• Title/Summary/Keyword: bovine hemoglobin

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Thermodynamic Elucidation of Binding Isotherms for Hemoglobin & Globin of Human and Bovine upon Interaction with Dodecyl Trimethyl Ammonium Bromide

  • Bordbar, A.K.;Nasehzadeh, A.;Ajloo, D.;Omidiyan, K.;Naghibi, H.;Mehrabi, M.;Khajehpour, H.;Rezaei-Tavirani, M.;Moosavi-Movahedi, A.A.
    • Bulletin of the Korean Chemical Society
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    • v.23 no.8
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    • pp.1073-1077
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    • 2002
  • Binding of dodecyl trimethylammonium bromide (DTAB) to human and bovine hemoglobin and globin samples has been investigated in 50 mM glycine buffer pH = 10, I = 0.0318 and 300 K by equilibrium dialysis and temperature scanning spectrophotometry techniques and method for calculation of average hydrophobicity. The binding data has been analyzed, in terms of binding capacity concept $({\theta})$, Hill coefficient (nH) and intrinsic Gibbs free energy of binding $({\Delta}Gbv).$ The results of binding data, melting point (Tm) and average hydrophobicity show that human hemoglobin has more structural stability than bovine hemoglobin sample. Moreover the results of binding data analysis represent the systems with two and one sets of binding sites for hemoglobin and globin, respectively. It seems that the destabilization of hemoglobin structure due to removal of heme group, is responsible of such behavior. The results indicating the removal of heme group from hemoglobin caused the depletion of first binding set as an electrostatic site upon interaction with DTAB and exposing the hydrophobic patches for protein.

Electrochemical Investigation of Bovine Hemoglobin at an Acetylene Black Paste Electrode in the Presence of Sodium Dodecyl Sulfate

  • Zhan, Guoqing;Li, Chunya;Luo, Dengbai
    • Bulletin of the Korean Chemical Society
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    • v.28 no.10
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    • pp.1720-1724
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    • 2007
  • Electrochemical behaviors of bovine hemoglobin (Hb) at an acetylene black paste electrode based on the enhancement effect of sodium dodecyl sulfate (SDS) were investigated. In the optimal conditions, a very weak reduction peak was observed at an acetylene black paste electrode for hemoglobin in the absence of SDS. However, the reduction peak current increased remarkably after the addition of 4.0 × 10?4 mol L?1 SDS, suggesting that SDS exhibits obvious enhancement effect to the determination of hemoglobin. All the experimental parameters, such as pH value, concentration of SDS, accumulation time and accumulation potential were optimized for hemoglobin analysis. The proposed method possesses high sensitivity (detection limit is 3.0 × 10?9 mol L?1), wide linearity (6.0 × 10?9 to 6.0 × 10?7 mol L?1), rapid response and low cost. Finally, the method was successfully employed to determine hemoglobin in a spiked sample.

Characteristics of Synthesized Red Cells Using Bovine Hemoglobin as Oxygen Carrier

  • Cho, Eng-Haeng;Hah, Jong-Sik;Kim, Ku-Ja
    • The Korean Journal of Physiology
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    • v.26 no.1
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    • pp.37-44
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    • 1992
  • The encapsulation of the Purified bovine hemoglobin with Phospholipids obtained from egg folk was performed using a rotary vacuum evaporator. The prepared Hb-containing liposome (hemosome) had good properties as artificial red blood cell. The shape and size of the hemosomes were measured by a phase contrast microscope and image analyzer. The function of the hemosome as oxygen carrier was tested by measuring oxygen saturation curve with blood gas analyzer and infusing it into rats. The prepared hemosome was 1.184 + 0.423 ${\mu}m$ in diameter and round in shape. $P_{50}$ value of the hemosome solution was 28 mmHg. The synthesized red cells seem to function as oxygen carrier, because the severely bled rats were prolonged in their life by transfusion of the hemosome solution containing bovine hemoglobin.

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Separation and flux characteristics in cross-flow ultrafiltration of bovine serum albumin and bovine hemoglobin solutions

  • Hsiao, Ruey-Chang;Hung, Chia-Lin;Lin, Su-Hsia;Juang, Ruey-Shin
    • Membrane and Water Treatment
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    • v.2 no.2
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    • pp.91-103
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    • 2011
  • The flux behavior in the separation of equimolar bovine serum albumin (BSA) and bovine hemoglobin (HB) in aqueous solutions by cross-flow ultrafiltration (UF) was investigated, in which polyacylonitrile membrane with a molecular weight cut-off (MWCO) of 100 kDa was used. BSA and HB have comparable molar mass (67,000 vs. 68,000) but different isoelectric points (4.7 vs. 7.1). The effects of process variables including solution pH (6.5, 7.1, and 7.5), total protein concentration (1.48 and 7.40 ${\mu}M$), transmembrane pressure (69, 207, and 345 kPa), and solution ionic strength (with or without 0.01 M NaCl) on the separation were examined. It was shown that the ionic strength had a negligible effect on separation performance under the conditions studied. Although BSA and HB are not rigid bodies, the flux decline in the present cross-flow UF did not result from the mechanism of cake filtration with compression. In this regard, the specific cake resistance when pseudo steady-state was reached was evaluated and discussed.

Subtilisin QK, a Fibrinolytic Enzyme, Inhibits the Exogenous Nitrite and Hydrogen Peroxide Induced Protein Nitration, inVitro and inVivo

  • Ko, Ju-Ho;Yan, Junpeng;Zhu, Lei;Qi, Yipeng
    • BMB Reports
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    • v.38 no.5
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    • pp.577-583
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    • 2005
  • Subtilisin QK, which is newly identified as a fibrinolytic enzyme from Bacillus subtilis QK02, has the ability of preventing nitrotyrosine formation in bovine serum albumin induced by nitrite, hydrogen peroxide and hemoglobin in vitro verified by ELISA, Western-blot and spectrophotometer assay. Subtilisin QK also attenuates the fluorescence emission spectra of bovine serum albumin in the course of oxidation caused by nitrite, hydrogen peroxide and hemoglobin. Furthermore, subtilisin QK could suppress the transformation of oxy-hemoglobin to met-hemoglobin caused by sodium nitrite, but not the heat-treated subtilisn QK. Compared with some other fibrinolytic enzymes and inactivated subtilisin QK treated by phenylmethylsulfonylfluoride, the ability of inhibiting met-hemoglobin formation of subtilisin QK reveals that the anti-oxidative ability of subtilisin QK is not concerned with its fibrinolytic function. Additionally, nitrotyrosine formation in proteins from brain, heart, liver, kidney, and muscle of mice that is intramuscular injected the mixture of nitrite, hydrogen peroxide and hemoglobin is attenuated by subtilisin QK. Subtilisin QK can also protect Human umbilical vein endothelial cell (ECV-304) from the damage caused by nitrite and hydrogen peroxide.

QCM biosensor for the determination of haptoglobin (헵토글로빈 측정을 위한 수정미소저울 바이오센서 개발)

  • Kim, Sung-Il;Lee, Hyeon-Jeong;Park, Soon-Hyuk;Bhang, Hyun-Soo;Lee, Jin-Ah;Choi, Suk-Jung
    • Journal of Sensor Science and Technology
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    • v.16 no.2
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    • pp.132-141
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    • 2007
  • The purpose this research is to develop QCM (quartz crystal microbalance) biosensor for the determination of haptoglobin. Haptoglobin is an acute-phase protein with a hemoglobin-binding activity and has a potential to be used as a biomarker for infection or cancer. Haptoglobin level in milk has been used for the diagnosis of cow mastitis. In this study, anti-bovine haptoglobin antibody or bovine hemoglobin was chemically immobilized on the surface of the QCM, and the resulting sensor chips were tested for their response to samples containing haptoglobin at different concentrations. Concentration dependent frequency change was observed with both of the sensor chips. Especially, the sensor chip containing anti-bovine haptoglobin antibody showed sufficient sensitivity in the concentrations typically observed in the cows with mastitis.

Pharmacokinetics of PEG-Hemoglobin SB1, a Hemoglobin-Based Oxygen Carrier, after Its Intravenous Administration in Beagle Dogs

  • Kwon, Oh-Seung;Chung, Uoo-Tae;Chung, Youn-Bok
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.259-264
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    • 2004
  • The purpose of the present study was to investigate the pharmacokinetics of PEG-hemoglobin SB 1, a modified bovine hemoglobin with polyethylene glycol, after its single and multiple administration in beagle dogs. For this purpose, the analytical method of free hemoglobin in the plasma was developed and validated. Excellent linearity ($r^2$=0.999) was observed in the calibration curve data, with the limit of quantification of 0.005 g/dL. The precision and the deviation of the theoretical values for accuracy were always within $\pm$15% in both the between-and the within-day results. The method was tested by measuring the plasma concentrations following intravenous administration to beagle dogs and was shown to be suitable for pharmacokinetic studies. In a single dose study, the plasma half-life (t$_{1}$2/) increased and the total body clearance (Cl$_{t}$) decreased with the dose (i.e., 0.017 to 0.75 gHb/kg as PEG-hemoglobin SB1) in both sexes. The volume of distribution at steady-state (Vd$_{ss}$ ) showed no difference with the dose. In contrast, the values of t$_{1}$2/, CL$_{t}$ and the area under the plasma concentration-time curve (AUC) after the multiple dose were significantly different from those of the single dose administration. The values of t$_{1}$2/ in the multiple administration were about two times higher-than that of the single dose. As a result, t$_{1}$2/ of hemoglobin after the administration of PEG-hemoglobin SB1 was about 15-30 h, indicating the PEG modification of the hemoglobin lead to a prolongation of plasma concentration of the protein. Therefore, these observations suggested that the PEG modification of hemoglobin is potentially applicable in the hemoglobin-based therapeutics.tics.

Inhibitory Effects of Total Extract and Flavonols from Hardy Rubber Tree (Eucommia ulmoides Oliver) Leaves on the Glycation of Hemoglobin

  • Kim, Hye-Young;Kim, Kyong;Lee, Myung-Ki
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.603-605
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    • 2006
  • Our previous study demonstrated that aqueous ethanolic extract of hardy rubber tree (HRT; Eucommia ulmoides Oliver) leaves potently inhibited glycation, and that two known flavonols and one new flavonol were the active compounds in protein glycation in vitro using the model system of bovine serum albumin and fructose. The aim of this study was to examine the effectiveness of HRT extract and its flavonols against the glycation of hemoglobin (Hb) in primary cultured rat erythrocytes. The extract at $40-400\;{\mu}g/mL$ and the flavonols at $0.2-20\;{\mu}g/mL$ significantly inhibited Hb glycation in a concentration-dependent manner. They were more effective than aminoguanidine, a well-known inhibitor of glycation. Three flavonols seemed to be responsible for the inhibitory activity of the extract, furthermore, the extract of HRT leaves showed advantages over single isolated flavonols in the inhibition of Hb glycation.

Effects of Nitric Oxide Scavenger and Inhibitor on the Development of Bovine IVM/IVF Embryos (Nitric Oxide 화합물 첨가가 소 체외수정란의 체외발육에 미치는 효과)

  • Jang H. Y.;Kim J. T.;Park C. K.;Cheong H. T.;Kim C. I.;Yang B. K.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.161-166
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    • 2004
  • This study was designed to evaluate the effects of nitric oxide scavenger (hemoglobin) and inhibitor (L-nitro-L-arginine methyl ester; L-NAME) with or without cumulus cell on the development of bovine IVM/IVF embryos. When CR/sub 1aa/ medium were supplemented with different dosage (lug/m, 5ug/m and 10ug/ml) of hemoglobin at 48hrs for in vitro culture, the proportion of embryos developing beyond morulae stage in 0, 1ug/ml and 5ug/ml with or without cumulus cell were 23.8%, 33.3 % and 26.8%, and 39.5%, 54.8% and 48.8%, respectively. There was a significantly difference the developmental rate of 1ug/ml hemoglobin intact cumulus cells to any other groups (P<0.05). On the other hand, when added to hemoglobin at 96 hrs, 1 ug/ml hemoglobin with cumulus cell group was significantly increased the percentage of developing into morulae and blastocysts to any other groups (P<0.05), and similar trend that of added at 48hrs. The overall means of the percentage of developing into morulae and blstocysts in 1ug/ml hemoglobin group was significantly increased than those of any other groups (P<0.05) and cumulus co-culture with hemoglobin was increased the in vitro developing rate of IVM/IVF embryos. In CR/sub 1aa/ medium treated with L-NAME 0, 10, 50 and 100mM, the developmental rate of morula plus blastocysts were 55.6%, 64.9%, 58.8% and 66.7%, respectively. The L-NAME did not affect the developmental rate and the cell numbers of blastocysts in all treated groups. These results indicate that hemoglobin and cumulus co-culture can increase the proportion of embryos that developed into morulae and blastocysts, but cell numbers of blastocysts were not affect in all groups.