2-dihydroxy-9, 10-anthraquinone-3-aminomethyl-N, N-diacetate-Ferrous(III)) were used to study the sonodynamic and sonocatalytic damage of bovine serum albumin (BSA) molecules according to the hyperchromic effect of UV-vis spectra and quenching effect of intrinsic fluorescence. Meanwhile, some influencing factors such as ultrasonic irradiation time, anthraquinone derivants concentration and ionic strength on the damage of BSA molecules were also considered. The results show that the synergetic effect of anthraquinone derivants and ultrasonic irradiation can efficiently damage the BSA molecules. Finally, some special radical scavengers were used to determine the kind of generated reactive oxygen species (ROS) in the presence of three anthraquinone derivants under ultrasonic irradiation. The results show that the ROS, at least, including singlet oxygen ({sup 1}O{sub 2}) and hydroxyl radicals (OH) are generated during the sonodynamic and sonocatalytic processes. It is wished that this paper could offer some valuable references for the application of anthraquinone derivants in sonodynamic therapy (SDT) and sonocatalytic therapy (SCT) for tumor treatment. - Highlights: Black-Right-Pointing-Pointer Anthraquinone derivants were used to study the sonodynamic and sonocatalytic damage to BSA. Black-Right-Pointing-Pointer The generations of ROS during sonodynamic and sonocatalytic process were estimated. Black-Right-Pointing-Pointer Some quenchers were used to determine the kind of the ROS.

  4. Investigation of the binding affinity in vitamin B12-Bovine serum albumin system using various spectroscopic methods

    Science.gov (United States)

    Makarska-Bialokoz, Magdalena

    2017-09-01

    The binding affinity between vitamin B12 (VitB12) and bovine serum albumin (BSA) has been investigated in aqueous solution at pH = 7.4, employing UV-vis absorption and steady-state, synchronous and three-dimensional fluorescence spectra techniques. Representative effects noted for BSA intrinsic fluorescence resulting from the interactions with VitB12 confirm the formation of π-π stacked non-covalent and non-fluorescent complexes in the system VitB12-BSA. All the determined parameters, the binding, fluorescence quenching and bimolecular quenching rate constants (of the order of 104 L mol- 1, 103 L mol- 1 and 1011 L mol- 1 s- 1, respectively), as well as Förster resonance energy transfer parameters validate the mechanism of static quenching. The interaction with VitB12 induces folding of the polypeptide chains around Trp residues of BSA, resulting in a more hydrophobic surrounding. Presented outcomes suggest that the addition of VitB12 can lead to the more organized BSA conformation and its more folded tertiary structure, what could influence the physiological functions of bovine serum albumin, notably in case of its overuse or abnormal metabolism.

  5. Titania nanotube delivery fetal bovine serum for enhancing MC3T3-E1 activity and osteogenic gene expression

    International Nuclear Information System (INIS)

    Peng, Jing; Zhang, Xinming; Li, Zhaoyang; Liu, Yunde; Yang, Xianjin

    2015-01-01

    Titania nanotube (TNT) delivery of fetal bovine serum (FBS) was conducted on titanium (Ti) to enhance bone tissue repair. Scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS) showed FBS increased the tube wall thickness and decreased the tube diameter. Attenuated total reflectance Fourier transform infrared further confirmed that FBS completely covered the TNT and changed the surface composition. Water contact angle tests showed TNT/FBS possessed hydrophilic properties. Compared to original Ti, the TNT/FBS group had more attached osteoblasts after 2 h and enhanced filopodia growth at 0.5 h. Significantly, more osteoblasts were also observed on TNT/FBS after 7 d culturing. FBS was released steadily from TNT; about 70% of FBS had been released at 3 d and 90% at 5 d, as shown by the bicinchoninic acid method. TNT/FBS also enhanced subsequent osteoblast differentiation and gene expression; the quantum real-time polymerase chain reaction test showed that TNT/FBS up-regulated alkaline phosphatase and osteocalcin gene expression at 7 d and 14 d. Therefore, TNT/FBS delivered sustained in situ nutrition and enhanced osteoblast activity and osteogenic gene expression. - Highlights: • Fetal Bovine Serum (FBS) was filled in titania nanotube (TNT) structures. • FBS provided sustained-release in situ nutrition for surface osteoblast growth. • TNT/FBS enhanced osteoblast activity and osteogenic gene expression

  6. Titania nanotube delivery fetal bovine serum for enhancing MC3T3-E1 activity and osteogenic gene expression

    Energy Technology Data Exchange (ETDEWEB)

    Peng, Jing, E-mail: pengjingtd@163.com [Airport College, Civil Aviation University of China, Tianjin 300300 (China); Zhang, Xinming, E-mail: xinmingmail@163.com [Tianjin Product Quality Inspection Technology Research Institute, Tianjin 300384 (China); School of Materials Science and Engineering, Tianjin University, Tianjin 300072 (China); Li, Zhaoyang [School of Materials Science and Engineering, Tianjin University, Tianjin 300072 (China); Liu, Yunde [School of Medical Laboratory, Tianjin Medical University, Tianjin 300203 (China); Yang, Xianjin [School of Materials Science and Engineering, Tianjin University, Tianjin 300072 (China)

    2015-11-01

    Titania nanotube (TNT) delivery of fetal bovine serum (FBS) was conducted on titanium (Ti) to enhance bone tissue repair. Scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS) showed FBS increased the tube wall thickness and decreased the tube diameter. Attenuated total reflectance Fourier transform infrared further confirmed that FBS completely covered the TNT and changed the surface composition. Water contact angle tests showed TNT/FBS possessed hydrophilic properties. Compared to original Ti, the TNT/FBS group had more attached osteoblasts after 2 h and enhanced filopodia growth at 0.5 h. Significantly, more osteoblasts were also observed on TNT/FBS after 7 d culturing. FBS was released steadily from TNT; about 70% of FBS had been released at 3 d and 90% at 5 d, as shown by the bicinchoninic acid method. TNT/FBS also enhanced subsequent osteoblast differentiation and gene expression; the quantum real-time polymerase chain reaction test showed that TNT/FBS up-regulated alkaline phosphatase and osteocalcin gene expression at 7 d and 14 d. Therefore, TNT/FBS delivered sustained in situ nutrition and enhanced osteoblast activity and osteogenic gene expression. - Highlights: • Fetal Bovine Serum (FBS) was filled in titania nanotube (TNT) structures. • FBS provided sustained-release in situ nutrition for surface osteoblast growth. • TNT/FBS enhanced osteoblast activity and osteogenic gene expression.

  7. Immunization of cattle against Schistosome bovis (including pathophysiological studies on schistosome infection in bovines)

    International Nuclear Information System (INIS)

    Hussain, M.F.

    1978-12-01

    Bovine schistosomiasis caused by S. bovis constitutes a serious veterinary problem in the Sudan, yet very little is known about the epidemiology, pathogenesis and immunology of the disease. Over the past 5 years, work on these aspects has been conducted at Khartoum and several outlying areas of the White Nile Province in Sudan. In studies involving over 1,000 cattle, it was found that almost 100% of animals are infected by 2 years of age but that the prevalence falls to less than 60% over the following 7 years. There was also a marked reduction in the intensity of infection with increasing age, indicating the development of a high degree of acquired resistance. This was confirmed experimentally by challenging animals from an endemic area with massive numbers of cercariae. These animals completely resisted the challenge whereas animals never previously exposed either died or became moribund due to the severe haemorrhagic diarrhoea resulting from the passage of schistosome eggs through the gut wall. Attempts were made to vaccinate calves using irradiated organisms. These gave 70-80% protection against a challenge infection and this was sufficient to allow these animals to gain weight and remain clinically healthy. Animals not given the vaccine deteriorated. The efficacy of the vaccine was then tested under field conditions and found to give a high level of protection against S. bovis. These animals were also less susceptible to intercurrent infections

  8. Circulating microRNAs in serum from cattle challenged with Bovine Viral Diarrhea Virus

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) is an RNA virus that is often associated with respiratory disease in cattle. MicroRNAs have been proposed as indicators of exposure to respiratory pathogens. The objective of this study was to identify microRNAs in cattle that had been challenged with a non-cytopat...

  9. Calcium and magnesium concentrations in uterine fluid and blood serum during the estrous cycle in the bovine

    Directory of Open Access Journals (Sweden)

    Sayed Mortaza Alavi-Shoushtari

    2012-06-01

    Full Text Available To investigate uterine and serum Ca++ and Mg++ variations during the estrous cycle in the bovine, 66 genital tracts and blood samples were collected from Urmia abattoir, Urmia, Iran. The phase of the estrous cycle was determined by examination of the structures present on ovaries and uterine tonicity. Of the collected samples, 17 were pro-estrus, 12 estrus, 14 metestrus and 23 diestrus. The uterine fluid was collected by gentle scraping of the uterine mucosa with a curette. The mean ± SEM concentration of serum Ca++ in pro-estrus, estrus, metestrus and diestrus was 5.77 ± 0.69, 8.87 ± 1.83, 10.95 ± 1.52, 11.09 ± 1.08 mg dL-1, and the mean concentration of uterine fluid Ca++ was 4.40 ± 0.72, 3.15 ± 0.67, 5.89 ± 0.88, 8.63 ± 0.97 mg dL-1, respectively. The mean concentration of serum Mg++ in pro-estrus, estrus, metestrus and diestrus was 3.53 ± 0.30, 4.20 ± 0.52, 3.49 ± 0.38, 3.39 ± 0.29 mg dL-1, and mean concentration of uterine fluid Mg++ was 5.27 ± 0.42, 4.92 ± 0.60, 5.56 ± 0.30, 5.88 ± 0.36 mg dL-1, respectively. The serum and uterine fluid Ca++ in pro-estrus were significantly different from those of the metestrus and diestrus. In all stages of estrous cycle the mean concentration of serum Ca++ was higher than that in the uterine fluid. The difference between serum and uterine fluid Ca++ in estrus, metestrus and diestrus was significant. There was no significant difference between serum Mg++ content nor was it different from uterine fluid Mg++ content at any stages of estrous cycle. In all stages of estrous cycle the uterine fluid Mg++ was higher than that of the serum. These results suggest that during the estrous cycle in the cow, Ca++ is passively secreted in uterine fluids and is mostly dependent on blood serum Ca++ variations but Mg++ is secreted independently and does not follow variations in the serum concentrations.

  10. A sensitive double-antibody enzyme-linked immunosorbant assay for bovine myeloperoxidase and its application to serum and neutrophil extracts.

    Science.gov (United States)

    Cooray, R; Petersson, C G; Moberg, L; Båge, R

    1995-10-01

    The heme enzyme myeloperoxidase (MPO), with a spectral A430/A280 ratio > 0.78, was purified from isolated bovine neutrophils. Using highly specific anti-MPO monoclonal and anti-MPO polyclonal antibodies raised against MPO, a specific and sensitive double-antibody enzyme-linked immunosorbant assay (ELISA) was developed to measure bovine MPO in serum and neutrophil extracts. The ELISA shows good precision and accuracy, with intra- and interassay coefficients of variation of < 10% for MPO concentrations ranging from 0.5 to 50 ng/ml. The accuracy of the ELISA for measuring MPO in bovine serum was further confirmed by the similarity between the standard curve and curves obtained with successive dilutions of MPO-rich serum samples. The mean analytical recovery of MPO from serum was approximately 90%. Long delays between blood sampling and serum preparation were found to affect the level of MPO in the serum. Mean MPO values in the serum of healthy adult cows were 6.5 ng/ml, with a range of 3.5-15.3 ng/ml. In dairy cows with acute mastitis, mean serum MPO values were approximately 30 ng/ml, with a range of 6.0-59.6 ng/ml, and the elevation was markedly higher than the normal values (P = 0.0001). In isolated neutrophils from healthy cattle, MPO concentrations were found to be 7 x 10(-4) ng, with a range of 6.5-8.3 x 10(-4) ng/neutrophil. The ELISA was used to study the distribution of MPO in the bovine neutrophil granules; it was found to be localized to one distinct compartment.

  11. Automated column liquid chromatographic determination of amoxicillin and cefadroxil in bovine serum and muscle tissue using on-line dialysis for sample preparation

    NARCIS (Netherlands)

    Snippe, N; van de Merbel, N C; Ruiter, F P; Steijger, O M; Lingeman, H; Brinkman, U A

    1994-01-01

    A fully automated method is described for the determination of amoxicillin and cefadroxil in bovine serum and muscle tissue. The method is based on the on-line combination of dialysis and solid-phase extraction for sample preparation, and column liquid chromatography with ultraviolet detection. In

  12. Pulse radiolysis investigation of the reaction of the electronic adduct of bovine serum albumin with oxygen. Polychromatic kinetics of the reaction with adsorbed oxygen

    International Nuclear Information System (INIS)

    Pribush, A.G.

    1986-01-01

    The method of pulse radiolysis was used to investigate the reaction of the electronic adduct of bovine serum albumin with oxygen. It was suggested that the disappearance of the electronic adduct of the protein occurs in the course of its interaction with oxygen adsorbed on the globular protein molecule

  13. Persistent Bovine Viral Diarrhea Virus infection in domestic and wild small ruminants and camelids including the mountain goat (Oreamnos americanus

    Directory of Open Access Journals (Sweden)

    Danielle Darracq Nelson

    2016-01-01

    Full Text Available Bovine viral diarrhea virus (BVDV is a Pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus.

  14. Persistent Bovine Viral Diarrhea Virus Infection in Domestic and Wild Small Ruminants and Camelids Including the Mountain Goat (Oreamnos americanus)

    Science.gov (United States)

    Nelson, Danielle D.; Duprau, Jennifer L.; Wolff, Peregrine L.; Evermann, James F.

    2016-01-01

    Bovine viral diarrhea virus (BVDV) is a pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus). PMID:26779126

  15. Persistent Bovine Viral Diarrhea Virus Infection in Domestic and Wild Small Ruminants and Camelids Including the Mountain Goat (Oreamnos americanus).

    Science.gov (United States)

    Nelson, Danielle D; Duprau, Jennifer L; Wolff, Peregrine L; Evermann, James F

    2015-01-01

    Bovine viral diarrhea virus (BVDV) is a pestivirus best known for causing a variety of disease syndromes in cattle, including gastrointestinal disease, reproductive insufficiency, immunosuppression, mucosal disease, and hemorrhagic syndrome. The virus can be spread by transiently infected individuals and by persistently infected animals that may be asymptomatic while shedding large amounts of virus throughout their lifetime. BVDV has been reported in over 40 domestic and free-ranging species, and persistent infection has been described in eight of those species: white-tailed deer, mule deer, eland, mousedeer, mountain goats, alpacas, sheep, and domestic swine. This paper reviews the various aspects of BVDV transmission, disease syndromes, diagnosis, control, and prevention, as well as examines BVDV infection in domestic and wild small ruminants and camelids including mountain goats (Oreamnos americanus).

  16. Spectroscopic analyses and studies on respective interaction of cyanuric acid and uric acid with bovine serum albumin and melamine

    Science.gov (United States)

    Chen, Dandan; Wu, Qiong; Wang, Jun; Wang, Qi; Qiao, Heng

    2015-01-01

    In this work, the fluorescence quenching was used to study the interaction of cyanuric acid (CYA) and uric acid (UA) with bovine serum albumin (BSA) at two different temperatures (283 K and 310 K). The bimolecular quenching constant (Kq), apparent quenching constant (Ksv), effective binding constant (KA) and corresponding dissociation constant (KD), binding site number (n) and binding distance (r) were calculated by adopting Stern-Volmer, Lineweaver-Burk, Double logarithm and overlap integral equations. The results show that CYA and UA are both able to obviously bind to BSA, but the binding strength order is BSA + CYA < BSA + UA. And then, the interactions of CYA and UA with melamine (MEL) under the same conditions were also studied by using similar methods. The results indicates that both CYA and UA can bind together closely with melamine (MEL). It is wished that these research results would facilitate the understanding the formation of kidney stones and gout in the body after ingesting excess MEL.

  17. Comparative studies on the interaction of caffeic acid, chlorogenic acid and ferulic acid with bovine serum albumin

    Science.gov (United States)

    Li, Shuang; Huang, Kelong; Zhong, Ming; Guo, Jun; Wang, Wei-zheng; Zhu, Ronghua

    2010-10-01

    The substitution of the hydrogen on aromatic and esterification of carboxyl group of the phenol compounds plays an important role in their bio-activities. In this paper, caffeic acid (CaA), chlorogenic acid (ChA) and ferulic acid (FA) were selected to investigate the binding to bovine serum albumin (BSA) using UV absorption spectroscopy, fluorescence spectroscopy and synchronous fluorescence spectroscopy. It was found that the methoxyl group substituting for the 3-hydroxyl group of CaA decreased the affinity for BSA and the esterification of carboxyl group of CaA with quinic acid increased the affinities. The affinities of ChA and FA with BSA were more sensitive to the temperature than that of CaA with BSA. Synchronous fluorescence spectroscopy and time-resolved fluorescence indicated that the Stern-Volmer plots largely deviated from linearity at high concentrations and were caused by complete quenching of the tyrosine fluorescence of BSA.

  18. Kinetics of Glycoxidation of Bovine Serum Albumin by Methylglyoxal and Glyoxal and its Prevention by Various Compounds

    Directory of Open Access Journals (Sweden)

    Izabela Sadowska-Bartosz

    2014-04-01

    Full Text Available The aim of this study was to compare several methods for measurement of bovine serum albumin (BSA modification by glycoxidation with reactive dicarbonyl compounds (methylglyoxal ‒ MGO and glyoxal ‒ GO, for studies of the kinetics of this process and to compare the effects of 19 selected compounds on BSA glycation by the aldehydes. The results confirm the higher reactivity of MGO with respect to GO and point to the usefulness of AGE, dityrosine and N′-formylkynurenine fluorescence for monitoring glycation and evaluation of protection against glycation. Different extent of protection against glycation induced by MGO and GO was found for many compounds, probably reflecting effects on various stages of the glycation process. Polyphenols (genistein, naringin and ellagic acid were found to protect against aldehyde-induced glycation; 1-cyano-4-hydroxycinnamic acid was also an effective protector.

  19. The fluorescence spectroscopic study on the interaction between imidazo[2,1-b]thiazole analogues and bovine serum albumin

    Science.gov (United States)

    Yu, Xianyong; Yang, Ying; Shiyu, Lu; Yao, Qing; Heting, Liu; Xiaofang, Li; Pinggui, Yi

    2011-12-01

    The interaction between imidazo[2,1-b]thiazole (IMTZ) and bovine serum albumin (BSA) was analyzed by fluorescence and ultraviolet spectroscopy at 302 and 310 K under simulative physiological conditions. The results show that IMTZ can effectively quench the intrinsic fluorescence of BSA via static and dynamic quenching. The binding constant, binding sites of IMTZ with BSA were calculated. According to the Förster non-radiation energy transfer theory, the average binding distance between IMTZ and BSA was obtained. What's more, the synchronous fluorescence spectra indicated that the conformation of BSA has been changed. The results provided the information for the binding of IMTZ to BSA, and the influences of substituent group on the interaction were also discussed.

  20. Fabrication of curcumin-loaded bovine serum albumin (BSA)-dextran nanoparticles and the cellular antioxidant activity.

    Science.gov (United States)

    Fan, Yuting; Yi, Jiang; Zhang, Yuzhu; Yokoyama, Wallace

    2018-01-15

    Bovine serum albumin (BSA)-dextran conjugate was prepared with glycation. Self-assembly nanoparticles were synthesized with a green, and facile approach. The effects of dry-heating time on the fabrication and characteristics of BSA-dextran conjugate nanoparticles were examined. Stable nanoparticles (nanoparticles were stable in a wide pH range (pH 2.0-7.0). The particle size of nanoparticles increased to 115nm after curcumin incorporation and was stable even after one-month storage. TEM results demonstrated that curcumin-loaded nanoparticles displayed a spherical structure and were homogeneously dispersed. Curcumin in BSA-dextran nanoparticle showed better stability, compared to free curcumin. In addition, BSA-dextran nanoparticles can improve the cellular antioxidant activity of curcumin in Caco-2 cells. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Biomimetic synthesis of hollow calcium carbonate with the existence of the agar matrix and bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Jianhua, E-mail: fjh2008@126.com; Wu, Gang; Qing, Chengsong

    2016-01-01

    Proteins play important roles in the process of biomineralization. Vaterite and calcite have been synthesized by the reaction of Na{sub 2}CO{sub 3} and CaCl{sub 2} in the bovine serum albumin (BSA) and agar system. The samples have been characterized by Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). The shape of CaCO{sub 3} crystal has been analyzed by scanning electronic microscopy (SEM). The results show that calcite is a single product in the absence of BSA, but the product is a mixture of calcite and vaterite in the presence of BSA. The spheral shell of CaCO{sub 3} crystal was obtained when the concentration of BSA increased to 9.0 mg/mL. - Highlights: • Biomimetic synthesis of hollow calcium carbonate • Calcification mechanisms in the presence of both protein and polysaccharides • Biomineralization under the action of protein and polysaccharides.

  2. Preparation and characterization of nanosized P(NIPAM-MBA) hydrogel particles and adsorption of bovine serum albumin on their surface.

    Science.gov (United States)

    Zhu, Xiaoli; Gu, Xiangling; Zhang, Lina; Kong, Xiang-Zheng

    2012-09-24

    Thermosensitive polymer hydrogel particles with size varying from 480 to 620 nm were prepared through precipitation copolymerization of N-isopropylacrylamide with N,N'-methylenebisacrylamide (MBA) in water with ammonium persulfate as the initiator. Only polymer hydrogels without any coagula were obtained when MBA concentration in the monomer mixture was kept between 2.5 and 10.0 wt%; with increased MBA concentration, the monomer conversion was enhanced, the size of the hydrogels was increased, and their shrinking was lessened when heated from 25°C to 40°C. Bovine serum albumin adsorption on the surface of the hydrogels of different MBA content was measured at different pH levels and under different temperatures. The results demonstrated that the adsorption of the protein on the hydrogels could be controlled by adjusting the pH, the temperature of adsorption, and the crosslinking in the hydrogels. The results were interpreted, and the mechanisms of the polymerization were proposed.

  3. In-vitro displacement interaction of atenolol and amlodipine on binding with bovine serum albumin when co-administered

    Directory of Open Access Journals (Sweden)

    Md. Ashraful Alam, Md. Abdul Awal, Mahbub Mostofa, Md. Kamrul Islam and Nusrat Subhan

    2007-06-01

    Full Text Available The binding of atenolol (selective β1-blocker and amlodipine (calcium channel blocker to bovine serum albumin (BSA was studied by equilibrium dialysis method in order to have an insight into the binding chemistry of these two to BSA. Free atenolol concentration was increased due to addition of amlodipine which reduced the binding of the compounds to BSA. However, the free fraction was increased to a level as it was expected from direct competitive displacement while the free atenolol concentration was increased according to increasing the amlodipine concentration when only the BSA was present. The result obtained when the binding site was blocked by sufficient amount of amlodipine was that the increment of free concentration of atenolol was prominent. When no amlodipine was added the free concentration of atenolol was only 28% whereas this release was 93 % to 98.01% when amlodipine was added with increasing concentration.

  4. Biosorption of bovine serum albumin by Ulva lactuca biomass from industrial wastewater: Equilibrium, kinetic and thermodynamic study

    Energy Technology Data Exchange (ETDEWEB)

    Rathinam, Aravindhan [Chemical Laboratory, Central Leather Research Institute, Adyar, Chennai 600020 (India); Zou, Linda, E-mail: linda.zou@unisa.edu.au [SA Water Centre for Water Management and Reuse, University of South Australia, Mawson Lakes Campus, Adelaide, SA 5095 (Australia)

    2010-12-15

    Batch biosorption experiments have been carried out for the removal of bovine serum albumin (BSA) from simulated industrial wastewater onto Ulva lactuca seaweed. Various vital parameters influencing the biosorption process such as initial concentration of BSA, pH of the solution, adsorbent dosage and temperature have been determined. The biosorption kinetics follows a pseudo-second order kinetic model. Equilibrium isotherm studies demonstrate that the biosorption followed the Freundlich isotherm model, which implies a heterogeneous sorption phenomenon. Various thermodynamic parameters such as changes in enthalpy, free energy and entropy have been calculated. The positive value of {Delta}H{sup o} and the negative value of {Delta}G{sup o} show that the sorption process is endothermic and spontaneous. The positive value of change in entropy {Delta}S{sup o} shows increased randomness at the solid-liquid interface during the biosorption of BSA onto U. lactuca seaweed.

  5. Biosorption of bovine serum albumin by Ulva lactuca biomass from industrial wastewater: equilibrium, kinetic and thermodynamic study.

    Science.gov (United States)

    Rathinam, Aravindhan; Zou, Linda

    2010-12-15

    Batch biosorption experiments have been carried out for the removal of bovine serum albumin (BSA) from simulated industrial wastewater onto Ulva lactuca seaweed. Various vital parameters influencing the biosorption process such as initial concentration of BSA, pH of the solution, adsorbent dosage and temperature have been determined. The biosorption kinetics follows a pseudo-second order kinetic model. Equilibrium isotherm studies demonstrate that the biosorption followed the Freundlich isotherm model, which implies a heterogeneous sorption phenomenon. Various thermodynamic parameters such as changes in enthalpy, free energy and entropy have been calculated. The positive value of ΔH° and the negative value of ΔG° show that the sorption process is endothermic and spontaneous. The positive value of change in entropy ΔS° shows increased randomness at the solid-liquid interface during the biosorption of BSA onto U. lactuca seaweed. Copyright © 2010 Elsevier B.V. All rights reserved.

  6. Biosorption of bovine serum albumin by Ulva lactuca biomass from industrial wastewater: Equilibrium, kinetic and thermodynamic study

    International Nuclear Information System (INIS)

    Rathinam, Aravindhan; Zou, Linda

    2010-01-01

    Batch biosorption experiments have been carried out for the removal of bovine serum albumin (BSA) from simulated industrial wastewater onto Ulva lactuca seaweed. Various vital parameters influencing the biosorption process such as initial concentration of BSA, pH of the solution, adsorbent dosage and temperature have been determined. The biosorption kinetics follows a pseudo-second order kinetic model. Equilibrium isotherm studies demonstrate that the biosorption followed the Freundlich isotherm model, which implies a heterogeneous sorption phenomenon. Various thermodynamic parameters such as changes in enthalpy, free energy and entropy have been calculated. The positive value of ΔH o and the negative value of ΔG o show that the sorption process is endothermic and spontaneous. The positive value of change in entropy ΔS o shows increased randomness at the solid-liquid interface during the biosorption of BSA onto U. lactuca seaweed.

  7. STUDIES ON THE INTERACTION BETWEEN TRIPTOLIDE AND BOVINE SERUM ALBUMIN (BSA) BY SPECTROSCOPIC AND MOLECULAR MODELING METHODS.

    Science.gov (United States)

    Wang, Haidong; Shi, Hailang; Pang, Jie; Song, Xingfa; Xu, Caiyun; Sun, Zengxian

    2016-01-01

    Triptolide is a major active constituent isolated from Tripterygiumwilfordii Hook F, a Chinese herbal medicine. This study investigated the intermolecular interaction between triptolide and bovine serum albumin (BSA). The fluorescence, circular dichroism (CD) and molecular docking methods were used to investigate the intermolecular interaction between triptolide and BSA. The binding constant, the number of binding sites, binding subdomain and the thermodynamic parameters were measured. The results of this experiment revealed that the intrinsic fluorescence of BSA was effectively quenched by triptolide via static quenching. The experimental results of synchronous fluorescence and CD spectra showed that the conformation of BSA was changed in the presence of triptolide. It indicated that triptolide could spontaneously bind on site II (subdomain IIIA) of BSA mainly via hydrogen bonding interactions and Van der Waals force.

  8. From guest to ligand - A study on the competing interactions of antitumor drug resveratrol with {beta}-cyclodextrin and bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Li, Xudong [College of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, Shandong Province 252059 (China); Department of Clinical Laboratory, Liaocheng People' s Hospital, Liaocheng, Shandong Province 252000 (China); Li, Hui [College of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, Shandong Province 252059 (China); Liu, Min, E-mail: liumin_panpan@163.com [College of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, Shandong Province 252059 (China); Li, Guangqian; Li, Linwei [College of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, Shandong Province 252059 (China); Sun, Dezhi, E-mail: sundezhisdz@163.com [College of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng, Shandong Province 252059 (China)

    2011-07-10

    Graphical abstract: Thermodynamic behavior of the interaction between bovine serum albumin and antitumor drug resveratrol delivered by {beta}-cyclodextrin in buffer solutions (pH 7.40) have been investigated by ITC combined with UV, FS and circular dichroism at 298.15 K. The results indicated that the affinity of resveratrol with the host ({beta}-cyclodextrin) was evidently weaker than that of the drug with the both classes of binding sites on the protein molecule. Highlights: {yields} Supramolecular complex of a drug with BSA could form in aqueous medium. {yields} A set of thermodynamic parameters were determined. {yields} Affinity of the drug to {beta}-CD is weaker than that of it to the protein. {yields} The molecular conformation of BSA was (slightly) changed by the drug. - Abstract: Interaction between bovine serum albumin (BSA) and resveratrol (RES) included by {beta}-cyclodextrin ({beta}-CD) in Tris-HCl aqueous buffer solutions (pH 7.4) has been investigated by isothermal titration calorimetry (ITC) combined with ultraviolet, fluorescence and circular dichroism spectra analyses. The results indicate that there are two classes of ligand binding sites. The first class of binding is mainly driven by enthalpy, while the second one is driven by both enthalpy and entropy. The secondary structure of BSA in the aqueous system was slightly changed with addition of the drug. Thermodynamic parameters, i.e., equilibrium constants, standard enthalpy changes and the entropy effects for the binding process of RES with BSA were calculated based on the calorimetric data. In fact, due to the poor solubility of RES in aqueous buffer medium, these parameters could not be determined by the employed experimental method without the existence of the CD.

  9. Bovine serum prolactin, growth hormone, and triiodothyronine levels during late pregnancy and early lactation

    International Nuclear Information System (INIS)

    Johke, Tetsu; Hodate, Kouichi

    1977-01-01

    Changes of serum prolactin (PRL), growth hormone (GH), and triiodothyronine (T3) levels in Holstein dairy cows during late pregnancy and early lactation were determined by radioimmunoassays. The cows calved in September. Mean serum PRL ranged 9.9 ng/ml to 12.4 ng/ml during the 3rd to 7th day prepartum. It increased to a peak value of 87.3 ng/ml on the day of parturition from 57.5 ng/ml on the day before calving, and decreased to 13.9 ng/ml within 3 days postpartum. During the 2nd to 30th day prepartum, serum GH varied between 2.4 and 4.0 ng/ml and increased to 8.0 ng/ml on the day of calving. During a week postpartum, serum GH ranged from 5.2 to 8.2 ng/ml. Mean serum GH during a week postpartum was higher than that of prepartum (P < 0.001). Mean serum T3 level during 5 days prepartum varied 104.2 to 108.8 ng/dl and decreased to 93 ng/dl on the day of calving. During 5 days postpartum, mean T3 level varied 73.0 to 83.8 ng/dl. Contrary to GH, mean serum T3 level during 5 day postpartum was significantly lower than that of prepartum (P < 0.001). (auth.)

  10. Genomic and antigenic characterization of bovine parainfluenza-3 viruses in the United States including modified live virus vaccine (MLV) strains and field strains from cattle.

    Science.gov (United States)

    Fulton, R W; Neill, J D; Saliki, J T; Landis, C; Burge, L J; Payton, M E

    2017-05-02

    This study investigated the genetic and antigenic characterization of parainfluenza-3 virus (PI3V) of cattle. Using molecular tests including real time PCR and viral genome sequencing, PI3V strains could be separated into PI3V types, including PI3V A, PI3V B, and PI3V C. Isolates from cattle with bovine respiratory disease clinical signs and commercial vaccines in the U.S. with MLV PI3V were typed using these molecular tests. All the MLV vaccine strains tested were PI3V A. In most cases PI3V field strains from calves receiving MLV vaccines were types heterologous to the vaccine type A. Also antigenic differences were noted as PI3V C strains had lower antibody levels than PI3V A in serums from cattle receiving MLV PI3V A vaccines. This study further demonstrates there is genetic variability of U.S. PI3V strains and also antigenic variability. In addition, isolates from cattle with BRD signs and receiving MLV vaccines may have heterologous types to the vaccines, and molecular tests should be performed to differentiate field from vaccine strains. Potentially the efficacy of current PI3V A vaccines should be evaluated with other types such a PI3V B and PI3V C. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Comparative epigenetic influence of autologous versus fetal bovine serum on mesenchymal stem cells through in vitro osteogenic and adipogenic differentiation

    International Nuclear Information System (INIS)

    Fani, Nesa; Ziadlou, Reihane; Shahhoseini, Maryam; Baghaban Eslaminejad, Mohamadreza

    2016-01-01

    Mesenchymal stem cells (MSCs) derived from bone marrow (BM) represents a useful source of adult stem cells for cell therapy and tissue engineering. MSCs are present at a low frequency in the BM; therefore expansion is necessary before performing clinical studies. Fetal bovine serum (FBS) as a nutritional supplement for in vitro culture of MSCs is a suitable additive for human cell culture, but not regarding subsequent use of these cells for clinical treatment of human patients due to the risk of viral and prion transmission as well as xenogeneic immune responses after transplantation. Recently, autologous serum (AS) has been as a supplement to replace FBS in culture medium. We compared the effect of FBS versus AS on the histone modification pattern of MSCs through in vitro osteogenesis and adipogenesis. Differentiation of stem cells under various serum conditions to a committed state involves global changes in epigenetic patterns that are critically determined by chromatin modifications. Chromatin immunoprecipitation (ChIP) coupled with real-time PCR showed significant changes in the acetylation and methylation patterns in lysine 9 (Lys9) of histone H3 on the regulatory regions of stemness (Nanog, Sox2, Rex1), osteogenic (Runx2, Oc, Sp7) and adipogenic (Ppar-γ, Lpl, adiponectin) marker genes in undifferentiated MSCs, FBS and AS. All epigenetic changes occurred in a serum dependent manner which resulted in higher expression level of stemness genes in undifferentiated MSCs compared to differentiated MSCs and increased expression levels of osteogenic genes in AS compared to FBS. Adipogenic genes showed greater expression in FBS compared to AS. These findings have demonstrated the epigenetic influence of serum culture conditions on differentiation potential of MSCs, which suggest that AS is possibly more efficient serum for osteogenic differentiation of MSCs in cell therapy purposes. - Highlights: • Bone marrow derived MSC could proliferate in AS as well as in FBS

  12. Comparative epigenetic influence of autologous versus fetal bovine serum on mesenchymal stem cells through in vitro osteogenic and adipogenic differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Fani, Nesa [Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran (Iran, Islamic Republic of); Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran (Iran, Islamic Republic of); Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Ziadlou, Reihane [Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran (Iran, Islamic Republic of); Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran (Iran, Islamic Republic of); Shahhoseini, Maryam, E-mail: m.shahhoseini@royaninstitute.org [Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran (Iran, Islamic Republic of); Baghaban Eslaminejad, Mohamadreza, E-mail: eslami@royaninstitute.org [Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran (Iran, Islamic Republic of)

    2016-06-10

    Mesenchymal stem cells (MSCs) derived from bone marrow (BM) represents a useful source of adult stem cells for cell therapy and tissue engineering. MSCs are present at a low frequency in the BM; therefore expansion is necessary before performing clinical studies. Fetal bovine serum (FBS) as a nutritional supplement for in vitro culture of MSCs is a suitable additive for human cell culture, but not regarding subsequent use of these cells for clinical treatment of human patients due to the risk of viral and prion transmission as well as xenogeneic immune responses after transplantation. Recently, autologous serum (AS) has been as a supplement to replace FBS in culture medium. We compared the effect of FBS versus AS on the histone modification pattern of MSCs through in vitro osteogenesis and adipogenesis. Differentiation of stem cells under various serum conditions to a committed state involves global changes in epigenetic patterns that are critically determined by chromatin modifications. Chromatin immunoprecipitation (ChIP) coupled with real-time PCR showed significant changes in the acetylation and methylation patterns in lysine 9 (Lys9) of histone H3 on the regulatory regions of stemness (Nanog, Sox2, Rex1), osteogenic (Runx2, Oc, Sp7) and adipogenic (Ppar-γ, Lpl, adiponectin) marker genes in undifferentiated MSCs, FBS and AS. All epigenetic changes occurred in a serum dependent manner which resulted in higher expression level of stemness genes in undifferentiated MSCs compared to differentiated MSCs and increased expression levels of osteogenic genes in AS compared to FBS. Adipogenic genes showed greater expression in FBS compared to AS. These findings have demonstrated the epigenetic influence of serum culture conditions on differentiation potential of MSCs, which suggest that AS is possibly more efficient serum for osteogenic differentiation of MSCs in cell therapy purposes. - Highlights: • Bone marrow derived MSC could proliferate in AS as well as in FBS

  13. Characterization of 6-mercaptopurine binding to bovine serum albumin and its displacement from the binding sites by quercetin and rutin

    Energy Technology Data Exchange (ETDEWEB)

    Ehteshami, Mehdi [Nutrition Research Center, School of Health and Nutrition, Tabriz University of Medical Sciences, Tabriz 51644-14766 (Iran, Islamic Republic of); Rasoulzadeh, Farzaneh [Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz 51644-14766 (Iran, Islamic Republic of); Mahboob, Soltanali [Nutrition Research Center, School of Health and Nutrition, Tabriz University of Medical Sciences, Tabriz 51644-14766 (Iran, Islamic Republic of); Rashidi, Mohammad-Reza, E-mail: rashidi@tbzmed.ac.ir [Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz 51644-14766 (Iran, Islamic Republic of)

    2013-03-15

    Binding of a drug to the serum albumins as major serum transport proteins can be influenced by other ligands leading to alteration of its pharmacological properties. In the present study, binding characteristics of 6-mercaptopurine (6-MP) with bovine serum albumin (BSA) together with its displacement from its binding site by quercetin and rutin have been investigated by the spectroscopic method. According to the binding parameters, a static quenching component in overall dynamic quenching process is operative in the interaction between 6-MP and BSA. The binding of 6-MP to BSA occurred spontaneously due to entropy-driven hydrophobic interactions. The synchronous fluorescence spectroscopy study revealed that the secondary structure of BSA is changed in the presence of 6-MP and both Tyr and Trp residues participate in the interaction between 6-MP and BSA with the later one being more dominant. The binding constant value of 6-MP-BSA in the presence of quercetin and rutin increased. 6-MP was displaced by ibuprofen indicating that the binding site of 6-MP on albumin is site II. Therefore, the change of the pharmacokinetic and pharmacodynamic properties of 6-MP by quercetin and rutin through alteration of binding capacity of 6-MP to the serum albumin cannot be ruled out. In addition, the displacement study showed that 6-MP is located in site II of BSA. - Highlights: Black-Right-Pointing-Pointer Participation of both Tyr and particularly Trp residues in the interaction between 6-MP and BSA. Black-Right-Pointing-Pointer Involvement of a static quenching component in an overall dynamic quenching process. Black-Right-Pointing-Pointer Ability of quercetin and rutin to change the binding constants of 6-MP-BSA complex. Black-Right-Pointing-Pointer Binding of 6-MP to BSA through entropy-driven hydrophobic interactions.

  14. Effect of Human and Bovine Serum Albumin on kinetic Chemiluminescence of Mn (III-Tetrakis (4-Sulfonatophenyl Porphyrin-Luminol-Hydrogen Peroxide System

    Directory of Open Access Journals (Sweden)

    Sayed Yahya Kazemi

    2012-01-01

    Full Text Available The present work deals with an attempt to study the effect of human and bovine serum albumin on kinetic parameters of chemiluminescence of luminol-hydrogen peroxide system catalyzed by manganese tetrasulfonatophenyl porphyrin (MnTSPP. The investigated parameters involved pseudo-first-order rise and fall rate constant for the chemiluminescence burst, maximum level intensity, time to reach maximum intensity, total light yield, and values of the intensity at maximum CL which were evaluated by nonlinear least square program KINFIT. Because of interaction of metalloporphyrin with proteins, the CL parameters are drastically affected. The systems resulted in Stern-Volmer plots with values of 3.17×105 and 3.7×105M−1 in the quencher concentration range of 1.5×10−6 to 1.5×10−5 M for human serum albumin (HSA and bovine serum albumin (BSA, respectively.

  15. Study of Interactions of an Anticancer Drug Neratinib With Bovine Serum Albumin: Spectroscopic and Molecular Docking Approach

    Directory of Open Access Journals (Sweden)

    Tanveer A. Wani

    2018-03-01

    Full Text Available Binding of therapeutic agents to plasma proteins, particularly to serum albumin, provides valuable information in the drug development. This study was designed to evaluate the binding interaction of neratinib with bovine serum albumin (BSA. Neratinib blocks HER2 signaling and is effective in trastuzumab-resistant breast cancer treatment. Spectrofluorometric, UV spectrophotometric, and fourier transform infrared (FT-IR and molecular docking experiments were performed to study this interaction. The fluorescence of BSA is attributed to the presence of tryptophan (Trp residues. The fluorescence of BSA in presence of neratinib was studied using the excitation wavelength of 280 nm and the emission was measured at 300-500 nm at three different temperatures. Neratinib quenched the BSA intrinsic fluorescence by static mechanism. A complex formation occurred due to the interaction leading to BSA absorption shift. The fluorescence, UV- absorption, three dimensional fluorescence and FT-IR data showed conformational changes occurred in BSA after interaction with neratinib. The binding constant values decreased as the temperature increased suggesting an instable complex formation at high temperature. Site I (sub-domain IIA was observed as the principal binding site for neratinib. Hydrogen bonding and Van der Waals forces were suggested to be involved in the BSA-neratinib interaction due to the negative values of entropy and enthalpy changes.

  16. Probing Temperature- and pH-Dependent Binding between Quantum Dots and Bovine Serum Albumin by Fluorescence Correlation Spectroscopy

    Directory of Open Access Journals (Sweden)

    Zonghua Wang

    2017-04-01

    Full Text Available Luminescent quantum dots (QDs with unique optical properties have potential applications in bio-imaging. The interaction between QDs and bio-molecules is important to the biological effect of QDs in vivo. In this paper, we have employed fluorescence correlation spectroscopy (FCS to probe the temperature- and pH-dependent interactions between CdSe QDs with carboxyl (QDs-COOH and bovine serum albumin (BSA in buffer solutions. The results have shown that microscopic dissociation constant K′D is in the range of (1.5 ± 0.2 × 10−5 to (8.6 ± 0.1 × 10−7 M, the Hill coefficient n is from 0.4 to 2.3, and the protein corona thickness is from 3.0 to 9.4 nm. Variable-temperature measurements have shown both negative values of ∆H and ∆S for BSA adsorption on QDs-COOH, while pH has a profound effect on the adsorption. Additional, FCS measurement QDs-COOH and proteins in whole mice serum and plasma samples has also been conducted. Finally, simulation results have shown four favored QD binding sites in BSA.

  17. Molecular binding of toxic phenothiazinium derivatives, azures to bovine serum albumin: A comparative spectroscopic, calorimetric, and in silico study.

    Science.gov (United States)

    Das, Somnath; Islam, Md Maidul; Jana, Gopal Chandra; Patra, Anirudha; Jha, Pradeep K; Hossain, Maidul

    2017-07-01

    In this paper, the comparative binding behavior of antimalarial drug azure A, azure B and azure C with bovine serum albumin (BSA) has been studied. The interaction has been confirmed by multispectroscopic (UV, fluorescence, Fourier transform infrared (FT-IR), and circular dichroism) and molecular docking techniques. The experimental results show that azure B has the highest BSA binding affinity followed by azure A and azure C. The experimental evidence of binding showed a static quenching mechanism in the interaction azures with BSA. The isothermal titration calorimetry result reveals that the binding was exothermic with positive entropy contribution in each case. The thermodynamic parameters ΔH, ΔG, and ΔS at 25°C were calculated, which indicates that the weak van der Waals forces and hydrogen bonding rather than the hydrophobic effect played an important role in the interaction. According to the theory of Förster nonradiative energy transfer, the distance (r) between the donor (BSA) and acceptor azures found to be azures-BSA system. Overall, experimental studies characterize the interaction dynamics and energetics of the binding of three toxic analogs towards the physiologically relevant serum albumins. We hope, the outcome of this work will be most helpful for synthesizing a new type of phenothiazinium derivatives of the better therapeutic application. Copyright © 2017 John Wiley & Sons, Ltd.

  18. Influence of galloyl moiety in interaction of epicatechin with bovine serum albumin: a spectroscopic and thermodynamic characterization.

    Science.gov (United States)

    Pal, Sandip; Saha, Chabita; Hossain, Maidul; Dey, Subrata Kumar; Kumar, Gopinatha Suresh

    2012-01-01

    The health benefits stemming from green tea are well known, but the exact mechanism of its biological activity is not elucidated. Epicatechin (EC) and epicatechin gallate (ECG) are two dietary catechins ubiquitously present in green tea. Serum albumins functionally carry these catechins through the circulatory system and eliminate reactive oxygen species (ROS) induced injury. In the present study ECG is observed to have higher antioxidant activity; which is attributed to the presence of galloyl moiety. The binding affinity of these catechins to bovine serum albumin (BSA) will govern the efficacy of their biological activity. EC and ECG bind with BSA with binding constants 1.0 × 10(6) M(-1) and 6.6 × 10(7) M(-1), respectively. Changes in secondary structure of BSA on interaction with EC and ECG have been identified by circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy. Thermodynamic characterization reveals the binding process to be exothermic, spontaneous and entropy driven. Mixed binding forces (hydrophobic, electrostatic and hydrogen bonding) exist between ECG and BSA. Binding site for EC is primarily site-II in sub-domain IIIA of BSA and for ECG; it is site-I in sub-domain IIA. ECG with its high antioxidant activity accompanied by high affinity for BSA could be a model in drug designing.

  19. Influence of galloyl moiety in interaction of epicatechin with bovine serum albumin: a spectroscopic and thermodynamic characterization.

    Directory of Open Access Journals (Sweden)

    Sandip Pal

    Full Text Available The health benefits stemming from green tea are well known, but the exact mechanism of its biological activity is not elucidated. Epicatechin (EC and epicatechin gallate (ECG are two dietary catechins ubiquitously present in green tea. Serum albumins functionally carry these catechins through the circulatory system and eliminate reactive oxygen species (ROS induced injury. In the present study ECG is observed to have higher antioxidant activity; which is attributed to the presence of galloyl moiety. The binding affinity of these catechins to bovine serum albumin (BSA will govern the efficacy of their biological activity. EC and ECG bind with BSA with binding constants 1.0 × 10(6 M(-1 and 6.6 × 10(7 M(-1, respectively. Changes in secondary structure of BSA on interaction with EC and ECG have been identified by circular dichroism (CD and Fourier transform infrared (FT-IR spectroscopy. Thermodynamic characterization reveals the binding process to be exothermic, spontaneous and entropy driven. Mixed binding forces (hydrophobic, electrostatic and hydrogen bonding exist between ECG and BSA. Binding site for EC is primarily site-II in sub-domain IIIA of BSA and for ECG; it is site-I in sub-domain IIA. ECG with its high antioxidant activity accompanied by high affinity for BSA could be a model in drug designing.

  20. Unveiling the stimulatory effects of tartrazine on human and bovine serum albumin fibrillogenesis: Spectroscopic and microscopic study

    Science.gov (United States)

    Al-Shabib, Nasser Abdulatif; Khan, Javed Masood; Alsenaidy, Mohammad A.; Alsenaidy, Abdulrahman M.; Khan, Mohd Shahnawaz; Husain, Fohad Mabood; Khan, Mohammad Rashid; Naseem, Mohammad; Sen, Priyankar; Alam, Parvez; Khan, Rizwan Hasan

    2018-02-01

    Amyloid fibrils are playing key role in the pathogenesis of various neurodegenerative diseases. Generally anionic molecules are known to induce amyloid fibril in several proteins. In this work, we have studied the effect of anionic food additive dye i.e., tartrazine (TZ) on the amyloid fibril formation of human serum albumins (HSA) and bovine serum albumin (BSA) at pHs 7.4 and 3.5. We have employed various biophysical methods like, turbidity measurements, Rayleigh Light Scattering (RLS), Dynamic Light Scattering (DLS), intrinsic fluorescence, Congo red assay, far-UV CD, transmission electron microscopy (TEM) and atomic force microscopy (AFM) to decipher the mechanism of TZ-induce amyloid fibril formation in both the serum albumins at pHs 7.4 and 3.5. The obtained results suggest that both the albumins forms amyloid-like aggregates in the presence of 1.0 to 15.0 mM of TZ at pH 3.5, but no amyloid fibril were seen at pH 7.4. The possible cause of TZ-induced amyloid fibril formation is electrostatic and hydrophobic interaction because sulfate group of TZ may have interacted electrostatically with positively charged amino acids of the albumins at pH 3.5 and increased protein-protein and protein-TZ interactions leading to amyloid fibril formation. The TEM, RLS and DLS results are suggesting that BSA forms bigger size amyloids compared to HSA, may be due to high surface hydrophobicity of BSA.

  1. Interaction of norfloxacin with bovine serum albumin studied by different spectrometric methods; displacement studies, molecular modeling and chemometrics approaches

    Energy Technology Data Exchange (ETDEWEB)

    Naseri, Abdolhossein, E-mail: a_naseri@tabrizu.ac.ir [Departments of Analytical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz 51666-16471 (Iran, Islamic Republic of); Hosseini, Soheila [Departments of Analytical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz 51666-16471 (Iran, Islamic Republic of); Rasoulzadeh, Farzaneh [Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz 51644-14766 (Iran, Islamic Republic of); Rashidi, Mohammad-Reza [Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz 51644-14766 (Iran, Islamic Republic of); Zakery, Maryam; Khayamian, Taghi [Department of Chemistry, College of Chemistry, Isfahan University of Technology, Isfahan 84154 (Iran, Islamic Republic of)

    2015-01-15

    Serum albumins as major target proteins can bind to other ligands leading to alteration of their pharmacological properties. The mechanism of interaction between norfloxacin (NFLX) with bovine serum albumin (BSA) was investigated. Fuorescence quenching of serum albumin by this drug was found to be a static quenching process. The binding sites number, n, apparent binding constant, K, and thermodynamic parameters were calculated at different temperatures. The distance, r, between donor, BSA, and acceptor, NFLX, was calculated according to the Forster theory of non-radiation energy transfer. Also binding characteristics of NFLX with BSA together with its displacement from its binding site by kanamycin and effect of common metal ions on binding constant were investigated by the spectroscopic methods. The conformational change in the secondary structure of BSA upon interaction with NFLX was investigated qualitatively from synchronous fluorescence spectra, Fourier Transform Infrared (FTIR) and circular dichroism (CD) spectrometric methods. Molecular docking studies were performed to obtain information on the possible residues involved in the interaction process and changes in accessible surface area of the interacting residues. The results showed that the conformation of BSA changed in the presence of NFLX. For the first time, displacement studies were used for this interaction; displacement studies showed that NFLX was displaced by phenylbutazon and ketoprofen but was not displaced by ibuprofen indicating that the binding site of NFLX on albumin was site I. In addition a powerful chemometrics method, multivariate curve resolution-alternating least square, was used for resolution of spectroscopic augmented data obtained in two different titration modes in order to extract spectral information regardless of spectral overlapping of components. - Highlights: • Interaction between norfloxacin and BSA is studied by spectral methods. • Chemometrics methods are used to

  2. Effect of processing intensity on immunologically active bovine milk serum proteins

    NARCIS (Netherlands)

    Brick, Tabea; Ege, Markus; Boeren, Sjef; Böck, Andreas; Mutius, Von Erika; Vervoort, Jacques; Hettinga, Kasper

    2017-01-01

    Consumption of raw cow’s milk instead of industrially processed milk has been reported to protect children from developing asthma, allergies, and respiratory infections. Several heat-sensitive milk serum proteins have been implied in this effect though unbiased assessment of milk proteins in general

  3. Kinetics of local and systemic isoforms of serum amyloid A in bovine mastitic milk

    DEFF Research Database (Denmark)

    Jacobsen, Stine; Niewold, T.A.; Kornalijnslijper, E.

    2005-01-01

    The aim of the present study was to characterise the serum amyloid A (SAA) response to intramammary inoculation of Escherichia coli and to examine the distribution of hepatically and extrahepatically pruduced SAA isoforms in plasma and milk fra cows with mastitis....

  4. Purification, subunit characterization and ultrastructure of three soluble bovine lectins: conglutinin, mannose-binding protein and the pentraxin serum amyloid P-component

    DEFF Research Database (Denmark)

    Andersen, Ove; Friis, P; Holm Nielsen, E

    1992-01-01

    Conglutinin and mannose-binding protein (MBP) are members of the C-type lectins which are widely present in mammalian plasma. Serum amyloid P-component (SAP) is a member of the pentraxin family with lectin properties. A scheme for the partial purification of all three lectins by carbohydrate...... affinity chromatography and selective elution was developed. The purification was monitored by SDS-PAGE, Western blotting and electron microscopy. Binding of the lectins to Sephadex-iC3b, their collagenase sensitivity, and the size and antibody reactivity of their subunits was investigated....... The demonstration, by SDS-PAGE, of 25-kDa subunits, which were unaffected by collagenase treatment but bound to Sephadex-iC3b and antibodies to human SAP, indicated the existence of bovine SAP. Bovine conglutinin (BK) also showed calcium-dependent binding to Sephadex-iC3b, whereas bovine MBP did not. The binding...

  5. Characterization of erythrosine B binding to bovine serum albumin and bilirubin displacement.

    Science.gov (United States)

    Mathavan, Vinodaran M K; Boh, Boon Kim; Tayyab, Saad

    2009-08-01

    The interaction of crythrosine B (ErB), a commonly used dye for coloring foods and drinks, with bovine scrum albumin (BSA) was investigated both in the absence and presence of bilirubin (BR) using absorption and absorption difference spectroscopy. ErB binding to BSA was reflected from a significant red shift of 11 nm in the absorption maximum of ErB (527 nm) with the change in absorbance at lamdamax. Analysis of absorption difference spectroscopic titration results of BSA with increasing concentrations of ErB3 using Benesi-Hildebrand equation gave the association constant, K as 6.9 x 10(4) M(-1). BR displacing action of ErB was revealed by a significant blue shift in the absorption maximum, accompanied by a decrease in absorbance difference at lamdamax in the difference spectrum of BR-BSA complex upon addition of increasing concentrations of ErB. This was further substantiated by fluorescence spectroscopy, as addition of increasing concentrations of ErB to BR-BSA complex caused a significant decrease in fluoresccnce at 510 nm. The results suggest that ErB binds to a site in the vicinity of BR binding site on BSA. Therefore, intake of ErB may increase the risk of hyperbilirubinemia in the healthy subjects.

  6. Environment sensitive fluorescent analogue of biologically active oxazoles differentially recognizes human serum albumin and bovine serum albumin: Photophysical and molecular modeling studies

    Science.gov (United States)

    Maiti, Jyotirmay; Biswas, Suman; Chaudhuri, Ankur; Chakraborty, Sandipan; Chakraborty, Sibani; Das, Ranjan

    2017-03-01

    An environment sensitive fluorophore, 4-(5-(4-(dimethylamino)phenyl)oxazol-2-yl)benzoic acid (DMOBA), that closely mimics biologically active 2,5-disubstituited oxazoles has been designed to probe two homologous serum proteins, human serum albumin (HSA) and bovine serum albumin (BSA) by means of photophysical and molecular modeling studies. This fluorescent analogue exhibits solvent polarity sensitive fluorescence due to an intramolecular charge transfer in the excited state. In comparison to water, the steady state emission spectra of DMOBA in BSA is characterized by a greater blue shift ( 10 nm) and smaller Stokes' shift ( 5980 cm- 1) in BSA than HSA (Stokes'shift 6600 cm- 1), indicating less polar and more hydrophobic environment of the dye in the former than the latter. The dye-protein binding interactions are remarkably stronger for BSA than HSA which is evident from higher value of the association constant for the DMOBA-BSA complex (Ka 5.2 × 106 M- 1) than the DMOBA-HSA complex (Ka 1.0 × 106 M- 1). Fӧrster resonance energy transfer studies revealed remarkably less efficient energy transfer (8%) between the donor tryptophans in BSA and the acceptor DMOBA dye than that (30%) between the single tryptophan moiety in HSA and the dye, which is consistent with a much larger distance between the donor (tryptophan)-acceptor (dye) pair in BSA (34.5 Å) than HSA (25.4 Å). Site specific competitive binding assays have confirmed on the location of the dye in Sudlow's site II of BSA and in Sudlow's site I of HSA, respectively. Molecular modeling studies have shown that the fluorescent analogue is tightly packed in the binding site of BSA due to strong steric complementarity, where, binding of DMOBA to BSA is primarily dictated by the van der Waals and hydrogen bonding interactions. In contrast, in HSA the steric complementarity is less significant and binding is primarily guided by polar interactions and van der Waals interactions appear to be less significant in the

  7. Effect of Processing Intensity on Immunologically Active Bovine Milk Serum Proteins

    Science.gov (United States)

    Brick, Tabea; Ege, Markus; Böck, Andreas; von Mutius, Erika; Vervoort, Jacques

    2017-01-01

    Consumption of raw cow’s milk instead of industrially processed milk has been reported to protect children from developing asthma, allergies, and respiratory infections. Several heat-sensitive milk serum proteins have been implied in this effect though unbiased assessment of milk proteins in general is missing. The aim of this study was to compare the native milk serum proteome between raw cow’s milk and various industrially applied processing methods, i.e., homogenization, fat separation, pasteurization, ultra-heat treatment (UHT), treatment for extended shelf-life (ESL), and conventional boiling. Each processing method was applied to the same three pools of raw milk. Levels of detectable proteins were quantified by liquid chromatography/tandem mass spectrometry following filter aided sample preparation. In total, 364 milk serum proteins were identified. The 140 proteins detectable in 66% of all samples were entered in a hierarchical cluster analysis. The resulting proteomics pattern separated mainly as high (boiling, UHT, ESL) versus no/low heat treatment (raw, skimmed, pasteurized). Comparing these two groups revealed 23 individual proteins significantly reduced by heating, e.g., lactoferrin (log2-fold change = −0.37, p = 0.004), lactoperoxidase (log2-fold change = −0.33, p = 0.001), and lactadherin (log2-fold change = −0.22, p = 0.020). The abundance of these heat sensitive proteins found in higher quantity in native cow’s milk compared to heat treated milk, renders them potential candidates for protection from asthma, allergies, and respiratory infections. PMID:28858242

  8. Conjugation of ampicillin and enrofloxacin residues with bovine serum albumin and raising of polyclonal antibodies against them

    Directory of Open Access Journals (Sweden)

    B. Sampath Kumar

    2016-04-01

    Full Text Available Aim: The aim of this study is to test the potency of bovine serum albumin (BSA conjugated ampicillin (AMP and enrofloxacin (ENR antigens in eliciting an immune response in rats using indirect competitive enzyme-linked immunosorbent assay (icELISA. Materials and Methods: AMP and ENR antibiotics were conjugated with BSA by carbodiimide reaction using 1-ethyl-3-(3-dimethylaminopropyl carbodiimide (EDC as a cross-linker. The successful conjugation was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Sprague-Dawley rats were immunized with the conjugates and blood samples were collected serially at 15 days time interval after first immunization plus first booster, second booster, third booster, and the fourth sampling was done 1½ month after the third booster. The antibody titres in the antisera of each antibiotic in all the four immunization cycles (ICs were determined by an icELISA at various serum dilutions ranging from 1/100 to 1/6400. Results: Analysis of antibiotic-BSA conjugates by sodium dodecyl sulfate polyacrylamide gel electrophoresis and coomassie blue staining revealed high molecular weight bands of 85 kDa and 74 kDa for AMP-BSA and ENR-BSA respectively when compared to 68 kDa band of BSA. Both the antibiotic conjugates elicited a good immune response in rats but comparatively the response was more with AMP-BSA conjugate than ENR-BSA conjugate. Maximum optical density 450 value of 2.577 was recorded for AMP-BSA antisera, and 1.723 was recorded for ENR-BSA antisera at 1/100th antiserum dilution in third IC. Conclusion: AMP and ENR antibiotics proved to be good immunogens when conjugated to BSA by carbodiimide reaction with EDC as crosslinker. The polyclonal antibodies produced can be employed for detecting AMP and ENR residues in milk and urine samples.

  9. Interaction mechanisms between organic UV filters and bovine serum albumin as determined by comprehensive spectroscopy exploration and molecular docking.

    Science.gov (United States)

    Ao, Junjie; Gao, Li; Yuan, Tao; Jiang, Gaofeng

    2015-01-01

    Organic UV filters are a group of emerging PPCP (pharmaceuticals and personal care products) contaminants. Current information is insufficient to understand the in vivo processes and health risks of organic UV filters in humans. The interaction mechanism of UV filters with serum albumin provides critical information for the health risk assessment of these active ingredients in sunscreen products. This study investigates the interaction mechanisms of five commonly used UV filters (2-hydroxy-4-methoxybenzophenone, BP-3; 2-ethylhexyl 4-methoxycinnamate, EHMC; 4-methylbenzylidene camphor, 4-MBC; methoxydibenzoylmethane, BDM; homosalate, HMS) with bovine serum albumin (BSA) by spectroscopic measurements of fluorescence, circular dichroism (CD), competitive binding experiments and molecular docking. Our results indicated that the fluorescence of BSA was quenched by these UV filters through a static quenching mechanism. The values of the binding constant (Ka) ranged from (0.78±0.02)×10(3) to (1.29±0.01)×10(5) L mol(-1). Further exploration by synchronous fluorescence and CD showed that the conformation of BSA was demonstrably changed in the presence of these organic UV filters. It was confirmed that the UV filters can disrupt the α-helical stability of BSA. Moreover, the results of molecular docking revealed that the UV filter molecule is located in site II (sub-domain IIIA) of BSA, which was further confirmed by the results of competitive binding experiments. In addition, binding occurred mainly through hydrogen bonding and hydrophobic interaction. This study raises critical concerns regarding the transportation, distribution and toxicity effects of organic UV filters in human body. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Electrochemical microbead-based immunoassay using an (.eta..sup.5./sup.-cyclopentadienyl)tricarbonylmanganese redox marker bound to bovine serum albumin

    Czech Academy of Sciences Publication Activity Database

    Hromadová, Magdaléna; Salmain, M.; Fischer-Durand, N.; Pospíšil, Lubomír; Jaouen, G.

    2005-01-01

    Roč. 22, - (2005), s. 506-511 ISSN 0743-7463 R&D Projects: GA ČR GA203/03/0821; GA AV ČR IAA400400505; GA MŠk LC510 Grant - others:CNRS(FR) 12148; CNRS(FR) 2005-06-005-1 Institutional research plan: CEZ:AV0Z40400503 Keywords : electrochemistry * bovine serum albumin * antigen-antibody complex Subject RIV: CG - Electrochemistry Impact factor: 3.705, year: 2005

  11. Glycation does not modify bovine serum albumin (BSA)-induced reduction of rat aortic relaxation: The response to glycated and nonglycated BSA is lost in metabolic syndrome

    OpenAIRE

    Rubio-Ruiz, Maria Esther; D?az-D?az, Eulises; C?rdenas-Le?n, Mario; Arg?elles-Medina, Rabindranath; S?nchez-Canales, Patricia; Larrea-Gallo, Fernando; Soria-Castro, Elizabeth; Guarner-Lans, Ver?nica

    2008-01-01

    The effects of nonglycated bovine serum albumin (BSA) and advanced glycosylation end products of BSA (AGE-BSA) on vascular responses of control and metabolic syndrome (MS) rats characterized by hypertriglyceridemia, hypertension, hyperinsulinemia, and insulin resistance were studied. Albumin and in vitro prepared AGE-BSA have vascular effects; however, recent studies indicate that some effects of in vitro prepared AGEs are due to the conditions in which they were generated. We produced AGEs b...

  12. Comparison of microcalorimetry and haze formation to quantify the association of B-type procyanidins to poly-L-proline and bovine serum albumin

    OpenAIRE

    Watrelot, Aude; Renard, Catherine; Le Bourvellec, Carine

    2015-01-01

    Though many different methods have been applied to protein-tannin interactions, divergent results are often reported. To better understand the origin of these differences, we compare here haze/aggregates formation and thermodynamic parameters occurring for protein-procyanidin interactions. Proteins well referenced for interaction with polyphenols, namely a polypeptide of extended structure that resembles salivary proteins (poly-L-proline (PLP) and a standard globular protein (bovine serum alb...

  13. The production of cross-reactive autoantibodies that bind to bovine serum albumin in mice administered reducing sugars by subcutaneous injection

    OpenAIRE

    Park, Ji-Hun; Choi, Tae-Saeng

    2015-01-01

    Introduction In a previous study, we identified the formation of cross-reactive autoantibodies that bound to bovine serum albumin (BSA) in a D-galactose-induced aging mouse model. Aim of the study In this study, we investigated the effect of other reducing sugars (namely, glucose and fructose) on the formation of autoantibodies. The effects of concentration and route of administration on the formation of autoantibodies were examined in detail. Material and methods Three concentrations (100, 5...

  14. Effect of pH on the interaction of vitamin B12 with bovine serum albumin by spectroscopic approaches

    Science.gov (United States)

    Li, Daojin; Zhang, Tian; Xu, Chen; Ji, Baoming

    2011-12-01

    The interaction mechanism between vitamin B12 (B12, cyanocobalamin) and bovine serum albumin (BSA) has been investigated by fluorescence, synchronous fluorescence, ultraviolet-vis (UV) absorbance, and three-dimensional fluorescence. The intrinsic fluorescence of BSA was strongly quenched by the addition of B12 in different pH buffer solutions (pH 2.5, 3.5, 5.0, 7.4, and 9.0) and spectroscopic observations are mainly rationalized in terms of a static quenching process at lower concentration of B12 ( CB12/ CBSA B12 ( CB12/ CBSA > 5). The structural characteristics of B12 and BSA were probed, and their binding affinities were determined under different pH conditions. The results indicated that the binding abilities of B12 to BSA in the acidic and basic pH regions (pH 2.5, 3.5, 5.0, and 9.0) were lower than that at simulating physiological condition (pH 7.4). In addition, the efficiency of energy transfer from tryptophan fluorescence to B12 was found to depend on the binding distance r between the donor and acceptor calculated using Förster's theory. The effect of B12 on the conformation of BSA was analyzed using UV, synchronous fluorescence and three-dimensional fluorescence under different pH conditions. These results showed that the binding of B12 to BSA causes apparent change in the secondary and tertiary structures of BSA.

  15. Deciphering the complexation process of a fluoroquinolone antibiotic, levofloxacin, with bovine serum albumin in the presence of additives

    Science.gov (United States)

    Kaur, Amandeep; Khan, Imran Ahmd; Banipal, Parampaul Kaur; Banipal, Tarlok Singh

    2018-02-01

    The current work aims to explore the thermodynamic and conformational aspects for the binding of fluoroquinolone antibacterial drug, levofloxacin (LFC), with bovine serum albumin (BSA) using calorimetric, spectroscopic (UV-visible, fluorescence, circular dichroism, and 1H NMR), dynamic light scattering (DLS) and computational methods (molecular docking). The binding of LFC with BSA at two sequential sites with higher affinity ( 103 M- 1) at the first site has been explored by calorimetry whereas the binding at a single site with affinity of the order of 104 M- 1 has been observed from fluorescence spectroscopy. The calorimetric study in the presence of additives along with docking analysis reveals the significant role of electrostatic, hydrogen bonding, and hydrophobic interactions in the association process. The slight conformational changes in protein as well as the changes in the water network structure around the binding cavity of protein have been observed from spectroscopic and DLS measurements. The LFC induced quenching of BSA fluorescence was observed to be initiated mainly through the static quenching process and this suggests the formation of ground state LFC-BSA association complex. The stronger interactions of LFC in the cavity of Sudlow site I (subdomain IIA) of protein have been explored from site marker calorimetric and molecular docking study.

  16. Bovine serum albumin bioconjugated graphene oxide: Red blood cell adhesion and hemolysis studied by QCM-D

    Science.gov (United States)

    Cai, Bing; Hu, Kebang; Li, Chunming; Jin, Jing; Hu, Yuexin

    2015-11-01

    Graphene oxide (GO) had great potential in various applications especial biomedical materials. In this study, we improved the hemocompatibility especial hemolysis properties of GO nanosheets by grafting bovine serum albumin (BSA). The hemocompatibility of GO-g-BSA was improved. The hemolysis ratio of GO-g-BSA was lower than 0.2% and no visible hemoglobin release was observed. In a flowed condition, the interaction between GO and RBC was monitored real time by quartz crystal microbalance with dissipation (QCM-D) and the hemolysis rates of eluted RBC solution was determined. The balance between the adsorption and degradation of RBC on the surface of GO was a linear process. The GO-g-BSA surface decreased the adhesion of RBC in a flowed condition, maintained the morphology of RBC and reduced the hemolysis rate in the most effective manner. The inert of BSA resisted GO interacting with the lipid bilayers of RBC and the negative charge on the surface of BSA repelled the approach of negative charged RBC. The excellent hemocompatibility of the BSA modified GO might confer its great potentials for various biomedical applications.

  17. Study of the interaction between 8-azaguanine and bovine serum albumin using optical spectroscopy and molecular modeling methods.

    Science.gov (United States)

    Gong, Qiao-Ling; Hu, Xin-Gen; Fang, Guo-Yong; Li, Xin-Hua

    2012-02-01

    The interaction between 8-azaguanine (8-Azan) and bovine serum albumin (BSA) in Tris-HCl buffer solutions at pH 7.4 was investigated by means of fluorescence and ultraviolet-visible (UV-Vis) spectroscopy. At 298 K and 310 K, at a wavelength of excitation (λ (ex)) of 282 nm, the fluorescence intensity decreased significantly with increasing concentrations of 8-Azan. Fluorescence static quenching was observed for BSA, which was attributed to the formation of a complex between 8-Azan and BSA during the binding reaction. This was illuminated further by the UV-Vis absorption spectra and the decomposition of the fluorescence spectra. The thermodynamic parameters ∆G, ∆H, ∆S were calculated. The results showed that the forces acting between 8-Azan and BSA were typical hydrophobic forces, and that the interaction process was spontaneous. The interaction distance r between 8-Azan and BSA, evaluated according to fluorescence resonance energy transfer theory, suggested that there is a high possibility of energy transfer from BSA to 8-Azan. Theoretical investigations based on homology modeling and molecular docking suggested that binding between 8-Azan and BSA is dominated by hydrophilic forces and hydrogen bonding. The theoretical investigations provided a good structural basis to explain the phenomenon of fluorescence quenching between 8-Azan and BSA.

  18. Spectroscopic studies on the interaction of bovine serum albumin with Ginkgol C15:1 from Ginkgo biloba L

    Energy Technology Data Exchange (ETDEWEB)

    Fang, Yang-Yang [School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013 (China); School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang 212013 (China); Yang, Xiao-Ming, E-mail: XM_Yang1963@126.com [School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013 (China); Li, Yue-Ying [School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang 212013 (China); Feng, Chun-Lai [School of Pharmacy, Jiangsu University, Zhenjiang 212013 (China)

    2015-06-15

    The interaction between Ginkgol C15:1 (Ginkgol), a natural bioactive compound from Ginkgo biloba, and bovine serum albumin (BSA) was studied by fluorescence, UV–vis absorption, Fourier transform infrared (FT-IR) and circular dichroism (CD) spectroscopy under simulative physiological conditions. The results showed that the fluorescence quenching of BSA by Ginkgol was a static quenching procedure through forming a 1:1 ground-state Ginkgol–BSA complex with a binding constant of about 2.6×10{sup 3} L mol{sup −1}. The values of the thermodynamic parameters indicated that electrostatic and hydrophobic forces played important roles in the interaction of BSA with Ginkgol. The binding distance between BSA and Ginkgol was 3.37 nm, based on Föster’s non-radiative energy transfer theory, and subdomain IIA (Sudlow site I) was the primary binding site which was consistent with that results of molecular docking modeling. The results of UV–vis, CD, three-dimensional fluorescence and FT-IR spectra indicated that binding of Ginkgol to BSA induced conformational changes of BSA. - Highlights: • This is the first time to report the interaction between Ginkgol C15:1 and BSA. • Researching the binding properties of Ginkgol C15:1 and BSA in-depth. • From the aspect of BSA structure change, verified the anticancer activity of Ginkgol. • Molecular docking study explored the interaction of Ginkgol on BSA.

  19. Growth and development of rabbit oocytes in vitro: effect of fetal bovine serum concentration on culture medium.

    Science.gov (United States)

    Sugimoto, H; Kida, Y; Miyamoto, Y; Kitada, K; Matsumoto, K; Saeki, K; Taniguchi, T; Hosoi, Y

    2012-09-15

    The objective was to develop a culture system that produced blastocyst stage embryos from rabbit oocytes grown in vitro. Two experiments were performed. First, various concentrations of fetal bovine serum (FBS, 0, 0.05, 0.5 and 5%) were used in the culture medium for in vitro growth (IVG) of oocytes recovered from follicles 200 to 299 μm in diameter. Intracytoplasmic sperm injection (ICSI) was performed on mature oocytes obtained after IVG for 8 days and in vitro maturation for 14 to 16 h. Rates of survival and pronuclear formation after ICSI were higher for oocytes grown in a medium with 0.05% FBS compared to oocytes grown in a medium lacking FBS (97.6 vs. 76.9%, 97.5 vs. 70%, P medium containing 0.05% FBS than in the medium lacking FBS (9.5 vs. 17.9%, P medium for IVG improved developmental competence of rabbit oocytes grown in vitro. Copyright © 2012 Elsevier Inc. All rights reserved.

  20. Preparation and adsorption of bovine serum albumin-imprinted polyacrylamide hydrogel membrane grafted on non-woven polypropylene.

    Science.gov (United States)

    Zhao, Kongyin; Lin, Beibei; Cui, Wenkui; Feng, Lingzhi; Chen, Tian; Wei, Junfu

    2014-04-01

    Bovine serum albumin (BSA) imprinted polypropylene (PP) fiber-grafted polyacrylamide (PAM) hydrogel membrane (PP-g-PAM MIP) was prepared using non-woven PP fiber as matrix, BSA as template molecule, and acrylamide (AM) as functional monomer via UV radiation-reduced polymerization in an aqueous phase. SEM, FT-IR, DSC and TG were used to characterize the PP grafted PAM hydrogel. Influence factors on the adsorption capacity of PP-g-PAM MIP were investigated, such as monomer concentration, cross-linker concentration, template molecule amount and pH values in BSA solution. The adsorption and recognition properties of PP-g-PAM MIP were evaluated and the results showed that the PP-g-PAM MIP exhibited an obvious improvement in terms of adsorption capacity for BSA as compared with non-imprinted ones. PP-g-PAM MIPs could recognize the template protein using Lys, Ova, BHb, and Glo as control proteins, and the selectivity factor (β) was above 2.0. The imprinting efficiency of PP-g-PAM MIP tended to be stable after three cycles and maintained 76% of the initial value of the imprinting efficiency even after five repetitions, which was more excellent than that of PAM microsphere. The PP-g-PAM MIP is low cost and easy to be prepared, which would show its potential applications in the fields of extracting and testing required proteins from cells or particulate samples. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Spectroscopic studies of the reaction between bovine serum amine oxidase (copper-containing) and some hydrazides and hydrazines.

    Science.gov (United States)

    Morpurgo, L; Befani, O; Sabatini, S; Mondovì, B; Artico, M; Corelli, F; Massa, S; Stefancich, G; Avigliano, L

    1988-01-01

    The carbonyl cofactor of bovine serum amine oxidase, recently identified as pyrroloquinoline quinone [Ameyama, Hayashi, Matsushita, Shinagawa & Adachi (1984) Agric. Biol. Chem. 48, 561-565; Lobenstein-Verbeek, Jongejan, Frank & Duine (1984) FEBS Lett. 170, 305-309], reacts stoichiometrically and irreversibly with hydrazides of phenylacetic acid and of benzoic acid. With the phenylacetic hydrazides a reversible intermediate step was detected by competition with substrate, carbonylic reagents or phenylhydrazine, a typical inhibitor of the enzyme. All hydrazides form an intense broad band with maximum absorbance in a narrow wavelength range (350-360 nm), irrespective of the acyl group, suggesting that the transition is located on the organic cofactor. A different situation is found with some phenylhydrazines, where extended conjugation can occur between the cofactor and the phenyl pi-electron system via the azo group, as shown by the lower energy and higher intensity of the transition. In this case the transition is sensitive to substituents in the phenyl ring. The c.d. spectrum of the adducts is influenced by the type of hydrazide (derived from phenylacetic acid or benzoic acid), by pH and by NN-diethyldithiocarbamate binding to copper, probably as a result of shifts of equilibria between hydrazone-azo tautomers. PMID:3146976

  2. Mechanistic and conformational studies on the interaction of anti-inflammatory drugs, isoxicam and tenoxicam with bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Punith, Reeta; Katrahalli, Umesha; Kalanur, Shankara S. [Department of Chemistry, Karnatak University, Dharwad 580 003 (India); Jaldappagari, Seetharamappa, E-mail: jseetharam@yahoo.co [Department of Chemistry, Karnatak University, Dharwad 580 003 (India)

    2010-11-15

    The mechanism of interaction of the non-steroidal anti-inflammatory drugs, isoxicam (IXM) and tenoxicam (TXM) with bovine serum albumin (BSA) has been studied using spectroscopic techniques, viz., spectrofluorescence, circular dichroism (CD), UV-visible absorption and FT-IR under simulative physiological conditions. Stern-Volmer analysis of fluorescence quenching data shows the presence of the static quenching mechanism. Thermodynamic parameters (negative {Delta}H{sup 0} and positive {Delta}S{sup 0} values obtained in the present study) revealed that the hydrophobic interactions played a major role in the interaction of these drugs with BSA. The distance, r between the donor (BSA) and acceptor (IXM/TXM) was calculated based on the Forster's theory of non-radiation energy transfer and the values were observed to be 3.85 nm and 2.60 nm in IXM-BSA and TXM-BSA system, respectively. CD and FT-IR studies indicated that the binding of IXM/TXM to BSA induced conformational changes in BSA. The effect of common ions on the binding of IXM/TXM to BSA has been investigated.

  3. Affinity study on bovine serum albumin's peptides to amphiphilic gold nanoparticles: A test of epitopes and non-epitopes

    Science.gov (United States)

    Yuan, Ming; Li, Wanrong; Yang, Mingming; Huang, Xiufeng; Bai, Zhijun; Liu, Yushuang; Cai, Weijun; Wang, Yuqin; Zhang, Feng

    2017-09-01

    It is an inevitable event that nanoparticles (NPs) will encounter proteins/peptides in nano-medicine, so it has been significant to know their interaction mechanism before in vivo applications. Previously, a 105-amino-acid sequence had been reported as the binding site between bovine serum albumin (BSA) and amphiphilic polymer coated gold nanoparticles (AP-AuNPs) along with a mortise-tenon joint hypothesis. This article tested the affinity difference between two epitope peptide sequences such as: LGEYGFQNALIVR (S1), DAFLGSFLYEYSR (S2) and one non-epitope peptide sequence as: FDEHVKLVNELTEF (S3). With the photoluminescent amino acid residues, the fluorescence quenching method based on the nanometal surface energy transfer (NSET) principle was able to study the thermodynamics of the current binding system. The binding constants (Ka) were determined and followed the order as: Ka-S1 > Ka-S2 >> Ka-S3. Moreover, Hill constants indicated that cooperativity only presented in the interactions of AP-AuNP with either S1 or S2, but not for S3. Moreover, gel electrophoresis, surface plasmon resonance, atomic force microscopy and three dimensional fluorescence microscopy were all also used to comprehensively analyse the binding interaction mechanism. These results further provided useful information to better understand the mortise-tenon joint, which might find applications to nanofabrication and biomedicine.

  4. Application of MoS{sub 2} modified screen-printed electrodes for highly sensitive detection of bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Kukkar, Manil [Central Scientific Instruments Organisation (CSIR-CSIO), Sector 30-C, Chandigarh, 160030 (India); Academy of Scientific and Innovative Research, CSIR-CSIO, Sector 30-C, Chandigarh, 160030 (India); Sharma, Ashish [Central Scientific Instruments Organisation (CSIR-CSIO), Sector 30-C, Chandigarh, 160030 (India); UIET, Panjab University, Sector 14, Chandigarh, 160014 (India); Kumar, Parveen [Department of Biotechnology, Kurukshetra University, Kurukshetra, 136119 (India); Kim, Ki-Hyun, E-mail: kkim61@hanyang.ac.kr [Department of Civil & Environmental Engineering, Hanyang University, 222 Wangsimni-Ro, Seoul 04763 (Korea, Republic of); Deep, Akash, E-mail: dr.akashdeep@csio.res.in [Central Scientific Instruments Organisation (CSIR-CSIO), Sector 30-C, Chandigarh, 160030 (India); Academy of Scientific and Innovative Research, CSIR-CSIO, Sector 30-C, Chandigarh, 160030 (India)

    2016-10-05

    The present work reports the application of a new molybdenum disulphide (MoS{sub 2})-based electrochemical platform for highly sensitive quantitation of an iron-binding protein, bovine serum albumin (BSA). The gold screen-printed electrodes were modified with MoS{sub 2} nanoflakes, followed by bioconjugation with anti-BSA antibodies. Using the above bioelectrode, cyclic voltammetric analysis was carried out in the presence of a Fe{sup 3+}/Fe{sup 2+} redox probe which exhibited a linear response of peak current with varying concentrations of BSA up to 10 ng/mL (with a detection limit of 0.006 ng/mL). This study is novel in that it shows a considerable enhancement of signal during electrochemical sensing of a protein. - Highlights: • MoS{sub 2} nanoflakes have been used for an electrochemical immunosensor. • The sensor's response was proportional to the antigen concentration. • Highly sensitive and specific detection of iron-binding protein ‘BSA’ is achieved. • A wide linear range of detection of BSA is demonstrated.

  5. Preparation of carbon quantum dots with a high quantum yield and the application in labeling bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Pengpeng; Zhang, Changchang; Liu, Xiang, E-mail: liuxiang@ahut.edu.cn; Cui, Ping, E-mail: cokecp@sohu.com

    2016-04-15

    Graphical abstract: - Highlights: • Cheap carbon quantum dots (CQDs) with a high quantum yield were prepared. • The preparation process and surface functionalization on CQDs are rather facile. • Such functionalized CQDs can be attached to BSA covalently. • This predicts that some biomolecules can be labeled by the fluorescent CQDs. - Abstract: An economic and green approach of manufacturing carbon quantum dots (CQDs) with a high quantum yield (denoted with HQY-CQDs) and the application in labeling bovine serum albumin (BSA) were described in detail in this work. Firstly, the cheap resources of citric acid and glycine were pyrolysed in drying oven for preparing the CQDs. Then the product was immersed in tetrahydrofuran for 8 h. HQY-CQDs were obtained by removing tetrahydrofuran from the supernate and were evaluated that they possessed a much higher quantum yield compared with that without dealing with tetrahydrofuran and a wonderful photo-bleaching resistance. Such HQY-CQDs could be functionalized by N-hydroxysuccinimide and successively combined with BSA covalently. Thus fluorescent labeling on BSA was realized. The HQY-CQDs were demonstrated with transmission electron microscopy and the chemical modification with N-hydroxysuccinimide was proved by infrared and X-ray photoelectron spectra. Labeling BSA with the HQY-CQDs was confirmed by gel electrophoresis and fluorescence imaging.

  6. Studies on the antagonistic action between chloramphenicol and quinolones with presence of bovine serum albumin by fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Liu Baosheng, E-mail: lbs@hbu.edu.c [Key Laboratory of Medical Chemistry and Molecular Diagnosis, Ministry of Education, Center of Physics and Chemistry, Hebei University, Baoding 071002 (China); Zhao Fengli; Xue Chunli; Wang Jing; Lu Yunkai [Key Laboratory of Medical Chemistry and Molecular Diagnosis, Ministry of Education, Center of Physics and Chemistry, Hebei University, Baoding 071002 (China)

    2010-05-15

    Chloramphenicol (CHL) and quinolone drugs like ofloxacin (OFLX), lomefloxacin (LMX) and ciprofloxacin (CPFX) can all quench the fluorescence of bovine serum albumin (BSA) in the aqueous solution of pH=7.40. This quenching effect becomes more significant when CHL and quinolone drugs coexist. Based on this, further studies on the interactions between CHL and quinolone drugs using fluorescence spectrum are established. The results showed that the interaction between the drugs would increase the binding constant and binding stability of the drug and protein, thus reducing the amount of drugs transported to their targets. Therefore, free drug concentration at targets would decrease, reducing the efficacy of the drugs. It indicated that there exists antagonistic action between drugs. The results also showed that the quenching mechanism of BSA by the drugs is a static procedure. The number of binding sites is 1 in various systems. Due to the existence of the antagonistic action between drugs, the binding distance r is reduced. Studies utilizing synchronous spectra showed that the antagonistic action between the drugs would affect the conformation of BSA, making protein molecules extend and hydrophobic decrease. The order of antagonistic action between CHL and quinolone drugs is: CPFX>OFLX>LMX with presence of BSA.

  7. Use of 6α- and 6β-carboxymethyl testosterone-bovine serum albumin conjugates in radioimmunoassay for testosterone

    International Nuclear Information System (INIS)

    Jones, C.D.; Mason, N.R.

    1975-01-01

    The synthesis of 6α- and 6β-testosterone-bovine serum albumin (BSA) conjugates is described. 6β-Carboxymethyl-4-androstene-3,17-dione was prepared by a route analogous to that described earlier for 6β-carboxymethyl progesterone. Sodium borohydride reduction of the 3 and 17 keto groups and subsequent selective oxidation of the resulting 3β,17β-diol using MnO 2 provided 6β-carboxymethyl testosterone. Further acid catalyzed epimerization of the C-6 center gave the isomeric 6α-carboxymethyl testosterone. The 6α- and 6β-testosterone derivatives were attached to BSA via a mixed anhydride coupling employing tributylamine and i-butylchlorocarbonate. For each molecule of BSA, the 6α- and 6β-conjugates contained an average of 23 and 20 steroid residues, respectively. Antisera to the conjugates exhibited similar high specificities toward various steroids, the only incidence of serious cross-reaction being the expected case of dihydrotestosterone. (U.S.)

  8. Dietary intervention with serum-derived bovine immunoglobulins protects barrier function in a mouse model of colitis.

    Science.gov (United States)

    Pérez-Bosque, Anna; Miró, Lluïsa; Maijó, Mònica; Polo, Javier; Campbell, Joy; Russell, Louis; Crenshaw, Joe; Weaver, Eric; Moretó, Miquel

    2015-06-15

    Dietary supplementation with immunoglobulins from animal plasma has anti-inflammatory effects on intestinal and lung models of acute inflammation. Here, we aimed to establish whether dietary intervention with serum-derived bovine immunoglobulin (SBI) can prevent alterations in intestinal barrier function in a mouse model with a genetic predisposition to inflammatory bowel disease (IBD). Wild-type (WT) mice and mice lacking the mdr1a gene (KO) were fed diets supplemented with either SBI (2% wt/wt) or milk proteins (control diet), from day 21 (weaning) until day 56. The epithelial permeability of distal colon crypts was measured by confocal microscopy using a fluorescent marker. The expression of junctional epithelial E-cadherin and β-catenin proteins were determined by Western blot and zonula occludens-1 (ZO-1) by immunofluorescence. Mucins (MUC1, MUC2, MUC4), TFF3, cytokines (TNF-α, IFN-γ), and inducible nitric oxide synthase RNA expression were quantified by real-time PCR. SBI blocked the increase in colon crypt permeability and partially prevented the reduction in E-cadherin and ZO-1 expression that characterize the KO mouse model (both P genetic model of IBD.

  9. Spectroscopic and molecular docking approaches for investigating conformation and binding characteristics of clonazepam with bovine serum albumin (BSA).

    Science.gov (United States)

    Lou, Yan-Yue; Zhou, Kai-Li; Pan, Dong-Qi; Shen, Jia-Le; Shi, Jie-Hua

    2017-02-01

    Clonazepam, a type of benzodiazepine, is a classical drug used to prevent and treat seizures, panic disorder, movement disorder, among others. For further clarifying the distribution of clonazepam in vivo and the pharmacodynamic and pharmacokinetic mechanisms, the binding interaction between clonazepam and bovine serum albumin (BSA) was investigated using ultraviolet spectroscopy (UV), steady-state fluorescence spectroscopy, synchronous fluorescence spectroscopy, three-dimensional (3D) fluorescence spectroscopy, Fourier transform infrared spectroscopy (FT-IR) and molecular docking methods. The results well confirmed that clonazepam bound on the subdomain III A (Site II) of BSA through van der Waals force and hydrogen bonding interaction, and quenched the intrinsic fluorescence of BSA through a static quenching process. The number of binding sites (n) and binding constant (K b ) of clonazepam-BSA complex were about 1 and 7.94×10 4 M -1 at 308K, respectively. The binding process of clonazepam with BSA was spontaneous and enthalpy-driven process due to ΔG 0 T|ΔS 0 | over the studied temperature range. Meanwhile, the binding interaction of clonazepam with BSA resulted in the slight change in the conformation of BSA and the obvious change in the conformation of clonazepam, implying that the flexibility of clonazepam also played an important role in increasing the stability of the clonazepam-BSA complex. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Influence of bovine serum albumin in Hanks' solution on the corrosion and stress corrosion cracking of a magnesium alloy.

    Science.gov (United States)

    Harandi, Shervin Eslami; Banerjee, Parama Chakraborty; Easton, Christopher D; Singh Raman, R K

    2017-11-01

    It is essential for any temporary implant to possess adequate strength to maintain their mechanical integrity under the synergistic effects of mechanical loading characteristics of human body and the corrosive physiological environment. Such synergistic effects can cause stress corrosion cracking (SCC). The aim of the present study is to investigate the effect of the addition of bovine serum albumin (BSA) to Hanks' solution in corrosion and SCC susceptibility of AZ91D magnesium alloy. The electrochemical impedance spectroscopy (EIS) results indicated that the addition of BSA increased corrosion resistance of the alloy during the first 48h of immersion and then decreased it rapidly. The energy-dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectroscopy (XPS) analyses indicated adsorption of BSA on the alloy surface during initial hours of immersion. However, with the increasing immersion time, BSA chelated with the corrosion products causing disruption of the protective film; thus, it accelerated the corrosion of the alloy. Both the mechanical data and fractographic evidence have confirmed susceptibility of the alloy to SCC. However, in the presence of BSA, the alloy suffered greater SCC which was attributed to its increased susceptibility towards localized corrosion. Copyright © 2017. Published by Elsevier B.V.

  11. Bovine serum albumin conformational changes upon adsorption on titania and on hydroxyapatite and their relation with biomineralization.

    Science.gov (United States)

    Serro, A P; Bastos, M; Pessoa, J Costa; Saramago, B

    2004-09-01

    The biocompatibility of implant materials used for substitution of bone tissue depends on its ability to induce the deposition of a hydroxyapatite layer when in contact with body fluids. In previous work, some of the authors found that bovine serum albumin (BSA) promotes calcium phosphate deposition if preadsorbed on hydroxyapatite and retards precipitation if preadsorbed on titania. In the present study, we investigated the adsorption of BSA upon particles of titania and hydroxyapatite in order to understand the different role played by the protein on the mineralization of both biomaterials. The adsorption isotherms were determined and the structural changes induced by adsorption at different surface coverages were investigated by circular dichroism spectroscopy and differential scanning microcalorimetry. At low surface coverages, the adsorbed BSA molecules lost part of their alpha-helix content. However, at high surface coverages, corresponding to the plateau values of the adsorption isotherms, the BSA molecules did not undergo structural rearrangements upon adsorption. In the latter circumstances, the availability of BSA calcium binding sites, which should be responsible for inducing mineralization, depends on the electrostatic interactions between BSA and the sorbent surface. A possible explanation for the different mineralization behavior of hydroxyapatite and titania is advanced. Copyright 2004 Wiley Periodicals, Inc.

  12. Bioconjugation of gold-polymer core-shell nanoparticles with bovine serum amine oxidase for biomedical applications.

    Science.gov (United States)

    Venditti, I; Hassanein, T F; Fratoddi, I; Fontana, L; Battocchio, C; Rinaldi, F; Carafa, M; Marianecci, C; Diociaiuti, M; Agostinelli, E; Cametti, C; Russo, M V

    2015-10-01

    Core-shell gold nanoparticles [AuNPs], stabilized with a hydrophilic polymer, poly(3-dimethylammonium-1-propyne hydrochloride) [PDMPAHCl], have been used for the immobilization of bovine serum amine oxidase [BSAO]. The functionalized surface of the hybrid nanoparticles is pH responsive, due to the presence of aminic groups that carry out a double role: on one hand they act as ligands for the gold nanoparticle surface, allowing the colloidal stabilization and, on the other hand, they give a hydrophilic characteristic to the whole colloidal suspension. The core-shell nanoparticles [Au@PDMPAHCl] have been characterized by using UV-vis and X-ray photoelectron spectroscopy, DLS, ζ-potential measurements and by FE-TEM microscopy. BSAO enzyme can be loaded by non-covalent immobilization onto Au@PDMPAHCl nanoparticles up to 70% in weight, depending on the pH values of the environmental medium. Activity tests on Au@PDMPAHCl-BSAO bioconjugates confirm an enzymatic activity up to 40%, with respect to the free enzyme activity. Moreover, our results show that loading and enzymatic activity are rather interrelated characteristics and that, under appropriate polymer concentration and pH conditions, a satisfactory compromise can be reached. These results, as a whole, indicate that Au@PDMPAHCl-BSAO bioconjugate systems are promising for future biomedical applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Bovine Serum Albumin (BSA) coated iron oxide magnetic nanoparticles as biocompatible carriers for curcumin-anticancer drug.

    Science.gov (United States)

    Nosrati, Hamed; Sefidi, Naser; Sharafi, Ali; Danafar, Hossein; Kheiri Manjili, Hamidreza

    2018-02-01

    The bovine serum albumin-coated magnetic nanoparticles (F@BSA NPs) were prepared as curcumin (CUR) carriers through desolvation and chemical co-precipitation process. The characteristics of CUR loaded F@BSA NPs (F@BSA@CUR NPs) were determined by X-ray diffraction (XRD), thermogravimetric analysis (TGA), fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), and vibrating-sampling magnetometry (VSM) techniques. It was found that the synthesized F@BSA@CUR NPs were spherical in shape with an average size of 56 ± 11.43 nm (mean ± SD (n = 33)), ζ-potential of -10.1 mV, and good magnetic responsivity. Meanwhile, the drug content of the nanoparticles was 6.88%. These F@BSA@CUR NPs also demonstrated sustained release of CUR at 37 °C in different buffer solutions. Cellular toxicity of F@BSA@CUR NPs was studied on HFF2 cell line. Also, the cytotoxicity of F@BSA@CUR NPs towards MCF-7 breast cancer cells was investigated. The results revealed that F@BSA@CUR NPs have significant cytotoxicity activity on MCF-7 cell line. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Probing into the binding interaction between medroxyprogesterone acetate and bovine serum albumin (BSA): spectroscopic and molecular docking methods.

    Science.gov (United States)

    Fang, Fang; Pan, Dong-Qi; Qiu, Min-Jie; Liu, Ting-Ting; Jiang, Min; Wang, Qi; Shi, Jie-Hua

    2016-09-01

    To further understand the mechanism of action and pharmacokinetics of medroxyprogesterone acetate (MPA), the binding interaction of MPA with bovine serum albumin (BSA) under simulated physiological conditions (pH 7.4) was studied using fluorescence emission spectroscopy, synchronous fluorescence spectroscopy, circular dichroism and molecular docking methods. The experimental results reveal that the fluorescence of BSA quenches due to the formation of MPA-BSA complex. The number of binding sites (n) and the binding constant for MPA-BSA complex are ~1 and 4.6 × 10(3)  M(-1) at 310 K, respectively. However, it can be concluded that the binding process of MPA with BSA is spontaneous and the main interaction forces between MPA and BSA are van der Waals force and hydrogen bonding interaction due to the negative values of ΔG(0) , ΔH(0) and ΔS(0) in the binding process of MPA with BSA. MPA prefers binding on the hydrophobic cavity in subdomain IIIA (site II'') of BSA resulting in a slight change in the conformation of BSA, but BSA retaining the α-helix structure. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  15. Thimerosal changes protein conformation and increase the rate of fibrillation in physiological conditions: Spectroscopic studies using bovine serum albumin (BSA).

    Science.gov (United States)

    Santos, João César N; da Silva, Isabella M; Braga, Taniris C; de Fátima, Ângelo; Figueiredo, Isis M; Santos, Josué Carinhanha Caldas

    2018-02-21

    The interaction between bovine serum albumin (BSA) and thimerosal (TM), an organomercury compound widely employed as a preservative in vaccines, was investigated simulating physiological conditions and using different spectroscopic techniques. The results, employing molecular fluorescence showed the interaction occurs by static quenching through electrostatic forces (ΔH  0), spontaneously (ΔG = -4.40 kJ mol -1 ) and with a binding constant of 3.24 × 10 3  M -1 . Three-dimensional fluorescence studies indicated that TM causes structural changes in the polypeptide chain of the BSA, confirmed by circular dichroism that showed an increase in α-helix (from 43.9 to 47.8%) content after interaction process. Through synchronized fluorescence and employing bilirubin as a protein site marker, it was confirmed the preferential interaction of TM in the subdomain IB of BSA. The interaction mechanism proposed in this work is based on the reaction of TM with BSA through of free Cys34 residue, forming the adduct BSA-HgEt with the thiosalicylic acid release, which possibly interacts electrostatically with positive side chain amino acids of the modified protein. Finally, it was proven that both TM and EtHgCl accelerate the protein fibrillation kinetics in 42 and 122%, respectively, indicating the toxicity of these compounds in biological systems. Copyright © 2017. Published by Elsevier B.V.

  16. Voltammetric behavior of Mammeisin (MA) at a glassy carbon electrode and its interaction with Bovine Serum Albumin (BSA).

    Science.gov (United States)

    Leuna, Jules-Blaise Mabou; Sop, Sergeot Kungo; Makota, Suzanne; Njanja, Evangeline; Ebelle, Thiery Christophe; Azebaze, Anatole Guy; Ngameni, Emmanuel; Nassi, Achille

    2018-02-01

    The electrochemical oxidation of Mammeisin (MA) was studied in a solution containing acetone and 0.1M phosphate buffer +0.1M KCl (pH=5.3) at a glassy carbon electrode (GCE), using cyclic (CV) and square wave voltammetry (SWV). MA showed a quasi-reversible process, which is pH dependent and that involves the exchange of two electrons and two protons. The oxidation product was adsorbed by the electrode surface to form a film that blocks active sites over repetitive cyclic. Moreover, the interaction of MA and bovine serum albumin (BSA) was studied by CV and SWV at different pHs (5.4, 7.2, 9.5). As a result of the affinity binding with BSA, electrochemically inactive complex was formed. In addition, the oxidation potential of MA in the presence of BSA depends on the pH. The diffusion coefficients of both free and bound MA were estimated from the cyclic voltammetry data using the method developed by Randles-Sevich (D f =9.85×10 -5 cm 2 s -1 and D b =1.27×10 -9 cm 2 s -1 ) and the binding constant of MA-BSA complex, K=3.47×10 2 Lmol -1 , was obtained. Copyright © 2017. Published by Elsevier B.V.

  17. Spectroscopic Investigations of the Binding Interaction of a New Indanedione Derivative with Human and Bovine Serum Albumins

    Directory of Open Access Journals (Sweden)

    Mihaela Hillebrand

    2009-04-01

    Full Text Available Binding of a newly synthesized indanedione derivative, 2-(2-hydroxy-3-ethoxybenzylidene-1,3-indanedione (HEBID, to human and bovine serum albumins (HSA and BSA, under simulated physiological conditions was monitored by fluorescence spectroscopy. The binding parameters (binding constants and number of binding sites and quenching constants were determined according to literature models. The quenching mechanism was assigned to a Förster non-radiative energy transfer due to the HEBID-SA complex formation. A slightly increased affinity of HEBID for HSA was found, while the number of binding sites is approximately one for both albumins. The molecular distance between donor (albumin and acceptor (HEBID and the energy transfer efficiency were estimated, in the view of Förster’s theory. The effect of HEBID on the protein conformation was investigated using circular dichroism and synchronous fluorescence spectroscopies. The results revealed partial unfolding in the albumins upon interaction, as well as changes in the local polarity around the tryptophan residues

  18. Water soluble peripheral and non-peripheral tetrasubstituted zinc phthalocyanines: Synthesis, photochemistry and bovine serum albumin binding behavior

    Energy Technology Data Exchange (ETDEWEB)

    Çakır, Volkan; Çakır, Dilek [Department of Chemistry, Faculty of Sciences, Karadeniz Technical University, 61080 Trabzon (Turkey); Pişkin, Mehmet [Department of Food Technology, Çanakkale Vocational School of Technical Sciences, Çanakkale Onsekiz Mart University, 17100 Çanakkale (Turkey); Durmuş, Mahmut [Gebze Institute of Technology, Department of Chemistry, PO Box 141, Gebze 41400, Kocaeli (Turkey); Bıyıklıoğlu, Zekeriya, E-mail: zekeriya_61@yahoo.com [Department of Chemistry, Faculty of Sciences, Karadeniz Technical University, 61080 Trabzon (Turkey)

    2014-10-15

    The new peripherally and non-peripherally tetra-1,3-bis[3-(diethylamino)phenoxy]propan-2-ol substituted zinc(II) phthalocyanine complexes (2a and 3a) and their quaternized amphiphilic derivatives (2b and 3b) have been synthesized and characterized using UV–vis, FT-IR, {sup 1}H-NMR, {sup 13}C-NMR, MS spectroscopic data and elemental analysis for the first time. The quaternized complexes (2b and 3b) show excellent solubility in water and DMSO which makes them potential photosensitizers for use in photodynamic therapy (PDT) of cancer. The photochemical (singlet oxygen generation and photodegradation under light irradiation) properties of these novel phthalocyanines are investigated in dimethylsulfoxide (DMSO) for non-quaternized complexes and in DMSO, phosphate buffered solution (PBS) or PBS+triton X-100 (TX) solution for quaternized complexes. In this study, the effects of the aggregation of the molecules, quaternization, position of the substituents (peripherally or non-peripherally) and nature of the solvents (DMSO, PBS or PBS+triton X-100) on the photochemical parameters of the zinc (II) phthalocyanines are also reported. In addition, a spectroscopic investigation of the binding behavior of the quaternized zinc(II) phthalocyanine complexes to bovine serum albumin (BSA) is also presented in this work. - Highlights: • Synthesis of zinc phthalocyanines is performed. • Photophysical and photochemical properties of phthalocyanines are studied. • Photodynamic therapy studies are performed.

  19. Spectroscopic analysis on structure-affinity relationship in the interactions of different oleanane-type triterpenoids with bovine serum albumin.

    Science.gov (United States)

    Hou, Jia; Wang, Zhenzhong; Yue, Ying; Li, Qian; Shao, Shijun

    2015-09-01

    Oleanane-type triterpenoids serve as an important group of plant secondary metabolites with a variety of biological activities and the C-3 position substitution pattern is a significant structural feature for their biological activities. Three selected oleanane-type triterpenoids (glycyrrhizin, glycyrrhetinic acid, and carbenoxolone) bearing different substituents (glucuronic acid dimer, hydroxyl, and succinyl groups) at the C-3 position were studied for their affinities to bind bovine serum albumin (BSA) by steady-state fluorescence, synchronous, three-dimensional fluorescence and ultraviolet-visible (UV-vis) absorption spectra. The binding mechanism of the triterpenoids to BSA is due to the formation of the triterpenoids-BSA complex and the binding affinity is strongest for carbenoxolone and ranked in the order carbenoxolone > glycyrrhetinic acid > glycyrrhizin. The thermodynamic parameters calculated at different temperatures showed that triterpenoids binding to BSA primarily depended on hydrophobic interaction and hydrogen bonding. The distance between the bound triterpenoid and BSA was determined on the basis of the Förster's energy transfer theory. Displacement experiments using phenylbutazone and ibuprofen showed the binding site of triterpenoids on BSA at subdomain IIA (Sudlow's site I). The effect of triterpenoids on BSA conformation was analyzed by UV-vis absorption, and synchronous and three-dimensional fluorescence spectra. These results revealed that the C-3 position substitution pattern significantly affects the structure-affinity relationships of oleanane-type triterpenoid binding to BSA and further affects the bioavailability of triterpenoids in the blood circulatory system. Copyright © 2014 John Wiley & Sons, Ltd.

  20. Label-free detection of bovine serum albumin based on an in-fiber Mach-Zehnder interferometric biosensor.

    Science.gov (United States)

    Li, Zhengyong; Liao, Changrui; Chen, Danni; Song, Jun; Jin, Wei; Peng, Gang-Ding; Zhu, Feng; Wang, Ying; He, Jun; Wang, Yiping

    2017-07-24

    We propose and experimentally verify an innovative label-free optical fiber biosensor based on a Mach-Zehnder interferometer for bovine serum albumin (BSA) concentration detection. The proposed fiber biosensor utilized a micro-cavity within a single-mode fiber to induce Mach-Zehnder interference. A remarkable feature of this biosensor is that external media can directly interact with the fiber core signal through microfluidic channels connected to the micro-cavity and sensor surface. The device was fabricated by means of femtosecond laser micromachining and chemical etching. A fiber interferometer of this type exhibits an ultrahigh refractive index sensitivity of -10,055 nm/RIU and a detection limit of 3.5 × 10 -5 RIU. Different concentrations of BSA with an infinitesimally small refractive index difference can be clearly differentiated in situ by the interferential spectra of the structure. Experiments demonstrated the biosensor exhibited a BSA solution concentration sensitivity of -38.9 nm/(mg/mL) and a detection limit of 2.57 × 10 -4 mg/mL, respectively. Moreover, this biosensor is a sub-microliter dose and ultrasensitive at the low concentrations detected in BSA, which make it a promising for biochemical applications such as DNA hybridization, cancer screenings, medicine examination and environmental engineering, etc.

  1. Electrolyte effect on the phase behavior of silica nanoparticles with lysozyme and bovine-serum-albumin proteins

    Science.gov (United States)

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

    2015-05-01

    Small-angle neutron scattering (SANS) and dynamic light scattering (DLS) studies have been carried out to investigate the effect of an electrolyte on the phase behavior of anionic silica nanoparticles with two globular proteins—cationic lysozyme [molecular weight (MW) 14.7 kDa] and anionic bovine serum albumin (MW 66.4 kDa). The results are compared with our earlier published work on similar systems without any electrolyte [I. Yadav, S. Kumar, V. K. Aswal, and J. Kohlbrecher, Phys. Rev. E 89, 032304 (2014), 10.1103/PhysRevE.89.032304]. Both the nanoparticle-protein systems transform to two phase at lower concentration of protein in the presence of an electrolyte. The autocorrelation function in DLS suggests that the diffusion coefficient (D) of a nanoparticle-protein system decreases in approaching two phase with the increase in protein concentration. This variation in D can be attributed to increase in attractive interaction and/or overall increase in the size. Further, these two contributions (interaction and structure) are determined from the SANS data. The changes in the phase behavior of nanoparticle-protein systems in the presence of an electrolyte are explained in terms of modifications in both the repulsive and attractive components of interaction between nanoparticles. In a two-phase system individual silica nanoparticles coexist along with their fractal aggregates.

  2. Hyaluronic acid-coated bovine serum albumin nanoparticles loaded with brucine as selective nanovectors for intra-articular injection

    Directory of Open Access Journals (Sweden)

    Chen Z

    2013-10-01

    Full Text Available Zhipeng Chen,* Juan Chen,* Li Wu, Weidong Li, Jun Chen, Haibo Cheng, Jinhuo Pan, Baochang CaiDepartment of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, People's Republic of China*These authors contributed equally to this workObjective: To evaluate the potential of hyaluronic acid (HA-coated bovine serum albumin nanoparticles (BSANPs as a novel chondrocyte-targeting drug-delivery nanomedicine.Methods: The HA-BSANPs were characterized by dynamic light scattering, transmission electron microscopy, differential scanning calorimetry, and X-ray diffraction. Fluorescence imaging was used to visualize the distribution of nanoparticles after intra-articular injection. The chondrocyte-targeting efficiency and cellular uptake mechanism of HA-BSANPs were investigated using endocytic inhibitors.Results: HA-BSANPs were successfully prepared with HA coating the surface and amorphous drug in the core. Compared with BSANPs, HA-BSANPs exhibited improved uptake by chondrocytes through a receptor-mediated active uptake mechanism. The endocytosis process of BSANPs and HA-BSANPs involved clathrin-mediated endocytosis, caveolae-mediated endocytosis, and macropinocytosis. No apparent thickening or hyperplasia of the synovium was observed in either BSANPs or HA-BSANPs. The HA-BSANPs could reside in the articular cavity of rats for more than 14 days, which was significantly longer than BSANPs.Conclusion: HA-BSANPs are a promising carrier for articular-related diseases due to elongated articular residence and improved chondrocytic accumulation.Keywords: chondrocyte, intra-articular injection, hyaluronic acid, BSA, nanoparticles

  3. Studies on the antagonistic action between chloramphenicol and quinolones with presence of bovine serum albumin by fluorescence spectroscopy

    International Nuclear Information System (INIS)

    Liu Baosheng; Zhao Fengli; Xue Chunli; Wang Jing; Lu Yunkai

    2010-01-01

    Chloramphenicol (CHL) and quinolone drugs like ofloxacin (OFLX), lomefloxacin (LMX) and ciprofloxacin (CPFX) can all quench the fluorescence of bovine serum albumin (BSA) in the aqueous solution of pH=7.40. This quenching effect becomes more significant when CHL and quinolone drugs coexist. Based on this, further studies on the interactions between CHL and quinolone drugs using fluorescence spectrum are established. The results showed that the interaction between the drugs would increase the binding constant and binding stability of the drug and protein, thus reducing the amount of drugs transported to their targets. Therefore, free drug concentration at targets would decrease, reducing the efficacy of the drugs. It indicated that there exists antagonistic action between drugs. The results also showed that the quenching mechanism of BSA by the drugs is a static procedure. The number of binding sites is 1 in various systems. Due to the existence of the antagonistic action between drugs, the binding distance r is reduced. Studies utilizing synchronous spectra showed that the antagonistic action between the drugs would affect the conformation of BSA, making protein molecules extend and hydrophobic decrease. The order of antagonistic action between CHL and quinolone drugs is: CPFX>OFLX>LMX with presence of BSA.

  4. Fluorescent copper(II complexes: The electron transfer mechanism, interaction with bovine serum albumin (BSA and antibacterial activity

    Directory of Open Access Journals (Sweden)

    Madhumita Hazra

    2017-01-01

    Full Text Available Dinuclear copper(II complexes with formula [Cu2(L2(N32] (1 and [Cu2(L2(NCS2] (2 HL = (1-[(3-methyl-pyridine-2-ylimino-methyl]-naphthalen-2-ol were synthesized by controlling the molar ratio of Cu(OAC2·6H2O, HL, sodium azide (1 and ammonium thiocyanate (2. The end on bridges appear exclusively in azide and thiocyanate to copper complexes. The electron transfer mechanism of copper(II complexes is examined by cyclic voltammetry indicating copper(II complexes are Cu(II/Cu(I couple. The interactions of copper(II complexes towards bovine serum albumin (BSA were examined with the help of absorption and fluorescence spectroscopic tools. We report a superficial solution-based route for the synthesis of micro crystals of copper complexes with BSA. The antibacterial activity of the Schiff base and its copper complexes were investigated by the agar disc diffusion method against some species of pathogenic bacteria (Escherichia coli, Vibrio cholerae, Streptococcus pneumonia and Bacillus cereus. It has been observed that the antibacterial activity of all complexes is higher than the ligand.

  5. Using resonance light scattering and UV/vis absorption spectroscopy to study the interaction between gliclazide and bovine serum albumin.

    Science.gov (United States)

    Zhang, Qiu-Ju; Liu, Bao-Sheng; Li, Gai-Xia; Han, Rong

    2016-08-01

    At different temperatures (298, 310 and 318 K), the interaction between gliclazide and bovine serum albumin (BSA) was investigated using fluorescence quenching spectroscopy, resonance light scattering spectroscopy and UV/vis absorption spectroscopy. The first method studied changes in the fluorescence of BSA on addition of gliclazide, and the latter two methods studied the spectral change in gliclazide while BSA was being added. The results indicated that the quenching mechanism between BSA and gliclazide was static. The binding constant (Ka ), number of binding sites (n), thermodynamic parameters, binding forces and Hill's coefficient were calculated at three temperatures. Values for the binding constant obtained using resonance light scattering and UV/vis absorption spectroscopy were much greater than those obtained from fluorescence quenching spectroscopy, indicating that methods monitoring gliclazide were more accurate and reasonable. In addition, the results suggest that other residues are involved in the reaction and the mode 'point to surface' existed in the interaction between BSA and gliclazide. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

  6. Tribological investigation of diamond-like carbon coated micro-dimpled surface under bovine serum and osteoarthritis oriented synovial fluid

    Science.gov (United States)

    Ghosh, Subir; Choudhury, Dipankar; Roy, Taposh; Mamat, Azuddin Bin; Masjuki, H. H.; Pingguan-Murphy, Belinda

    2015-06-01

    Osteoarthritis-oriented synovial fluid (OASF), i.e., that typical of a patient with osteoarthritis, has different physical and biological characteristics than bovine serum (BS), a lubricant widely used in biotribological investigations. Micro-dimpled and diamond-like carbon- (DLC) coated surfaces are key emerging interfaces for orthopedic implants. In this study, tribological performances of dimpled surfaces, with and without DLC coating, have been investigated under both BS and OASF. The friction tests were performed utilizing a pin on a disk tribometer, whereas contact pressure, speed, and temperature were simulated to a ‘medium walking gait’ of hip joint conditions. The mechanical properties of the specimen and the physical properties of the lubricant were characterized before the friction test. Raman analysis was conducted to identify the coating condition both before and after the test. The DLC-coated dimpled surface showed maximum hardness and residual stress. A DLC-coated dimpled surface under an OASF lubricated condition yielded a lower friction coefficient and wear compared to those of plain and dimpled specimens. The higher graphitization of coated materials with increasing load was confirmed by Raman spectroscopy.

  7. Tribological investigation of diamond-like carbon coated micro-dimpled surface under bovine serum and osteoarthritis oriented synovial fluid

    International Nuclear Information System (INIS)

    Ghosh, Subir; Roy, Taposh; Pingguan-Murphy, Belinda; Choudhury, Dipankar; Bin Mamat, Azuddin; Masjuki, H H

    2015-01-01

    Osteoarthritis-oriented synovial fluid (OASF), i.e., that typical of a patient with osteoarthritis, has different physical and biological characteristics than bovine serum (BS), a lubricant widely used in biotribological investigations. Micro-dimpled and diamond-like carbon- (DLC) coated surfaces are key emerging interfaces for orthopedic implants. In this study, tribological performances of dimpled surfaces, with and without DLC coating, have been investigated under both BS and OASF. The friction tests were performed utilizing a pin on a disk tribometer, whereas contact pressure, speed, and temperature were simulated to a ‘medium walking gait’ of hip joint conditions. The mechanical properties of the specimen and the physical properties of the lubricant were characterized before the friction test. Raman analysis was conducted to identify the coating condition both before and after the test. The DLC-coated dimpled surface showed maximum hardness and residual stress. A DLC-coated dimpled surface under an OASF lubricated condition yielded a lower friction coefficient and wear compared to those of plain and dimpled specimens. The higher graphitization of coated materials with increasing load was confirmed by Raman spectroscopy. (paper)

  8. Preparation, characterization and cytotoxic evaluation of bovine serum albumin nanoparticles encapsulating 5-methylmellein: A secondary metabolite isolated from Xylaria psidii.

    Science.gov (United States)

    Arora, Divya; Kumar, Amit; Gupta, Prasoon; Chashoo, Gousia; Jaglan, Sundeep

    2017-12-01

    In this study, 5-methylmellein (5-MM) loaded bovine serum albumin nanoparticles (BSA NPs) were developed using desolvation technique. The developed nanoparticles were characterized for their mean particle size, polydispersity, zeta potential, loading efficiency, X-ray diffractometry (XRD), differential scanning calorimetry (DSC) and release profile. The developed nanoparticles were spherical in shape under transmission electron microscopy (TEM) and atomic force microscopy (AFM). The developed 5-MM loaded BSA NPs demonstrated a mean particle size with a diameter of 154.95 ± 4.44 nm. The results from XRD and DSC studies demonstrated that the crystal state of the 5-MM was converted to an amorphous state in polymeric matrix. The encapsulation and loading efficiency was found to be 73.26 ± 4.48% and 7.09 ± 0.43%. The in vitro cytotoxicity in human prostate cancer cell line (PC-3), human colon cancer cells (HCT-116) and human breast adenocarcinoma cell line (MCF-7) cells demonstrated enhanced cytotoxicity of 5-MM BSA NPs as compared to native 5-MM after 72-h treatment. The enhancement in cytotoxicity of 5-MM BSA NPs was also supported by increase in cellular apoptosis, mitochondrial membrane potential loss and generation of high reactive oxygen species (ROS). In conclusion, these findings collectively indicated that BSA nanoparticles may serve as promising drug delivery system for improving the efficacy of 5-methylmellein. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Bovine serum albumin-GABA-His-Pro-NH2: an immunogen for production of higher affinity antisera for TRH

    International Nuclear Information System (INIS)

    Youngblood, W.W.; Moray, L.J.; Busby, W.H.; Kizer, J.S.

    1983-01-01

    Coupling the synthesize hapten, GABA-His-Pro-NH 2 to bovine serum albumin at a molar ratio of 18 : 1 by means of water-soluble carbodiimide produced an immunogen which stimulated the rapid production in New Zealand white rabbits of antisera with an affinity (2.42+-0.3x10 9 l/mol) for TRH, some 8-fold higher than that of antisera (0.33+-0.03x10 9 l/mol) raised by immunization with a conjugate produced by the currently accepted bis-diazotized-benzidine bridging technique. These higher affinity antibodies when used in a standard TRH radioimmunoassay permitted the detection of less than 1/pg of TRH per assay tube and showed an extremely low affinity for the two major metabolites of TRH, p-Glu-His-Pro-COOH and His-Pro diketopiperazine (4.84x10 4 and 4.0x10 4 l/mol, respectively). Application of this newer radioimmunoassay to the measurement of TRH in brain tissue yielded measurements of TRH content similar to those determined by current RIA methods. Chromatography of whole crude brain extracts revealed one major immunoreactive peak corresponding to authentic TRH. It is concluded that immunization of rabbits with this hapten rapidly produces antisera with a high affinity for TRH suitable for the development of a very sensitive TRH radioimmunoassay. (Auth.)

  10. Tribological investigation of diamond-like carbon coated micro-dimpled surface under bovine serum and osteoarthritis oriented synovial fluid

    Science.gov (United States)

    Ghosh, Subir; Choudhury, Dipankar; Roy, Taposh; Bin Mamat, Azuddin; Masjuki, H H; Pingguan-Murphy, Belinda

    2015-01-01

    Osteoarthritis-oriented synovial fluid (OASF), i.e., that typical of a patient with osteoarthritis, has different physical and biological characteristics than bovine serum (BS), a lubricant widely used in biotribological investigations. Micro-dimpled and diamond-like carbon- (DLC) coated surfaces are key emerging interfaces for orthopedic implants. In this study, tribological performances of dimpled surfaces, with and without DLC coating, have been investigated under both BS and OASF. The friction tests were performed utilizing a pin on a disk tribometer, whereas contact pressure, speed, and temperature were simulated to a ‘medium walking gait’ of hip joint conditions. The mechanical properties of the specimen and the physical properties of the lubricant were characterized before the friction test. Raman analysis was conducted to identify the coating condition both before and after the test. The DLC-coated dimpled surface showed maximum hardness and residual stress. A DLC-coated dimpled surface under an OASF lubricated condition yielded a lower friction coefficient and wear compared to those of plain and dimpled specimens. The higher graphitization of coated materials with increasing load was confirmed by Raman spectroscopy. PMID:27877803

  11. Interaction of bovine serum albumin with N-acyl amino acid based anionic surfactants: Effect of head-group hydrophobicity.

    Science.gov (United States)

    Ghosh, Subhajit; Dey, Joykrishna

    2015-11-15

    The function of a protein depends upon its structure and surfactant molecules are known to alter protein structure. For this reason protein-surfactant interaction is important in biological, pharmaceutical, and cosmetic industries. In the present work, interactions of a series of anionic surfactants having the same hydrocarbon chain length, but different amino acid head group, such as l-alanine, l-valine, l-leucine, and l-phenylalanine with the transport protein, bovine serum albumin (BSA), were studied at low surfactant concentrations using fluorescence and circular dichroism (CD) spectroscopy, and isothermal titration calorimetry (ITC). The results of fluorescence measurements suggest that the surfactant molecules bind simultaneously to the drug binding site I and II of the protein subdomain IIA and IIIA, respectively. The fluorescence as well as CD spectra suggest that the conformation of BSA goes to a more structured state upon surfactant binding at low concentrations. The binding constants of the surfactants were determined by the use of fluorescence as well as ITC measurements and were compared with that of the corresponding glycine-derived surfactant. The binding constant values clearly indicate a significant head-group effect on the BSA-surfactant interaction and the interaction is mainly hydrophobic in nature. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. Kinetics of Glycoxidation of Bovine Serum Albumin by Glucose, Fructose and Ribose and Its Prevention by Food Components

    Directory of Open Access Journals (Sweden)

    Izabela Sadowska-Bartosz

    2014-11-01

    Full Text Available The aim of this study was to compare the kinetics of the glycoxidation of bovine serum albumin (BSA as a model protein by three sugars: glucose, fructose and ribose, using fluorometric measurements of the content of advanced glycation end products (AGEs, protein-bound fructosamine, dityrosine, N'-formylkynurenine, kynurenine, tryptophan, the content of advanced oxidation protein products (AOPP, protein carbonyl groups, as well as thiol groups. Moreover, the levels of glycoalbumin and AGEs were determined by using an enzyme-linked immunosorbent assay. Based on the kinetic results, the optimal incubation time for studies of the modification of the glycoxidation rate by additives was chosen, and the effects of 25 compounds of natural origin on the glycoxidation of BSA induced by various sugars were examined. The same compounds were found to have different effects on glycoxidation induced by various sugars, which suggests caution in extrapolation from experiments based on one sugar to other sugars. From among the compounds tested, the most effective inhibitors of glycoxidation were: polyphenols, pyridoxine and 1-cyano-4-hydroxycinnamic acid.

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

    Science.gov (United States)

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

    2011-07-01

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

  14. Effect of ionic micellar medium on kinetics and mechanism of oxidation of bovine serum albumin: A pulse radiolysis study

    Energy Technology Data Exchange (ETDEWEB)

    Joshi, Ravi, E-mail: rjudrin@yahoo.co [Radiation and Photochemistry Division, Chemistry Group, Bhabha Atomic Research Center, Mumbai 400085 (India); Mukherjee, Tulsi [Radiation and Photochemistry Division, Chemistry Group, Bhabha Atomic Research Center, Mumbai 400085 (India)

    2010-09-15

    Effect of protein-micelle interaction on bovine serum albumin (BSA) oxidation by trichloromethyl peroxyl radical (CCl{sub 3}O{sub 2}{sup .}) in anionic sodium dodecyl sulfate (SDS) and cationic cetyltrimethyl ammonium bromide (CTAB) micellar media has been studied using nanosecond pulse radiolysis technique. Viscosity measurement and light scattering studies have suggested that SDS and CTAB micelles produce BSA-micelle aggregates of different sizes and polydispersity. Oxidation kinetics and transients have been affected both by anionic SDS and cationic CTAB micelles but in a different manner. Tryptophanyl-CCl{sub 3}O{sub 2}{sup .} adduct radical to tyrosyl radical transformation in BSA has been observed in anionic SDS micelles but not in cationic CTAB micelles. Similar studies have also been done with tryptophan and tyrosine amino acids, which undergo oxidation in BSA. The study suggests that Coulombic and hydrophobic interactions between micelles and protein affect the structure of the protein to shield its functional amino acids, like tryptophan and tyrosine, to neutral oxidizing radical.

  15. Role of pH-induced structural change in protein aggregation in foam fractionation of bovine serum albumin

    Directory of Open Access Journals (Sweden)

    Rui Li

    2016-03-01

    Full Text Available For reducing protein aggregation in foam fractionation, the role of pH-induced structural change in the interface-induced protein aggregation was analyzed using bovine serum albumin (BSA as a model protein. The results show that the decrease in pH from 7.0 to 3.0 gradually unfolded the BSA structure to increase the molecular size and the relative content of β-sheet and thus reduced the stability of BSA in the aqueous solution. At the isoelectric point (pH 4.7, BSA suffered the lowest level in protein aggregation induced by the gas–liquid interface. In the pH range from 7.0 to 4.7, most BSA aggregates were formed in the defoaming process while in the pH range from 4.7 to 3.0, the BSA aggregates were formed at the gas–liquid interface due to the unfolded BSA structure and they further aggregated to form insoluble ones in the desorption process.

  16. Effects of Multiwalled Carbon Nanotube Surface Modification and Purification on Bovine Serum Albumin Binding and Biological Responses

    Directory of Open Access Journals (Sweden)

    Wei Bai

    2016-01-01

    Full Text Available Carboxylation of multiwalled carbon nanotubes (MWCNTs has been used to improve solubility in aqueous systems and for further functionalization with biologically active moieties for biomedical uses. An important consideration is that oxidation debris is generated during the process of carboxylation, which can be removed by base washing. We hypothesized that surface modification as well as purification by debris removal may alter physicochemical properties of MWCNTs and their ability to bind proteins. We utilized pristine MWCNT, carboxylated MWCNTs (F-MWCNTs, and base-washed carboxylated MWCNTs (BW-F-MWCNTs to examine formation of a bovine serum albumin (BSA protein corona and impact on biological responses. We found that carboxylation increased the capability of F-MWCNTs to bind BSA, and base washing further increased this binding. Functionalization increased cellular uptake by rat aortic endothelial cells (RAEC and mouse macrophages (RAW264.7, while base washing showed results similar to the functionalized analog. Interestingly, BSA binding downregulated mRNA levels of interleukin-6 (IL-6 and heme oxygenase 1 (Hmox1 in RAEC cells but upregulated the expression of IL-6 and Hmox1 in RAW264.7 cells. Overall, our study demonstrated that surface modification as well as further purification impacted the interaction of MWCNTs with proteins and subsequent cellular responses.

  17. Insight into the Interaction between the HIV-1 Integrase Inhibitor Elvitegravir and Bovine Serum Albumin: A Spectroscopic Study

    Directory of Open Access Journals (Sweden)

    Ali Saber Abdelhameed

    2015-01-01

    Full Text Available The interaction between the anti-HIV drug Elvitegravir (EVG and bovine serum albumin (BSA was investigated by fluorescence spectroscopy and UV-visible absorption spectra. The mechanism for quenching the fluorescence of BSA by EVG is discussed. It was found that EVG can quench the intrinsic fluorescence of BSA through a static quenching procedure. The quenching type, association constant, and number of binding sites were investigated. The binding constant of EVG with BSA was calculated at different temperatures based on fluorescence quenching results. The thermodynamic parameters ΔHθ, ΔGθ, and ΔSθ were determined. The positive ΔSθ and negative ΔHθ and ΔGθ values showed that a spontaneous interaction may involve both roles of hydrophobic interaction and hydrogen bonding. The interaction of BSA with EVG was also confirmed by UV absorption spectra. The average distance, r, between donor (BSA and acceptor (EVG was obtained according to Förster’s theory of nonradiation energy transfer. Synchronous fluorescence and three-dimensional fluorescence spectra were used to investigate the conformational change of BSA molecules that occur upon addition of EVG and showed, upon binding, a possibility of increasing hydrophobicity around tryptophan residues of BSA.

  18. Microplasma jet treatment of bovine serum albumin coatings for controlling enzyme and cell attachmenttype="fn" rid="FN1">

    Science.gov (United States)

    Szili, Endre J.; Becker, Stefanie; Short, Robert D.; Al-Bataineh, Sameer A.

    2017-08-01

    We investigated a new approach to control protein and cell attachment inside 96-well polystyrene plates. The wells were first coated with bovine serum albumin (BSA) to inhibit cell and protein attachment. The BSA-coated wells were then treated with a helium microplasma jet for increasing times that resulted in gradual removal of BSA from the surface. It was found that the amount of enzyme and cell attachment could be controlled in the wells where BSA was only partially removed by the microplasma jet. In addition to the surface coverage of BSA, the new surface chemistry induced by the microplasma jet treatment also had an important role in the control of enzyme and cell attachment. In summary, microplasma jet treatment of BSA-coated polystyrene wells is a simple and effective method for controlling enzyme and cell attachment. This might find use for high-throughput screening of new cell culture platforms where control over the level protein, enzyme or cell adherence is needed in order to maintain a specific cell function.

  19. A sol-gel derived pH-responsive bovine serum albumin molecularly imprinted poly(ionic liquids) on the surface of multiwall carbon nanotubes

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Mingming, E-mail: liumm@mail.hzau.edu.cn [Key Laboratory of Arable Land Conservation (Middle and Lower Reaches of Yangtse River), Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070 (China); Pi, Jiangyan; Wang, Xiaojie; Huang, Rong; Du, Yamei; Yu, Xiaoyang; Tan, Wenfeng; Liu, Fan [Key Laboratory of Arable Land Conservation (Middle and Lower Reaches of Yangtse River), Ministry of Agriculture, College of Resources and Environment, Huazhong Agricultural University, Wuhan 430070 (China); Shea, Kenneth J., E-mail: kjshea@uci.edu [Department of Chemistry, University of California-Irvine, Irvine, CA 92697 (United States)

    2016-08-17

    A pH-responsive surface molecularly imprinted poly(ionic liquids) (MIPILs) was prepared on the surface of multiwall carbon nanotubes (MWCNTs) by a sol-gel technique. The material was synthesized using a 3-aminopropyl triethoxysilane modified multiwall carbon nanotube (MWCNT-APTES) as the substrate, bovine serum albumin (BSA) as the template molecule, an alkoxy-functionalized IL 1-(3-trimethoxysilyl propyl)-3-methyl imidazolium chloride ([TMSPMIM]Cl) as both the functional monomer and the sol-gel catalyst, and tetraethoxysilane (TEOS) as the crosslinking agent. The molecular interaction between BSA and [TMSPMIM]Cl was quantitatively evaluated by UV–vis spectroscopy prior to polymerization so as to identify an optimal template/monomer ratio and the most suitable pH value for the preparation of the MWCNTs@BSA-MIPILs. This strategy was found to be effective to overcome the problems of trial-and-error protocol in molecular imprinting. The optimum synthesis conditions were as follows: template/monomer ratio 7:20, crosslinking agent content 2.0–2.5 mL, temperature 4 °C and pH 8.9 Tris–HCl buffer. The influence of incubation pH on adsorption was also studied. The result showed that the imprinting effect and selectivity improved significantly with increasing incubation pH from 7.7 to 9.9. This is mainly because the non-specific binding from electrostatic and hydrogen bonding interactions decreased greatly with the increase of pH value, which made the specific binding affinity from shape selectivity strengthened instead. The polymers synthesized under the optimal conditions were then characterized by BET surface area measurement, FTIR, thermogravimetric analysis (TGA) and scanning electron microscopy (SEM). The adsorption capacity, imprinting effect, selective recognition and reusability were also evaluated. The as-prepared MWCNTs@BSA-MIPILs were also found to have a number of advantages including high surface area (134.2 m{sup 2} g{sup −1}), high adsorption

  20. A sol-gel derived pH-responsive bovine serum albumin molecularly imprinted poly(ionic liquids) on the surface of multiwall carbon nanotubes

    International Nuclear Information System (INIS)

    Liu, Mingming; Pi, Jiangyan; Wang, Xiaojie; Huang, Rong; Du, Yamei; Yu, Xiaoyang; Tan, Wenfeng; Liu, Fan; Shea, Kenneth J.

    2016-01-01

    A pH-responsive surface molecularly imprinted poly(ionic liquids) (MIPILs) was prepared on the surface of multiwall carbon nanotubes (MWCNTs) by a sol-gel technique. The material was synthesized using a 3-aminopropyl triethoxysilane modified multiwall carbon nanotube (MWCNT-APTES) as the substrate, bovine serum albumin (BSA) as the template molecule, an alkoxy-functionalized IL 1-(3-trimethoxysilyl propyl)-3-methyl imidazolium chloride ([TMSPMIM]Cl) as both the functional monomer and the sol-gel catalyst, and tetraethoxysilane (TEOS) as the crosslinking agent. The molecular interaction between BSA and [TMSPMIM]Cl was quantitatively evaluated by UV–vis spectroscopy prior to polymerization so as to identify an optimal template/monomer ratio and the most suitable pH value for the preparation of the MWCNTs@BSA-MIPILs. This strategy was found to be effective to overcome the problems of trial-and-error protocol in molecular imprinting. The optimum synthesis conditions were as follows: template/monomer ratio 7:20, crosslinking agent content 2.0–2.5 mL, temperature 4 °C and pH 8.9 Tris–HCl buffer. The influence of incubation pH on adsorption was also studied. The result showed that the imprinting effect and selectivity improved significantly with increasing incubation pH from 7.7 to 9.9. This is mainly because the non-specific binding from electrostatic and hydrogen bonding interactions decreased greatly with the increase of pH value, which made the specific binding affinity from shape selectivity strengthened instead. The polymers synthesized under the optimal conditions were then characterized by BET surface area measurement, FTIR, thermogravimetric analysis (TGA) and scanning electron microscopy (SEM). The adsorption capacity, imprinting effect, selective recognition and reusability were also evaluated. The as-prepared MWCNTs@BSA-MIPILs were also found to have a number of advantages including high surface area (134.2 m 2  g −1 ), high adsorption capacity

  1. Calorimetric and spectroscopic properties of small globular proteins (bovine serum albumin, hemoglobin) after free radical generation

    International Nuclear Information System (INIS)

    Farkas, N.; Belagyi, J.; Lorinczy, D.

    2003-01-01

    Mild oxidation of -SH-containing proteins (serum albumin, hemoglobin) by Ce(IV)-ions in the presence of the spin trap phenyl-tert-butylnitrone (PBN) resulted in the appearance of strongly immobilized nitroxide free radicals which evidences the formation of thiyl radicals on the thiol site of the proteins. In hydroxyl free radical generating system a fraction of strongly immobilized nitroxide radicals was also detected in these proteins, which implies that the oxidation of a fraction of the thiol groups was also involved in the free radical reaction. According to the differential scanning calorimetry (DSC) experiments the melting processes of the proteins were calorimetrically irreversible, therefore the two-state kinetic model was used to evaluate the experiments. The results support the view that site-specific interaction of SH-containing proteins with hydroxyl and thiyl free radicals is able to modify the internal dynamics of proteins and affect the conformation of large molecules

  2. A sol-gel derived pH-responsive bovine serum albumin molecularly imprinted poly(ionic liquids) on the surface of multiwall carbon nanotubes.

    Science.gov (United States)

    Liu, Mingming; Pi, Jiangyan; Wang, Xiaojie; Huang, Rong; Du, Yamei; Yu, Xiaoyang; Tan, Wenfeng; Liu, Fan; Shea, Kenneth J

    2016-08-17

    A pH-responsive surface molecularly imprinted poly(ionic liquids) (MIPILs) was prepared on the surface of multiwall carbon nanotubes (MWCNTs) by a sol-gel technique. The material was synthesized using a 3-aminopropyl triethoxysilane modified multiwall carbon nanotube (MWCNT-APTES) as the substrate, bovine serum albumin (BSA) as the template molecule, an alkoxy-functionalized IL 1-(3-trimethoxysilyl propyl)-3-methyl imidazolium chloride ([TMSPMIM]Cl) as both the functional monomer and the sol-gel catalyst, and tetraethoxysilane (TEOS) as the crosslinking agent. The molecular interaction between BSA and [TMSPMIM]Cl was quantitatively evaluated by UV-vis spectroscopy prior to polymerization so as to identify an optimal template/monomer ratio and the most suitable pH value for the preparation of the MWCNTs@BSA-MIPILs. This strategy was found to be effective to overcome the problems of trial-and-error protocol in molecular imprinting. The optimum synthesis conditions were as follows: template/monomer ratio 7:20, crosslinking agent content 2.0-2.5 mL, temperature 4 °C and pH 8.9 Tris-HCl buffer. The influence of incubation pH on adsorption was also studied. The result showed that the imprinting effect and selectivity improved significantly with increasing incubation pH from 7.7 to 9.9. This is mainly because the non-specific binding from electrostatic and hydrogen bonding interactions decreased greatly with the increase of pH value, which made the specific binding affinity from shape selectivity strengthened instead. The polymers synthesized under the optimal conditions were then characterized by BET surface area measurement, FTIR, thermogravimetric analysis (TGA) and scanning electron microscopy (SEM). The adsorption capacity, imprinting effect, selective recognition and reusability were also evaluated. The as-prepared MWCNTs@BSA-MIPILs were also found to have a number of advantages including high surface area (134.2 m(2) g(-1)), high adsorption capacity (55.52

  3. Probing the interaction of a new synthesized CdTe quantum dots with human serum albumin and bovine serum albumin by spectroscopic methods

    International Nuclear Information System (INIS)

    Bardajee, Ghasem Rezanejade; Hooshyar, Zari

    2016-01-01

    A novel CdTe quantum dots (QDs) were prepared in aqueous phase via a facile method. At first, poly (acrylic amide) grafted onto sodium alginate (PAAm-g-SA) were successfully synthesized and then TGA capped CdTe QDs (CdTe-TGA QDs) were embed into it. The prepared CdTe-PAAm-g-SA QDs were optimized and characterized by transmission electron microscopy (TEM), thermo-gravimetric (TG) analysis, Fourier transform infrared (FT-IR), UV–vis and fluorescence spectroscopy. The characterization results indicated that CdTe-TGA QDs, with particles size of 2.90 nm, were uniformly dispersed on the chains of PAAm-g-SA biopolymer. CdTe-PAAm-g-SA QDs also exhibited excellent UV–vis absorption and high fluorescence intensity. To explore biological behavior of CdTe-PAAm-g-SA QDs, the interactions between CdTe-PAAm-g-SA QDs and human serum albumin (HSA) (or bovine serum albumin (BSA)) were investigated by cyclic voltammetry, FT-IR, UV–vis, and fluorescence spectroscopic. The results confirmed the formation of CdTe-PAAm-g-SA QDs-HSA (or BSA) complex with high binding affinities. The thermodynamic parameters (ΔG < 0, ΔH < 0 and ΔS < 0) were indicated that binding reaction was spontaneous and van der Waals interactions and hydrogen-bond interactions played a major role in stabilizing the CdTe-PAAm-g-SA QDs-HSA (or BSA) complexes. The binding distance between CdTe-PAAm-g-SA QDs and HSA (or BSA)) was calculated about 1.37 nm and 1.27 nm, respectively, according to Forster non-radiative energy transfer theory (FRET). Analyzing FT-IR spectra showed that the formation of QDs-HSA and QDs-BSA complexes led to conformational changes of the HSA and BSA proteins. All these experimental results clarified the effective transportation and elimination of CdTe-PAAm-g-SA QDs in the body by binding to HSA and BSA, which could be a useful guideline for the estimation of QDs as a drug carrier. - Highlights: • The CdTe quantum dots coated with polyacrylamide grafted onto sodium alginate. • The

  4. Probing the interaction of a new synthesized CdTe quantum dots with human serum albumin and bovine serum albumin by spectroscopic methods

    Energy Technology Data Exchange (ETDEWEB)

    Bardajee, Ghasem Rezanejade, E-mail: rezanejad@pnu.ac.ir; Hooshyar, Zari

    2016-05-01

    A novel CdTe quantum dots (QDs) were prepared in aqueous phase via a facile method. At first, poly (acrylic amide) grafted onto sodium alginate (PAAm-g-SA) were successfully synthesized and then TGA capped CdTe QDs (CdTe-TGA QDs) were embed into it. The prepared CdTe-PAAm-g-SA QDs were optimized and characterized by transmission electron microscopy (TEM), thermo-gravimetric (TG) analysis, Fourier transform infrared (FT-IR), UV–vis and fluorescence spectroscopy. The characterization results indicated that CdTe-TGA QDs, with particles size of 2.90 nm, were uniformly dispersed on the chains of PAAm-g-SA biopolymer. CdTe-PAAm-g-SA QDs also exhibited excellent UV–vis absorption and high fluorescence intensity. To explore biological behavior of CdTe-PAAm-g-SA QDs, the interactions between CdTe-PAAm-g-SA QDs and human serum albumin (HSA) (or bovine serum albumin (BSA)) were investigated by cyclic voltammetry, FT-IR, UV–vis, and fluorescence spectroscopic. The results confirmed the formation of CdTe-PAAm-g-SA QDs-HSA (or BSA) complex with high binding affinities. The thermodynamic parameters (ΔG < 0, ΔH < 0 and ΔS < 0) were indicated that binding reaction was spontaneous and van der Waals interactions and hydrogen-bond interactions played a major role in stabilizing the CdTe-PAAm-g-SA QDs-HSA (or BSA) complexes. The binding distance between CdTe-PAAm-g-SA QDs and HSA (or BSA)) was calculated about 1.37 nm and 1.27 nm, respectively, according to Forster non-radiative energy transfer theory (FRET). Analyzing FT-IR spectra showed that the formation of QDs-HSA and QDs-BSA complexes led to conformational changes of the HSA and BSA proteins. All these experimental results clarified the effective transportation and elimination of CdTe-PAAm-g-SA QDs in the body by binding to HSA and BSA, which could be a useful guideline for the estimation of QDs as a drug carrier. - Highlights: • The CdTe quantum dots coated with polyacrylamide grafted onto sodium alginate. • The

  5. Development of bovine serum albumin-water partition coefficients predictive models for ionogenic organic chemicals based on chemical form adjusted descriptors.

    Science.gov (United States)

    Ding, Feng; Yang, Xianhai; Chen, Guosong; Liu, Jining; Shi, Lili; Chen, Jingwen

    2017-10-01

    The partition coefficients between bovine serum albumin (BSA) and water (K BSA/w ) for ionogenic organic chemicals (IOCs) were different greatly from those of neutral organic chemicals (NOCs). For NOCs, several excellent models were developed to predict their logK BSA/w . However, it was found that the conventional descriptors are inappropriate for modeling logK BSA/w of IOCs. Thus, alternative approaches are urgently needed to develop predictive models for K BSA/w of IOCs. In this study, molecular descriptors that can be used to characterize the ionization effects (e.g. chemical form adjusted descriptors) were calculated and used to develop predictive models for logK BSA/w of IOCs. The models developed had high goodness-of-fit, robustness, and predictive ability. The predictor variables selected to construct the models included the chemical form adjusted averages of the negative potentials on the molecular surface (V s-adj - ), the chemical form adjusted molecular dipole moment (dipolemoment adj ), the logarithm of the n-octanol/water distribution coefficient (logD). As these molecular descriptors can be calculated from their molecular structures directly, the developed model can be easily used to fill the logK BSA/w data gap for other IOCs within the applicability domain. Furthermore, the chemical form adjusted descriptors calculated in this study also could be used to construct predictive models on other endpoints of IOCs. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Cooperative cytotoxic activity of Zn and Cu in bovine serum albumin-conjugated ZnS/CuS nano-composites in PC12 cancer cells

    Science.gov (United States)

    Wang, Hua-Jie; Yu, Xue-Hong; Wang, Cai-Feng; Cao, Ying

    2013-11-01

    Series of self-assembled and mono-dispersed bovine serum albumin (BSA)-conjugated ZnS/CuS nano-composites with different Zn/Cu ratios had been successfully synthesized by a combination method of the biomimetic synthesis and ion-exchange strategy under the gentle conditions. High-resolution transmission electron microscopy observation, Fourier transform infrared spectra and zeta potential analysis demonstrated that BSA-conjugated ZnS/CuS nano-composites with well dispersity had the hierarchical structure and BSA was a key factor to control the morphology and surface electro-negativity of final products. The real-time monitoring by atomic absorption spectroscopy and powder X-ray diffraction revealed that the Zn/Cu ratio of nano-composites could be controlled by adjusting the ion-exchange time. In addition, the metabolic and morphological assays indicated that the metabolic proliferation and spread of rat pheochromocytoma (PC12) cells could be inhibited by nano-composites, with the high anti-cancer activity at a low concentration (4 ppm). What were more important, Zn and Cu in nano-composites exhibited a positive cooperativity at inhibiting cancer cell functions. The microscope observation and biochemical marker analysis clearly revealed that the nano-composites-included lipid peroxidation and disintegration of membrane led to the death of PC12 cells. Summarily, the present study substantiated the potential of BSA-conjugated ZnS/CuS nano-composites as anti-cancer drug.

  7. Cooperative cytotoxic activity of Zn and Cu in bovine serum albumin-conjugated ZnS/CuS nano-composites in PC12 cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Hua-Jie, E-mail: wanghuajie972001@163.com; Yu, Xue-Hong; Wang, Cai-Feng; Cao, Ying, E-mail: caoying1130@sina.com [Henan Normal University, Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, College of Chemistry and Chemical Engineering (China)

    2013-11-15

    Series of self-assembled and mono-dispersed bovine serum albumin (BSA)-conjugated ZnS/CuS nano-composites with different Zn/Cu ratios had been successfully synthesized by a combination method of the biomimetic synthesis and ion-exchange strategy under the gentle conditions. High-resolution transmission electron microscopy observation, Fourier transform infrared spectra and zeta potential analysis demonstrated that BSA-conjugated ZnS/CuS nano-composites with well dispersity had the hierarchical structure and BSA was a key factor to control the morphology and surface electro-negativity of final products. The real-time monitoring by atomic absorption spectroscopy and powder X-ray diffraction revealed that the Zn/Cu ratio of nano-composites could be controlled by adjusting the ion-exchange time. In addition, the metabolic and morphological assays indicated that the metabolic proliferation and spread of rat pheochromocytoma (PC12) cells could be inhibited by nano-composites, with the high anti-cancer activity at a low concentration (4 ppm). What were more important, Zn and Cu in nano-composites exhibited a positive cooperativity at inhibiting cancer cell functions. The microscope observation and biochemical marker analysis clearly revealed that the nano-composites-included lipid peroxidation and disintegration of membrane led to the death of PC12 cells. Summarily, the present study substantiated the potential of BSA-conjugated ZnS/CuS nano-composites as anti-cancer drug.

  8. Dual effect of fetal bovine serum on early development depends on stage-specific reactive oxygen species demands in pigs.

    Directory of Open Access Journals (Sweden)

    Seong-Eun Mun

    Full Text Available Despite the application of numerous supplements to improve in vitro culture (IVC conditions of mammalian cells, studies regarding the effect of fetal bovine serum (FBS on mammalian early embryogenesis, particularly in relation to redox homeostasis, are lacking. Herein, we demonstrated that early development of in vitro-produced (IVP porcine embryos highly depends on the combination of FBS supplementation timing and embryonic reactive oxygen species (ROS requirements. Interestingly, FBS significantly reduced intracellular ROS levels in parthenogenetically activated (PA embryos regardless of the developmental stage. However, the beneficial effect of FBS on early embryogenesis was found only during the late phase (IVC 4-6 days treatment group. In particular, developmental competence parameters, such as blastocyst formation rate, cellular survival, total cell number and trophectoderm proportion, were markedly increased by FBS supplementation during the late IVC phase. In addition, treatment with FBS elevated antioxidant transcript levels during the late IVC phase. In contrast, supplementation with FBS during the entire period (1-6 days or during the early IVC phase (1-2 days greatly impaired the developmental parameters. Consistent with the results from PA embryos, the developmental competence of in vitro fertilization (IVF or somatic cell nuclear transfer (SCNT embryos were markedly improved by treatment with FBS during the late IVC phase. Moreover, the embryonic stage-specific effects of FBS were reversed by the addition of an oxidant and were mimicked by treatment with an antioxidant. These findings may increase our understanding of redox-dependent early embryogenesis and contribute to the large-scale production of high-quality IVP embryos.

  9. Theoretical model to investigate the alkyl chain and anion dependent interactions of gemini surfactant with bovine serum albumin.

    Science.gov (United States)

    Vishvakarma, Vijay K; Kumari, Kamlesh; Patel, Rajan; Dixit, V S; Singh, Prashant; Mehrotra, Gopal K; Chandra, Ramesh; Chakrawarty, Anand Kumar

    2015-05-15

    Surfactants are used to prevent the irreversible aggregation of partially refolded proteins and they also assist in protein refolding. We have reported the design and screening of gemini surfactant to stabilize bovine serum albumin (BSA) with the help of computational tool (iGEMDOCK). A series of gemini surfactant has been designed based on bis-N-alkyl nicotinate dianion via varying the alkyl group and anion. On changing the alkyl group and anion of the surfactant, the value of Log P changes means polarity of surfactant can be tuned. Further, the virtual screening of the gemini surfactant has been carried out based on generic evolutionary method. Herein, thermodynamic data was studied to determine the potential of gemini surfactant as BSA stabilizer. Computational tools help to find out the efficient gemini surfactant to stabilize the BSA rather than to use the surfactant randomly and directionless for the stabilization. It can be confirmed through the experimental techniques. Previously, researcher synthesized one of the designed and used gemini surfactant to stabilize the BSA and their interactions were confirmed through various techniques and computational docking. But herein, the authors find the most competent gemini surfactant to stabilize BSA using computational tools on the basis of energy score. Different from the single chain surfactant, the gemini surfactants exhibit much stronger electrostatic and hydrophobic interactions with the protein and are thus effective at much lower concentrations. Based on the present study, it is expected that gemini surfactants may prove useful in the protein stabilization operations and may thus be effectively employed to circumvent the problem of misfolding and aggregation. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Isoelectric Bovine Serum Albumin: Robust Blocking Agent for Enhanced Performance in Optical-Fiber Based DNA Sensing.

    Science.gov (United States)

    Wang, Ruoyu; Zhou, Xiaohong; Zhu, Xiyu; Yang, Chao; Liu, Lanhua; Shi, Hanchang

    2017-02-24

    Surface blocking is a well-known process for reducing unwanted nonspecific adsorption in sensor fabrication, especially important in the emerging field where DNA/RNA applied. Bovine serum albumin (BSA) is one of the most popular blocking agents with an isoelectric point at pH 4.6. Although it is widely recognized that the adsorption of a blocking agent is strongly affected by its net charge and the maximum adsorption is often observed under its isoelectric form, BSA has long been perfunctorily used for blocking merely in neutral solution, showing poor blocking performances in the optical-fiber evanescent wave (OFEW) based sensing toward DNA target. To meet this challenge, we first put forward the view that isoelectric BSA (iep-BSA) has the best blocking performance and use an OFEW sensor platform to demonstrate this concept. An optical-fiber was covalently modified with amino-DNA, and further coupled with the optical system to detect fluorophore labeled complementary DNA within the evanescent field. A dramatic improvement in the reusability of this DNA modified sensing surface was achieved with 120 stable detection cycles, which ensured accurate quantitative bioassay. As expected, the iep-BSA blocked OFEW system showed enhanced sensing performance toward target DNA with a detection limit of 125 pM. To the best of our knowledge, this is the highest number of regeneration cycles ever reported for a DNA immobilized optical-fiber surface. This study can also serve as a good reference and provide important implications for developing similar DNA-directed surface biosensors.

  11. Enhanced extraction of bovine serum albumin with aqueous biphasic systems of phosphonium- and ammonium-based ionic liquids

    Science.gov (United States)

    Pereira, Matheus M.; Pedro, Sónia N.; Quental, Maria V.; Lima, Álvaro S.; Coutinho, João A. P.; Freire, Mara G.

    2016-01-01

    Novel aqueous biphasic systems (ABS) composed of phosphonium- or ammonium-based ionic liquids (ILs), combined with a buffered aqueous solution of potassium citrate/citric acid (pH=7.0), were investigated for the extraction of proteins. For that purpose, the phase diagrams, tie-lines and tie-line lengths were determined at 25ºC, and the performance of these ABS for the extraction of bovine serum albumin (BSA) was then evaluated. The obtained results reveal that, with the exception of the more hydrophobic ILs, most of the systems investigated allow the complete extraction of BSA for the IL-rich phase in a single-step. These remarkable extraction efficiencies are far superior to those afforded by more conventional extraction systems previously reported. The composition of the biphasic systems, i.e., the amount of phase-forming components, was also investigated aiming at reducing the overall costs of the process without losing efficiency on the protein extraction. It is shown that the extraction efficiencies of BSA are maintained at 100% up to high protein concentrations (at least up to 10 g.L-1). The recovery of the BSA from the IL-rich phase by dialysis is also shown in addition to the demonstration of the IL recyclability and reusability, at least for 3 times. In the sequential three-step extractions (BSA recovery/IL reusability), the extraction efficiencies of BSA for the IL-rich phase were maintained at 100%. For the improved ABS, the preservation of the protein native conformation was confirmed by Size Exclusion High-Performance Liquid Chromatography (used also as the quantification method) and by Fourier Transform Infra-Red spectroscopy. According to the results herein reported, ABS composed of phosphonium- or ammonium-based ILs and a biodegradable organic salt represent an alternative and remarkable platform for the extraction of BSA and may be extended to other proteins of interest. PMID:25865275

  12. Effect of bovine serum albumin on the functionality and structure of catanionic surfactant at air-buffer interface

    Energy Technology Data Exchange (ETDEWEB)

    Maiti, Kajari; Bhattacharya, Subhash C.; Moulik, Satya P. [Centre for Surface Science, Department of Chemistry, Jadavpur University, Kolkata 700 032 (India); Panda, Amiya K., E-mail: akpanda1@yahoo.com [Department of Chemistry, University of North Bengal, Darjeeling 734 013 (India)

    2013-03-01

    Interaction of bovine serum albumin (BSA) with the solvent spread monolayer of a catanionic surfactant, octadecyltrimethylammonium dodecylsulfate, (C{sub 18}TA{sup +}DS{sup -}) at the air-buffer interface was investigated by measuring the surface pressure with time and change in surface area. Dipalmitoylphosphatidylcholine (DPPC) was used as reference. Kinetics of BSA desorption from the interface to the buffer subphase, that of C{sub 18}TA{sup +}DS{sup -} and DPPC through their interaction with BSA, were also studied at different BSA concentrations (in the subphase) and surface pressures. Surface pressure ({pi})-area (A) isotherms (at pH = 5.4, {mu} = 0.01, T = 298 K) revealed that the coacervate/DPPC monolayer becomes expanded in the presence of BSA at low {pi} while their protein bound species are released into the subphase at high {pi}. Film morphology, studied by epifluorescence microscopy (EFM) and atomic force microscopy (AFM), reveals that the sizes of the domains of both DPPC and coacervate decrease in the presence of BSA. Presence of BSA in the coacervate and DPPC monolayer was supported from AFM data analysis. Highlights: Black-Right-Pointing-Pointer Effect of BSA on the functionality and structure of C{sub 18}TA{sup +}DS{sup -}/DPPC at the air-buffer interface was studied. Black-Right-Pointing-Pointer BSA molecules coadsorb at lower surface pressure, while they abstract amphiphiles at higher surface pressure into the bulk. Black-Right-Pointing-Pointer Kinetic studies of adsorption/desorption of BSA at/from the interface were performed. Black-Right-Pointing-Pointer Organized amphiphiles are perturbed in the presence of BSA.

  13. Characterizing the binding interaction between antimalarial artemether (AMT) and bovine serum albumin (BSA): Spectroscopic and molecular docking methods.

    Science.gov (United States)

    Shi, Jie-Hua; Pan, Dong-Qi; Wang, Xiou-Xiou; Liu, Ting-Ting; Jiang, Min; Wang, Qi

    2016-09-01

    Artemether (AMT), a peroxide sesquiterpenoides, has been widely used as an antimalarial for the treatment of multiple drug-resistant strains of plasmodium falciparum malaria. In this work, the binding interaction of AMT with bovine serum albumin (BSA) under the imitated physiological conditions (pH7.4) was investigated by UV spectroscopy, fluorescence emission spectroscopy, synchronous fluorescence spectroscopy, Fourier transform infrared spectroscopy (FT-IR), circular dichroism (CD), three-dimensional fluorescence spectroscopy and molecular docking methods. The experimental results indicated that there was a change in UV absorption of BSA along with a slight red shift of absorption wavelength, indicating that the interaction of AMT with BSA occurred. The intrinsic fluorescence of BSA was quenched by AMT due to the formation of AMT-BSA complex. The number of binding sites (n) and binding constant of AMT-BSA complex were about 1 and 2.63×10(3)M(-1) at 298K, respectively, suggesting that there was stronger binding interaction of AMT with BSA. Based on the analysis of the signs and magnitudes of the free energy change (ΔG(0)), enthalpic change (ΔH(0)) and entropic change (ΔS(0)) in the binding process, it can be concluded that the binding of AMT with BSA was enthalpy-driven process due to |ΔH°|>|TΔS°|. The results of experiment and molecular docking confirmed the main interaction forces between AMT and BSA were van der Waals force. And, there was a slight change in the BSA conformation after binding AMT but BSA still retains its secondary structure α-helicity. However, it had been confirmed that AMT binds on the interface between sub-domain IIA and IIB of BSA. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Preparation of Bovine Serum Albumin (BSA) nanoparticles by desolvation using a membrane contactor: a new tool for large scale production.

    Science.gov (United States)

    Yedomon, B; Fessi, H; Charcosset, C

    2013-11-01

    Albumin nanoparticles are attractive drug delivery systems as they can be prepared under soft conditions and incorporate several kinds of molecules. The aim of this study was to upscale the desolvation process for preparing Bovine Serum Albumin (BSA) nanoparticles using a membrane contactor. At a first step, the BSA nanoparticles were prepared at small scale using a syringe pump. BSA nanoparticles of 139 nm in size, with a polydispersity index of 0.046, were obtained at the optimal conditions: pH 8.2, 100 mg mL(-1) BSA albumin solution (2 mL), and 1 mL min(-1) flow rate of ethanol addition (8 mL). The upscaling with a membrane contactor was achieved by permeating ethanol through the pores of a Shirasu Porous Glass (SPG Technology Co., Japan) membrane and circulating the aqueous phase tangentially to the membrane surface. By increasing the pressure of the ethanol from 1 to 2.7 bars, a progressive decrease in nanoparticle size was obtained with a high nanoparticles yield (around 94-96%). In addition, the flow rate of the circulating phase did not affect the BSA nanoparticle characteristics. At the optimal conditions (pH 8.2, 100 mg mL(-1) BSA albumin solution, pressure of ethanol 2.7 bars, flow rate of the circulating phase 30.7 mL s(-1)), the BSA nanoparticles showed similar characteristics to those obtained with the syringe pump. Large batches of BSA nanoparticles were prepared up to 10 g BSA. The BSA nanoparticles were stable at least during 2 months at 4 °C, and their characteristics were reproducible. It was then concluded that the membrane contactor technique could be a suitable method for the preparation of albumin nanoparticles at large scale with properties similar to that obtained at small scale. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Spectroscopic and molecular docking studies of binding interaction of gefitinib, lapatinib and sunitinib with bovine serum albumin (BSA).

    Science.gov (United States)

    Shen, Guo-Feng; Liu, Ting-Ting; Wang, Qi; Jiang, Min; Shi, Jie-Hua

    2015-12-01

    The binding interactions of three kinds of tyrosine kinase inhibitors (TKIs), such as gefitinib, lapatinib and sunitinib, with bovine serum albumin (BSA) were studied using ultraviolet spectrophotometry, fluorescence spectroscopy, circular dichroism (CD), Fourier transform infrared spectroscopy (FT-IR) and molecular docking methods. The experimental results showed that the intrinsic fluorescence quenching of BSA induced by the three TKIs resulted from the formation of stable TKIs-BSA complexes through the binding interaction of TKIs with BSA. The stoichiometry of three stable TKIs-BSA complexes was 1:1 and the binding constants (Kb) of the three TKIs-BSA complexes were in the order of 10(4)M(-1) at 310 K, indicating that there was a strong binding interaction of the three TKIs with BSA. Based on the analysis of the signs and magnitudes of the free energy change (ΔG(0)), enthalpic change (ΔH(0)) and entropic change (ΔS(0)) in the binding process, it can be deduced that the binding process of the three TKIs with BSA was spontaneous and enthalpy-driven process, and the main interaction forces between the three TKIs and BSA were van der Waals force and hydrogen bonding interaction. Moreover, from the results of CD, FT-IR and molecular docking, it can be concluded that there was a significant difference between the three TKIs in the binding site on BSA, lapatinib was located on site II (m) of BSA while gefitinib and sunitinib were bound on site I of BSA, and there were some changes in the BSA conformation when binding three TKIs to BSA but BSA still retains its secondary structure α-helicity. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Binding interaction of ramipril with bovine serum albumin (BSA): Insights from multi-spectroscopy and molecular docking methods.

    Science.gov (United States)

    Shi, Jie-Hua; Pan, Dong-Qi; Jiang, Min; Liu, Ting-Ting; Wang, Qi

    2016-11-01

    The binding interaction between a typical angiotensin-converting enzyme inhibitor (ACEI), ramipril, and a transport protein, bovine serum albumin (BSA), was studied in vitro using UV-vis absorption spectroscopy, steady-state fluorescence spectroscopic titration, synchronous fluorescence spectroscopy, three dimensional fluorescence spectroscopy, circular dichroism and molecular docking under the imitated physiological conditions (pH=7.4). The experimental results suggested that the intrinsic fluorescence of BSA was quenched by ramipril thought a static quenching mechanism, indicating that the stable ramipril-BSA complex was formed by the intermolecular interaction. The number of binding sites (n) and binding constant of ramipril-BSA complex were about 1 and 3.50×10 4 M -1 at 298K, respectively, suggesting that there was stronger binding interaction of ramipril with BSA. The thermodynamic parameters together with molecular docking study revealed that both van der Waal's forces and hydrogen bonding interaction dominated the formation of the ramipril-BSA complex and the binding interaction of BSA with ramipril is enthalpy-driven processes due to |ΔH°|>|TΔS°| and ΔG°BSA was calculated to be 3.56nm based on Förster's non-radiative energy transfer theory. The results of the competitive displacement experiments and molecular docking confirmed that ramipril inserted into the subdomain IIA (site I) of BSA, resulting in a slight change in the conformation of BSA but BSA still retained its secondary structure α-helicity. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Direct binding of ethanol to bovine serum albumin: a fluorescent and 13C NMR multiplet relaxation study.

    Science.gov (United States)

    Avdulov, N A; Chochina, S V; Daragan, V A; Schroeder, F; Mayo, K H; Wood, W G

    1996-01-09

    Molecular mechanisms of ethanol interaction with proteins are not well-understood. In the present study, direct interaction of ethanol with hydrophobic binding sites on fatty acid free bovine serum albumin (BSA) was determined using the fluorescent probe 1-anilinonaphthalene-8-sulfonic acid (1,8-ANS), cis-parinaric acid, and 13C NMR. The affinity of ethanol for BSA (Kd) was (5.21 +/- 0.31) x 10(-2) mol. Ethanol (25-200 mmol) competitively inhibited 1,8-ANS binding to BSA in a concentration-dependent manner with a Ki (concentration of ethanol that decreased 1,8-ANS binding by 50%) of 658 mmol. Preincubation of BSA with ethanol significantly decreased cis-parinaric acid binding to BSA, indicating interaction of ethanol with hydrophobic fatty acid-binding site(s) on BSA. Furthermore, ethanol was found to act on three of the five fatty acid-binding sites on BSA. These data indicated selectivity in the interaction of ethanol with hydrophobic sites on BSA. 13C NMR multiplet relaxation was used to characterize the interaction of ethanol with binding sites on BSA. Detailed analysis of [13C]ethanol relaxation data obtained in the presence of increasing BSA concentrations (25-200 mg/mL) led to the conclusion that the ethanol methyl group, as opposed to its hydroxyl group, binds in a hydrophobic pocket(s) on the protein. Ethanol-induced changes in activity of certain proteins may result from direct binding of ethanol to specific hydrophobic binding sites and/or displacement of endogenous ligands from those sites.

  18. A combined binding mechanism of nonionic ethoxylated surfactants to bovine serum albumin revealed by fluorescence and circular dichroism.

    Science.gov (United States)

    Iovescu, Alina; Băran, Adriana; Stîngă, Gabriela; Cantemir-Leontieş, Anca Ruxandra; Maxim, Monica Elisabeta; Anghel, Dan Florin

    2015-12-01

    The study systematically investigates aqueous mixtures of fixed bovine serum albumin (BSA) and various ethoxylated nonionic surfactants belonging to a homologous series or not. Mono-disperse tetra-(C12E4), hexa-(C12E6) and octa-ethyleneglycol mono-n-dodecyl ether (C12E8), and poly-disperse eicosa-ethyleneglycol mono-n-tetradecyl ether (C14EO20) are respectively employed. Fluorescence and circular dichroism measurements are performed at surfactant/protein molar ratios (rm)s lower and higher than one. We aim to get new insights into the binding mechanism of these species and to differentiate among the interaction abilities of these surfactants. The relative magnitude of the binding thermodynamic parameters by fluorescence, and the increase of α-helix prove that hydrogen bonding drives the interaction next to the hydrophobic attraction. C12En (n=4,6,8) develop more H bonds with the albumin than C14EO20 owing to a zigzag conformation of their short ethyleneoxide chains. Among the homologous surfactants, C12E6 has a slightly stronger interaction with BSA due to a maximal number of H bonds at a minimal hindering. Static fluorescence and dynamic fluorescence indicate an inter-conversion between the tryptophan (Trp) rotamers which happens around the surfactants critical micellar concentration. For C14EO20, the meander conformation of the polar group determines a less evident conversion of the Trp rotamers and smaller α-helix rise. Binding isotherms of the homologous surfactants and the fluorescence quenching mechanism by C12E6 are also provided. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Influence of mixed electrolytes and pH on adsorption of bovine serum albumin in hydrophobic interaction chromatography.

    Science.gov (United States)

    Hackemann, Eva; Hasse, Hans

    2017-10-27

    Using salt mixtures instead of single salts can be beneficial for hydrophobic interaction chromatography (HIC). The effect of electrolytes on the adsorption of proteins, however, depends on the pH. Little is known on that dependence for mixed electrolytes. Therefore, the effect of the pH on protein adsorption from aqueous solutions containing mixed salts is systematically studied in the present work for a model system: the adsorption of bovine serum albumin (BSA) on the mildly hydrophobic resin Toyopearl PPG-600M. The pH is adjusted to 4.0, 4.7 or 7.0 using 25mM sodium phosphate or sodium citrate buffer. Binary and ternary salt mixtures of sodium chloride, ammonium chloride, sodium sulfate and ammonium sulfate as well as the pure salts are used at overall ionic strengths between 1500 and 4200mM. The temperature is always 25°C. The influence of the mixed electrolytes on the adsorption behavior of BSA changes completely with varying pH. Positive as well as negative cooperative effects of the mixed electrolytes are observed. The results are analyzed using a mathematical model which was recently introduced by our group. In that model the influence of the electrolytes is described by a Taylor series expansion in the individual ion molarities. After suitable parametrization using a subset of the data determined in the present work, the model successfully predicts the influence of mixed electrolytes on the protein adsorption. Furthermore, results for BSA from the present study are compared to literature data for lysozyme, which are available for the same adsorbent, temperature and salts. By calculating the ratio of the loading of the adsorbent for both proteins particularly favorable separation conditions can be selected. Hence, a model-based optimization of solvents for protein separation is possible. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Isolation and characterization of Wharton’s jelly-derived multipotent mesenchymal stromal cells obtained from bovine umbilical cord and maintained in a defined serum-free three-dimensional system

    Directory of Open Access Journals (Sweden)

    Cardoso Tereza C

    2012-05-01

    bovine-derived UC-WJ cells was included in the culture which demonstrated the immunossupression profile typically observed among isolated mesenchymal cells from other species. After classified the UC-WJ cells as mesenchymal stromal phenotype the in vitro 3D cultures was performed using the AlgiMatrix® protocol. Based on the size of spheroids (283,07 μm ± 43,10 μm we found that three weeks of culture was the best period to growth the UC-WJ cells on 3D dimension. The initial cell density was measured and the best value was 1.5 × 106 cells/well. Conclusions We described for the first time the isolation and characterization of UC-WJ cells in a serum-free condition and maintenance of primitive mesenchymal phenotype. The culture was stable under 60 consecutive passages with no genetic abnormalities and proliferating ratios. Taken together all results, it was possible to demonstrate an easy way to isolate and culture of bovine-derived UC-WJ cells under 2D and 3D serum-free condition, from fetal adnexa with a great potential in cell therapy and biotechnology.

  1. Interactions between {beta}-carboline alkaloids and bovine serum albumin: Investigation by spectroscopic approach

    Energy Technology Data Exchange (ETDEWEB)

    Nafisi, Shohreh, E-mail: drshnafisi@gmail.com [Department of Chemistry, Islamic Azad University, Central Tehran Branch (IAUCTB), Tehran (Iran, Islamic Republic of); Panahyab, Ataollah [Department of Chemistry, Islamic Azad University, Central Tehran Branch (IAUCTB), Tehran (Iran, Islamic Republic of); Bagheri Sadeghi, Golshan [Department of Biology, Islamic Azad University, Science and Research Branch, Tehran (Iran, Islamic Republic of)

    2012-09-15

    {beta}-Carboline alkaloids are present in medicinal plants such as Peganum harmala L. that have been used as folk medicine in anticancer therapy. BSA is the major soluble protein constituent of the circulatory system, and has many physiological functions including the transport of a variety of compounds. This study is the first attempt to investigate the binding of {beta}-carboline alkaloids to BSA by using a constant protein concentration and varying drug concentrations at pH 7.2. FTIR and UV-Vis spectroscopic methods were used to analyze the binding modes of {beta}-carboline alkaloids, the binding constants and the effects of drug complexation on BSA stability and conformation. Spectroscopic evidence showed that {beta}-carboline alkaloids bind BSA via hydrophobic interaction and van der Waals contacts along with H-bonding with the -NH groups, with overall binding constants of K{sub harmine-BSA}=2.04 Multiplication-Sign 10{sup 4} M{sup -1}, K{sub tryptoline-BSA}=1.2 Multiplication-Sign 10{sup 4} M{sup -1}, K{sub harmaline-BSA}=5.04 Multiplication-Sign 10{sup 3} M{sup -1}, K{sub harmane-BSA}=1.41 Multiplication-Sign 10{sup 3} M{sup -1} and K{sub harmalol-BSA}=1.01 Multiplication-Sign 10{sup 3} M{sup -1}, assuming that there is one drug molecule per protein. The BSA secondary structure was altered with a major decrease of {alpha}-helix from 64% (free protein) to 59% (BSA-harmane), 56% (BSA-harmaline and BSA-harmine), 55% (BSA-tryptoline), 54% (BSA-harmalol) and {beta}-sheet from 15% (free protein) to 6-8% upon {beta}-carboline alkaloids complexation, inducing a partial protein destabilization. - Highlights: Black-Right-Pointing-Pointer We model the binding of {beta}-carboline alkaloids to BSA by using the spectroscopic methods. Black-Right-Pointing-Pointer We investigate the effects of drug complexation on BSA stability and conformation. Black-Right-Pointing-Pointer A partial protein destabilization occurred at high alkaloids concentration. Black

  2. Savinase action on bovine serum albumin (BSA) monolayers demonstrated with measurements at the air-water interface and liquid Atomic Force Microscopy (AFM) imaging

    DEFF Research Database (Denmark)

    Balashev, Konstantin; Callisen, Thomas H; Svendsen, Allan

    2011-01-01

    We studied the enzymatic action of Savinase on bovine serum albumin (BSA) organized in a monolayer spread at the air/water interface or adsorbed at the mica surface. We carried out two types of experiments. In the first one we followed the degradation of the protein monolayer by measuring...... the surface pressure and surface area decrease versus time. In the second approach we applied AFM imaging of the supported BSA monolayers adsorbed on mica solid supports and extracted information for the enzyme action by analyzing the obtained images of the surface topography in the course of enzyme action...

  3. Surface functionalization of zirconium dioxide nano-adsorbents with 3-aminopropyl triethoxysilane and promoted adsorption activity for bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Gen; Wu, Chaochao [Department of Chemistry, College of Sciences, Northeastern University, Shenyang 110819 (China); Zhang, Xia, E-mail: xzhang@mail.neu.edu.cn [Department of Chemistry, College of Sciences, Northeastern University, Shenyang 110819 (China); Liu, Yufeng, E-mail: liuyufeng@bjmu.edu.cn [College of Pharmacy, Liaoning University, Shenyang 110036 (China); Meng, Hao; Xu, Junli; Han, Yide; Xu, Xinxin; Xu, Yan [Department of Chemistry, College of Sciences, Northeastern University, Shenyang 110819 (China)

    2016-06-15

    Surface functionalization of zirconium dioxide (ZrO{sub 2}) nano-adsorbents was carried out by using 3-aminopropyl triethoxysilane (APTES) as the modifier. The addition amount of APTES was varied to determine the optimum modification extent, and the bulk ZrO{sub 2} microparticles were also modified by APTES for comparison. Some means, such as TEM, XRD, FT-IR, XPS and TG-DSC were used to character these ZrO{sub 2} particles. The results showed that the APTES molecules were chemically immobilized on the surface of ZrO{sub 2} nanoparticles via Zr−O−Si bonds, and the nano-ZrO{sub 2} samples showed larger special surface area. In the adsorption of bovine serum albumin (BSA), nano-ZrO{sub 2} samples exhibited enhanced adsorption activity, and APTES modified nano-ZrO{sub 2} with proper APTES content presented the best adsorption property. Under the same adsorption conditions, the equilibrium adsorption capacity of BSA on APTES-ZrO{sub 2}-2 was almost 2.3 times as that on pristine nano-ZrO{sub 2} and 3.0 times as on bulk ZrO{sub 2} microparticles. The increased adsorption capacity of APTES-ZrO{sub 2} nano-adsorbents can be attributed to the chemical interaction between amino and carboxyl groups at APTES-ZrO{sub 2}/BSA interface. The pH-dependent experiments showed that the optimum pH value for the adsorption and desorption was 5.0 and 9.0, respectively, which suggested that the adsorption and release of BSA could be controlled simply by adjusting the solution pH condition. - Highlights: • APTES chemically immobilized on ZrO{sub 2} nanoparticles via Zr−O−Si bond. • Enhanced adsorption capacity of BSA was observed on APTES-ZrO{sub 2}. • Chemical adsorption character of BSA on APTES-ZrO{sub 2}. • Adsorption/release of BSA on APTES-ZrO{sub 2} accomplished by adjusting pH value.

  4. Interaction mechanisms of ionic liquids [Cnmim]Br (n=4, 6, 8, 10) with bovine serum albumin

    International Nuclear Information System (INIS)

    Yan Hua; Wu Junyong; Dai Guoliang; Zhong Aiguo; Chen Hao; Yang Jianguo; Han Deman

    2012-01-01

    It is important to study the interaction of ionic liquids (ILs) with protein for the applications of ILs in biochemical process, and help the researchers to choose and design the better ILs to serve as a solvent. In this work, the interaction between 1-alkyl-3-methylimidazolium bromide [C n mim]Br (n=4, 6, 8, 10) and bovine serum albumin (BSA) was systematically investigated for the first time by multi-spectroscopic approach (fluorescence, UV–vis and FT-IR spectroscopy) and density functional theory (DFT). [C n mim]Br (n=4, 6, 8, 10) can bind to BSA by H-bond interaction between their cationic headgroups and Asp/Glu amino acid residue at the surface of BSA, and hydrophobic interaction between their hydrocarbon chains and the hydrophobic amino acid residues in the interior of BSA. On the basis of thermodynamic parameters and the similar structure of [C n mim]Br (n=4, 6, 8, 10), it can be inferred that the hydrophobic interaction plays a major role in the interaction of [C 10 mim]Br with BSA, while the hydrogen bond and van der Waals force play a major role in the interaction of [C n mim]Br (n=4, 6, 8) with BSA. Synchronous fluorescence and FT-IR spectra indicate that [C 10 mim]Br could markedly change the secondary structure of BSA, while [C n mim]Br (n=4, 6, 8) could slightly change the secondary structure of BSA. The results allowed us to understand (i) the effect of the alkyl chain length of the cation on the mechanism of ILs–protein interaction and (ii) the effect of the alkyl chain length of the cation on the protein secondary structure. - Highlights: ► Interaction of [C n mim]Br with BSA was investigated by spectroscopy and DFT. ► Interaction mechanisms and BSA secondary structure were discussed. ► Effects of the alkyl chain length of cation were presented.

  5. Interaction of NAEn-s-n gemini surfactants with bovine serum albumin: A structure-activity probe

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Houchen [Chemical Synthesis and Pollution Control Key Laboratory of Sichuan Province, China West Normal University, Nanchong, Sichuan 637009 (China); Jiang, Xiaohui, E-mail: jxh2314508@163.com [Chemical Synthesis and Pollution Control Key Laboratory of Sichuan Province, China West Normal University, Nanchong, Sichuan 637009 (China); Zhou, Limei; Cheng, Zhenjun; Yin, Wenmin [Chemical Synthesis and Pollution Control Key Laboratory of Sichuan Province, China West Normal University, Nanchong, Sichuan 637009 (China); Duan, Ming; Liu, Pingli [State Key Laboratory of Oil and Gas Reservoir Geology and Exploitation, Southwest Petroleum University, Chengdu, Sichuan 610500 (China); Jiang, Xiaomin [Southwest Electric Power Design Institute, Chengdu, Sichuan 61002 (China)

    2013-02-15

    Gemini surfactants, {alpha},{omega}-bis (3-(alkyloxylacyl) pyridinium) propane/butane/hexane dibromide (designated as NAEn-s-n), have been prepared and the interactions of NAEn-s-n with bovine serum albumin (BSA) were investigated by fluorescence, UV-vis and FTIR spectroscopies. The intrinsic fluorescence of BSA was significantly quenched by NAEn-s-n through static quenching. NAEn-s-n combined mainly with Trp-212 in BSA by van der Waals force, hydrogen bonding, electrostatic and hydrophobic interaction. The binding process was spontaneous, exothermic and enthalpy driven. The synchronous and tridimensional fluorescence revealed the changed conformation of the peptide backbone and the altered microenvironment of tryptophan and tyrosine residues in BSA. The red-shift in the IR spectrum of the BSA amide I peak, the blue-shift of amide II peak, as well as the appearance of a new peak at around 1514 cm{sup -1} suggested unfolding of the protein secondary structure upon the addition of NAEn-s-n. The lengths of spacer and hydrophobic chain greatly influenced the interaction. With the lengthening alkyl chain in NAEn-s-n, the binding constant of BSA-NAEn-4-n increased, while the thermodynamic parameters and the {alpha}-helix content of BSA decreased. This indicated an enhancement of hydrophobic interaction between BSA and NAEn-s-n. However, these values (i.d. binding constant, {alpha}-helix content etc.) fluctuated with methylene numbers in the spacer of NAEn-s-n, which might be due to the different spatial arrangement of the spacer of the gemini surfactants. This investigation may shed new light on the understanding of structure-activity correlation. Highlights: Black-Right-Pointing-Pointer NAEn-s-n is a serial of newly synthesized cationic gemini surfactants. Black-Right-Pointing-Pointer NAEn-s-n binds in hydrophobic domain II and III of BSA by hydrophobic forces. Black-Right-Pointing-Pointer The concentrations of NAEn-s-n and BSA affect the binding constants and the

  6. Interaction study on bovine serum albumin physically binding to silver nanoparticles: Evolution from discrete conjugates to protein coronas

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Jun; Zhong, Ruibo; Li, Wanrong; Liu, Yushuang; Bai, Zhijun; Yin, Jun; Liu, Jingran; Gong, Pei [Agricultural Nanocenter, School of Life Science, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot 010018 (China); Zhao, Xinmin, E-mail: zhao.xinmin@hotmail.com [School of Foreign Language, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot 010018 (China); Zhang, Feng, E-mail: fengzhang1978@hotmail.com [Agricultural Nanocenter, School of Life Science, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot 010018 (China)

    2015-12-30

    Graphical abstract: With the non-uniform coating of amphiphilic polymer, the silver nanoparticles (AgNPs) can form protein coronas which can become discrete protein–nanoparticle conjugates when controlling the protein–nanoparticle molar ratios. The protein's conformational changes upon binding NPs was also studied by both circular dichroism and three-dimensional fluorescence spectroscopy. - Highlights: • The amphiphilic polymer coating can not only transfer hydrophobic NPs into water soluble, but also providing a thick shell responsible for the strong physisorption to proteins without significantly changing their spatial conformations. • NP with discrete proteins can be simply obtained by a simple mixing procedure followed by a gel electrophoresis separation, and the resulting conjugates are robust enough to resist common separation techniques like gel electrophoresis. • In combination with the universal amphiphilic polymer coating strategy and the physisorption mediated protein–NP conjugation, proteins like BSA can be effectively conjugated to different materials such as noble metal, semiconductor and magnetic NPs. • In contrast to chemical coupling methods, the physisorption mediated protein–NP conjugation holds facile, robust and reversible advantages, which may find wide applications in nano-biomedicine field. - Abstract: The nanostructures formed by inorganic nanoparticles together with organic molecules especially biomolecules have attracted increasing attention from both industries and researching fields due to their unique hybrid properties. In this paper, we systemically studied the interactions between amphiphilic polymer coated silver nanoparticles and bovine serum albumins by employing the fluorescence quenching approach in combination with the Stern-Volmer and Hill equations. The binding affinity was determined to 1.30 × 10{sup 7} M{sup −1} and the interaction was spontaneously driven by mainly the van der Waals force and

  7. Determination of trace elements in human blood serum and in the standard reference material ''Bovine Liver'' by instrumental neutron activation analysis

    International Nuclear Information System (INIS)

    Behne, D.; Juergensen, H.

    1978-01-01

    Ag, Br, Co, Cr, Cs, Fe, Na, Rb, Sb, Se and Zn were determined in biological materials by gamma-spectrometry of long-lived radionuclides after long-time irradiation with thermal neutrons. Blood was taken from a vein in the arm of the test person via an indwelling plastic cannula. The serum was separated from other blood components by centrifugation and stored at a temperature of -20 deg C. Ampoules of high purity silica quartz were chosen as irradiation containers. The vials were cleaned by etching with 40% hydrofluoric acid. For the analysis 300 μl of serum were taken. The ampoules were irradiated for 10 days at a thermal neutron flux density of 5.10 13 n.cm -2 .sec -1 . They were then cleaned by etching for 5 min with hydrofluoric acid. The gamma-ray spectra of the irradiated samples were measured twice. From the first spectra, which were obtained 10 days after irradiation, the concentrations of Br and Na were calculated. From the gamma-peaks after a decay time of 3 months Ag, Co, Cr, Cs, Fe, Rb, Sc, Se and Zn were determined. The accuracy and precision of the procedure were tested, using the standard reference material ''Bovine Liver'' and identical serum samples. In the case of blood serum the method proved to be suitable for the determination of Br, Cs, Fe, Na, Rb, Se and Zn. By analysing samples from several subjects information about element levels in human serum was obtained. (T.G.)

  8. Theoretical and experimental studies on freezing point depression and vapor pressure deficit as methods to measure osmotic pressure of aqueous polyethylene glycol and bovine serum albumin solutions.

    Science.gov (United States)

    Kiyosawa, Keitaro

    2003-05-01

    For survival in adverse environments where there is drought, high salt concentration or low temperature, some plants seem to be able to synthesize biochemical compounds, including proteins, in response to changes in water activity or osmotic pressure. Measurement of the water activity or osmotic pressure of simple aqueous solutions has been based on freezing point depression or vapor pressure deficit. Measurement of the osmotic pressure of plants under water stress has been mainly based on vapor pressure deficit. However, differences have been noted for osmotic pressure values of aqueous polyethylene glycol (PEG) solutions measured by freezing point depression and vapor pressure deficit. For this paper, the physicochemical basis of freezing point depression and vapor pressure deficit were first examined theoretically and then, the osmotic pressure of aqueous ethylene glycol and of PEG solutions were measured by both freezing point depression and vapor pressure deficit in comparison with other aqueous solutions such as NaCl, KCl, CaCl(2), glucose, sucrose, raffinose, and bovine serum albumin (BSA) solutions. The results showed that: (1) freezing point depression and vapor pressure deficit share theoretically the same physicochemical basis; (2) theoretically, they are proportional to the molal concentration of the aqueous solutions to be measured; (3) in practice, the osmotic pressure levels of aqueous NaCl, KCl, CaCl(2), glucose, sucrose, and raffinose solutions increase in proportion to their molal concentrations and there is little inconsistency between those measured by freezing point depression and vapor pressure deficit; (4) the osmotic pressure levels of aqueous ethylene glycol and PEG solutions measured by freezing point depression differed from the values measured by vapor pressure deficit; (5) the osmotic pressure of aqueous BSA solution measured by freezing point depression differed slightly from that measured by vapor pressure deficit.

  9. The effects of hydroxyapatite/calcium phosphate glass scaffold and its surface modification with bovine serum albumin on 1-wall intrabony defects of beagle dogs: a preliminary study

    International Nuclear Information System (INIS)

    Um, Yoo-Jung; Jung, Ui-Won; Chae, Gyung-Joon; Kim, Chang-Sung; Cho, Kyoo-Sung; Kim, Chong-Kwan; Choi, Seong-Ho; Lee, Yong-Keun

    2008-01-01

    The purpose of this study was to evaluate the effects of biphasic hydroxyapatite/calcium phosphate glass (HA/CPG) scaffold and its surface modification with bovine serum albumin (BSA) on periodontal regeneration. 1-wall intrabony defects were surgically created on five beagle dogs. HA/CPG scaffolds, with a hydroxyapatite (HA)/calcium phosphate glass (CPG) ratio of 95:5 by weight (%) and surface modification done by 2% bovine serum albumin, were used. The control group received surgical flap operation, and the experimental groups were filled with HA/CPG scaffolds and HA/CPG(BSA) scaffolds. The animals were sacrificed eight weeks after surgery. Histological findings revealed better space maintenance in the experimental groups than the control group, and showed new bone formation intermittently in between the residual material particles. The newly formed bone was mostly woven bone and the residual particles were undergoing resorption. Cementum regeneration was observed with limited root resorption in all the groups. Histometric analysis also revealed greater mean values in new bone formation, cementum regeneration and bone area than the control group in both experimental groups. However, similar findings were presented between HA/CPG and HA/CPG(BSA). The result of the present study revealed the newly fabricated HA/CPG scaffold to have a potential use as a bone substitute material.

  10. The effects of hydroxyapatite/calcium phosphate glass scaffold and its surface modification with bovine serum albumin on 1-wall intrabony defects of beagle dogs: a preliminary study

    Energy Technology Data Exchange (ETDEWEB)

    Um, Yoo-Jung; Jung, Ui-Won; Chae, Gyung-Joon; Kim, Chang-Sung; Cho, Kyoo-Sung; Kim, Chong-Kwan; Choi, Seong-Ho [Department of Periodontology, Research Institute for Periodontal Regeneration, College of Dentistry, Yonsei University, 134 Shinchon-Dong, Seodaemun-gu, Seoul 120-752 l (Korea, Republic of); Lee, Yong-Keun [Department and Research Institute of Dental Biomaterials and Bioengineering, College of Dentistry, Yonsei University, 134 Shinchon-Dong, Seodaemun-gu, Seoul 120-750 (Korea, Republic of)], E-mail: shchoi726@yuhs.ac

    2008-12-15

    The purpose of this study was to evaluate the effects of biphasic hydroxyapatite/calcium phosphate glass (HA/CPG) scaffold and its surface modification with bovine serum albumin (BSA) on periodontal regeneration. 1-wall intrabony defects were surgically created on five beagle dogs. HA/CPG scaffolds, with a hydroxyapatite (HA)/calcium phosphate glass (CPG) ratio of 95:5 by weight (%) and surface modification done by 2% bovine serum albumin, were used. The control group received surgical flap operation, and the experimental groups were filled with HA/CPG scaffolds and HA/CPG(BSA) scaffolds. The animals were sacrificed eight weeks after surgery. Histological findings revealed better space maintenance in the experimental groups than the control group, and showed new bone formation intermittently in between the residual material particles. The newly formed bone was mostly woven bone and the residual particles were undergoing resorption. Cementum regeneration was observed with limited root resorption in all the groups. Histometric analysis also revealed greater mean values in new bone formation, cementum regeneration and bone area than the control group in both experimental groups. However, similar findings were presented between HA/CPG and HA/CPG(BSA). The result of the present study revealed the newly fabricated HA/CPG scaffold to have a potential use as a bone substitute material.

  11. Pengaruh penggantian Bovine Serum Albumin (BSA dengan putih telur pada pengencer CEP-2 terhadap kualitas semen sapi Peranakan Ongole pada suhu penyimpanan 3-5oC

    Directory of Open Access Journals (Sweden)

    Nisa’us Sholikah

    2016-04-01

    Full Text Available Purpose of this research was to investigate the effect of Bovine Serum Albumin substitution by albumen on CEP-2 to semen quality Ongole CrossBred bull stored at 3-5oC. Research was conducted at Research Centre Beef Cattle Laboratory, Grati, Pasuruan on February 2016. Semen diluent was divided into two groups, there were P0 (90% CEP-2 + 10% Egg Yolk (EY; and P1 (90% CEP + 0.4% albumen + 10% EY. Data of the research were analyzed using paired design t test. The result showed that after eight days chilled preservation, percentage of motility P1 (47.4±10.9% was higher than P0 (47±9.2%. Percentage of viability P0 (83.1±1.9% was higher than P1 (81.3±1.5%. Percentage of abnormality P1 (3.6±0.4% was lower than P0 (3.8±0.3%. Total motile sperm count after six days chilled preservation was significantly higher in all treatments compared to the standard criteria of SNI 40% motile sperm/ml. The conclusion of this research was 0,4% albumen could replace the BSA capability on CEP-2. Keywords: CEP-2, Bovine Serum Albumin, chilled preservation, semen quality

  12. The effects of allantoin, arginine and NaCl on thermal melting and aggregation of ribonuclease, bovine serum albumin and lysozyme.

    Science.gov (United States)

    Arakawa, Tsutomu; Maluf, Nasib Karl

    2018-02-01

    Allantoin is widely used as a skin care agent and readily forms crystals, which were recently shown to bind endotoxins and high molecular weight aggregates in cell culture harvests. Here, we have investigated the effects of allantoin on thermal stability and aggregation of protein using ribonuclease, bovine serum albumin and lysozyme using temperature-regulated circular dichroism (CD) and differential scanning microcalorimetry (DSC). Ribonuclease showed no change in thermal stability and aggregation by the addition of allantoin. While allantoin showed no effects on the thermal stability of bovine serum albumin, it enhanced aggregation. Similarly, allantoin showed no stabilizing effects on lysozyme, but it strongly suppressed aggregation. Such suppressed aggregation resulted in reversibility of thermal unfolding of lysozyme. These effects of allantoin were then compared with those of NaCl and arginine hydrochloride. Arginine was similar to allantoin at low concentrations, where both solvent additives can be compared due to limited solubility of allantoin. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Studying the electromagnetic-induced changes of the secondary structure of bovine serum albumin and the bioprotective effectiveness of trehalose by Fourier transform infrared spectroscopy.

    Science.gov (United States)

    Magazù, Salvatore; Calabrò, Emanuele

    2011-06-02

    Samples of bovine serum albumin in H(2)O and D(2)O solutions, in the absence or presence of trehalose, were exposed separately to a static magnetic field at 200 mT and to a 50 Hz electromagnetic field at 1.8 mT, studying the relative effects on the secondary structure of the protein by means of Fourier transform infrared spectroscopy. The spectra acquired in the mid-infrared region after 2 and 4 h of exposures to the static magnetic field showed a decrease in amide A and amide I band intensities for the protein in bidistilled aqueous solutions that was also evidenced after exposures to a 50 Hz electromagnetic field. These results led us to conclude that electromagnetic fields of low intensities can affect the C═O and C-N stretching vibrations and N-H plane bending of peptide linkages. Furthermore, mid-infrared spectra of bovine serum albumin in trehalose aqueous solutions were not significantly modified after the exposures, confirming the hypothesis of the possible bioprotective effectiveness of trehalose against electromagnetic fields.

  14. Preoperative Serum Interleukin-6 Is a Potential Prognostic Factor for Colorectal Cancer, including Stage II Patients

    Directory of Open Access Journals (Sweden)

    Kazuyoshi Shiga

    2016-01-01

    Full Text Available Aims. To evaluate the prognostic significance of serum interleukin-6 (IL-6 in colorectal cancer (CRC. Patients and Methods. Preoperative serum IL-6 was measured in 233 CRC patients and 13 healthy controls. Relationships between IL-6 and various clinicopathological factors were evaluated, and the overall survival (OS and disease-free survival (DFS rates according to IL-6 status were calculated for all patients and according to disease stage. Results. The mean IL-6 level was 6.6 pg/mL in CRC patients and 2.6 pg/mL in healthy controls. Using a cutoff of 6.3 pg/mL, obtained using receiver operating characteristic curve analysis, 57 patients had a high IL-6 level. The mean value was higher for stage II disease than for stage III disease. IL-6 status correlated with C-reactive protein (CRP and carcinoembryonic antigen levels, obstruction, and pT4 disease. The OS differed according to the IL-6 status for all patients, whereas the DFS differed for all patients and for those with stage II disease. The Cox proportional hazards model showed that pT4 disease was an independent risk factor for recurrence in all CRC patients; IL-6, CRP, and pT4 were significant risk factors in stage II patients. Conclusions. The preoperative IL-6 level influences the risk of CRC recurrence.

  15. Gas chromatography-mass spectrometry/mass spectrometry analysis to determine natural and post-administration levels of oestrogens in bovine serum and urine

    Energy Technology Data Exchange (ETDEWEB)

    Biddle, S. [HFL, Newmarket Road, Fordham, Cambridgeshire (United Kingdom)]. E-mail: sbiddle@hfl.co.uk; Teale, P. [HFL, Newmarket Road, Fordham, Cambridgeshire (United Kingdom); Robinson, A. [HFL, Newmarket Road, Fordham, Cambridgeshire (United Kingdom); Bowman, J. [HFL, Newmarket Road, Fordham, Cambridgeshire (United Kingdom); Houghton, E. [HFL, Newmarket Road, Fordham, Cambridgeshire (United Kingdom)

    2007-03-14

    A novel analytical approach has been developed and shown to be capable of detecting the isomers of oestradiol in the low ppt (pg mL{sup -1}) range in bovine serum and urine. Following extractive derivatisation the analytes were detected as their 3-pentafluorobenzoyl 17-trimethylsilyl ether derivatives by gas chromatography-mass spectrometry/mass spectrometry (GC-MS/MS), using electron capture negative ion chemical ionisation. The isomers of oestradiol were quantified in both blank and post-administration urine and serum samples, with a view to setting action/threshold levels for these compounds, to allow discrimination between normal samples and samples from animals treated with growth promoting ear implants. A non-parametric statistical assessment of the data resulted in proposed action levels (with a false positive probability of 1 in 1000) of 1.6 and 2.7 ng mL{sup -1} for 17{alpha}-oestradiol, in male and female urine, respectively, and 40 and 44 pg mL{sup -1} for 17{beta}-oestradiol, in male and female urine, respectively. An action level of 20 pg mL{sup -1} was proposed for 17{alpha}- and 17{beta}-oestradiol in male serum. In female serum the proposed action levels were 40 and 20 pg mL{sup -1} for 17{alpha}- and 17{beta}-oestradiol, respectively.

  16. Gas chromatography-mass spectrometry/mass spectrometry analysis to determine natural and post-administration levels of oestrogens in bovine serum and urine

    International Nuclear Information System (INIS)

    Biddle, S.; Teale, P.; Robinson, A.; Bowman, J.; Houghton, E.

    2007-01-01

    A novel analytical approach has been developed and shown to be capable of detecting the isomers of oestradiol in the low ppt (pg mL -1 ) range in bovine serum and urine. Following extractive derivatisation the analytes were detected as their 3-pentafluorobenzoyl 17-trimethylsilyl ether derivatives by gas chromatography-mass spectrometry/mass spectrometry (GC-MS/MS), using electron capture negative ion chemical ionisation. The isomers of oestradiol were quantified in both blank and post-administration urine and serum samples, with a view to setting action/threshold levels for these compounds, to allow discrimination between normal samples and samples from animals treated with growth promoting ear implants. A non-parametric statistical assessment of the data resulted in proposed action levels (with a false positive probability of 1 in 1000) of 1.6 and 2.7 ng mL -1 for 17α-oestradiol, in male and female urine, respectively, and 40 and 44 pg mL -1 for 17β-oestradiol, in male and female urine, respectively. An action level of 20 pg mL -1 was proposed for 17α- and 17β-oestradiol in male serum. In female serum the proposed action levels were 40 and 20 pg mL -1 for 17α- and 17β-oestradiol, respectively

  17. Differential expression of miRNA-423-5p in serum from cattle challenged with bovine viral diarrhea virus

    Science.gov (United States)

    Bovine viral diarrhea virus (BVDV) is an RNA virus that causes respiratory disease in cattle. MicroRNAs have been proposed as indicators of exposure to respiratory pathogens. However, microRNA profiles in cattle exposed to BVDV are currently nonexistent and few studies have been reported; therefore,...

  18. Genetic diversity of bovine papillomavirus types, including two putative new types, in teat warts from dairy cattle herds.

    Science.gov (United States)

    Lunardi, Michele; de Camargo Tozato, Claudia; Alfieri, Alice Fernandes; de Alcântara, Brígida Kussumoto; Vilas-Boas, Laurival Antonio; Otonel, Rodrigo Alejandro Arellano; Headley, Selwyn Arlington; Alfieri, Amauri Alcindo

    2016-06-01

    Teat papillomatosis affects dairy cows worldwide. Milking can become difficult due to teat warts, and maintaining affected cows in the herds may diminish economic profit in the dairy industry. Currently, 13 bovine papillomavirus (BPV) types have been fully characterized, and numerous putative BPV types have been identified through partial L1 gene PCR. In order to identify the viral types present in warts on the udders of dairy cows, 40 teat lesions from 24 cows from 13 cattle farms in three States of Brazil were evaluated by PV L1 gene PCR. The warts that were evaluated contained sequences from BPVs 6-10, the putative BPV types BAPV9 and BAPV4, and two unreported putative papillomavirus (PV) types, named BPV/BR-UEL6 and BPV/BR-UEL7. In addition, mixed infections and coinfections were identified, since more than one lesion was observed on the udders of 13 cows. Phylogenetic analysis showed that BPV/BR-UEL6 is closely related to BPVs belonging to the genus Xipapillomavirus, while BPV/BR-UEL7 clustered with the previously reported strains Cervus timorensis and Pudu puda PVs, which represent a putative new PV type, and it was only distantly related to xi-, epsilon-, delta- and dyoxi-PVs. These results provide information that will assist in the understanding of the association of BPVs 6, 7, 8, 9, and 10, as well as putative BPV types BAPV4 and BAPV9, with mammary papillomatosis. This is the first characterization of putative novel PV types BPV/BR-UEL6 and BPV/BR-UEL7 in teat warts of dairy cows, highlighting the high genetic diversity of BPVs associated with teat papillomatosis.

  19. Immune responses induced in rabbits after oral administration of bovine serum albumin in combination with different adjuvants (herb extracts, aluminium hydroxide and platinum nanoparticles

    Directory of Open Access Journals (Sweden)

    G. Bižanov

    2016-12-01

    Full Text Available The aim of the current study was to evaluate the immunostimulatory activity of 10 different herbal extracts from Vitex agnus-castus, Vinca major, Aloe arborescens and the polyherbal product containing extracts from Sambucus nigra, Primula versis, Pinus alba, Gentiana lutea, Cetraria islandica, Eucaliptus globulus, Citrus limon and aluminium hydroxide, as well as platinum nanoparticles. Rabbits were immunized three times orally with bovine serum albumin (BSA in combination with the components mentioned above. BSA-specific IgA antibodies in saliva and IgG antibodies in serum were examined by ELISA. It was found that the rabbits immunized with BSA in combination with either platinum nanoparticles or aluminium hydroxide had higher titres of BSA-specific IgA antibodies in their saliva at day 56 of observation. Likewise, rabbits treated with BSA and Vinca major or Aloe arborescens extracts showed higher levels of BSA-specific IgG antibodies in the serum at the end of observation. These results suggest that some plant extracts, aluminium hydroxide and platinum nanoparticles components could be used as oral adjuvants or as immunomodulators for rabbits.

  20. Continuous recording of long-chain acyl-coenzyme A synthetase activity using fluorescently labeled bovine serum albumin

    DEFF Research Database (Denmark)

    Demant, Erland J.F.; Nystrøm, Birthe T.

    2001-01-01

    acyl-Coenzyme A, synthetase, activity assay, fluorescence recording, fatty acid probe, serum albumin, hydroxycoumarin, detergent, micelles, Pseudomonas fragi, rat liver microsomes......acyl-Coenzyme A, synthetase, activity assay, fluorescence recording, fatty acid probe, serum albumin, hydroxycoumarin, detergent, micelles, Pseudomonas fragi, rat liver microsomes...

  1. Interaction of bovine serum albumin and human blood plasma with PEO-tethered surfaces: Influence of PEO chain length, grafting density, and temperature : Influence of PEO chain length, grafting density, and temperature

    NARCIS (Netherlands)

    Norde, Willem; Gage, D.

    2004-01-01

    Solid surfaces are modified by grafting poly(ethylene oxide), PEO, to influence their interaction with indwelling particles, in particular molecules of bovine serum albumin and human plasma proteins. As a rule, the grafted PEO layers suppress protein adsorption. The suppression is most effective

  2. Pulse radiolytic study of the oxidation reaction of uric acid in presence of bovine serum albumin: evidence of possible complex formation in the transient state

    International Nuclear Information System (INIS)

    Adhikari, S.; Gopinathan, C.

    1996-01-01

    The pulse radiolytic and spectrophotometric study of uric acid in presence of bovine serum albumin (BSA) has been carried out. In the spectrophotometric study there is no evidence for ground state interaction between BSA and uric acid. The reaction of CCl 3 OO . radical with uric acid produces a transient having absorption maximum at 330 nm and that with BSA produces transient having absorption maximum at 410 nm. In a composition of equal concentration of uric acid and BSA the CCl 3 OO . radical produces a transient absorption spectrum which shows two peaks at 330 nm and 350 nm and a shoulder at 410 nm. The peak at 350 nm is ascribed due to weak complex formation between BSA and uric acid radicals. The rate constant of CCl 3 OO . radical with uric acid increases with the increase in BSA concentration which is explained as protection of BSA by uric acid from radical attack. (author). 4 refs., 2 figs., 1 tab

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

    Science.gov (United States)

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

    2006-01-01

    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.

  4. Study of the interaction between fluoxetine hydrochloride and bovine serum albumin in the imitated physiological conditions by multi-spectroscopic methods

    Energy Technology Data Exchange (ETDEWEB)

    Katrahalli, Umesha [Department of Chemistry, Karnatak University, Dharwad 580 003 (India); Jaldappagari, Seetharamappa, E-mail: j_seetharam@rediffmail.co [Department of Chemistry, Karnatak University, Dharwad 580 003 (India); Kalanur, Shankara S. [Department of Chemistry, Karnatak University, Dharwad 580 003 (India)

    2010-02-15

    The mechanism of interaction of an antidepressant, fluoxetine hydrochloride (FLX) with bovine serum albumin (BSA) has been studied by different spectroscopic techniques under physiological conditions. FLX was found to quench the intrinsic fluorescence of protein by static quenching mechanism. The binding constant 'K' was found to be 7.06x10{sup 3} M{sup -1} at 296 K. The value of 'n' close to unity revealed that the BSA has a single class of binding site for FLX. Based on thermodynamic parameters, hydrogen bonding and van der Waals forces were proposed to operate between BSA and FLX. The change in conformation of protein was noticed upon its interaction with the drug. From displacement studies it was concluded that the FLX bound to protein at site I. The effects of various common metals ions on the binding were also investigated.

  5. Using response surface methodology to optimize process parameters and cross-linking agents for production of combined-cross-linked bovine serum albumin gels.

    Science.gov (United States)

    Gan, Chee-Yuen; Alkarkhi, Abbas F M; Easa, Azhar Mat

    2009-04-01

    D-optimal design was employed to optimize the mixture of cross-linking agents formulation: microbial transglutaminase (MTGase) and ribose, and the processing parameters (i.e. incubation and heating time) in the mixture in order to obtain combined-cross-linked bovine serum albumin gels that have high gel strength, pH close to neutral and yet medium in browning. Analysis of variance (ANOVA) showed that the contribution of quadratic term to the model over the linear was significant for pH and L* value, whereas linear model was significant for gel strength. Optimization study using response surface methodology (RSM) was performed to the mixture components and process variables and the optimum conditions obtained were: MTGase of 1.34-1.43 g/100 mL, ribose of 1.07-1.16 g/100 mL, incubation time of 5 h at 40 degrees C and heating time of 3 h at 90 degrees C.

  6. The Formation of Polycomplexes of Poly(Methyl Vinyl Ether-Co-Maleic Anhydride and Bovine Serum Albumin in the Presence of Copper Ions

    Directory of Open Access Journals (Sweden)

    Karahan Mesut

    2014-09-01

    Full Text Available The binary and ternary complex formations of poly(methyl vinyl ether-co-maleic anhydride (PMVEMA with copper ions and with bovine serum albumin (BSA in the presence of copper ions in phosphate buffer solution at pH = 7 were examined by the techniques of UV-visible, fluorescence, dynamic light scattering, atomic force microscopy measurements. In the formation of binary complexes of PMVEMA-Cu(II, the addition of copper ions to the solution of PMVEMA in phosphate buffer solution at pH = 7 forms homogeneous solutions when the molar ratio of Cu(II/MVEMA is 0.5. Then the formations of ternary complexes of PMVEMA-Cu(II-BSA were examined. Study analysis revealed that the toxicities of polymer-metal and polymer-metal-protein mixture solutions depend on the nature and ratio of components in mixtures.

  7. Effect of Moderate UVC Irradiation on Bovine Serum Albumin and Complex with Antimetabolite 5-Fluorouracil: Fluorescence Spectroscopic and Molecular Modelling Studies

    Directory of Open Access Journals (Sweden)

    Shanmugavel Chinnathambi

    2015-01-01

    Full Text Available The interaction of antimetabolite 5-fluorouracil (5FU with bovine serum albumin (BSA under UVC (253.7 nm irradiation was investigated in the present study using UV-Vis spectroscopy, steady state/time resolved fluorescence spectroscopic techniques. The stability of protein was found to be very strong when BSA gets bind to 5FU and moreover it is compared with the free BSA under UVC irradiation. From the fluorescence spectroscopic study, the stability of the complex was found to acquire 2-fold stronger than free protein. From the molecular modelling studies, we came to know the hydrogen bonds between BSA and antimetabolite 5FU are strong, up to 70.4 J/m2 under UVC irradiation.

  8. Synthesis and characterization of mononuclear copper(II complex of tetradentate N2S2 donor set and the study of DNA and bovine serum albumin binding

    Directory of Open Access Journals (Sweden)

    Sandipan Sarkar

    2014-12-01

    Full Text Available One mononuclear copper(II complex, containing neutral tetradentate NSSN-type ligands, of formulation [Cu II(L 1Cl]ClO 4 (1, was synthesized and isolated in pure form [where L 1˭ 1,3-bis(3-pyridylmethylthiopropane]. Green-colored copper(II complex was characterized by physicochemical, spectroscopic methods and conductivity measurement. These experimental data matched well with the proposed structure of the complex. Biological activity of the complex (1 toward calf thymus DNA and bovine serum albumin has been examined systematically and groove-binding behavior of the Copper(II complex 1 with calf thymus DNA has been observed from the spectral study.

  9. The standard molar enthalpy of formation of a new copper(II) Schiff-base complex and its interaction with bovine serum albumin

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Jin-Qi [Hunan Provincial Key Laboratory of Xiangnan Rare-Precious Metals Compounds and Applications, Department of Chemistry and Life Science, Xiangnan University, Chenzhou 423000, Hunan (China); Li, Chuan-Hua, E-mail: lichuanhua0526@126.com [Hunan Provincial Key Laboratory of Xiangnan Rare-Precious Metals Compounds and Applications, Department of Chemistry and Life Science, Xiangnan University, Chenzhou 423000, Hunan (China); Dong, Jia-Xin [School of Chemistry and Pharmaceutical Sciences, Guangxi Normal University, Guilin 541004 (China); Qu, Wei; Pan, Lan; Peng, Meng-La; Xie, Ming-An; Tao, Xu; Yu, Cheng-Mao; Zhu, Yi; Zhang, Ping-Hua; Tang, Chun-Guang [Hunan Provincial Key Laboratory of Xiangnan Rare-Precious Metals Compounds and Applications, Department of Chemistry and Life Science, Xiangnan University, Chenzhou 423000, Hunan (China); Li, Qiang-Guo, E-mail: liqiangguo@163.com [Hunan Provincial Key Laboratory of Xiangnan Rare-Precious Metals Compounds and Applications, Department of Chemistry and Life Science, Xiangnan University, Chenzhou 423000, Hunan (China)

    2014-12-20

    Highlights: • A new copper(II) Valen Schiff-base complex was synthesized and characterized. • The standard molar enthalpy of formation of the title complex was obtained. • The interaction between the complex and bovine serum albumin was investigated. - Abstract: A new copper(II) Schiff-base complex [Cu(HL)·NO{sub 3}·MeOH] was prepared by using equivalent molar of Valen Schiff-base ligand [H{sub 2}L=N,N′-ethylene-bis(3-methoxysalicylideneimine)] and Cu(NO{sub 3}){sub 2}·3H{sub 2}O. The structure of the complex was confirmed by single-crystal X-ray diffraction. Based on an ideal and feasible thermochemical cycle, the standard molar enthalpy of formation of the complex was estimated to be: Δ{sub f}H{sub m}{sup θ} [Cu(HL)·NO{sub 3}·MeOH(s), 298.15 K] = –(945.40 ± 2.44) kJ mol{sup −1} by an advanced solution-reaction isoperibol calorimeter. In particular, the interaction between the complex and bovine serum albumin (BSA) was investigated using the fluorescence quenching method. Fluorescence quenching data showed that the quenching mechanism of BSA treated by the complex was static quenching, which was highly accord with the non-radioactive energy transfer theory. And some relevant parameters such as binding sites, binding distance and intermolecular forces between the complex and BSA were also obtained by analyzing the fluorescence spectral data.

  10. Polymorphisms in the bovine leptin promoter associated with serum leptin concentration, growth, feed intake, feeding behavior, and measures of carcass merit.

    Science.gov (United States)

    Nkrumah, J D; Li, C; Yu, J; Hansen, C; Keisler, D H; Moore, S S

    2005-01-01

    Leptin is the hormone product of the obese gene synthesized and secreted predominantly by white adipocytes. It functions as a lipostatic signal regulating BW, food intake, energy expenditure, reproduction, and certain immune system functions. Although previous studies have identified polymorphisms in the coding regions of the leptin gene in cattle that show considerable associations with feed intake, milk quality and quantity, and carcass fatness, no such associations have been reported for the leptin promoter. The current study reports associations between SNP in the 5' untranslated promoter region of the bovine leptin gene with serum leptin concentration, growth, BW, feed intake, feeding behavior, and carcass merit in hybrid cattle (n = 150). The study showed that animals with the TT genotype of a less frequent cytosine/thymine (C/ T) substitution (UASMS2; frequency of thymine allele equals 0.21) detected at position 528 in the bovine leptin promoter (GenBank Accession No. AB070368) show 48 and 39% increases in serum leptin concentration (P < 0.001), 39 and 31% increases in backfat thickness (P < 0.001), and 13 and 9% increase in marbling score (P = 0.01), compared with CC or CT genotypes, respectively. Animals with the TT genotype also show significantly higher feed intake (P < 0.001), growth rate, metabolic BW (P < 0.05), and live weight at slaughter (P < 0.10). Animals with the GG genotype of a more frequent cytosine/guanine (C/G) substitution (UASMS3; frequency of G allele equals 0.59) at position 1759 in the bovine leptin promoter (GenBank Accession No. AB070368) also show higher feed intake (P = 0.001), growth rate (P < 0.10), and BW (P < 0.01). The thymine allele of UASMS2 and the guanine allele of UASMS3 were separately associated with higher feeding duration (P < 0.05). The two SNP show significant linkage disequilibrium and could also be relevant in predicting other characteristics, such as milk yield and quality in cattle. These results, however

  11. Zhen-wu-tang attenuates cationic bovine serum albumin-induced inflammatory response in membranous glomerulonephritis rat through inhibiting AGEs/RAGE/NF-κB pathway activation.

    Science.gov (United States)

    Wu, Junbiao; Liu, Bihao; Liang, Chunling; Ouyang, Hui; Lin, Jin; Zhong, Yanchun; He, Yu; Zhou, Jie; Zhou, Yuan; Zhou, Jiuyao

    2016-04-01

    Zhen-wu-tang (ZWT), a traditional Chinese compound formula recorded in the Treatise on Febrile Diseases, has significant inhibitory effects on inflammatory damage and oxidative lesions in rats, but its mechanism of action remains unclear. The aim of the present study was to explore whether the anti-inflammatory and anti-oxidative effects of ZWT were mediated by the AGEs/RAGE/NF-κB signaling pathway in rats with cationic bovine serum albumin (C-BSA)-induced membranous glomerulonephritis (MGN). We found that ZWT significantly reduced the production of malondialdehyde (MDA), but enhanced the superoxide dismutase (SOD) activity. The ELISA results showed that ZWT not only reduced the serum levels of AGEs but also decreased the release of inflammatory mediators (TNF-α, IL-1β, and IL-6). Meanwhile, HE staining showed that pathological kidney injury was alleviated by ZWT. In addition, ZWT suppressed the expression of RAGE1 and NF-κB p65, as well as the nuclear translocation of NF-κB p65. The accumulation of AGEs, oxidative lesions and inflammation damage were reduced by an AGE inhibitor. Thus, the present study demonstrates that AGEs play a role in the pathogenesis of MGN and that AGE inhibition could reduce the inflammatory reactions and oxidative lesions in MGN. In general, ZWT attenuated MGN, in part, by inhibiting the AGEs/RAGE/NF-κB pathway. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Potential mechanisms of effects of serum-derived bovine immunoglobulin/protein isolate therapy in patients with diarrhea-predominant irritable bowel syndrome.

    Science.gov (United States)

    Valentin, Nelson; Camilleri, Michael; Carlson, Paula; Harrington, Sean C; Eckert, Deborah; O'Neill, Jessica; Burton, Duane; Chen, Jun; Shaw, Audrey L; Acosta, Andres

    2017-03-01

    Serum-derived bovine immunoglobulin/protein isolate (SBI), an oral nutritional therapy, is efficacious in diverse diarrheal diseases. In an open-label study in 15 patients with irritable bowel syndrome-diarrhea (IBS-D), we evaluated effects of SBI (5.0 g, twice a day) for 8 weeks on safety, on bowel function and abdominal pain, tryptophan metabolism (K:T ratio), intestinal permeability ( 13 C-mannitol and lactulose excretion), bile acid synthesis (fasting serum FGF-19 and C4), duodenal and stool microbiome, and the expression of 90 genes related to inflammation, immune function, and tight junctions in duodenal mucosa. Statistical analysis (paired tests, baseline vs. treatment) was based on intention to treat (ITT) principles. One of 15 Caucasian patients (13F, 2M, age 40.3 ± 2.3y, BMI 34.3 ± 3.0 kg/m 2 ) withdrew without completing studies. There were improvements in stools/day (decrease, P  D patients is associated with improved bowel function; the mechanism of benefit is unclear, though there were microbiota structure differences in duodenal brushings. Further studies in patients with low-grade inflammation and intestinal barrier dysfunction at baseline are indicated. © 2017 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.

  13. Human Serum is as Efficient as Fetal Bovine Serum in Supporting Proliferation and Differentiation of Human Multipotent Stromal (Mesenchymal) Stem Cells In Vitro and In Vivo

    DEFF Research Database (Denmark)

    Aldahmash, Abdullah; Haack-Sørensen, Mandana; Al-Nbaheen, May

    2011-01-01

    pose a health risk for patients. METHODS: We carried out a side-by-side comparison of the effects of allogenic pooled human serum (HuS) versus FBS on hMSC proliferation and differentiation in vitro and in vivo. As a model for hMSC, we employed telomerase-immortalized hMSC; hMSC-TERT cell line. RESULTS....... FBS. hMSC-TERT or primary bone marrow derived hMSC induced to osteoblastic or adipocytic differentiation in the presence of HuS or FBS showed comparable levels of gene expression and protein production of osteoblastic markers (CBFA1/Runx2, alkaline phosphastase, collagen type I and osteocalcin...... subcutaneously in immune deficient mice. hMSC maintained in HuS vs. FBS formed comparable heterotopic bone. DISCUSSION: Human serum can support proliferation and differentiation of hMSC in vitro and can maintain their bone forming capacity in vivo. The use of human serum in cell cultures of hMSC intended...

  14. A combined spectroscopic and molecular docking study on site selective binding interaction of Toluidine blue O with Human and Bovine serum albumins

    Energy Technology Data Exchange (ETDEWEB)

    Selva Sharma, Arumugam [Department of Chemistry, Bharathiar University, Coimbatore 641046 (India); Anandakumar, Shanmugam [Department of Bioinformatics, Bharathiar University, Coimbatore 641046 (India); Ilanchelian, Malaichamy, E-mail: chelian73@yahoo.com [Department of Chemistry, Bharathiar University, Coimbatore 641046 (India)

    2014-07-01

    In the present investigation the interaction of a biologically active photodynamic therapeutic agent Toluidine blue O (TBO) with Serum albumins viz Human serum albumin (HSA) and Bovine serum albumin (BSA) was studied using absorption, emission, circular dichroism spectroscopy and molecular docking experiments. The emission titration experiments between HSA/BSA and TBO revealed the existence of strong interactions between TBO and the proteins. The site competitive experiment of HSA and BSA showed that the primary binding site of TBO is located in site I of HSA/BSA involving hydrophobic, hydrogen bonding and electrostatic interaction. To ascertain the results of site competitive experiments, molecular docking was utilized to characterize the binding models of TBO–HSA/BSA complexes. From the molecular docking studies, free energy calculations were undertaken to examine the energy contributions and the role of various amino acid residues of HSA/BSA in TBO binding. The existence of Forster Resonance Energy Transfer (FRET) between the ligand and the protein was utilized to calculate the donor–acceptor distance of TBO and protein. The TBO induced conformational changes of HSA/BSA was established using synchronous emission, three dimensional emission and circular dichroism studies. - Highlights: • Site selective binding interaction of TBO with HSA and BSA were investigated. • TBO quenches the intrinsic fluorescence of HSA/BSA by static quenching process. • Computational studies of TBO with HSA/BSA substantiate the experimental findings. • 3D and CD spectral studies of TBO–HSA/BSA revealed structural changes in protein. • The distance (r) between TBO and HSA/BSA were estimated from FRET theory.

  15. The production of cross-reactive autoantibodies that bind to bovine serum albumin in mice administered reducing sugars by subcutaneous injection.

    Science.gov (United States)

    Park, Ji-Hun; Choi, Tae-Saeng

    2015-01-01

    In a previous study, we identified the formation of cross-reactive autoantibodies that bound to bovine serum albumin (BSA) in a D-galactose-induced aging mouse model. In this study, we investigated the effect of other reducing sugars (namely, glucose and fructose) on the formation of autoantibodies. The effects of concentration and route of administration on the formation of autoantibodies were examined in detail. Three concentrations (100, 500, and 1,000 mg/kg) of reducing sugars were tested. The effects of different routes of administration (subcutaneous, oral, and intraperitoneal) on the formation of autoantibodies were also analysed. The immunoreactivities of serum samples from mice treated with reducing sugars were analysed by an enzyme-linked immunosorbent assay (ELISA) using BSA or mouse serum albumin antigens (MSA). Repeated subcutaneous administration of all reducing sugars lead to autoantibody formation in a concentration-dependent manner. However, these autoantibodies did not cross-react with MSA, and simultaneous treatment of aminoguanidine with reducing sugars did not show any inhibitory effects on the formation of autoantibodies. No autoantibodies were detected after oral or intraperitoneal administration of reducing sugars. Immunohistochemistry data showed that the target antigen(s) of the autoantibodies were present only in the skin tissue of mice treated with reducing sugars. Our results show that administration of reducing sugars by subcutaneous injection leads to the formation of autoantibodies that cross-react with BSA; the formation and target antigen(s) of the autoantibodies may originate from within the skin tissue treated with the reducing sugars.

  16. Sensitivity, specificity and predictive probability values of serum agglutination test titres for the diagnosis of Salmonella Dublin culture-positive bovine abortion and stillbirth.

    Science.gov (United States)

    Sánchez-Miguel, C; Crilly, J; Grant, J; Mee, J F

    2017-12-12

    The objective of this study was to determine the diagnostic value of maternal serology for the diagnosis of Salmonella Dublin bovine abortion and stillbirth. A retrospective, unmatched, case-control study was carried out using twenty year's data (1989-2009) from bovine foetal submissions to an Irish government veterinary laboratory. Cases (n = 214) were defined as submissions with a S. Dublin culture-positive foetus from a S. Dublin unvaccinated dam where results of maternal S. Dublin serology were available. Controls (n = 415) were defined as submissions where an alternative diagnosis other than S. Dublin was made in a foetus from an S. Dublin unvaccinated dam where the results of maternal S. Dublin serology were available. A logistic regression model was fitted to the data: the dichotomous dependent variable was the S. Dublin foetal culture result, and the independent variables were the maternal serum agglutination test (SAT) titre results. Salmonella serology correctly classified 87% of S. Dublin culture-positive foetuses at a predicted probability threshold of 0.44 (cut-off at which sensitivity and specificity are at a maximum, J = 0.67). The sensitivity of the SAT at the same threshold was 73.8% (95% CI: 67.4%-79.5%), and the specificity was 93.2% (95% CI: 90.3%-95.4%). The positive and negative predictive values were 84.9% (95% CI: 79.3%-88.6%) and 87.3% (95% CI: 83.5%-91.3%), respectively. This study illustrates that the use of predicted probability values, rather than the traditional arbitrary breakpoints of negative, inconclusive and positive, increases the diagnostic value of the maternal SAT. Veterinary laboratory diagnosticians and veterinary practitioners can recover from the test results, information previously categorized, particularly from those results declared to be inconclusive. © 2017 Blackwell Verlag GmbH.

  17. Is Serum Hypovitaminosis D Associated with Chronic Widespread Pain Including Fibromyalgia? A Meta-analysis of Observational Studies.

    Science.gov (United States)

    Hsiao, Ming-Yen; Hung, Chen-Yu; Chang, Ke-Vin; Han, Der-Sheng; Wang, Tyng-Guey

    2015-01-01

    Chronic widespread pain (CWP) is a global musculoskeletal disorder leading to disability and a reduced quality of life. Low levels of serum vitamin D has long been proposed to be associated with CWP, but previous research remains inconclusive. To determine whether hypovitaminosis D was independently associated with CWP. Meta-analysis of observational study. Electronic databases were searched for studies published up to November 2014 comparing the prevalence of hypovitaminosis D and serum vitamin D levels between participants with and without CWP. The crude and adjusted odds ratios (ORs) of hypovitaminosis D with CWP were calculated. Subgroup analysis according to gender, threshold of hypovitaminosis, and definition of patients was performed, as well as meta-regression to test the linear relationship between crude ORs and the latitude of study locations. Twelve studies were included, comprising 1,854 patients with CWP. The patient group showed a significantly higher risk of hypovitaminosis D than the control group (crude OR, 1.63; 95% CI, 1.20-2.23). The association was slightly attenuated after adjusting confounders, with a pooled adjusted OR of 1.41 (95% CI, 1.00-2.00). There was an increase in ORs of hypovitaminosis D using a lower diagnostic value of serum vitamin D (8 and 10 ng/mL). The subgroup analysis according to gender and definition of CWP did not reveal significant between-group differences. The meta-regression showed no linear relationship between latitude and the crude ORs. There was a positive crude association between hypovitaminosis D and CWP, and the association was likely to remain after adjusting confounding factors. Use of a cut-off value of hypovitaminosis D (8-10 ng/mL) could better define the population with and without CWP. Further prospective follow-up studies are warranted to clarify the causal relationship between hypovitaminosis D and CWP.

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

    Energy Technology Data Exchange (ETDEWEB)

    Perkins, J; Parida, S; Clavijo, A

    2007-05-14

    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.

  19. Fretting wear behaviour of hydroxyapatite-titanium composites in simulated body fluid, supplemented with 5 g l-1 bovine serum albumin

    Science.gov (United States)

    Kumar, Alok; Biswas, Krishanu; Basu, Bikramjit

    2013-10-01

    Damaged articulating joints can be repaired or replaced with synthetic biomaterials, which can release wear debris due to articulation, leading to the osteolysis. In a recent work, it has been shown that it is possible to achieve a better combination of flexural strength/fracture toughness as well as in vitro bioactivity and cytocompatibility properties in spark plasma sintered hydroxyapatite-titanium (HA-Ti) composites. Although hydroxyapatite and titanium are well documented for their good biocompatibility, nanosized hydroxyapatite (HA) and titanium (Ti) particles can cause severe toxicity to cells. In order to address this issue, fretting wear study of HA-Ti composites under dry and wet (1× SBF, supplemented with 5 g l-1 bovine serum albumin (BSA)) condition was performed to assess the wear resistance as well as wear debris formation, in vitro. The experimental results reveal one order of magnitude lower wear rate for HA-10 wt% Ti (7.5 × 10-5 mm3 N-1 m-1) composite than monolithic HA (3.9 × 10-4 mm3 N-1 m-1) in simulated body fluid. The difference in the tribological properties has been analyzed in the light of phase assemblages and mechanical properties. Overall, the results suggest the potential use of HA-Ti composites over existing HA-based biocomposites in orthopedic as well as dental applications.

  20. Spectroscopic analyses on interaction of Amantadine-Salicylaldehyde, Amantadine-5-Chloro-Salicylaldehyde and Amantadine-o-Vanillin Schiff-Bases with bovine serum albumin (BSA)

    Science.gov (United States)

    Wang, Zhiqiu; Gao, Jingqun; Wang, Jun; Jin, Xudong; Zou, Mingming; Li, Kai; Kang, Pingli

    2011-12-01

    In this work, three Tricyclo [3.3.1.1(3,7)] decane-1-amine (Amantadine) Schiff-Bases, Amantadine-Salicylaldehyde (AS), Amantadine-5-Chloro-Salicylaldehyde (AS-5-C) and Amantadine-o-Vanillin (AS-o-V), were synthesized by direct heating reflux method in ethanol solution and characterized by infrared spectrum and elementary analysis. Fluorescence quenching was used to study the interaction of these Amantadine Schiff-Bases (AS, AS-5-C and AS-o-V) with bovine serum albumin (BSA). According to fluorescence quenching calculations the bimolecular quenching constant ( Kq), apparent quenching constant ( KSV), effective binding constant ( KA) and corresponding dissociation constant ( KD), binding site number ( n) and binding distance ( r) were obtained. The results show that these Amantadine Schiff-Bases can obviously bind to BSA molecules and the binding strength order is AS < AS-5-C = AS-o-V. Synchronous fluorescence spectroscopy reveals that these Amantadine Schiff-Bases adopt different way to bind with BSA molecules. That is, the AS and AS-5-C are accessibility to tryptophan (Trp) residues more than the tyrosine (Tyr) residues, while the AS-o-V is equally close to the Tyr and Trp residues.

  1. Transmission electron microscopy and time resolved optical spectroscopy study of the electronic and structural interactions of ZnO nanorods with bovine serum albumin.

    Science.gov (United States)

    Klaumünzer, M; Weichsel, U; Mačković, M; Spiecker, E; Peukert, W; Kryschi, C

    2013-08-22

    The adsorption behavior and electronic interactions of bovine serum albumin (BSA) with ZnO nanorod surfaces were investigated using high-resolution transmission electron microscopy as well as stationary and time-resolved optical spectroscopy techniques. Transmission electron microscopy shows that ZnO nanorod surfaces are surrounded by a homogeneous amorphous BSA film with thicknesses between ~2.5 and 5.0 nm. The electronic structure and adsorption geometry of BSA were examined using high-angle annular dark field scanning transmission electron microscopy combined with electron energy loss spectroscopy. The adsorption process was observed to result into an unfolded conformation of BSA becoming predominantly bound in the side-on orientation at the ZnO surface. This adsorption mode of the BSA molecules allows for a strong interaction with surface states of the ZnO nanorods. This is obvious from its efficient quenching of the defect-center photoluminescence of ZnO. Complementary information of electronic interactions across the ZnO nanorod interface was obtained from femtosecond transient absorption spectroscopy experiments. The rise dynamics of the measured transients revealed altered hole trapping dynamics and, thus, indicated to heterogeneous charge transfer as emerging from adsorbed BSA molecules to defect centers of the ZnO interface.

  2. Ultrasonic-assisted fabrication and characterization of PVC-SiO2nanocomposites having bovine serum albumin as a bio coupling agent.

    Science.gov (United States)

    Mallakpour, Shadpour; Nazari, Hossein Yazdan

    2017-11-01

    In this work, SiO 2 nanoparticles (NPs) were modified with bovine serum albumin (BSA) under ultrasound irradiations as a green and fast route to achieve their good dispersion. Subsequently, different weight percentages of the modified NPs (3, 6, and 9wt%) were incorporated in poly(vinyl chloride) (PVC) as the matrix. Thermogravimetric analysis of the SiO 2 -BSA NPs indicated that 12wt% of the modifier was loaded on the surface of SiO 2 NPs. Encapsulation of the SiO 2 -BSA resulted in a meaningful improvement in the optical, mechanical and thermal characteristics of the prepared PVC nanocomposites (NCs). X-ray diffraction (XRD) patterns for the PVC/SiO 2 -BSA NCs showed a crystalline behavior for the NC with 6wt% of the SiO 2 -BSA originated from the phosphate buffer on the NPs. Water contact angle of the PVC/SiO 2 -BSA NCs showed that the hydrophilicity enhanced with increasing of the NPs contents. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Zinc Phthalocyanine Labelled Polyethylene Glycol: Preparation, Characterization, Interaction with Bovine Serum Albumin and Near Infrared Fluorescence Imaging in Vivo

    Directory of Open Access Journals (Sweden)

    Tianjun Liu

    2012-05-01

    Full Text Available Zinc phthalocyanine labelled polyethylene glycol was prepared to track and monitor the in vivo fate of polyethylene glycol. The chemical structures were characterized by nuclear magnetic resonance and infrared spectroscopy. Their light stability and fluorescence quantum yield were evaluated by UV-Visible and fluorescence spectroscopy methods. The interaction of zinc phthalocyanine labelled polyethylene glycol with bovine serum albumin was evaluated by fluorescence titration and isothermal titration calorimetry methods. Optical imaging in vivo, organ aggregation as well as distribution of fluorescence experiments for tracking polyethylene glycol were performed with zinc phthalocyanine labelled polyethylene glycol as fluorescent agent. Results show that zinc phthalocyanine labelled polyethylene glycol has good optical stability and high emission ability in the near infrared region. Imaging results demonstrate that zinc phthalocyanine labelled polyethylene glycol can track and monitor the in vivo process by near infrared fluorescence imaging, which implies its potential in biomaterials evaluation in vivo by a real-time noninvasive method.

  4. The effect of Cu2+ or Fe3+ on the noncovalent binding of rutin with bovine serum albumin by spectroscopic analysis.

    Science.gov (United States)

    Li, Daojin; Zhu, Mei; Xu, Chen; Chen, Jianjun; Ji, Baoming

    2011-01-01

    The interaction of rutin to bovine serum albumin (BSA) in aqueous solution was investigated by fluorescence spectra and ultraviolet-visible (UV-vis) spectra at pH 7.40. There are also some metal ions present in blood plasma, thus the research about the effect of metal ions on the interaction of drugs with proteins is crucial. Therefore, we have studied the effect of Cu2+ or Fe3+ on the interaction between rutin and BSA by using spectroscopic technique at pH 7.40, for the first time. The results of fluorescence titration indicated that rutin could quench the intrinsic fluorescence of BSA in a static quenching way. The binding sites number (n), the binding constant (K) and the spatial-distance (r) of rutin with BSA without or with Cu2+ or Fe3+ at 310 K were calculated. The result showed that the presence of Cu2+ or Fe3+ increased the binding constant and changed the binding distance between the acceptor and the donor, which possibly results from the formation of metal ions-BSA complex. The effect of rutin on the conformation of BSA was also analyzed using UV under experimental conditions. Furthermore, the fluorescence displacement experiments indicated that rutin is situated within subdomain IIA of BSA. Copyright © 2010 Elsevier B.V. All rights reserved.

  5. Multi-spectroscopic and molecular modeling approaches to elucidate the binding interaction between bovine serum albumin and darunavir, a HIV protease inhibitor

    Science.gov (United States)

    Shi, Jie-Hua; Zhou, Kai-Li; Lou, Yan-Yue; Pan, Dong-Qi

    2018-01-01

    Darunavir (DRV), a second-generation HIV protease inhibitor, is widely used across the world as an important component of HIV therapy. The interaction of DRV with bovine serum albumin (BSA), a major carrier protein, has been studied under simulated physiological conditions (pH 7.4) by multi-spectroscopic techniques in combination with molecular modeling. Fluorescence data revealed that the intrinsic fluorescence of BSA was quenched by DRV in terms of a static quenching procedure due to the formation of the DRV-BSA complex. The results indicated the presence of single weak affinity binding site ( 103 M- 1, 310 K) on protein. The thermodynamic parameters, namely enthalpy change (ΔH0), entropy change (ΔS0) and Gibbs free energy change (ΔG0) were calculated, which signified that the binding reaction was spontaneous, the main binding forces were hydrogen bonding and van der Waals forces. Importantly, competitive binding experiments with three site probes, phenylbutazone (in sub-domain IIA, site I), ibuprofen (in sub-domain IIIA, site II) and artemether (in the interface between sub-domain IIA and IIB, site II'), suggested that DRV was preferentially bound to the hydrophobic cavity in site II' of BSA, and this finding was validated by the docking results. Additionally, synchronous fluorescence, three-dimensional fluorescence and Resonance Rayleigh Scattering (RRS) spectroscopy gave qualitative information on the conformational changes of BSA upon adding DRV, while quantitative data were obtained with Fourier transform infrared spectroscopy (FT-IR).

  6. Binding investigation on the interaction between Methylene Blue (MB)/TiO2 nanocomposites and bovine serum albumin by resonance light-scattering (RLS) technique and fluorescence spectroscopy.

    Science.gov (United States)

    Li, Yuesheng; Zhang, Yue; Sun, Shaofa; Zhang, Aiqing; Liu, Yi

    2013-11-05

    The interaction between Methylene Blue (MB)/TiO2 nanocomposites and bovine serum albumin (BSA) was investigated by resonance light scattering (RLS), fluorescence, three-dimension spectra and UV-vis absorbance spectroscopy. Several factors which may influence the RLS intensity were also investigated before characterizing MB/TiO2-BSA complex. It was proved that the mechanism of MB/TiO2 nanocomposites binding to BSA was mainly a result of the formation of MB/TiO2-BSA complex. The binding constant of MB/TiO2-BSA is 0.762 × 10(-5) L mol(-1) at 298K. By calculating the binding constant at different temperature, the thermodynamic parameters ΔH, ΔG, and ΔS can be observed and deduced that the hydrophobic interactions played an important role to stabilize the complex. The distance r (3.73 nm) between donor (BSA) and acceptor (MB/TiO2) was obtained according to fluorescence resonance energy transfer (FRET). The binding site for MB/TiO2 on BSA was mainly located in sub-domain IIA. The UV-vis absorbance, circular dichroism and three dimension fluorescence have also been used to investigate the effect of MB/TiO2 on the conformation of BSA. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Understanding thermodynamics of drug partitioning in micelles and delivery to proteins: Studies with naproxen, diclofenac sodium, tetradecyltrimethylammonium bromide, and bovine serum albumin

    International Nuclear Information System (INIS)

    Talele, Paurnima; Choudhary, Sinjan; Kishore, Nand

    2016-01-01

    Highlights: • Interactions of non-steroidal anti-inflammatory drugs studied with TTAB micelles, monomers. • Thermodynamics of drug-surfactant interactions and partitioning in micelles addressed. • Mechanism of drug partitioning addressed based on energetics of interactions. • Partitioning in micelles depends on functional groups on drugs. • Such studies are needed for target oriented synthesis and efficient drug delivery. - Abstract: The use of surfactants in drug delivery has offered several advantages. Quantitative knowledge of the interactions of drugs with micellar systems is essential for deriving guidelines to design efficient drug delivery systems. In this work we have quantitatively addressed the mechanism of interaction of naproxen and diclofenac sodium with the micelles and monomers of tetradecyltrimethylammonium bromide (TTAB) based on thermodynamic studies by using isothermal titration calorimetry. The mechanism of interaction of the drugs with TTAB is based on identification of the nature of interactions of the former with the surfactant micelles and monomers. The values of partitioning constant (which is same as equilibrium constant for the reaction of drugs with the surfactant micelles), enthalpy, entropy and stoichiometry of partitioning have been determined and discussed in terms of possible intermolecular interactions. Further, the interaction of the drug naproxen with bovine serum albumin, when delivered from the micellar media has also been addressed in terms of binding constant, enthalpy and entropy of binding. The results are important in developing improved strategies for effective drug delivery systems.

  8. The Binding of Four Licorice Flavonoids to Bovine Serum Albumin by Multi-Spectroscopic and Molecular Docking Methods: Structure-Affinity Relationship

    Science.gov (United States)

    Hou, J.; Liang, Q.; Shao, S.

    2017-03-01

    Flavanones are the main compound of licorice, and the C'-4 position substitution is a significant structural feature for their biological activity. The ability of three selected flavanones (liquiritigenin, liquiritin, and liquiritin apioside) bearing different substituents (hydroxyl groups, glucose, and glucose-apiose sugar moiety) at the C'-4 position and a chalcone ( isoliquiritigenin, an isomer of liquiritigenin) to bind bovine serum albumin (BSA) was studied by multispectroscopic and molecular docking methods under physiological conditions. The binding mechanism of fl avonoids to BSA can be explained by the formation of a flavonoids-BSA complex, and the binding affinity is the strongest for isoliquiritigenin, followed by liquiritin apioside, liquiritin, and liquiritigenin. The thermodynamic analysis and the molecular docking indicated that the interaction between flavonoids and BSA was dominated by the hydrophobic force and hydrogen bonds. The competitive experiments as well as the molecular docking results suggested the most possible binding site of licorice flavonoids on BSA at subdomain IIA. These results revealed that the basic skeleton structure and the substituents at the C'-4 position of flavanones significantly affect the structure-affinity relationships of the licorice flavonoid binding to BSA.

  9. Characterizing the binding interaction of fungicide boscalid with bovine serum albumin (BSA): A spectroscopic study in combination with molecular docking approach.

    Science.gov (United States)

    Lou, Yan-Yue; Zhou, Kai-Li; Shi, Jie-Hua; Pan, Dong-Qi

    2017-08-01

    Boscalid, a carboxamide fungicide, is used in the treatment of grey mould and powdery mildew, widely applied to a variety of crops and fruits such as rice, wheat, grapes and pears. It will become a potential risk for health due to its widely application and residue in crops and fruits. In this study, the binding interaction between boscalid and bovine serum albumin (BSA) was characterized using steady-state fluorescence spectroscopy, ultraviolet spectroscopy (UV), synchronous fluorescence spectroscopy, 3D fluorescence spectroscopy, Fourier transform infrared spectroscopy (FT-IR) and molecular docking to ascertain the store, transport and distribution of boscalid in vivo. The experimental results indicated that the fluorescence of BSA was quenched due to the forming the static boscalid-BSA complex with the binding constant of 4.57×10 3 M -1 at 298 K and boscalid bound on the subdomain III A (site II) of BSA through van der Waals force and hydrogen bonding interaction. The binding process of boscalid with BSA was spontaneous and enthalpy-driven process based on ΔG 0 T|ΔS 0 | over the studied temperature range. Meanwhile, the obvious change in the conformation of boscalid was observed while the slight change in the conformation of BSA when binding boscalid to the BSA, implying that the flexibility of boscalid contributes to increasing the stability of the boscalid-BSA complex. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Interaction of bovine serum albumin (BSA) with novel gemini surfactants studied by synchrotron radiation scattering (SR-SAXS), circular dichroism (CD), and nuclear magnetic resonance (NMR).

    Science.gov (United States)

    Gospodarczyk, W; Szutkowski, K; Kozak, M

    2014-07-24

    The interaction of three dicationic (gemini) surfactants-3,3'-[1,6-(2,5-dioxahexane)]bis(1-dodecylimidazolium) chloride (oxyC2), 3,3'-[1,16-(2,15-dioxahexadecane)]bis(1-dodecylimidazolium) chloride (oxyC12), and 1,4-bis(butane)imidazole-1-yl-3-dodecylimidazolium chloride (C4)--with bovine serum albumin (BSA) has been studied by the use of small-angle X-ray scattering (SAXS), circular dichroism (CD), and (1)H nuclear magnetic resonance diffusometry. The results of CD studies show that the conformation of BSA was changed dramatically in the presence of all studied surfactants. The greater decrease (from 56 to 24%) in the α-helical structure of BSA was observed for oxyC2 surfactant. The radii of gyration estimated from SAXS data varied between 3 and 26 nm for the BSA/oxyC2 and BSA/oxyC12 systems. The hydrodynamic radius of the BSA/surfactant system estimated from NMR diffusometry varies between 5 and 11 nm for BSA/oxyC2 and 5 and 8 nm for BSA/oxyC12.

  11. Antimicrobial and cell viability measurement of bovine serum albumin capped silver nanoparticles (Ag/BSA) loaded collagen immobilized poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) film.

    Science.gov (United States)

    Bakare, Rotimi; Hawthrone, Samantha; Vails, Carmen; Gugssa, Ayele; Karim, Alamgir; Stubbs, John; Raghavan, Dharmaraj

    2016-03-01

    Bacterial infection of orthopedic devices has been a major concern in joint replacement procedures. Therefore, this study is aimed at formulating collagen immobilized poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) film loaded with bovine serum albumin capped silver nanoparticles (Ag/BSA NPs) to inhibit bacterial growth while retaining/promoting osteoblast cells viability. The nanoparticles loaded collagen immobilized PHBV film was characterized for its composition by X-ray Photoelectron Spectroscopy and Anodic Stripping Voltammetry. The extent of loading of Ag/BSA NPs on collagen immobilized PHBV film was found to depend on the chemistry of the functionalized PHBV film and the concentration of Ag/BSA NPs solution used for loading nanoparticles. Our results showed that more Ag/BSA NPs were loaded on higher molecular weight collagen immobilized PHEMA-g-PHBV film. Maximum loading of Ag/BSA NPs on collagen immobilized PHBV film was observed when 16ppm solution was used for adsorption studies. Colony forming unit and optical density measurements showed broad antimicrobial activity towards Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa at significantly lower concentration i.e., 0.19 and 0.31μg/disc, compared to gentamicin and sulfamethoxazole trimethoprim while MTT assay showed that released nanoparticles from Ag/BSA NPs loaded collagen immobilized PHBV film has no impact on MCTC3-E1 cells viability. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. Bovine Serum Albulmin Protein-Templated Silver Nanocluster (BSA-Ag13): An Effective Singlet Oxygen Generator for Photodynamic Cancer Therapy.

    Science.gov (United States)

    Yu, Yong; Geng, Junlong; Ong, Edward Yong Xi; Chellappan, Vijila; Tan, Yen Nee

    2016-10-01

    This paper reports a novel synthesis approach of bovine serum albumin (BSA) protein-templated ultrasmall (BSA-Ag 13 NC (i.e., 13 Ag atoms per cluster) is successfully synthesized for the first time by using NaOH-dissolved NaBH 4 solution as the controlling reducing agent. The ubiquitous size of BSA-Ag 13 NC results in unique behaviors of its photoexcited states as characterized by the ultrafast laser spectroscopy using time-correlated single photon counting and transient absorption techniques. In particular, triply excited states can be largely present in the excited BSA-Ag 13 NC and readily sensitized molecular oxygen to produce singlet oxygen ( 1 O 2 ) with a high quantum efficiency (≈1.26 using Rose Bengal as a standard). This value is much higher than its Au analogue (i.e., ≈0.07 for BSA-Au 25 NC) and the commonly available photosensitizers. Due to the good cellular uptake and inherent biocompatibility imparted by the surface protein, BSA-Ag 13 NC can be applied as an effective PDT agent in generating 1 O 2 to kill cancer cell as demonstrated in this study. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Effect of apatite formation of biphasic calcium phosphate ceramic (BCP) on osteoblastogenesis using simulated body fluid (SBF) with or without bovine serum albumin (BSA).

    Science.gov (United States)

    Huang, Li; Zhou, Bo; Wu, Huayu; Zheng, Li; Zhao, Jinmin

    2017-01-01

    Although biphasic calcium phosphate ceramic (BCP) holds promise in therapy of bone defect, surface mineralization prior to implantation may improve the bioactivity to better integrate with the host. Immersion in simulated body fluid (SBF) and bovine serum albumin-simulated body fluid (BSA-SBF) are common methods to form apatite interface layer. This study was intended to investigate the effect of SBF and BSA-SBF treatment on the bioactivity of BCP in vitro. In this study, osteoblasts were grown on BCP with or without treatment of SBF or BSA-SBF, and detected with general observation, scanning electron microscope (SEM), cell proliferation assay, morphology observation, viability assay, alkaline phosphatase (ALP) activity assay, and osteogenic specific gene expression of alkaline phosphatase (ALPL), bone gamma-carboxyglutamate (gla) protein (BGLAP), bone morphogenetic protein 2 (BMP2), bone sialoprotein (BSP), type I collagen (COLI) and runt-related transcription factor 2 (RUNX2) after culture of 2, 5 and 8days. As the results shown, BCP pre-incubated in SBF and BSA-SBF up-regulated ALP activity and osteogenic related genes and proteins, which testified the positive effect of SBF and BSA-SBF. Especially, BSA-SBF enhanced the cell growth significantly. This study indicated that treatment by BSA-SBF is of importance for BCP before clinical application. Copyright © 2016. Published by Elsevier B.V.

  14. Fundamental studies on the intermediate layer of a bipolar membrane. Part II. Effect of bovine serum albumin (BSA) on water dissociation at the interface of a bipolar membrane.

    Science.gov (United States)

    Fu, Rong Qiang; Xu, Tong Wen; Yang, Wei Hua; Pan, Zhong Xiao

    2004-10-15

    This paper investigates the behavior of bovine serum albumin (BSA) during water dissociation on a bipolar membrane (BPM). BSA-modified BPM is prepared by immersing polyethylene anion exchange membrane in different concentration solutions of BSA, then casting the solution of sulfonated poly(phenylene oxide) (SPPO) in dimethyl formamide. The modification of BSA was evidenced by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). The junction thickness was evaluated by electrochemical impedance spectroscopy (EIS). The results showed that the typical I-V curves for bipolar membranes were heavily affected by the BSA modifications: the more the adsorbed amount of BSA, the larger the potential drop across a bipolar membrane. The new phenomena is underlined by the intrinsic properties of BSA molecules: steric effects give rise to an increase in the thickness of the depletion layer, amphoteric properties weaken the electric field of the junction, and hydrophobicity makes the junction less wet. All of these cause negative effects on water dissociation on a bipolar membrane.

  15. Effect of urea on heat-induced gelation of bovine serum albumin (BSA) studied by rheology and small angle neutron scattering (SANS)

    Science.gov (United States)

    Nnyigide, Osita Sunday; Oh, Yuna; Song, Hyeong Yong; Park, Eun-kyoung; Choi, Soo-Hyung; Hyun, Kyu

    2017-05-01

    This paper reports the effects of urea on the heat-induced gelation of bovine serum albumin (BSA), which was studied by the tube inversion method, rheological measurements, and small-angle neutron scattering (SANS). An increase in the urea concentration accelerated the rate of gelation because the protein molecules have already been unfolded to some extent during sample preparation in the urea solution. In addition, the BSA solution in the presence of urea underwent a sol-gel-sol transition during the time sweep test at a constant temperature of 80oC. On the other hand, the BSA solution without urea turned into a hard and brittle gel that did not return to the solution state during isothermal heating at a constant temperature of 80oC. Aggregation and re-bonding of the denatured and unfolded protein chains led to gel formation. Urea added to the protein denatures its tertiary and secondary structures by simultaneously disrupting the hydrogen bonds, hydrophobic interactions, and altering the solvent properties. Furthermore, urea induces thermoreversible chemical interactions in BSA solutions leading to the formation of a gel with dynamic properties under these experimental conditions.

  16. Study of conformational changes and protein aggregation of bovine serum albumin in presence of Sb(III) and Sb(V).

    Science.gov (United States)

    Verdugo, Marcelo; Ruiz Encinar, Jorge; Costa-Fernández, José Manuel; Menendez-Miranda, Mario; Bouzas-Ramos, Diego; Bravo, Manuel; Quiroz, Waldo

    2017-01-01

    Antimony is a metalloid that affects biological functions in humans due to a mechanism still not understood. There is no doubt that the toxicity and physicochemical properties of Sb are strongly related with its chemical state. In this paper, the interaction between Sb(III) and Sb(V) with bovine serum albumin (BSA) was investigated in vitro by fluorescence spectroscopy, and circular dichroism (CD) under simulated physiological conditions. Moreover, the coupling of the separation technique, asymmetric flow field-flow fractionation, with elemental mass spectrometry to understand the interaction of Sb(V) and Sb(III) with the BSA was also used. Our results showed a different behaviour of Sb(III) vs. Sb(V) regarding their effects on the interaction with the BSA. The effects in terms of protein aggregates and conformational changes were higher in the presence of Sb(III) compared to Sb(V) which may explain the differences in toxicity between both Sb species in vivo. Obtained results demonstrated the protective effect of GSH that modifies the degree of interaction between the Sb species with BSA. Interestingly, in our experiments it was possible to detect an interaction between BSA and Sb species, which may be related with the presence of labile complex between the Sb and a protein for the first time.

  17. Evaluation of sericin as a fetal bovine serum-replacing cryoprotectant during freezing of human mesenchymal stromal cells and human osteoblast-like cells.

    Science.gov (United States)

    Verdanova, Martina; Pytlik, Robert; Kalbacova, Marie Hubalek

    2014-04-01

    A reliable, cryoprotective, xeno-free medium suitable for different cell types is highly desirable in regenerative medicine. There is danger of infection or allergic reaction with the use of fetal bovine serum (FBS), making it problematic for medical applications. The aim of the present study was to develop an FBS-free cryoprotective medium for human mesenchymal stromal cells (hMSCs; primary cells) and immortalized human osteoblasts (SAOS-2 cell line). Furthermore, we endeavored to eliminate or reduce the presence of dimethyl sulfoxide (DMSO) in the medium. Sericin, a sticky protein derived from the silkworm cocoon, was investigated as a substitute for FBS and DMSO in the freezing medium. Cell viability (24 hours after thawing, both hMSC and SAOS-2) and colony-forming ability (2 weeks after thawing, only for hMSCs) were both determined. The FBS-free medium with 1% sericin in 10% DMSO was found to be a suitable freezing medium for primary hMSCs, in contrast to immortalized human osteoblasts. Surprisingly, the storage of hMSCs in a cultivation medium with only 10% DMSO also provided satisfactory results. Any drop in DMSO concentration led to significantly worse survival of cells, with little improvement in hMSC survival in the presence of sericin. Thus, sericin may substitute for FBS in the freezing medium for primary hMSCs, but cannot substitute for DMSO.

  18. Proteolytically-induced changes of secondary structural protein conformation of bovine serum albumin monitored by Fourier transform infrared (FT-IR) and UV-circular dichroism spectroscopy.

    Science.gov (United States)

    Güler, Günnur; Vorob'ev, Mikhail M; Vogel, Vitali; Mäntele, Werner

    2016-05-15

    Enzymatically-induced degradation of bovine serum albumin (BSA) by serine proteases (trypsin and α-chymotrypsin) in various concentrations was monitored by means of Fourier transform infrared (FT-IR) and ultraviolet circular dichroism (UV-CD) spectroscopy. In this study, the applicability of both spectroscopies to monitor the proteolysis process in real time has been proven, by tracking the spectral changes together with secondary structure analysis of BSA as proteolysis proceeds. On the basis of the FTIR spectra and the changes in the amide I band region, we suggest the progression of proteolysis process via conversion of α-helices (1654 cm(-1)) into unordered structures and an increase in the concentration of free carboxylates (absorption of 1593 and 1402 cm(-1)). For the first time, the correlation between the degree of hydrolysis and the concentration of carboxylic groups measured by FTIR spectroscopy was revealed as well. The far UV-CD spectra together with their secondary structure analysis suggest that the α-helical content decreases concomitant with an increase in the unordered structure. Both spectroscopic techniques also demonstrate that there are similar but less spectral changes of BSA for the trypsin attack than for α-chymotrypsin although the substrate/enzyme ratio is taken the same. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Effect of urea on bovine serum albumin in aqueous and reverse micelle environments investigated by small angle X-ray scattering, fluorescence and circular dichroism

    International Nuclear Information System (INIS)

    Itri, Rosangela; Caetano, Wilker; Barbosa, Leandro R.S.; Baptista, Mauricio S.

    2004-01-01

    The influence that urea has on the conformation of water-soluble globular protein, bovine serum albumin (BSA), exposed directly to the aqueous solution as compared to the condition where the macromolecule is confined in the Aerosol-OT (AOT - sodium bis-2-ethylhexyl sulfosuccinate)/n-hexane/water reverse micelle (RM) is addressed. Small angle X-ray scattering (SAXS), tryptophan (Trp) fluorescence emission and circular dichroism (CD) spectra of aqueous BSA solution in the absence and in the presence of urea (3M and 5M) confirm the known denaturing effect of urea in proteins. The loss of the globular native structure is observed by the increase in the protein maximum dimension and gyration radius, through the Trp emission increase and maximum red-shift as well as the decrease in helix content. In RMs, the Trp fluorescence and CD spectra show that BSA is mainly located in its interfacial region independently of the micellar size. Addition of urea in this BSA/RM system also causes changes in the Trp fluorescence (emission decrease and maximum red-shift) and in the BSA CD spectra (decrease in helix content), which are compatible with the denaturation of the protein and Trp exposition to a more apolar environment in the RM. The fact that urea causes changes in the protein structure when it is located in the interfacial region (evidenced by CD) is interpreted as an indication that the direct interaction of urea with the protein is the major factor to explain its denaturing effect. (author)

  20. Phenytoin-Bovine Serum Albumin interactions - modeling plasma protein - drug binding: A multi-spectroscopy and in silico-based correlation

    Science.gov (United States)

    Suresh, P. K.; Divya, Naik; Nidhi, Shah; Rajasekaran, R.

    2018-03-01

    The study focused on the analysis of the nature and site of binding of Phenytoin (PHT) -(a model hydrophobic drug) with Bovine Serum Albumin (BSA) (a model protein used as a surrogate for HSA). Interactions with defined amounts of Phenytoin and BSA demonstrated a blue shift (hypsochromic -change in the microenvironment of the tryptophan residue with decrease in the polar environment and more of hydrophobicity) with respect to the albumin protein and a red shift (bathochromic -hydrophobicity and polarity related changes) in the case of the model hydrophobic drug. This shift, albeit lower in magnitude, has been substantiated by a fairly convincing, Phenytoin-mediated quenching of the endogenous fluorophore in BSA. Spectral shifts studied at varying pH, temperatures and incubation periods (at varying concentrations of PHT with a defined/constant BSA concentration) showed no significant differences (data not shown). FTIR analysis provided evidence of the interaction of PHT with BSA with a stretching vibration of 1737.86 cm- 1, apart from the vibrations characteristically associated with the amine and carboxyl groups respectively. Our in vitro findings were extended to molecular docking of BSA with PHT (with the different ionized forms of the drug) and the subsequent LIGPLOT-based analysis. In general, a preponderance of hydrophobic interactions was observed. These hydrophobic interactions corroborate the tryptophan-based spectral shifts and the fluorescence quenching data. These results substantiates our hitherto unreported in vitro/in silico experimental flow and provides a basis for screening other hydrophobic drugs in its class.

  1. Synthesis and characterization of magnetite nanoparticles having different cover layer and investigation of cover layer effect on the adsorption of lysozyme and bovine serum albumin.

    Science.gov (United States)

    Shah, Muhammad Tariq; Alveroglu, Esra

    2017-12-01

    In this study, differently coated superparamagnetic Fe 3 O 4 (magnetite) nanoparticles were synthesized, characterized and used for lysozyme (Ly) and bovine serum albumin (BSA) adsorption. SiO 2 , carbon nanotubes (CNTs) and graphene were used for covering the readily synthesized magnetite nanoparticles to elucidate the effect of cover layer on the protein adsorption kinetics and capacities of nanostructure. XRD, FTIR, AFM, SEM, VSM and fluorescence measurements were used for the characterization of the samples and investigating the adsorption kinetics of Ly and BSA by these nanoparticles. The average particle size of the Fe 3 O 4 nanoparticles are approximately found as 10nm and VSM measurement shows that the Fe 3 O 4 particles have superparamagnetic behavior with no hysteresis and remnant. The adsorption kinetic of proteins on nanosized material is followed via fluorescence method. All the nanostructures with different cover layers obey pseudo first order kinetics and SiO 2 coated nanoparticles show the fastest kinetics and capabilities for Ly and BSA adsorption. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Exploring the affinity binding of alkylmaltoside surfactants to bovine serum albumin and their effect on the protein stability: A spectroscopic approach.

    Science.gov (United States)

    Hierrezuelo, J M; Carnero Ruiz, C

    2015-08-01

    Steady-state and time-resolved fluorescence together with circular dichroism (CD) spectroscopic studies was performed to examine the interactions between bovine serum albumin (BSA) and two alkylmaltoside surfactants, i.e. n-decyl-β-D-maltoside (β-C10G2) and n-dodecyl-β-D-maltoside (β-C12G2), having identical structures but different tail lengths. Changes in the intrinsic fluorescence of BSA from static as well as dynamic measurements revealed a weak protein-surfactant interaction and gave the corresponding binding curves, suggesting that the binding mechanism of surfactants to protein is essentially cooperative in nature. The behavior of both surfactants is similar, so that the differences detected were attributed to the more hydrophobic nature of β-C12G2, which favors the adsorption of micelle-like aggregates onto the protein surface. These observations were substantially demonstrated by data derived from synchronous, three-dimensional and anisotropy fluorescence experiments. Changes in the secondary structure of the protein induced by the interaction with surfactants were analyzed by CD to determine the contents of α-helix and β-strand. It was noted that whereas the addition of β-C10G2 appears to stabilize the secondary structure of the protein, β-C12G2 causes a marginal denaturation of BSA for a protein:surfactant molar ratio as high as 1 to 100. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Penetrable silica microspheres for immobilization of bovine serum albumin and their application to the study of the interaction between imatinib mesylate and protein by frontal affinity chromatography.

    Science.gov (United States)

    Ma, Liyun; Li, Jing; Zhao, Juan; Liao, Han; Xu, Li; Shi, Zhi-guo

    2016-01-01

    In the current study, novel featured silica, named penetrable silica, simultaneously containing macropores and mesopores, was immobilized with bovine serum albumin (BSA) via Schiff base method. The obtained BSA-SiO2 was employed as the high-performance liquid chromatographic (HPLC) stationary phase. Firstly, D- and L-tryptophan were used as probes to investigate the chiral separation ability of the BSA-SiO2 stationary phase. An excellent enantioseparation factor was obtained up to 4.3 with acceptable stability within at least 1 month. Next, the BSA-SiO2 stationary phase was applied to study the interaction between imatinib mesylate (IM) and BSA by frontal affinity chromatography. A single type of binding site was found for IM with the immobilized BSA, and the hydrogen-bonding and van der Waals interactions were expected to be contributing interactions based on the thermodynamic studies, and this was a spontaneous process. Compared to the traditional silica for HPLC stationary phase, the proposed penetrable silica microsphere possessed a larger capacity to bond more BSA, minimizing column overloading effects and enhancing enantioseparation ability. In addition, the lower running column back pressure and fast mass transfer were meaningful for the column stability and lifetime. It was a good substrate to immobilize biomolecules for fast chiral resolution and screening drug-protein interactions.

  4. Green synthesis of zero-valent Fe-nanoparticles: Catalytic degradation of rhodamine B, interactions with bovine serum albumin and their enhanced antimicrobial activities.

    Science.gov (United States)

    Khan, Zaheer; Al-Thabaiti, Shaeel Ahmad

    2018-03-01

    Biomimetic method was used for the synthesis of Fe-nanoparticles (FeNPs). FeCl 3 and Hibiscus sabdariffa, Roselle flower aqueous extract (HBS) were employed in the present studies. The FeNPs have been characterized by using UV-visible spectroscopy, transmission electron microscope (TEM), and energy dispersion X-ray spectroscopy (EDS). The average particles diameter was found to be 18 nm. The as prepared FeNPs were used as a catalyst to the oxidative degradation of rhodamine B (RB) in presence of NaBH 4 . The effects of various quencher on the degradation rates were examined by employing ammonium oxalate (AO), benzoquinone (BQ), isopropyl alcohol (IPA), and potassium iodide (KI). The interactions of FeNPs with bovine serum albumin (BSA) have been determined and discussed. Adsorption of FeNPs into the core of BSA changes the tryptophan environment from hydrophobic to hydrophilic (from folding to partially folded and/or unfolded). Tryptophan residues, indole moieties of BSA were responsible to complex formation with FeNPs in excited states via electrostatic, van der Waals, hydrogen bonding, hydrophobic and hydrophilic interactions with static quenching. The antimicrobial activities of FeNPs have been determined against human pathogens. Hibiscus sabdariffa flower extract shows mild antimicrobial activities against all target pathogenic organisms. FeNPs have potential antimicrobial activity against both bacterial strains and candida fungus even at low concentration, and retains potential application in biomedical industries. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. The modifier effects of chymotrypsin and trypsin enzymes on fluorescence lifetime distribution of "N-(1-pyrenyl)maleimide-bovine serum albumin" complex

    Science.gov (United States)

    Özyiğit, İbrahim Ethem; Karakuş, Emine; Pekcan, Önder

    2016-02-01

    Chymotrypsin and trypsin are the well known proteolytic enzymes, both of which are synthesized in the pancreas as their precursors - the inactive forms; chymotrypsinogen and trypsinogen - and then are released into the duodenum to cut proteins into smaller peptides. In this paper, the effects of activities of chymotrypsin and trypsin enzymes on fluorescence lifetime distributions of the substrat bovine serum albumin (BSA) modified with N-(1-pyrenyl)maleimide (PM) were examined. In the labeling study of BSA with PM, it is aimed to attach PM to the single free thiol (Cys34) and to all the free amine groups in accessible positions in order to produce excimers of pyrene planes of the possible highest amount to form the lifetime distributions in the widest range, that may show specifically distinguishing changes resulting from the activities of the proteases. The time resolved spectrofluorometer was used to monitor fluorescence decays, which were analyzed by using the exponential series method (ESM) to obtain the changes of lifetime distributions. After the exposure of the synthesized substrat PM-BSA to the enzymes, the fluorescence lifetime distributions exhibited different structures which were attributed to the different activities of the proteases.

  6. Study of fluorescence interaction and conformational changes of bovine serum albumin with histamine H₁ -receptor--drug epinastine hydrochloride by spectroscopic and time-resolved fluorescence methods.

    Science.gov (United States)

    Ariga, Girish G; Naik, Praveen N; Nandibewoor, Sharanappa T; Chimatadar, Shivamurti A

    2015-11-01

    The fluorescence, ultraviolet (UV) absorption, time resolved techniques, circular dichroism (CD), and infrared spectral methods were explored as tools to investigate the interaction between histamine H1 drug, epinastine hydrochloride (EPN), and bovine serum albumin (BSA) under simulated physiological conditions. The experimental results showed that the quenching of the BSA by EPN was static quenching mechanism and also confirmed by lifetime measurements. The value of n close to unity indicated that one molecule of EPN was bound to protein molecule. The binding constants (K) at three different temperatures were calculated (7.1 × 10(4), 5.5 × 10(4), and 3.9 × 10(4) M(-1)). Based on the thermodynamic parameters (ΔH(0), ΔG(0), and ΔS(0)), the nature of binding forces operating between drug and protein was proposed. The site of binding of EPN in the protein was proposed to be Sudlow's site I based on displacement experiments using site markers viz, warfarin, ibuprofen, and digitoxin. Based on the Förster's theory of non-radiation energy transfer, the binding average distance, r between the donor (BSA) and acceptor (EPN) was evaluated and found to be 4.48 nm. The UV-visible, synchronous fluorescence, CD, and three-dimensional fluorescence spectral results revealed the changes in secondary structure of the protein upon its interaction with EPN. © 2015 Wiley Periodicals, Inc.

  7. Viral and Bacterial Pathogens in Bovine Respiratory Disease in Finland

    Directory of Open Access Journals (Sweden)

    Soveri T

    2004-12-01

    Full Text Available Pathogens causing bovine respiratory tract disease in Finland were investigated. Eighteen cattle herds with bovine respiratory disease were included. Five diseased calves from each farm were chosen for closer examination and tracheobronchial lavage. Blood samples were taken from the calves at the time of the investigation and from 86 calves 3–4 weeks later. In addition, 6–10 blood samples from animals of different ages were collected from each herd, resulting in 169 samples. Serum samples were tested for antibodies to bovine parainfluenza virus-3 (PIV-3, bovine respiratory syncytial virus (BRSV, bovine coronavirus (BCV, bovine adenovirus-3 (BAV-3 and bovine adenovirus-7 (BAV-7. About one third of the samples were also tested for antibodies to bovine virus diarrhoea virus (BVDV with negative results. Bacteria were cultured from lavage fluid and in vitro susceptibility to selected antimicrobials was tested. According to serological findings, PIV-3, BAV-7, BAV-3, BCV and BRSV are common pathogens in Finnish cattle with respiratory problems. A titre rise especially for BAV-7 and BAV-3, the dual growth of Mycoplasma dispar and Pasteurella multocida, were typical findings in diseased calves. Pasteurella sp. strains showed no resistance to tested antimicrobials. Mycoplasma bovis and Mannheimia haemolytica were not found.

  8. Kinetics of early in vitro development of bovine in vivo- and in vitro-derived zygotes produced and/or cultured in chemically defined or serum-containing media

    DEFF Research Database (Denmark)

    Holm, P; Booth, P J; Callesen, H

    2002-01-01

    The kinetics of the in vitro development of early embryos from bovine zygotes derived in vitro and in vitro were compared, investigating the effect of serum during in vitro maturation and fertilization (IVM-IVF) and in culture. Zygotes were collected from superovulated heifers or produced in vitro...... from immature oocytes with or without serum supplementation, and cultured subsequently in defined culture medium (SOFaaci) with or without serum supplementation. Time-lapse images were recorded every 0.5 h throughout the culture period. More in vivo- than in vitro-derived zygotes developed...... at first cleavage (defined serum: 20-22% versus in vivo-derived: 8 P zygotes were 1-5 h shorter compared with those of in vitro-derived zygotes cultured under similar conditions (P

  9. Removal of transmissible spongiform encephalopathy prion from large volumes of cell culture media supplemented with fetal bovine serum by using hollow fiber anion-exchange membrane chromatography.

    Science.gov (United States)

    Chou, Ming Li; Bailey, Andy; Avory, Tiffany; Tanimoto, Junji; Burnouf, Thierry

    2015-01-01

    Cases of variant Creutzfeldt-Jakob disease in people who had consumed contaminated meat products from cattle with bovine spongiform encephalopathy emphasize the need for measures aimed at preventing the transmission of the pathogenic prion protein (PrPSc) from materials derived from cattle. Highly stringent scrutiny is required for fetal bovine serum (FBS), a growth-medium supplement used in the production of parenteral vaccines and therapeutic recombinant proteins and in the ex vivo expansion of stem cells for transplantation. One such approach is the implementation of manufacturing steps dedicated to removing PrPSc from materials containing FBS. We evaluated the use of the QyuSpeed D (QSD) adsorbent hollow-fiber anion-exchange chromatographic column (Asahi Kasei Medical, Tokyo, Japan) for the removal of PrPSc from cell culture media supplemented with FBS. We first established that QSD filtration had no adverse effect on the chemical composition of various types of culture media supplemented with 10% FBS or the growth and viability characteristics of human embryonic kidney (HEK293) cells, baby hamster kidney (BHK-21) cells, African green monkey kidney (Vero) cells, and Chinese hamster ovary (CHO-k1) cells propagated in the various culture-medium filtrates. We used a 0.6-mL QSD column for removing PrPSc from up to 1000 mL of Dulbecco's modified Eagle's medium containing 10% FBS previously spiked with the 263K strain of hamster-adapted scrapie. The Western blot analysis, validated alongside an infectivity assay, revealed that the level of PrPSc in the initial 200mL flow-through was reduced by 2.5 to > 3 log10, compared with that of the starting material. These results indicate that QSD filtration removes PrPSc from cell culture media containing 10% FBS, and demonstrate the ease with which QSD filtration can be implemented in at industrial-scale to improve the safety of vaccines, therapeutic recombinant proteins, and ex vivo expanded stem cells produced using growth

  10. Removal of transmissible spongiform encephalopathy prion from large volumes of cell culture media supplemented with fetal bovine serum by using hollow fiber anion-exchange membrane chromatography.

    Directory of Open Access Journals (Sweden)

    Ming Li Chou

    Full Text Available Cases of variant Creutzfeldt-Jakob disease in people who had consumed contaminated meat products from cattle with bovine spongiform encephalopathy emphasize the need for measures aimed at preventing the transmission of the pathogenic prion protein (PrPSc from materials derived from cattle. Highly stringent scrutiny is required for fetal bovine serum (FBS, a growth-medium supplement used in the production of parenteral vaccines and therapeutic recombinant proteins and in the ex vivo expansion of stem cells for transplantation. One such approach is the implementation of manufacturing steps dedicated to removing PrPSc from materials containing FBS. We evaluated the use of the QyuSpeed D (QSD adsorbent hollow-fiber anion-exchange chromatographic column (Asahi Kasei Medical, Tokyo, Japan for the removal of PrPSc from cell culture media supplemented with FBS. We first established that QSD filtration had no adverse effect on the chemical composition of various types of culture media supplemented with 10% FBS or the growth and viability characteristics of human embryonic kidney (HEK293 cells, baby hamster kidney (BHK-21 cells, African green monkey kidney (Vero cells, and Chinese hamster ovary (CHO-k1 cells propagated in the various culture-medium filtrates. We used a 0.6-mL QSD column for removing PrPSc from up to 1000 mL of Dulbecco's modified Eagle's medium containing 10% FBS previously spiked with the 263K strain of hamster-adapted scrapie. The Western blot analysis, validated alongside an infectivity assay, revealed that the level of PrPSc in the initial 200mL flow-through was reduced by 2.5 to > 3 log10, compared with that of the starting material. These results indicate that QSD filtration removes PrPSc from cell culture media containing 10% FBS, and demonstrate the ease with which QSD filtration can be implemented in at industrial-scale to improve the safety of vaccines, therapeutic recombinant proteins, and ex vivo expanded stem cells produced

  11. Validity of a commercial kit for detection of antibodies in bovine serum in an endemic area for fasciolosis

    Directory of Open Access Journals (Sweden)

    Aline Nunes Simões

    Full Text Available Abstract Fasciolosis is caused by Fasciola hepatica that affects the bile ducts and liver parenchyma of ruminants, which can result in economic loss. This study aimed to carry out the validity of the commercial kit ELISA® indirect front of the simple fecal sedimentation test used as the standard. 143 samples were collected blood and feces of cattle from Jerome, south of the Espírito Santo. Serum samples were left at -80 °C and used to perform the ELISA kit IDEXX®. All animals to stool examinations were also positive to the ELISA (22 and negative samples to test stool (121, 52 animals reacted positively against the antibody research. The frequency of fasciolosis was 15.4% in the stool examinations and 51.8% by ELISA. The validity was calculated by sensitivity (100%, specificity (57%, positive predictive value (29% and negative predictive value (100%, and the correlation between the tests calculated using the kappa index of 0.35. The better sensitivity of the ELISA commercial kit should not be a separately evaluated, since the cost benefit and the technical facility must be considered.

  12. Modulation of glycosylation by. cap alpha. -d-tocopherol delivered to hepatoma cell line via human lipoproteins versus bovine serum

    Energy Technology Data Exchange (ETDEWEB)

    Yogeeswaran, G.; Triau, J.E.; Koul, O.

    1987-05-01

    Vitamin E (E) has been shown to cause a generalized reduction in the metabolism and cell surface expression of glycoconjugates (GC) in K3T3 cells. Such changes in K3T3 cells were brought about by E administered via serum (FBS-E). A 6% agarose chromatography of FBS-E demonstrated that E is associated predominantly with albumin, liposomes, and to a small extent with LDL, HDL and micelles. Since the physiological carriers for E are lipoproteins (LP), the authors have examined the relative efficacy of receptor-mediated transfer of E from LP and nonspecific transfer from FBS-E to HepG2 cells. Plasma from 3 human subjects who ingested 1200 IU E/day for 5 days was used for LP-E isolation. During a 48 hrs culture in log-phase, the cells incorporated 3.7 ..mu..g/culture (23%) from FBS-E at 2 ..mu..g/ml, whereas similar amounts were incorporated from LDL-E and HDL-E administered at lower concentrations. E-treated HepG2 cultures via FBS-E methods showed a 39% and 29% reduction in incorporation of (/sup 3/H)-galactose (GAL) and (/sup 3/H)-N-acetylmannosamine (NAM), respectively into cellular and shed GC. Similar reduction in glycosylation of GC was detected by GAL and NAM incorporation in cells treated with a lower amount of LDL-E or HDL-E. These results indicate that E-dependent modulation of glycosylation is more sensitive to LP-E than to FBS-E.

  13. Serum levels of IGF-1 are related to human skin characteristics including the conspicuousness of facial pores.

    Science.gov (United States)

    Sugiyama-Nakagiri, Y; Naoe, A; Ohuchi, A; Kitahara, T

    2011-04-01

    Conspicuous facial pores are one type of serious aesthetic defects for many women. However, the mechanism(s) that underlie the conspicuousness of facial pores remains unclear. We previously characterized the epidermal architecture around facial pores that correlates with the appearance of those pores in various ethnic groups including Japanese. The goal of this study was to evaluate the possible relationships between facial pore size, the severity of impairment of epidermal architecture around facial pores and sebum output levels to investigate the possible role of IGF-1 in the pathogenesis of conspicuous facial pores. The subjects consisted of 38 healthy Japanese women (aged 22-41 years). IGF-1 was measured using immunoradiometric assay. Surface replicas were collected to compare pore sizes of cheek skin and horizontal cross-section images of cheek skin were obtained non-invasively from the same subjects using in vivo confocal laser scanning microscopy and the severity of impairment of epidermal architecture around facial pores was determined. The skin surface lipids of each subject were collected from their cheeks and lipid classes were determined using gas chromatography/flame ionization detection. The serum level of IGF-1 correlated significantly with total pore area (R = 0.36, P facial pores (R = 0.43, P pore area (R = 0.32, P facial skin characteristics including facial pore size and with the severity of impairment of epidermal architecture around facial pores. © 2010 The Authors. Journal compilation © 2010 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  14. In vitro study on binding interaction of quinapril with bovine serum albumin (BSA) using multi-spectroscopic and molecular docking methods.

    Science.gov (United States)

    Shi, Jie-Hua; Pan, Dong-Qi; Jiang, Min; Liu, Ting-Ting; Wang, Qi

    2017-08-01

    The binding interaction between quinapril (QNPL) and bovine serum albumin (BSA) in vitro has been investigated using UV absorption spectroscopy, steady-state fluorescence spectroscopic, synchronous fluorescence spectroscopy, 3D fluorescence spectroscopy, Fourier transform infrared spectroscopy, circular dichroism, and molecular docking methods for obtaining the binding information of QNPL with BSA. The experimental results confirm that the quenching mechanism of the intrinsic fluorescence of BSA induced by QNPL is static quenching based on the decrease in the quenching constants of BSA in the presence of QNPL with the increase in temperature and the quenching rates of BSA larger than 10 10  L mol -1  s -1 , indicating forming QNPL-BSA complex through the intermolecular binding interaction. The binding constant for the QNPL-BSA complex is in the order of 10 5  M -1 , indicating there is stronger binding interaction of QNPL with BSA. The analysis of thermodynamic parameters together with molecular docking study reveal that the main binding forces in the binding process of QNPL with BSA are van der Waal's forces and hydrogen bonding interaction. And, the binding interaction of BSA with QNPL is an enthalpy-driven process. Based on Förster resonance energy transfer, the binding distance between QNPL and BSA is calculated to be 2.76 nm. The results of the competitive binding experiments and molecular docking confirm that QNPL binds to sub-domain IIA (site I) of BSA. It is confirmed there is a slight change in the conformation of BSA after binding QNPL, but BSA still retains its secondary structure α-helicity.

  15. Static and dynamic interaction of a naturally occurring photochromic molecule with bovine serum albumin studied by UV-visible absorption and fluorescence spectroscopy.

    Science.gov (United States)

    Gentili, Pier Luigi; Ortica, Fausto; Favaro, Gianna

    2008-12-25

    In this work, the interaction of a naturally occurring chromene, flindersine (FL), and bovine serum albumin (BSA) has been investigated by UV-vis absorption and fluorescence spectroscopy, time-resolved lifetime measurements, steady state photochemistry, and semiempirical calculations. The interplay of FL with tryptophan (Trp) has been studied in parallel. The interaction of FL with BSA causes fluorescence quenching of BSA through both static and dynamic quenching mechanisms. FL binds BSA with a stoichiometry that varies from 1.09:1 to 0.80:1 as the temperature increases from 293 to 308 K. The reaction is characterized by negative enthalpy (deltaH degrees = -193 kJ mol(-1)) and negative entropy (deltaS degrees = -550 J K(-1) mol(-1)), indicating that the predominant forces in the FL-BSA complex are hydrogen bonding and van der Waals forces. The binding distance between the protein and the photochrome was calculated as 2.5 nm, according to the Foerster theory on resonance energy transfer. The effect of FL concentration on the BSA fluorescence was analyzed according to the maximum entropy method. FL also quenches the emission of Trp with a mechanism that, based on the experimental evidence, excludes both static and dynamic effects. An alternative relaxation pathway, consisting in an electron transfer from a prefluorescent state of Trp to FL, is put forward. The photobehavior of FL is affected by the interplay with BSA but not with Trp. When FL is complexed with BSA, it becomes a more fluorescent and more reactive species. Semiempirical calculations of the lowest optically active electronic transitions of hypothetical FL photoproducts suggest the most likely structure for the photoproduct.

  16. Phenytoin-Bovine Serum Albumin interactions - modeling plasma protein - drug binding: A multi-spectroscopy and in silico-based correlation.

    Science.gov (United States)

    Suresh, P K; Divya, Naik; Nidhi, Shah; Rajasekaran, R

    2018-03-15

    The study focused on the analysis of the nature and site of binding of Phenytoin (PHT) -(a model hydrophobic drug) with Bovine Serum Albumin (BSA) (a model protein used as a surrogate for HSA). Interactions with defined amounts of Phenytoin and BSA demonstrated a blue shift (hypsochromic -change in the microenvironment of the tryptophan residue with decrease in the polar environment and more of hydrophobicity) with respect to the albumin protein and a red shift (bathochromic -hydrophobicity and polarity related changes) in the case of the model hydrophobic drug. This shift, albeit lower in magnitude, has been substantiated by a fairly convincing, Phenytoin-mediated quenching of the endogenous fluorophore in BSA. Spectral shifts studied at varying pH, temperatures and incubation periods (at varying concentrations of PHT with a defined/constant BSA concentration) showed no significant differences (data not shown). FTIR analysis provided evidence of the interaction of PHT with BSA with a stretching vibration of 1737.86cm -1 , apart from the vibrations characteristically associated with the amine and carboxyl groups respectively. Our in vitro findings were extended to molecular docking of BSA with PHT (with the different ionized forms of the drug) and the subsequent LIGPLOT-based analysis. In general, a preponderance of hydrophobic interactions was observed. These hydrophobic interactions corroborate the tryptophan-based spectral shifts and the fluorescence quenching data. These results substantiates our hitherto unreported in vitro/in silico experimental flow and provides a basis for screening other hydrophobic drugs in its class. Copyright © 2017. Published by Elsevier B.V.

  17. Oral Serum-Derived Bovine Immunoglobulin/Protein Isolate Has Immunomodulatory Effects on the Colon of Mice that Spontaneously Develop Colitis.

    Science.gov (United States)

    Pérez-Bosque, Anna; Miró, Lluïsa; Maijó, Mònica; Polo, Javier; Campbell, Joy M; Russell, Louis; Crenshaw, Joe D; Weaver, Eric; Moretó, Miquel

    2016-01-01

    Dietary immunoglobulin concentrates prepared from animal plasma can modulate the immune response of gut-associated lymphoid tissue (GALT). Previous studies have revealed that supplementation with serum-derived bovine immunoglobulin/protein isolate (SBI) ameliorates colonic barrier alterations in the mdr1a-/- genetic mouse model of IBD. Here, we examine the effects of SBI on mucosal inflammation in mdr1a-/- mice that spontaneously develop colitis. Wild type (WT) mice and mice lacking the mdr1a gene (KO) were fed diets supplemented with either SBI (2% w/w) or milk proteins (Control diet), from day 21 (weaning) until day 56. Leucocytes in mesenteric lymph nodes (MLN) and in lamina propria were determined, as was mucosal cytokine production. Neutrophil recruitment and activation in MLN and lamina propria of KO mice were increased, but were significantly reduced in both by SBI supplementation (p stress (p < 0.05) and SBI supplementation reduced this variable (p < 0.05). The Tact/Treg lymphocyte ratios in MLN and lamina propria were also increased in KO animals, but SBI prevented these changes (both p < 0.05). In the colon of KO mice, there was an increased production of mucosal pro-inflammatory cytokines such as IL-2 (2-fold), IL-6 (26-fold) and IL-17 (19-fold), and of chemokines MIP-1β (4.5-fold) and MCP-1 (7.2-fold). These effects were significantly prevented by SBI (p < 0.05). SBI also significantly increased TGF-β secretion in the colon mucosa, suggesting a role of this anti-inflammatory cytokine in the modulation of GALT and the reduction of the severity of the inflammatory response during the onset of colitis.

  18. Study on the Effect of Adding Bovine Serum Albumin to Semen Diluent on the Viability of Awassi Ram Semen Preserved at 5oC

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    Osama Ibrahim Azawi

    2011-10-01

    Full Text Available This study was aimed to test the effect of adding bovine serum albumin (BSA to Tris-fructose-egg yolk diluent to increase the ram sperm storage period at 5ºC. Semen samples from six mature Awassi rams used in this study. Semen samples were diluted by Tris-glucose-egg yolk. Diluted semen sample was divided into two parts. The first part BSA was added 10 mg ml-1 and the second part considered as a control group without any addition. The diluted semen samples were cooled and preserved at 5oC for 5 days. Cooled diluted semen samples were examined for individual motility, percent of live sperm, sperm abnormalities and acrosomal defects every 24 h until 5 days. Results of the present study showed an increased viability of spermatozoa diluted in the Tris diluent containing BSA stored at 5oC for 120 h of preservation compared with the control group. Spermatozoa motility and viability declined gradually in BSA Tris containing diluent when preserved at 5oC from 0 h to 120 h to 54.0±0.9% and 59.0±0.9 %, respectively compared to control of Tris diluent without BSA which was declined markedly to 35.9±1.9 %, 40.9±1.8 %, respectively. In conclusion, this study demonstrated that supplementation with BSA of ram semen diluents during semen preservation at 5oC, may exert beneficial effects on the quality of the Awassi ram semen.

  19. [Investigation on damage of bovine serum albumin (BSA) catalyzed by nano-sized silicon dioxide (SiO2) under ultrasonic irradiation using spectral methods].

    Science.gov (United States)

    Wang, Jun; Ding, Na; Zhang, Zhao-hong; Guo, Ying; Wang, Shi-xian; Xu, Rui; Zhang, Xiang-dong

    2009-04-01

    The damage of bovine serum albumin (BSA) molecules under ultrasonic irradiation in the presence of nano-sized silicon dioxide (SiO2) particles was studied by UV-Vis and fluorescence spectra. In addition, the influences of ultrasonic irradiation time, nano-sized SiO2 addition amount, solution acidity (pH) and ultrasonic irradiation power on the damage of BSA molecules in aqueous solution were also detected. For BSA solution of 1.0 x 10(-5) mol x L(-1) at (37.0+/-0.2) degrees C, the UV-Vis spectra of BSA solutions showed that the absorption peaks of BSA displayed obvious hyperchromic effect with the increase in some influence factors such as ultrasonic irradiation time, nano-sized SiO2 addition amount, pH value and ultrasonic irradiation power. However, the fluorescence spectra of BSA solutions showed the phenomenon of fluorescence quenching with the increase in ultrasonic irradiation time, nano-sized SiO2 addition amount, pH value and ultrasonic irradiation power. Moreover, the possible mechanism behind the damage of BSA molecule in the presence of nano-sized SiO2 powders under ultrasonic irradiation was discussed. It was considered that the damage of BSA molecules was attributed to the formation of *OH radicals resulting from the sonoluminescence and high-heat excitation of ultrasonic cavitation. The research results could be of great significance to using sonocatalytic method to treat tumour in clinic application and for developing nano-sized drug in the future.

  20. Antioxidative effects of magnetized extender containing bovine serum albumin on sperm oxidative stress during long-term liquid preservation of boar semen

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Sang-Hee; Park, Choon-Keun, E-mail: parkck@kangwon.ac.kr

    2015-08-21

    Magnetized water is defined as water that has passed through a magnet and shows increased permeability into cells and electron-donating characteristics. These attributes can protect against membrane damage and remove reactive oxygen species (ROS) in mammalian cells. We explored the effects of improved magnetized semen extenders containing bovine serum albumin (BSA) as antioxidants on apoptosis in boar sperm. Ejaculated semen was diluted in magnetized extender (0G and 6000G) with or without BSA (0G + BSA and 6000G + BSA), and sperm were analyzed based on viability, acrosome reaction, and H{sub 2}O{sub 2} level of live sperm using flow cytometry. Sperm were then preserved for 11 days at 18 °C. We found that viability was significantly higher in 6000G + BSA than under the other treatments (P < 0.05). The acrosome reaction was significantly lower in the 6000G + BSA group compared with the other treatments (P < 0.05). Live sperm with high intracellular H{sub 2}O{sub 2} level were significantly lower in the 6000G + BSA group than under other treatments (P < 0.05). Based on our results, magnetized extenders have antioxidative effects on the liquid preservation of boar sperm. - Highlights: • Magnetized water is water that has been passed through a magnetic field. • Magnetized extender improve viability and decrease oxidative stress of boar sperm for preservation. • Ejaculated semen diluted with magnetized extender can improve liquid preservation period.

  1. 1,3-Bis(2-chloroethyl-1-nitrosourea-loaded bovine serum albumin nanoparticles with dual magnetic resonance–fluorescence imaging for tracking of chemotherapeutic agents

    Directory of Open Access Journals (Sweden)

    Wei KC

    2016-08-01

    Full Text Available Kuo-Chen Wei,1 Feng-Wei Lin,2 Chiung-Yin Huang,1 Chen-Chi M Ma,3 Ju-Yu Chen,1 Li-Ying Feng,1 Hung-Wei Yang2 1Department of Neurosurgery, Chang Gung Memorial Hospital, School of Medicine, Chang Gung University, Taoyuan, 2Institute of Medical Science and Technology, National Sun Yat-sen University, Kaohsiung, 3Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan, Republic of China Abstract: To date, knowing how to identify the location of chemotherapeutic agents in the human body after injection is still a challenge. Therefore, it is urgent to develop a drug delivery system with molecular imaging tracking ability to accurately understand the distribution, location, and concentration of a drug in living organisms. In this study, we developed bovine serum albumin (BSA-based nanoparticles (NPs with dual magnetic resonance (MR and fluorescence imaging modalities (fluorescein isothiocyanate [FITC]-BSA-Gd/1,3-bis(2-chloroethyl-1-nitrosourea [BCNU] NPs to deliver BCNU for inhibition of brain tumor cells (MBR 261-2. These BSA-based NPs are water dispersible, stable, and biocompatible as confirmed by XTT cell viability assay. In vitro phantoms and in vivo MR and fluorescence imaging experiments show that the developed FITC-BSA-Gd/BCNU NPs enable dual MR and fluorescence imaging for monitoring cellular uptake and distribution in tumors. The T1 relaxivity (R1 of FITC-BSA-Gd/BCNU NPs was 3.25 mM-1 s-1, which was similar to that of the commercial T1 contrast agent (R1 =3.36 mM-1 s-1. The results indicate that this multifunctional drug delivery system has potential bioimaging tracking of chemotherapeutic agents ability in vitro and in vivo for cancer therapy. Keywords: drug tracking, fluorescence imaging, MR imaging, BSA nanoparticles, cancer therapy

  2. Oral Serum-Derived Bovine Immunoglobulin/Protein Isolate Has Immunomodulatory Effects on the Colon of Mice that Spontaneously Develop Colitis.

    Directory of Open Access Journals (Sweden)

    Anna Pérez-Bosque

    Full Text Available Dietary immunoglobulin concentrates prepared from animal plasma can modulate the immune response of gut-associated lymphoid tissue (GALT. Previous studies have revealed that supplementation with serum-derived bovine immunoglobulin/protein isolate (SBI ameliorates colonic barrier alterations in the mdr1a-/- genetic mouse model of IBD. Here, we examine the effects of SBI on mucosal inflammation in mdr1a-/- mice that spontaneously develop colitis. Wild type (WT mice and mice lacking the mdr1a gene (KO were fed diets supplemented with either SBI (2% w/w or milk proteins (Control diet, from day 21 (weaning until day 56. Leucocytes in mesenteric lymph nodes (MLN and in lamina propria were determined, as was mucosal cytokine production. Neutrophil recruitment and activation in MLN and lamina propria of KO mice were increased, but were significantly reduced in both by SBI supplementation (p < 0.05. The increased neutrophil recruitment and activation observed in KO mice correlated with increased colon oxidative stress (p < 0.05 and SBI supplementation reduced this variable (p < 0.05. The Tact/Treg lymphocyte ratios in MLN and lamina propria were also increased in KO animals, but SBI prevented these changes (both p < 0.05. In the colon of KO mice, there was an increased production of mucosal pro-inflammatory cytokines such as IL-2 (2-fold, IL-6 (26-fold and IL-17 (19-fold, and of chemokines MIP-1β (4.5-fold and MCP-1 (7.2-fold. These effects were significantly prevented by SBI (p < 0.05. SBI also significantly increased TGF-β secretion in the colon mucosa, suggesting a role of this anti-inflammatory cytokine in the modulation of GALT and the reduction of the severity of the inflammatory response during the onset of colitis.

  3. Binding of antioxidant flavonol morin to the native state of bovine serum albumin: Effects of urea and metal ions on the binding

    Energy Technology Data Exchange (ETDEWEB)

    Singha Roy, Atanu; Dinda, Amit Kumar; Chaudhury, Susmitnarayan; Dasgupta, Swagata, E-mail: swagata@chem.iitkgp.ernet.in

    2014-01-15

    In consideration of the various medicinal aspects of the flavonoid polyphenols, the interaction of morin with bovine serum albumin (BSA) has been investigated using multi-spectroscopic approaches. The pKa{sub 1} of morin being 5.09, which is below physiological pH, binding studies provide important insights into its potential use as a biotherapeutic. The binding was performed under different pH (5, 7 and 9) conditions and in absence and presence of Cu(II) and Fe(III) ions. It is observed that the presence of metal ions affect the binding of morin towards BSA. The binding with BSA results in a motional restriction of morin in solution that causes an increase in anisotropy (r), rotational correlation time (t{sub r}) and steady-state lifetime (t{sub av}) of the ligand. Urea causes denaturation of BSA resulting in the release of morin from the protein core as determined from both the steady-state fluorescence and anisotropy (r) measurements. The possibility of non-radiative energy transfer from the donor tryptophan to the acceptor morin is detected following the Förster's theory. The site marker displacement studies along with the molecular docking results indicated that morin binds to the hydrophobic pocket of site 1 (subdomain IIA) near Trp 213 of BSA. -- Highlights: • Binding mainly occurs through the electrostatic forces with partial hydrophobic association. • Negative ΔG° indicates the spontaneity of the complexation between morin and BSA. • Morin binds near Trp 213 (site 1, subdomain IIA) of BSA only in its native state. • Lifetime of morin increases as a function of BSA. • Motional restriction of morin occurs in the presence of BSA.

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

    Science.gov (United States)

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

    2014-03-01

    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.

  5. Bovine serum albumin-catalyzed deprotonation of [1-(13)C]glycolaldehyde: protein reactivity toward deprotonation of the alpha-hydroxy alpha-carbonyl carbon.

    Science.gov (United States)

    Go, Maybelle K; Malabanan, M Merced; Amyes, Tina L; Richard, John P

    2010-09-07

    Bovine serum albumin (BSA) in D(2)O at 25 degrees C and pD 7.0 was found to catalyze the deuterium exchange reactions of [1-(13)C]glycolaldehyde ([1-(13)C]GA) to form [1-(13)C,2-(2)H]GA and [1-(13)C,2,2-di-(2)H]GA. The formation of [1-(13)C,2-(2)H]GA and [1-(13)C,2,2-di-(2)H]GA in a total yield of 51 +/- 3% was observed at early reaction times, and at later times, [1-(13)C,2-(2)H]GA was found to undergo BSA-catalyzed conversion to [1-(13)C,2,2-di-(2)H]GA. The overall second-order rate constant for these deuterium exchange reactions [(k(E))(P)] equals 0.25 M(-1) s(-1). By comparison, (k(E))(P) values of 0.04 M(-1) s(-1) [Go, M. K., Amyes, T. L., and Richard, J. P. (2009) Biochemistry 48, 5769-5778] and 0.06 M(-1) s(-1) [Go, M. K., Koudelka, A., Amyes, T. L., and Richard, J. P. (2010) Biochemistry 49, 5377-5389] have been determined for the wild-type- and K12G mutant TIM-catalyzed deuterium exchange reactions of [1-(13)C]GA, respectively, to form [1-(13)C,2,2-di-(2)H]GA. These data show that TIM and BSA exhibit a modest catalytic activity toward deprotonation of the alpha-hydroxy alpha-carbonyl carbon. We suggest that this activity is intrinsic to many globular proteins, and that it must be enhanced to demonstrate meaningful de novo design of protein catalysts of proton transfer at alpha-carbonyl carbon.

  6. Interaction of bovine serum albumin and lysozyme with stainless steel studied by time-of-flight secondary ion mass spectrometry and X-ray photoelectron spectroscopy.

    Science.gov (United States)

    Hedberg, Yolanda S; Killian, Manuela S; Blomberg, Eva; Virtanen, Sannakaisa; Schmuki, Patrik; Odnevall Wallinder, Inger

    2012-11-27

    An in-depth mechanistic understanding of the interaction between stainless steel surfaces and proteins is essential from a corrosion and protein-induced metal release perspective when stainless steel is used in surgical implants and in food applications. The interaction between lysozyme (LSZ) from chicken egg white and bovine serum albumin (BSA) and AISI 316L stainless steel surfaces was studied ex situ by means of X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) after different adsorption time periods (0.5, 24, and 168 h). The effect of XPS measurements, storage (aging), sodium dodecyl sulfate (SDS), and elevated temperature (up to 200 °C) on the protein layers, as well as changes in surface oxide composition, were investigated. Both BSA and LSZ adsorption induced an enrichment of chromium in the oxide layer. BSA induced significant changes to the entire oxide, while LSZ only induced a depletion of iron at the utmost layer. SDS was not able to remove preadsorbed proteins completely, despite its high concentration and relatively long treatment time (up to 36.5 h), but induced partial denaturation of the protein coatings. High-temperature treatment (200 °C) and XPS exposure (X-ray irradiation and/or photoelectron emission) induced significant denaturation of both proteins. The heating treatment up to 200 °C removed some proteins, far from all. Amino acid fragment intensities determined from ToF-SIMS are discussed in terms of significant differences with adsorption time, between the proteins, and between freshly adsorbed and aged samples. Stainless steel-protein interactions were shown to be strong and protein-dependent. The findings assist in the understanding of previous studies of metal release and surface changes upon exposure to similar protein solutions.

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

    Science.gov (United States)

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

    2014-03-01

    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.

  8. Evaluation of royal jelly as an alternative to fetal bovine serum in cell culture using cell proliferation assays and live cell imaging.

    Science.gov (United States)

    Musa, Marahaini; Nasir, Nurul Fatihah Mohamad; Thirumulu, Kannan Ponnuraj

    2014-01-01

    Royal jelly is a nutritious substance produced by the young nurse bees and contains significant amounts of proteins which are important for cell growth and proliferation. The aim of this study was to evaluate the effect of royal jelly as an alternative to fetal bovine serum (FBS) in cell culture using cell proliferation assays and live cell imaging. MRC-5 cells were treated with various concentrations of royal jelly extract in MTT assay. The control groups were comprised of Alpha-Minimal Essential Medium (α-MEM) alone and α-MEM with 10% FBS. Subsequently, the cell proliferation was studied for 10 days using Alamar Blue assay and live cell imaging from 48 to 72 h. The population doubling time (PDT) was determined using trypan blue assay after live cell imaging. In MTT assay, 0.156 and 0.078 mg/ml of royal jelly produced higher cell viability compared to positive control group but were not significantly different (P > 0.05). In the Alamar Blue assay, 0.156 and 0.078 mg/ml of royal jelly produced greater percentage of reduction at day 3 even though no significant difference was found (P > 0.05). Based on live cell imaging, the PDT for positive, negative, 0.156 and 0.078 mg/ml of royal jelly groups were 29.09, 62.50, 41.67 and 41.67 h respectively. No significant difference was found in the PDT between all the groups (P > 0.05). Royal jelly does not exhibit similar ability like FBS to facilitate cell growth under the present test conditions.

  9. Authentication of animal origin of heparin and low molecular weight heparin including ovine, porcine and bovine species using 1D NMR spectroscopy and chemometric tools.

    Science.gov (United States)

    Monakhova, Yulia B; Diehl, Bernd W K; Fareed, Jawed

    2018-02-05

    High resolution (600MHz) nuclear magnetic resonance (NMR) spectroscopy is used to distinguish heparin and low-molecular weight heparins (LMWHs) produced from porcine, bovine and ovine mucosal tissues as well as their blends. For multivariate analysis several statistical methods such as principal component analysis (PCA), factor discriminant analysis (FDA), partial least squares - discriminant analysis (PLS-DA), linear discriminant analysis (LDA) were utilized for the modeling of NMR data of more than 100 authentic samples. Heparin and LMWH samples from the independent test set (n=15) were 100% correctly classified according to its animal origin. Moreover, by using 1 H NMR coupled with chemometrics and several batches of bovine heparins from two producers were differentiated. Thus, NMR spectroscopy combined with chemometrics is an efficient tool for simultaneous identification of animal origin and process based manufacturing difference in heparin products. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. The Prognostic Significance of Pretreatment Serum CEA Levels in Gastric Cancer: A Meta-Analysis Including 14651 Patients

    Science.gov (United States)

    Deng, Kai; Yang, Li; Hu, Bing; Wu, Hao; Zhu, Hong; Tang, Chengwei

    2015-01-01

    Background Carcinoembryonic antigen (CEA) is commonly used as a serum tumor marker in clinical practice; however, its prognostic value for gastric cancer patients remains uncertain. This meta-analysis was performed to assess the prognostic value of CEA and investigate CEA as a tumor marker. Methods PubMed, EMBASE and other databases were searched for potentially eligible studies. Forty-one studies reporting the prognostic effect of pretreatment serum CEA expression in gastric cancer patients were selected. Data on 14651 eligible patients were retrieved for the meta-analysis. Based on the data extracted from the available literature, the hazard ratio (HR) and 95% confidence interval (CI) for an adverse prognosis were estimated for gastric cancer patients with elevated pretreatment serum levels of CEA (CEA+) relative to patients with normal pretreatment CEA levels (CEA-). Results The CEA+ patients had a significantly poorer prognosis than the CEA- patients in terms of overall survival (OS: HR 1.716, 95% CI 1.594 - 1.848, P 0.05). In the pooled analyses of multivariate-adjusted HRs, the results suggested that pretreatment serum CEA may be an independent prognostic factor in gastric cancer (OS: HR 1.681, 95% CI 1.425 - 1.982; DSS: HR 1.900, 95% CI 1.441 - 2.505; DFS: HR 2.579, 95% CI 1.935 - 3.436). Conclusion/Significance The meta-analysis based on the available literature supported the association of elevated pretreatment serum CEA levels with a poor prognosis for gastric cancer and a nearly doubled risk of mortality in gastric cancer patients. CEA may be an independent prognostic factor for gastric cancer patients and may aid in determining appropriate treatment which may preferentially benefit the CEA+ patients. PMID:25879931

  11. Degeneration of retinal on bipolar cells induced by serum including autoantibody against TRPM1 in mouse model of paraneoplastic retinopathy.

    Directory of Open Access Journals (Sweden)

    Shinji Ueno

    Full Text Available The paraneoplastic retinopathies (PRs are a group of eye diseases characterized by a sudden and progressive dysfunction of the retina caused by an antibody against a protein in a neoplasm. Evidence has been obtained that the transient receptor potential melastatin 1 (TRPM1 protein was one of the antigens for the autoantibody against the ON bipolar cells in PR patients. However, it has not been determined how the autoantibody causes the dysfunction of the ON bipolar cells. We hypothesized that the antibody against TRPM1 in the serum of patients with PR causes a degeneration of retinal ON bipolar cells. To test this hypothesis, we injected the serum from the PR patient, previously shown to contain anti-TRPM1 antibodies by westerblot, intravitreally into mice and examined the effects on the retina. We found that the electroretinograms (ERGs of the mice were altered acutely after the injection, and the shape of the ERGs resembled that of the patient with PR. Immunohistochemical analysis of the eyes injected with the serum showed immunoreactivity against bipolar cells only in wild-type animals and not in TRPM1 knockout mice,consistent with the serum containing anti-TRPM1 antibodies. Histology also showed that some of the bipolar cells were apoptotic by 5 hours after the injection in wild type mice, but no bipolar cell death was found in TRPM1 knockout mice, . At 3 months, the inner nuclear layer was thinner and the amplitudes of the ERGs were still reduced. These results indicate that the serum of a patient with PR contained an antibody against TRPM1 caused an acute death of retinal ON bipolar cells of mice.

  12. Separation and purification of bovine serum albumin binders from Fructus polygoni orientalis using off-line two-dimensional complexation high-speed counter-current chromatography target-guided by ligand fishing.

    Science.gov (United States)

    Liu, Qi; Shi, Shuyun; Liu, Liangliang; Yang, Hua; Su, Wen; Chen, Xiaoqing

    2013-08-23

    In this study, off-line two-dimensional (2D) complexation high speed counter-current chromatography (HSCCC) was developed for the separation of bovine serum albumin (BSA) binders from the ethyl acetate extract of Fructus polygoni orientalis. Target-guided strategy of BSA functionalized iron oxide magnetic nanoparticles coupled with high performance liquid chromatography-tandem mass spectrometry ((BSA-Fe3O4 MNPs)-HPLC-MS/MS) experiment was proposed. In the orthogonal separation system, a Normal-Phase HSCCC with 0.01mol/L copper ion as complexation agent in the aqueous phase was employed for the first dimension and Recycling HSCCC, Reverse-Phase HSCCC with 0.1mol/L copper ion were used for the second dimension in parallel. Including two pairs of cis-trans isomers, seven BSA binders including 3,5,7-Trihydroxychromone (1), taxifolin (2), N-cis-paprazine (3), N-cis-feruloyltyramine (4), N-trans-paprazine (5), N-trans-feruloyltyramine (6) and an unidentified compound (7) were obtained. The purities of these seven compounds were all over 95.0% as determined by HPLC. The complexation HSCCC behaviors of seven compounds were also investigated by studying their relationship with copper ion. Results showed that the combinative method using (BSA-Fe3O4 MNPs)-HPLC and HSCCC is a quick, efficient, and reproductive technique to isolate potentially bioactive compounds from the complex mixture system of natural products. And the usage of off-line 2D-HSCCC and introduction of chelating metal ion into solvent system are effective ways to implement HSCCC separations in complex samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Custom-made PEI/exfoliated-MoS2nanocomposite ultrafiltration membranes for separation of bovine serum albumin and humic acid.

    Science.gov (United States)

    Saraswathi, Meenakshi Sundaram Sri Abirami; Rana, Dipak; Nagendran, Alagumalai; Alwarappan, Subbiah

    2018-02-01

    Herein, we report the effect of the exfoliated molybdenum disulfide (eMoS 2 ) nanosheets in improving the permeability and anti-fouling properties of the PEI ultra-filtration (UF) membrane using bovine serum albumin (BSA) and humic acid (HA) as model fouling agents. The PEI/eMoS 2 nanocomposite membranes were prepared via phase inversion method using three different eMoS 2 concentrations (0.5, 1 and 2wt%) designated as PEI-0.5, PEI-1 and PEI-2, respectively. Fourier transform infra-red spectroscopy employed to probe the surface functionalities on the membranes. Contact angle measurement, pure water flux, swelling rate and solute rejection studies confirmed the improved hydrophilicity of the PEI/eMoS 2 nanocomposite membranes than the individual entities. Flux recovery ratio (FRR), reversible and irreversible fouling results evidenced the improved fouling resistance of PEI/eMoS 2 modified membranes than the individual counterparts. SEM results evidenced that the nanoscale eMoS 2 significantly altered the membrane morphology by causing increased porosity and larger macrovoids formation on the surface as well as in the bulk of the membrane. PEI-1 membrane showed an increased pure water flux (52.54Lm -2 h -1 ) and water content (74.8%) whereas lesser contact angle (69.2°) and hydraulic resistance (1.85kPa/Lm -2 h -1 ). Resistance to fouling performance of PEI-1 membrane was evident from the FRR values of 95.3 and 90.2% and rejection values of 94.5 and 92.4% for BSA and HA respectively. PEI-2 membrane agglomerates with eMoS 2 and hindered the membrane permeability by blocking the macrovoids in the bulk which restricted the permeation and fouling resistance of the membrane. Amongst various nanocomposite membranes investigated, the PEI-1 membrane exhibited better hydrophilicity and fouling resistance properties due to the availability of the favorable surface and bulk characteristics. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Role of hydrogen-bonding and photoinduced electron transfer (PET) on the interaction of resorcinol based acridinedione dyes with Bovine Serum Albumin (BSA) in water

    Energy Technology Data Exchange (ETDEWEB)

    Kumaran, Rajendran, E-mail: kumaranwau@rediffmail.com [Department of Chemistry, Dwaraka Doss Goverdhan Doss, Vaishnav College (Autonomous), 833, Gokul Bagh, E.V.R. Periyar Road, Arumbakkam, Chennai 600106, Tamil Nadu (India); Vanjinathan, Mahalingam [Department of Chemistry, Dwaraka Doss Goverdhan Doss, Vaishnav College (Autonomous), 833, Gokul Bagh, E.V.R. Periyar Road, Arumbakkam, Chennai 600106, Tamil Nadu (India); Ramamurthy, Perumal [National Centre for Ultrafast Processes, University of Madras, Taramani Campus Chennai 600113, Tamil Nadu (India)

    2015-08-15

    Resorcinol based acridinedione (ADDR) dyes are a class of laser dyes and have structural similarity with purine derivatives, nicotinamide adenine dinucleotide (NADH) analogs. These dyes are classified into photoinduced electron transfer (PET) and non-photoinduced electron transfer dyes, and the photophysical properties of family of these dyes exhibiting PET behavior are entirely different from that of non-PET dyes. The PET process in ADDR dyes is governed by the solvent polarity such that an ADDR dye exhibits PET process through space in an aprotic solvent like acetonitrile and does not exhibit the same in protic solvents like water and methanol. A comparison on the fluorescence emission, lifetime and nature of interaction of various ADDR dyes with a large globular protein like Bovine Serum Albumin (BSA) was carried out in aqueous solution. The interaction of PET based ADDR dyes with BSA in water is found to be largely hydrophobic, but hydrogen-bonding interaction of BSA with dye molecule influences the fluorescence emission of the dye and shifts the emission towards red region. Fluorescence spectral studies reveal that the excited state properties of PET based ADDR dyes are largely influenced by the addition of BSA. The microenvironment around the dye results in significant change in the fluorescence lifetime and emission. Fluorescence enhancement with a red shift in the emission results after the addition of BSA to ADDR dyes containing free amino hydrogen in the 10th position of basic acridinedione dye. The amino hydrogen (N–H) in the 10th position of ADDR dye is replaced by methyl group (N–CH{sub 3}), a significant decrease in the fluorescence intensity with no apparent shift in the emission maximum was observed after the addition of BSA. The nature of interaction between ADDR dyes with BSA is hydrogen-bonding and the dye remains unbound even at the highest concentration of BSA. Circular Dichroism (CD) studies show that the addition of dye to BSA results in

  15. Role of hydrogen-bonding and photoinduced electron transfer (PET) on the interaction of resorcinol based acridinedione dyes with Bovine Serum Albumin (BSA) in water

    International Nuclear Information System (INIS)

    Kumaran, Rajendran; Vanjinathan, Mahalingam; Ramamurthy, Perumal

    2015-01-01

    Resorcinol based acridinedione (ADDR) dyes are a class of laser dyes and have structural similarity with purine derivatives, nicotinamide adenine dinucleotide (NADH) analogs. These dyes are classified into photoinduced electron transfer (PET) and non-photoinduced electron transfer dyes, and the photophysical properties of family of these dyes exhibiting PET behavior are entirely different from that of non-PET dyes. The PET process in ADDR dyes is governed by the solvent polarity such that an ADDR dye exhibits PET process through space in an aprotic solvent like acetonitrile and does not exhibit the same in protic solvents like water and methanol. A comparison on the fluorescence emission, lifetime and nature of interaction of various ADDR dyes with a large globular protein like Bovine Serum Albumin (BSA) was carried out in aqueous solution. The interaction of PET based ADDR dyes with BSA in water is found to be largely hydrophobic, but hydrogen-bonding interaction of BSA with dye molecule influences the fluorescence emission of the dye and shifts the emission towards red region. Fluorescence spectral studies reveal that the excited state properties of PET based ADDR dyes are largely influenced by the addition of BSA. The microenvironment around the dye results in significant change in the fluorescence lifetime and emission. Fluorescence enhancement with a red shift in the emission results after the addition of BSA to ADDR dyes containing free amino hydrogen in the 10th position of basic acridinedione dye. The amino hydrogen (N–H) in the 10th position of ADDR dye is replaced by methyl group (N–CH 3 ), a significant decrease in the fluorescence intensity with no apparent shift in the emission maximum was observed after the addition of BSA. The nature of interaction between ADDR dyes with BSA is hydrogen-bonding and the dye remains unbound even at the highest concentration of BSA. Circular Dichroism (CD) studies show that the addition of dye to BSA results in a

  16. Effects of Gold Salt Speciation and Structure of Human and Bovine Serum Albumins on the Synthesis and Stability of Gold Nanostructures.

    Science.gov (United States)

    Miranda, Érica G A; Tofanello, Aryane; Brito, Adrianne M M; Lopes, David M; Albuquerque, Lindomar J C; de Castro, Carlos E; Costa, Fanny N; Giacomelli, Fernando C; Ferreira, Fabio F; Araújo-Chaves, Juliana C; Nantes, Iseli L

    2016-01-01

    The present study aimed to investigate the influence of albumin structure and gold speciation on the synthesis of gold nanoparticles (GNPs). The strategy of synthesis was the addition of HAuCl4 solutions at different pH values (3-12) to solutions of human and bovine serum albumins (HSA and BSA) at the same corresponding pH values. Different pH values influence the GNP synthesis due to gold speciation. Besides the inherent effect of pH on the native structure of albumins, the use N-ethylmaleimide (NEM)-treated and heat-denaturated forms of HSA and BSA provided additional insights about the influence of protein structure, net charge, and thiol group approachability on the GNP synthesis. NEM treatment, heating, and the extreme values of pH promoted loss of the native albumin structure. The formation of GNPs indicated by the appearance of surface plasmon resonance (SPR) bands became detectable from 15 days of the synthesis processes that were carried out with native, NEM-treated and heat-denaturated forms of HSA and BSA, exclusively at pH 6 and 7. After 2 months of incubation, SPR band was also detected for all synthesis carried out at pH 8.0. The mean values of the hydrodynamic radius (RH) were 24 and 34 nm for GNPs synthesized with native HSA and BSA, respectively. X-ray diffraction (XRD) revealed crystallites of 13 nm. RH, XRD, and zeta potential values were consistent with GNP capping by the albumins. However, the GNPs produced with NEM-treated and heat-denaturated albumins exhibited loss of protein capping by lowering the ionic strength. This result suggests a significant contribution of non-electrostatic interactions of albumins with the GNP surface, in these conditions. The denaturation of proteins exposes hydrophobic groups to the solvent, and these groups could interact with the gold surface. In these conditions, the thiol blockage or oxidation, the latter probably favored upon heating, impaired the formation of a stable capping by thiol coordination with the

  17. Can serums be replaced by Mueller‑Hinton agar in germ tube test?

    African Journals Online (AJOL)

    2016-03-03

    Mar 3, 2016 ... various serums (i.e., human, rabbit, horse, and fetal bovine serum) used in the GTT and Mueller‑Hinton agar (MHA). Materials and Methods: Fifty species isolated from various clinical samples that were defined as C. albicans by both conventional and DNA sequence analysis methods were included in the ...

  18. Pasteurella multocida and bovine respiratory disease.

    Science.gov (United States)

    Dabo, S M; Taylor, J D; Confer, A W

    2007-12-01

    Pasteurella multocida is a pathogenic Gram-negative bacterium that has been classified into three subspecies, five capsular serogroups and 16 serotypes. P. multocida serogroup A isolates are bovine nasopharyngeal commensals, bovine pathogens and common isolates from bovine respiratory disease (BRD), both enzootic calf pneumonia of young dairy calves and shipping fever of weaned, stressed beef cattle. P. multocida A:3 is the most common serotype isolated from BRD, and these isolates have limited heterogeneity based on outer membrane protein (OMP) profiles and ribotyping. Development of P. multocida-induced pneumonia is associated with environmental and stress factors such as shipping, co-mingling, and overcrowding as well as concurrent or predisposing viral or bacterial infections. Lung lesions consist of an acute to subacute bronchopneumonia that may or may not have an associated pleuritis. Numerous virulence or potential virulence factors have been described for bovine respiratory isolates including adherence and colonization factors, iron-regulated and acquisition proteins, extracellular enzymes such as neuraminidase, lipopolysaccharide, polysaccharide capsule and a variety of OMPs. Immunity of cattle against respiratory pasteurellosis is poorly understood; however, high serum antibodies to OMPs appear to be important for enhancing resistance to the bacterium. Currently available P. multocida vaccines for use in cattle are predominately traditional bacterins and a live streptomycin-dependent mutant. The field efficacy of these vaccines is not well documented in the literature.

  19. Bovine serum albumin-Cu(II) hybrid nanoflowers: An effective adsorbent for solid phase extraction and slurry sampling flame atomic absorption spectrometric analysis of cadmium and lead in water, hair, food and cigarette samples

    Energy Technology Data Exchange (ETDEWEB)

    Yilmaz, Erkan [Department of Chemistry, Faculty of Sciences, Erciyes University, Kayseri 38039 (Turkey); Ocsoy, Ismail [Department of Analytical Chemistry, Faculty of Pharmacy, Erciyes University, Kayseri 38039 (Turkey); Nanotechnology Research Center (ERNAM), Erciyes University, Kayseri 38039 (Turkey); Ozdemir, Nalan [Department of Chemistry, Faculty of Sciences, Erciyes University, Kayseri 38039 (Turkey); Soylak, Mustafa, E-mail: soylak@erciyes.edu.tr [Department of Chemistry, Faculty of Sciences, Erciyes University, Kayseri 38039 (Turkey)

    2016-02-04

    Herein, the synthesis of bovine serum albumin-Cu(II) hybrid nanoflowers (BSA-NFs) through the building blocks of bovine serum albumin (BSA) and copper(II) ions in phosphate buffered saline (PBS) and their use as adsorbent for cadmium and lead ions are reported. The BSA-NFs, for the first time, were efficiently utilized as novel adsorbent for solid phase extraction (SPE) of cadmium and lead ions in water, food, cigarette and hair samples. The method is based on the separation and pre-concentration of Cd(II) and Pb(II) by BSA-NFs prior to determination by slurry analysis via flame atomic absorption spectrometry (FAAS). The analytes were adsorbed on BSA-NFs under the vortex mixing and then the ion-loaded slurry was separated and directly introduced into the flame AAS nebulizer by using a hand-made micro sample introduction system to eliminate a number of drawbacks. The effects of analytical key parameters, such as pH, amount of BSA-NFs, vortexing time, sample volume, and matrix effect of foreign ions on adsorbing of Cd(II) and Pb(II) were systematically investigated and optimized. The limits of detection (LODs) for Cd(II) and Pb(II) were calculated as 0.37 μg L{sup −1} and 8.8 μg L{sup −1}, respectively. The relative standard deviation percentages (RSDs) (N = 5) for Cd(II) and Pb(II) were 7.2%, and 5.0%, respectively. The accuracy of the developed procedure was validated by the analysis of certified reference materials (TMDA-53.3 Fortified Water, TMDA-70 Fortified Water, SPS-WW2 Waste Water, NCSDC-73349 Bush Branches and Leaves) and by addition/recovery analysis. The quantitative recoveries were obtained for the analysis of certified reference materials and addition/recovery tests. The method was successfully applied to the analysis of cadmium and lead in water, food, cigarette and hair samples. - Highlights: • The synthesis of bovine serum albumin-Cu(II) hybrid nanoflowers is reported. • The nanoflowers were utilized for solid phase microextraction of

  20. Seroprevalence of viral and bacterial diseases among the bovines in Himachal Pradesh, India

    OpenAIRE

    Shailja Katoch; Shweta Dohru; Mandeep Sharma; Vikram Vashist; Rajesh Chahota; Prasenjit Dhar; Aneesh Thakur; Subhash Verma

    2017-01-01

    Aim: The study was designed to measure the seroprevalence of viral and bacterial diseases: Infectious bovine rhinotracheitis, bovine viral diarrhea, bovine leukemia, bovine parainfluenza, bovine respiratory syncytial disease, brucellosis, and paratuberculosis among bovine of Himachal Pradesh during the year 2013-2015. Materials and Methods: The serum samples were collected from seven districts of state, namely, Bilaspur, Kangra, Kinnaur, Lahul and Spiti, Mandi, Sirmour, and Solan. The sam...

  1. Purification, subunit characterization and ultrastructure of three soluble bovine lectins: conglutinin, mannose-binding protein and the pentraxin serum amyloid P-component

    DEFF Research Database (Denmark)

    Andersen, Ove; Friis, P; Holm Nielsen, E

    1992-01-01

    affinity chromatography and selective elution was developed. The purification was monitored by SDS-PAGE, Western blotting and electron microscopy. Binding of the lectins to Sephadex-iC3b, their collagenase sensitivity, and the size and antibody reactivity of their subunits was investigated....... The demonstration, by SDS-PAGE, of 25-kDa subunits, which were unaffected by collagenase treatment but bound to Sephadex-iC3b and antibodies to human SAP, indicated the existence of bovine SAP. Bovine conglutinin (BK) also showed calcium-dependent binding to Sephadex-iC3b, whereas bovine MBP did not. The binding...... of BK was inhibitable with GlcNAc. A 3000-fold increase in BK activity (ELISA) was obtained in eluates from Sephadex-iC3b. SDS-PAGE analyses of BK and MBP revealed subunits with an Mr of 43 kDa and 30 kDa, respectively. These subunits were sensitive to collagenase treatment which reduced the Mr to 20 k...

  2. Synthesis and characterizations of ultra-small ZnS and Zn(1-x)Fe(x)S quantum dots in aqueous media and spectroscopic study of their interactions with bovine serum albumin.

    Science.gov (United States)

    Khani, Omid; Rajabi, Hamid Reza; Yousefi, Mohammad Hasan; Khosravi, Ali Azam; Jannesari, Mohammad; Shamsipur, Mojtaba

    2011-07-01

    This work reports a new experimental methodology for the synthesis of ultra small zinc sulfide and iron doped zinc sulfide quantum dots in aqueous media. The nanoparticles were obtained using a simple procedure based on the precipitation of ZnS in aqueous solution in the presence of 2-mercaptoethanol as a capping agent, at room temperature. The effect of Fe(3+) ion concentration as dopant on the optical properties of ZnS was studied. The size of quantum dots was determined to be about 1nm, using scanning tunneling microscopy. The synthesized nanoparticles were characterized by X-ray diffraction, UV-Vis absorption and photoluminescence emission spectroscopies. The presence and amount of iron impurity in the structure of Zn((1-x))Fe(x)S nanocrystals were confirmed by atomic absorption spectrometry. A blue shift in band-gap of ZnS was observed upon increasing incorporation of Fe(3+) ion in the iron doped zinc sulfide quantum dots. The photoluminescence investigations showed that, in the case of iron doped ZnS nanoparticles, the emission band of pure ZnS nanoparticles at 427nm shifts to 442nm with appearance of a new sharp emission band around 532nm. The X-ray diffraction analysis indicated that the iron doped nanoparticles are crystalline, with cubic zinc blend structure, having particle diameters of 1.7±022nm. Finally, the interaction of the synthesized nanoparticles with bovine serum albumin was investigated at pH 7.2. The UV-Vis absorption and fluorescence spectroscopic methods were applied to compare the optical properties of pure and iron doped ZnS quantum dots upon interaction with BSA. It was proved that, in both cases, the fluorescence quenching of BSA by the quantum dots is mainly a result of the formation of QDs-BSA complex in solution. In the steady-state fluorescence studies, the interaction parameters including binding constants (K(a)), number of binding sites (n), quenching constants ( [Formula: see text] ), and bimolecular quenching rate constants (k

  3. Anti-bovine antibody in human sera as a cause of nonspecificity in enzyme immunoassay.

    OpenAIRE

    Dise, T; Brunell, P A

    1987-01-01

    Much nonspecific activity in enzyme immunoassay is due to reactions between anti-bovine antibodies which are commonly present in human serum samples and residual calf serum in viral antigens. Calf serum is used in cultured cells which are used to produce viral antigens. Addition of bovine proteins to serum samples to be tested for viral antibodies reduces this activity.

  4. Synthesis and characterization of mononuclear copper(II) complex of tetradentate N2S2 donor set and the study of DNA and bovine serum albumin binding

    OpenAIRE

    Sandipan Sarkar; Biswajit Das

    2014-01-01

    One mononuclear copper(II) complex, containing neutral tetradentate NSSN-type ligands, of formulation [Cu II(L 1)Cl]ClO 4 (1), was synthesized and isolated in pure form [where L 1˭ 1,3-bis(3-pyridylmethylthio)propane]. Green-colored copper(II) complex was characterized by physicochemical, spectroscopic methods and conductivity measurement. These experimental data matched well with the proposed structure of the complex. Biological activity of the complex (1) toward calf thymus DNA and bovine s...

  5. Serum concentrations of haptoglobin and haptoglobin-matrix metalloproteinase 9 (Hp-MMP 9) complexes of bovine calves in a bacterial respiratory challenge model.

    Science.gov (United States)

    Hanthorn, Christy J; Dewell, Grant A; Dewell, Renee D; Cooper, Vickie L; Wang, Chong; Plummer, Paul J; Lakritz, Jeffrey

    2014-12-06

    Serum haptoglobin (Hp) and haptoglobin matrix metalloproteinase 9 complexes (Hp-MMP 9) have been identified as biomarkers with diagnostic potential in cattle with conditions resulting in an acute inflammatory response. The purpose of this study was to evaluate potential diagnostic applications of serum Hp and Hp-MMP 9 concentrations in calves with BRD and establish a timeline for their detection in calves experimentally challenged with Bibersteinia trehalosi and Mannheimia haemolytica. Thirty-five cross bred dairy calves were inoculated via tracheal catheterization with either a PCR confirmed leukotoxin negative B. trehalosi isolate, a PCR confirmed leukotoxin positive B. trehalosi isolate, a Mannheimia haemolytica isolate, a combination of leukotoxin negative B. trehalosi and M. haemolytica, or a negative control. Serum samples were collected throughout the study. Calves were euthanized and necropsy performed on day 10 post inoculation. M. haemolytica inoculated calves had increased lung involvement. Serum Hp and Hp- MMP 9 concentrations were elevated compared to the other treatment groups. Increases in serum Hp and Hp-MMP 9 concentrations for the M. haemolytica group were significantly different from other study groups on day 7 of the study. B. trehalosi inoculated calves did not have increased lung involvement compared to control calves, but the leukotoxin positive B. trehalosi group demonstrated increased serum Hp-MMP 9 concentrations from day 3 to the end of the study compared to the pre-inoculation concentrations. Serum Hp-MMP 9 concentration is a useful diagnostic tool for detecting early pulmonary inflammation in calves challenged with B. trehalosi and M. haemolytica. Serum Hp-MMP 9 may also be a useful tool in detecting subclinical pulmonary inflammation in challenged calves.

  6. Potential Anticarcinogenic Peptides from Bovine Milk

    Directory of Open Access Journals (Sweden)

    Giacomo Pepe

    2013-01-01

    Full Text Available Bovine milk possesses a protein system constituted by two major families of proteins: caseins (insoluble and whey proteins (soluble. Caseins (αS1, αS2, β, and κ are the predominant phosphoproteins in the milk of ruminants, accounting for about 80% of total protein, while the whey proteins, representing approximately 20% of milk protein fraction, include β-lactoglobulin, α-lactalbumin, immunoglobulins, bovine serum albumin, bovine lactoferrin, and lactoperoxidase, together with other minor components. Different bioactivities have been associated with these proteins. In many cases, caseins and whey proteins act as precursors of bioactive peptides that are released, in the body, by enzymatic proteolysis during gastrointestinal digestion or during food processing. The biologically active peptides are of particular interest in food science and nutrition because they have been shown to play physiological roles, including opioid-like features, as well as immunomodulant, antihypertensive, antimicrobial, antiviral, and antioxidant activities. In recent years, research has focused its attention on the ability of these molecules to provide a prevention against the development of cancer. This paper presents an overview of antitumor activity of caseins and whey proteins and derived peptides.

  7. Revised description and classification of atypical isolates of Pasteurella multocida from bovine lungs based on genotypic characterization to include variants previously classified as biovar 2 of Pasteurella canis and Pasteurella avium

    DEFF Research Database (Denmark)

    Christensen, Henrik; Angen, Øystein; Olsen, John E.

    2004-01-01

    Strains deviating in key phenotypic characters, mainly isolated from cases of bovine pneumonia in five European countries, were genotyped in order to examine their genotypic relationship with Pasteurella multocida. Twenty-two strains of Pasteurella avium biovar 2, including variants in indole......, xylose and mannitol, 18 strains of Pasteurella canis biovar 2 and variants of this taxon, five strains of P multocida subsp. septica showing variations in indole and ornithine decarboxylase, nine strains of P. multocida subsp. multocida showing variation in ornithine decarboxylase and mannitol, and type...... strains of the subspecies of P. multocida were included. Ribotyping was used to examine the relationship of the strains, and 13 types, each containing between one and 20 isolates, were observed. Identical ribotypes; were observed in some cases for P. avium biovar 2 and either P. canis biovar 2 or P...

  8. Effect of ambient temperature on viral replication and serum antibody titers following administration of a commercial intranasal modified-live infectious bovine rhinotracheitis-parainfluenza-3 virus vaccine to beef cattle housed in high- and moderate-ambient temperature environments.

    Science.gov (United States)

    Grissett, Gretchen P; White, Brad J; Anderson, David E; Larson, Robert E; Miesner, Matt D

    2014-12-01

    To evaluate the effect of ambient temperature on viral replication and serum antibody titers following administration of an intranasal modified-live infectious bovine rhinotracheitis (IBR)-parainfluenza-3 (PI3) virus vaccine to beef calves housed in high- (> 32°C) and moderate- (21°C) ambient temperature environments. 28 calves (mean weight, 206.8 kg). Calves were randomly allocated to 4 treatment groups (housed outdoors during high ambient temperature with [HAT; n = 10] or without [HAC; 4] vaccination or housed indoors in a moderate ambient temperature with [MAT; 10] or without [MAC; 4] vaccination). Rectal and nasal mucosal temperatures were recorded every 2 hours from 8 AM to 8 PM on days 0 (vaccination) and 1. Nasal swab specimens were obtained on days 0 through 7 for virus isolation. Serum samples were collected on days 0, 7, 14, and 28 for determination of antibody titers. Mean rectal temperature did not differ among the treatment groups. Mean nasal temperature for the HAT group was significantly higher than that for the MAT group at 6, 24, 30, 32, and 38 hours after vaccination. Viable IBR virus was isolated from all vaccinated calves on days 1 through 6. Two weeks after vaccination, vaccinated calves had anti-IBR antibody titers that were significantly greater than those for unvaccinated calves. Mean anti-IBR antibody titers did not differ significantly between the HAT and MAT groups. Results indicated that, following vaccination with an intranasal modified-live IBR-PI3 virus vaccine, IBR viral replication and serum antibody titers did not differ significantly between calves housed in high- and moderate-ambient temperature environments.

  9. Development of antiserum against bovine prolactin for radioimmunoassay

    International Nuclear Information System (INIS)

    Razdan, M.N.; Kaker, M.L.; Galhotra, M.M.

    1980-01-01

    Antiserum against bovine prolactin was raised in male rabbits and tested against crossreacting contaminants by immunodiffusion and immunoelectrophoresis. One sharp precipitin band against prolactin was observed. The antiserum showed a small faint band against bovine growth hormone. No crossreaction was observed against bovine follicle stimulating hormone, bovine luteinizing hormone, bovine thyroid stimulating hormone and normal calf serum. The crossreaction with bovine growth hormone vanished after treating the antiserum with normal calf serum. Antibody titre of the absorbed anti-prolactin serum determined by radioimmunoassay was high, which could bind 31.30% at 1:10,000 final dilution. Maximum binding was not inhibited by the addition of higher amounts of contaminants, suggesting that the absorbed antiserum was immunologically monospecific and could be used for radioimmunoassay work. (author)

  10. Bovine serum albumin-Cu(II) hybrid nanoflowers: An effective adsorbent for solid phase extraction and slurry sampling flame atomic absorption spectrometric analysis of cadmium and lead in water, hair, food and cigarette samples.

    Science.gov (United States)

    Yilmaz, Erkan; Ocsoy, Ismail; Ozdemir, Nalan; Soylak, Mustafa

    2016-02-04

    Herein, the synthesis of bovine serum albumin-Cu(II) hybrid nanoflowers (BSA-NFs) through the building blocks of bovine serum albumin (BSA) and copper(II) ions in phosphate buffered saline (PBS) and their use as adsorbent for cadmium and lead ions are reported. The BSA-NFs, for the first time, were efficiently utilized as novel adsorbent for solid phase extraction (SPE) of cadmium and lead ions in water, food, cigarette and hair samples. The method is based on the separation and pre-concentration of Cd(II) and Pb(II) by BSA-NFs prior to determination by slurry analysis via flame atomic absorption spectrometry (FAAS). The analytes were adsorbed on BSA-NFs under the vortex mixing and then the ion-loaded slurry was separated and directly introduced into the flame AAS nebulizer by using a hand-made micro sample introduction system to eliminate a number of drawbacks. The effects of analytical key parameters, such as pH, amount of BSA-NFs, vortexing time, sample volume, and matrix effect of foreign ions on adsorbing of Cd(II) and Pb(II) were systematically investigated and optimized. The limits of detection (LODs) for Cd(II) and Pb(II) were calculated as 0.37 μg L(-)(1) and 8.8 μg L(-)(1), respectively. The relative standard deviation percentages (RSDs) (N = 5) for Cd(II) and Pb(II) were 7.2%, and 5.0%, respectively. The accuracy of the developed procedure was validated by the analysis of certified reference materials (TMDA-53.3 Fortified Water, TMDA-70 Fortified Water, SPS-WW2 Waste Water, NCSDC-73349 Bush Branches and Leaves) and by addition/recovery analysis. The quantitative recoveries were obtained for the analysis of certified reference materials and addition/recovery tests. The method was successfully applied to the analysis of cadmium and lead in water, food, cigarette and hair samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Bovine Mastitis: Frontiers in Immunogenetics

    Science.gov (United States)

    Thompson-Crispi, Kathleen; Atalla, Heba; Miglior, Filippo; Mallard, Bonnie A.

    2014-01-01

    Mastitis is one of the most prevalent and costly diseases in the dairy industry with losses attributable to reduced milk production, discarded milk, early culling, veterinary services, and labor costs. Typically, mastitis is an inflammation of the mammary gland most often, but not limited to, bacterial infection, and is characterized by the movement of leukocytes and serum proteins from the blood to the site of infection. It contributes to compromised milk quality and the potential spread of antimicrobial resistance if antibiotic treatment is not astutely applied. Despite the implementation of management practises and genetic selection approaches, bovine mastitis control continues to be inadequate. However, some novel genetic strategies have recently been demonstrated to reduce mastitis incidence by taking advantage of a cow’s natural ability to make appropriate immune responses against invading pathogens. Specifically, dairy cattle with enhanced and balanced immune responses have a lower occurrence of disease, including mastitis, and they can be identified and selected for using the high immune response (HIR) technology. Enhanced immune responsiveness is also associated with improved response to vaccination, increased milk, and colostrum quality. Since immunity is an important fitness trait, beneficial associations with longevity and reproduction are also often noted. This review highlights the genetic regulation of the bovine immune system and its vital contributions to disease resistance. Genetic selection approaches currently used in the dairy industry to reduce the incidence of disease are reviewed, including the HIR technology, genomics to improve disease resistance or immune response, as well as the Immunity+™ sire line. Improving the overall immune responsiveness of cattle is expected to provide superior disease resistance, increasing animal welfare and food quality while maintaining favorable production levels to feed a growing population. PMID

  12. Spectroscopic analyses on interaction of o-Vanillin- D-Phenylalanine, o-Vanillin- L-Tyrosine and o-Vanillin- L-Levodopa Schiff Bases with bovine serum albumin (BSA)

    Science.gov (United States)

    Gao, Jingqun; Guo, Yuwei; Wang, Jun; Wang, Zhiqiu; Jin, Xudong; Cheng, Chunping; Li, Ying; Li, Kai

    2011-04-01

    In this work, three o-Vanillin Schiff Bases (o-VSB: o-Vanillin- D-Phenylalanine (o-VDP), o-Vanillin- L-Tyrosine (o-VLT) and o-Vanillin- L-Levodopa (o-VLL)) with alanine constituent were synthesized by direct reflux method in ethanol solution, and then were used to study the interaction to bovine serum albumin (BSA) molecules by fluorescence spectroscopy. Based on the fluorescence quenching calculation, the bimolecular quenching constant ( Kq), apparent quenching constant ( Ksv), effective binding constant ( KA) and corresponding dissociation constant ( KD) as well as binding site number ( n) were obtained. In addition, the binding distance ( r) was also calculated according to Foster's non-radioactive energy transfer theory. The results show that these three o-VSB can efficiently bind to BSA molecules, but the binding array order is o-VDP-BSA > o-VLT-BSA > o-VLL-BSA. Synchronous fluorescence spectroscopy indicates that the o-VDP is more accessibility to tryptophan (Trp) residues of BSA molecules than to tyrosine (Tyr) residues. Nevertheless, the o-VLT and o-VLL are more accessibility to Tyr residues than to Trp residues.

  13. 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.

    Science.gov (United States)

    Mizuno, Masayasu; Pikal, Michael J

    2013-10-01

    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 "α-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. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. The studies of density, apparent molar volume, and viscosity of bovine serum albumin, egg albumin, and lysozyme in aqueous and RbI, CsI, and DTAB aqueous solutions at 303.15 K.

    Science.gov (United States)

    Singh, Man; Chand, Hema; Gupta, K C

    2005-06-01

    Density (rho), apparent molar volume (V(phi)), and viscosity (eta) of 0.0010 to 0.0018% (w/v) of bovine serum albumin (BSA), egg albumin, and lysozyme in 0.0002, 0.0004, and 0.0008 M aqueous RbI and CsI, and (dodecyl)(trimethyl)ammonium bromide (DTAB) solutions were obtained. The experimental data were regressed against composition, and constants are used to elucidate the conformational changes in protein molecules. With salt concentration, the density of proteins is found to decrease, and the order of the effect of additives on density is observed as CsI > RbI > DTAB. The trend of apparent molar volume of proteins is found as BSA > egg-albumin > lysozyme for three additives. In general, eta values of BSA remain higher for all compositions of RbI than that of egg-albumin for CsI and DTAB. These orders of the data indicate the strength of intermolecular forces between proteins and salts, and are helpful for understanding the denaturation of proteins.

  15. Nanocomposite materials based on poly(vinyl chloride) and bovine serum albumin modified ZnO through ultrasonic irradiation as a green technique: Optical, thermal, mechanical and morphological properties.

    Science.gov (United States)

    Mallakpour, Shadpour; Darvishzadeh, Marzieh

    2018-03-01

    In this project, physicochemical properties of poly(vinyl chloride) (PVC) reinforced by ZnO nanoparticles (NPs) were studied. Firstly, ZnO NPs were modified with bovine serum albumin (BSA) as an organo-modifier and biocompatible substance through ultrasound irradiation as environmental friendly, low cost and rapid means. Nanocomposite (NC) films were prepared by loadings of various ratios of ZnO/BSA NPs (3, 6 and 9wt%) inside the PVC. Structural morphology and physical properties of the ZnO-BSA NPs and NC films were investigated via Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis (TGA), transmission electron microscopy and field emission scanning electron microscopy. According to the obtained information from the TGA, an increase in the thermal stability can be clearly observed. Also the results of contact angle analysis indicated with increasing percent of ZnO/BSA NPs into PVC the hydrophilic behaviors of NCs were increased. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Synthesis and Biophysical Insights into the Binding of a Potent Anti-Proliferative Non-symmetric Bis-isatin Derivative with Bovine Serum Albumin: Spectroscopic and Molecular Docking Approaches

    Directory of Open Access Journals (Sweden)

    Ali Saber Abdelhameed

    2017-06-01

    Full Text Available As part of the research endeavors to combat cancer, a non-symmetric bis-isatin derivative (compound 3 was synthesized and showed a significant anti-proliferative potency. The current study provides a comprehensive characterization of the interaction of compound 3 with the drug-transporting protein bovine serum albumin (BSA via the use of spectroscopic tools along with molecular docking studies. Fluorescence spectral measurements showed that the BSA intrinsic fluorescence can be significantly quenched by the addition of compound 3 and the formation of a non-fluorescent complex. Further measurements revealed a static type of quenching with Stern–Volmer and Linweaver–Burk constants of 105. The thermodynamic parameters of the binding were calculated to be ΔS° 105.09 ± 5.32 with ΔH° of −0.72 ± 0.71 and negative ΔG° values. In addition, synchronous fluorescence and 3D fluorescence spectroscopy suggested that compound 3 did not induce conformational changes in BSA. Site competition experiments revealed that compound 3 competes with warfarin within the BSA binding domain (Sudlow site I. This was further confirmed by the molecular docking results showing a binding energy of −25.93 kJ/mol for compound 3-BSA. Hence, the observed results in the present study assumed that the compound 3-BSA binding is spontaneous, involving electrostatic forces and hydrogen bonding.

  17. Radioimmunoassay of bovine growth hormone

    International Nuclear Information System (INIS)

    Dvorak, P.; Becka, S.; Krejci, P.; Chrpova, M.

    1978-01-01

    A radioimmunological method for quantitative determination of bovine growth hormone in blood plasma or serum for routine use was worked out. The antibody is also suitable for radioimmunoassay of bovine growth hormone. The chloramin T and lactoperoxidase methods were used. Standard b-STH isolated by one of the authors was labelled with I-125(Na 125 I, carrier free, Amersham). The I-125-hormone was separated on Sephadex G-50 or G-100 and stored in 0.05M phosphate pH 7.5 with 1% of human serum albumine at 25 deg C. The method was checked on 2500 samples. Rabbit antiserum to b-STH was prepared in sufficient amount to serve also for other laboratories interested in the method. (author)

  18. A probiotic mixture including galactooligosaccharides decreases fecal β-glucosidase activity but does not affect serum enterolactone concentration in men during a two-week intervention.

    Science.gov (United States)

    Kekkonen, Riina A; Holma, Reetta; Hatakka, Katja; Suomalainen, Tarja; Poussa, Tuija; Adlercreutz, Herman; Korpela, Riitta

    2011-05-01

    A high serum concentration of enterolactone, an enterolignan produced by colonic microbiota from precursors in cereals, vegetables, and fruits, is associated with reduced risk of acute coronary events. Probiotics and prebiotics modify colonic metabolism and may affect the serum enterolactone concentration. The effects of a probiotic mixture alone and with galactooligosaccharides (GOS) on serum enterolactone concentration and fecal metabolism were investigated in 18 healthy men. Participants received 3 interventions, each for 2 wk: 1) probiotics [Lactobacillus rhamnosus strains GG (LGG) and LC705, Propionibacterium freudenreichii ssp. shermanii JS, and Bifidobacterium breve Bb99, for a total amount of 2 × 10(10) CFU/d]; 2) probiotics and GOS 3.8 g/d; 3) probiotics, GOS, and rye bread (minimum 120 g/d). Serum enterolactone and fecal dry weight, enzyme activities, pH, SCFA, lactic acid bacteria, bifidobacteria, propionibacteria, and the strains LGG and LC705 were determined. The serum enterolactone concentration (nmol/L) tended to be decreased from baseline [mean (95% CI) 18.6 (10.8-26.4)] by probiotics alone [15.2 (7.8-22.7); P = 0.095], was not significantly affected by probiotics with GOS [21.5 (13.2-29.8)], and was increased by probiotics with GOS and rye bread [24.6 (15.4-33.7); P < 0.05]. Probiotics alone did not affect fecal β-glucosidase activity and bifidobacteria, but probiotics with GOS decreased β-glucosidase activity and increased bifidobacteria compared with baseline (P < 0.05) and with probiotics alone (P < 0.01). In conclusion, this probiotic mixture with or without GOS does not significantly affect serum enterolactone concentration. Because probiotics with GOS decreased fecal β-glucosidase activity but not serum enterolactone, the reduced fecal β-glucosidase, within the range of activities measured, does not seem to limit the formation of enterolactone.

  19. DETERMINATION OF SERUM ALBUMIN WITH ...

    African Journals Online (AJOL)

    The reaction of tribromoarsenazo(TB-ASA) with serum albumin in the presence of emulgent OP was studied by spectrophotometry. In a Britton-Robinson buffer solution at pH 2.9, tribromoarsenazo and bovine serum albumin can immediately form a red compound in the presence of emulgent OP with a maximum absorption ...

  20. Serum neutralization with different types and subtypes of bovine herpesvirus 1 and 5 Soroneutralização com diferentes tipos e subtipos de herpesvírus bovinos 1 e 5

    Directory of Open Access Journals (Sweden)

    Carine Lidiane Holz

    2010-07-01

    Full Text Available The serum neutralization (SN test is the gold standard method to measure neutralizing antibodies to bovine herpesviruses. However, in view of the further subdivisions of bovine herpesviruses in types/subtypes, defining which virus to use at challenge in SN tests may be difficult. In view of that, this study was carried out to re-evaluate (SN sensitivity with different types/subtypes of bovine herpesviruses types 1 (BoHV-1 and 5 (BoHV-5 as challenge viruses. Bovine sera (n=810 were collected from two distinct geographic regions and tested by SN with three type 1 viruses (BoHV-1.1 strains "Los Angeles" and "EVI123/98"; BoHV-1.2a strain "SV265/96" and three type 5 viruses (BoHV-5a strain "EVI88/95"; BoHV-5b strain "A663" and BoHV-5c "ISO97/95". SN tests were performed with a 1 hour incubation of the serum-virus mixtures at 37ºC against 100 TCID50 of each of the viruses. SN sensitivity varied greatly depending on the challenge virus used in the test. The highest sensitivity (327 positive/810 total sera tested; 40.37% was attained when the positive results to the six viruses were added together. No association could be found between any particular type or subtype of virus and the sensitivity of the test. When positive results to each single strain were considered, SN sensitivity varied from 41.7% to 81.7%, depending on the virus and the geographic region of origin of the sera. Variation was detected even when challenge viruses belonged to the same subtype, where disagreement between positive results reached 41%. These results indicate that one hour incubation SN tests against single viruses, as performed here, may display a significantly low sensitivity (p=0.05; performing SN tests against a number of different viruses may increase considerably SN sensitivity. Furthermore, the choice of virus used for challenge is critical in SN tests. In addition, sera from different geographic regions may give rise to disagreeing results with different strains of

  1. Detection of lipomannan in cattle infected with bovine tuberculosis

    Science.gov (United States)

    Early and rapid detection of bovine tuberculosis (bTB) is critical to controlling the spread of this disease in cattle and other animals. In this study, we demonstrate the development of an immunoassay for the direct detection of the bovine bTB biomarker, lipomannan (LM) in serum using a waveguide-...

  2. 9 CFR 113.216 - Bovine Rhinotracheitis Vaccine, Killed Virus.

    Science.gov (United States)

    2010-01-01

    ... Virus. 113.216 Section 113.216 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Killed Virus Vaccines § 113.216 Bovine Rhinotracheitis Vaccine, Killed Virus. Infectious Bovine...

  3. Spectroscopic and molecular docking studies on the charge transfer complex of bovine serum albumin with quinone in aqueous medium and its influence on the ligand binding property of the protein

    Science.gov (United States)

    Satheshkumar, Angupillai; Elango, Kuppanagounder P.

    2014-09-01

    The spectral techniques such as UV-Vis, 1H NMR and fluorescence and electrochemical experiments have been employed to investigate the interaction between 2-methoxy-3,5,6-trichloro-1,4-benzoquinone (MQ; a water soluble quinone) and bovine serum albumin (BSA) in aqueous medium. The fluorescence of BSA was quenched by MQ via formation of a 1:1 BSA-MQ charge transfer adduct with a formation constant of 3.3 × 108 L mol-1. Based on the Forster’s theory the binding distance between them is calculated as 2.65 nm indicating high probability of binding. For the first time, influence of quinone on the binding property of various types of ligands such as aspirin, ascorbic acid, nicotinimide and sodium stearate has also been investigated. The results indicated that the strong and spontaneous binding existing between BSA and MQ, decreased the intensity of binding of these ligands with BSA. Since Tryptophan (Trp) is the basic residue present in BSA, a comparison between binding property of Trp-MQ adduct with that of BSA-MQ with these ligands has also been attempted. 1H NMR titration study indicated that the Trp forms a charge transfer complex with MQ, which reduces the interaction of Trp with the ligands. Molecular docking study supported the fact that the quinone interacts with the Trp212 unit of the BSA and the free energy change of binding (ΔG) for the BSA-MQ complex was found to be -46 kJ mol-1, which is comparable to our experimental free energy of binding (-49 kJ mol-1) obtained from fluorescence study.

  4. LPS from bovine serum albumin drives TNF-α release during ex-vivo placenta perfusion experiments, contaminates the perfusion system but can be effectively removed by oxidative cleaning.

    Science.gov (United States)

    Vasanthan, T; Rochow, N; Mian, F; Codini, T; DeFrance, B; Fusch, G; Samiee-Zafarghandy, S; Fusch, C

    2014-12-01

    The dual ex-vivo perfusion of human placental tissue is useful to study inflammatory pathways. We found significant TNF-α release in negative controls similar in concentration to lipopolysaccharide (LPS) stimulated placentas. The aim of the current study was to (i) identify sources driving TNF-α release and (ii) develop an approach to control for it. (i) To determine sources leading to TNF-α release, solutions frequently circulated through the perfusion system and perfusion media with different bovine serum albumin (BSA) quality were exposed to mouse macrophage cell lines (RAW264.7) and subsequently measured for TNF-α expression. (ii) To assess memory effects and validate cleaning procedures, sham perfusion experiments were conducted either in the presence or absence of exogenous LPS, in new tubing that was contaminated, cleaned and analyzed for the effectiveness of LPS removal. Oxidative and acid-base cleaning were tested for their effectiveness to reduce LPS contamination. TNF-α release, observed in negative control experiments, was attributed to the use of LPS-contaminated BSA as well as inadequate cleaning of the perfusion system. Once introduced in the perfusion system, LPS accumulated and created a memory effect. Oxidative but not acid-base depyrogenation effectively reduced LPS levels to concentrations that were in accordance with FDA guidelines (placenta perfusion model could have confounding effects on experimental outcomes leading to misinterpretation of data. To circumvent LPS contamination LPS-free BSA and oxidative depyrogenation cleaning techniques should be implemented in future placental perfusion studies. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Solvent-dependent binding interactions of the organophosphate pesticide, chlorpyrifos (CPF), and its metabolite, 3,5,6-trichloro-2-pyridinol (TCPy), with Bovine Serum Albumin (BSA): A comparative fluorescence quenching analysis.

    Science.gov (United States)

    Dahiya, Vandana; Chaubey, Bhawna; Dhaharwal, Ashok K; Pal, Samanwita

    2017-06-01

    Analysis of the interaction of pesticides and their metabolites with the cellular proteins has drawn considerable attention in past several years to understand the effect of pesticides on environment and mankind. In this study, we have investigated the binding interaction of Bovine Serum Albumin (BSA) with a widely used organophosphorous insecticide chlorpyrifos (CPF), and its stable metabolite, 3,5,6-trichloro-2-pyridinol (TCPy) to provide a comparative analysis of the two molecules by employing various spectroscopic techniques viz., UV-vis absorption, Circular Dichroism (CD), and Fluorescence spectroscopy. The fluorescence quenching studies of BSA emission in two different solvents viz., water and methanol in presence of CPF and TCPy have led to the revelation of several interesting facts about the pesticide-protein interaction. It has been found that both the molecules cause static quenching of BSA emission as seen from the Stern-Volmer constant (K sv ) irrespective of the solvent used for the analysis. While TCPy is a stronger quencher in water, it exhibits comparable quenching capacity with CPF in methanol. The solvent dependent differential binding interaction of the two molecules finally indicates possibility of diverse bio-distribution of the pesticides within human body. The UV-vis and CD spectra of BSA in presence of the test molecules have unravelled that the molecules formed ground state complex that are highly reversible in nature and have minimal effect on the protein secondary structure. Furthermore it is also understood that structural changes of BSA in presence of CPF is significantly higher compared to that in presence of TCPY. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. A novel and facile synthesis of carbon quantum dots via salep hydrothermal treatment as the silver nanoparticles support: Application to electroanalytical determination of H2O2 in fetal bovine serum.

    Science.gov (United States)

    Jahanbakhshi, Mojtaba; Habibi, Biuck

    2016-07-15

    A simple, low-cost, and green process was used for the synthesis of carbon quantum dots (CQDs) through the hydrothermal treatment of salep as a novel bio-polymeric carbon source in presence of only pure water. The silver nanoparticles (AgNPs) were embedded on the surface of CQDs by ultra-violate (UV) irradiation to the CQDs and silver nitrate mixture solution. The as-synthesized CQDs and AgNPs decorated CQDs nanohybrid (AgNPs/CQDs) were characterized by UV-vis and photoluminescence spectroscopy, Fourier transform-infrared spectroscopy, transmission electron microscopy, atomic force microcopy, X-ray diffraction, and field emission scanning electron microscopy. Then, the AgNPs/CQDs nanohybrid was casted on the glassy carbon electrode in order to prepare an amperometric hydrogen peroxide (H2O2) sensor. The electrochemical investigations show that the AgNPs/CQDs nanohybrid possesses an excellent performance toward the H2O2 reduction. In the optimum condition, the linear range of H2O2 determination was achieved from 0.2 to 27.0μM with high sensitivity (1.5μA/µM) and the limit of detection was obtained about 80nM (S/N=3). Finally, the prepared nanohybrid modified electrode was effectively applied to the H2O2 detection in the disinfected fetal bovine serum samples, and the recovery was obtained about 98%. The achieved results indicate that the AgNPs/CQDs nanohybrid with high reproducibility, repeatability, and stability has a favorable capability in electrochemical sensors improvement. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Antiviral effects of bovine interferons on bovine respiratory tract viruses.

    OpenAIRE

    Fulton, R W; Downing, M M; Cummins, J M

    1984-01-01

    The antiviral effects of bovine interferons on the replication of bovine respiratory tract viruses were studied. Bovine turbinate monolayer cultures were treated with bovine interferons and challenged with several bovine herpesvirus 1 strains, bovine viral diarrhea virus, parainfluenza type 3 virus, goat respiratory syncytial virus, bovine respiratory syncytial virus, bovine adenovirus type 7, or vesicular stomatitis virus. Treatment with bovine interferons reduced viral yield for each of the...

  8. Seroprevalence of viral and bacterial diseases among the bovines in Himachal Pradesh, India

    Directory of Open Access Journals (Sweden)

    Shailja Katoch

    2017-12-01

    Full Text Available Aim: The study was designed to measure the seroprevalence of viral and bacterial diseases: Infectious bovine rhinotracheitis, bovine viral diarrhea, bovine leukemia, bovine parainfluenza, bovine respiratory syncytial disease, brucellosis, and paratuberculosis among bovine of Himachal Pradesh during the year 2013-2015. Materials and Methods: The serum samples were collected from seven districts of state, namely, Bilaspur, Kangra, Kinnaur, Lahul and Spiti, Mandi, Sirmour, and Solan. The samples were screened using indirect ELISA kits to measure the seroprevalence of viral and bacterial diseases. Results: The overall seroprevalence of infectious bovine rhinotracheitis was 24.24%, bovine viral diarrhea 1.52%, bovine leukemia 9.09%, bovine parainfluenza 57.58%, bovine respiratory syncytial disease 50%, brucellosis 19.69%, and paratuberculosis 9.09% in Himachal Pradesh. The seroprevalence of bovine rhinotracheitis, bovine leukemia, bovine parainfluenza, bovine respiratory syncytial disease, and paratuberculosis in the state varied significantly (p0.01. Multiple seropositivity has been observed in this study. Bovine parainfluenza virus 3 was observed commonly in mixed infection with almost all viruses and bacteria under study. Conclusion: The viral and bacterial diseases are prevalent in the seven districts of Himachal Pradesh investigated in the study. The