• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.63-69
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• 2001

This study was carried out to examine the effects of nitric oxide compounds (hemoglobin and L-NAME) on the development of porcine in vitro maturation (IVM) and in vitro fertilization (IVF) oocytes. Cumulus cell free embryos derived from porcine IVM/IVF oocytes were cultured in NCSU23 medium containing 1~5 $\mu\textrm{g}$/$m\ell$ hemoglobin added to 44 and 96hrs in culture times, and in NCSU23 medium containing 0, 10, 50 or 100mM L-NAME. The developmental rates beyond morulae stage in 0, 1 and 5 $\mu\textrm{g}$/$m\ell$ hemoglobin groups add to 44hrs in vitro culture times were 52.4%, 57.6% and 57.4%, respectively. The addition of hemoglobin groups made it slightly higher than the control group. The proportion of embryos developed to morulae and blastocysts in 1 $\mu\textrm{g}$/$m\ell$ hemoglobin add to 96hrs after in vitro culture (70.8%) was a little higher than those of 0 and 5 $\mu\textrm{g}$/$m\ell$ hemoglobin (66.2% and 62.8%). There was no significant difference in all groups (P〉0.05). The developmental rates beyond morulae stage in 0, 10, 50 and 100mM of L-NAME groups add to 96hrs after in vitro culture were 65.2%, 73.5%, 70.1% and 53.3%, respectively 10mM and 50mM L-NAME groups were significantly higher than in 0 and 100mM of L-NAME groups (P＜0.05). In conclusions, these results indicate that L-NAME (10mM, 50mM) can increase the proportion of embryos that develop into morulae and blastocysts but hemoglobin did not affect.

• 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.

• BMB Reports
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• v.35 no.4
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• pp.364-370
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• 2002

The effect of sodium n-dodecyl sulphate (SDS) on hemoglobin autoxidation was studied in the presence of a 100mM phosphate buffer (pH 7.0) by different methods. These included spectorphotometry, fluorescence technique, cyclic voltametry, differential scanning calorimetry, and densitometry. Spectroscopic studies showed that SDS concentrations up to 1 mM increased deoxy-, decreases oxy-, and had no significant effect on the met- conformation of hemoglobin. Therefore, a SDS concentration up to 1 mM increased the deoxy form of hemoglobin as the folded, compact state and decreases the oxy conformation. The turbidity measurements and differential scanning calorimetry techniques indicated a more stable conformation for hemoglobin in the presence of SDS up to 1mM. Electrochemical studies also confirmed a more difficult oxidation under these conditions. The induction of the deoxy form in the presence of SDS was confirmed by densitometry techniques. The compact structure of deoxyhemoglobin blocks the formation of met-conformation in low SDS concentrations.

• BMB Reports
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• v.29 no.3
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• pp.261-265
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• 1996

Sheep hemoglobin (SHb) was modified with methoxy-polyethylene glycol (mPEG) to develop a potential blood substitute. mPEG has been used to decrease antigenicity and immunogenicity of foreign proteins. When the mPEG was attached to SHb, the modified hemoglobins showed decreased electrophoretic mobility on SDS-PAGE and decreased free amino groups. When the remaining free amino groups of mPEG modified SHb were determined by TNBS free amino group titration methods. about 34% of total free amino groups were modified with mPEG. This mPEG-SHb conjugate of 34% amino groups modified showed no precipitation by double immunodiffusion with polyclonal antibodies against SHb. This modified hemoglobin still has oxygen transport activity. So this antigenicity decreased hemoglobin may be used in humans as a potential blood substitute.

• Analytical Science and Technology
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• v.36 no.6
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• pp.332-339
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• 2023

This study aimed to verify the hypothesis that urea denatures hemoglobin in the blood, thereby exposing active sites of enzymes and enhancing the chemiluminescence of the blood-luminol reaction. When blood was pretreated with urea, higher concentrations of pretreatment urea or longer pretreatment times resulted in enhanced chemiluminescence in the blood-luminol reaction, supporting the above hypothesis. However, the chemiluminescence was enhanced when blood was treated with luminol mixed with an 8 M urea solution, although the fact that the time for urea to denature hemoglobin was shorter compared to when blood was pretreated with urea and followed by luminol. In addition, the chemiluminescence was enhanced when a transition metal without hemoglobin was reacted with urea-containing luminol. Based on these results, it is anticipated that urea not only denatures hemoglobin but also plays a role in the luminol-hydrogen peroxide reaction.

• Journal of Environmental Health Sciences
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• v.32 no.2 s.89
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• pp.164-170
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• 2006

Ethylene oxide is a genotoxic carcinogen with widespread uses as industrial chemical intermediate and gaseous sterilant. 2-hydroxyethylated N-terminal valine in Hb is a good biomarker for biological monitoring of ethylene oxide exposure, because of its stability. For measuring the hemoglobin adduct formed by exposure of ethylene oxide, we studied the determination of (N-2-hydroxy-ethyl)valine(HEV) in hemoglobin adduct by using GC/MS. Firstly we synthesized HEV with 2-amino-ethanol and bromoisovaleric acid(BIVA) and confirmed it with GC/MS-FID. Its fragmentations were m/z 116(base ion, M+-45) and m/z 130(M+-31). For measuring HEV with higher sensitivity, we use derivatives which were PFPITH(pentafluorophenylisothiocianate) and TBDMS (tributyldimethylsilylation) by using Edman procedure. Its fragmentation were m/z 425(M+-57), m/z 383(M+-99) and m/z 172(M+-310) by using GC/MS. We did biological monitoring for mice inhalation exposure with 400 ppm ethylene oxide. The concentrations of hemoglobin adduct were $168{\pm}3.8\;and\;512{\pm}04$(nmol g-1 globin) at 0.5 hr/day 400 ppm ethylene oxide inhalation exposure group, and $631{\pm}17\;and\;2265{\pm}9.4$(nmol g-1 globin) at 1.0 hr/day 400 ppm ethylene oxide inhalation exposure for 1 and 4 weeks, respectively. We confirmed that (N-2-hydroxy-ethyl)valine(HEV) of hemoglobin was a good biomarker for biomonitoring of ethylene oxide exposure, and can measured with derivatives such as PFPITH(pentafluorophenylisothiocianate) and TBDMS(tributyldimethylsilylation) by using GC/MS.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.10 no.2
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• pp.124-139
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• 2000

This study was performed to investigate monoacetylbenzidine(MABZ) and benzidine(BZ) hemoglobin adducts among workers who worked at benzidine based dye manufacturing company, and exposed by benzidine and benzidine based dye. The hemoglobin adducts were compared with work environment assessment result for evaluating the usefulness of biological monitoring. The mean BZ hemoglobin adducts among the first synthesis worker's hemoglobin adducts were $40.69{\mu}gBZ/g$ Hb and those of dry and packing workers were $22.14{\mu}gBZ/g$ Hb. The mean of MABZ hemoglobin adducts among 1st synthesis workers were $255.84{\mu}gMABZ/g$ Hb, dispersion worker's hemoglobin adducts were $76.17{\mu}gMABZ/g$ Hb and synthesis worker's hemoglogin adducts were $28.66{\mu}gMABZ/g$ Hb. Work environment assessment results during past 3 years were $0.0065mg/m^3$ and $0.5659mg/m^3$ of benzidine based dye concentration in ambient air of drying and packing only. Dye producing process was categorized by the possibility of exposure to benzidine and benzidine based dye. BZ and MABZ hemoglobin adducts were $19.55{\mu}gBZ/g$ Hb, $119.80{\mu}gMABZ/g$ Hb among workers who exposed by benzidine dihydrochloride and $16.32{\mu}gBZ/g$ Hb, $316.56{\mu}gMABZ/g$ Hb among workers who exposed by benzidine based dye. BZ hemoglobin adducts were not detected among control group and MABZ hemoglobin adducts were $5.33{\mu}gMABZ/g$ Hb. The differences between control and other exposed group was statistically significant. But there was no statistically significant differences between benzidine dihydrochloride exposed process and benzidine based dye exposed group. BZ and MABZ hemoglobin adducts were $2.23{\mu}gBZ/g$ Hb, $76.17{\mu}gMABZ/g$ Hb and $3.46{\mu}gBZ/g$ Hb, $21.33{\mu}gMABZ/g$ Hb. So hemoglobin adducts of MABZ were 5 ~ 30 time higher than those of BZ(P<0.003). Above results indicate that work environment assessment didn't detected benzidine and benzidine based dye in ambient air but biological monitoring detected those of hemoglobin adducts. Two group's hemoglobin adducts exposed benzidine dihydrochloride and benzidine based dye were high level but wasn't statistically significant and those were not detected in control group.

• The Korean Journal of Food And Nutrition
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• v.13 no.3
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• pp.242-248
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• 2000

수산물 가공시 흘러나오는 폐수 중에 함유되어 있는 수용성 단백질을 chitosan에 흡착시키기 위하여 albumin과 hemoglobin, albumin-myoglobin 혼합단백질을 이용하여 chitosan과의 흡착 및 chitosan 제조조건에 따른 흡착효과와 chitosan의 수용성 단백질 흡착에 미치는 인자를 살펴본 결과는 다음과 같다. Chitosan위 탈아세틸화 조건을 60, 70, 80%로 달리하여, albumin, hemoglobin, albumin-myoglobin 혼합용액에 적용했을때 chitosan의 탈아세틸화도가 높을수록 chitosan과 단백질 사이의 흡착률은 높게 나타났다. 초음파 처리에 의하여 chitosan의 분자량이 작을수록 chitosan과 단백질 사이의 흡착률은 높게 나타났다. pH변화에 따른 chitosan과 수용성 단백질의 흡착률은 albumin 및 albumin-myoglobin 혼합용액에서는 pH 4.0에서, hemoglobin용액에서는 pH 7.0에서 흡착률이 높게 나타났다 chitosan과 수용성단백질과의 흡착에서 반응시간은 albumin및 albumin-myoglobin 혼합용액에서는 4시간, hemoglobin용액에서는 3시간까지 흡착률이 증가하였고, 그 이후의 시간이 경화하여도 흡착률의 증가는 거의 보이지 않았다. 수용성 단백질 용액에 NaCl 농도를 0.1M에서 1.0M로 증가시켜 첨가했을때 염의 농도가 높을수록 chitosan과 단백질 사이의 흡착이 잘 일어나지 않았다.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.5 no.4
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• pp.105-107
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• 1972

The effects of copper and salt on the hemoglobin patterns of Carassius carassius, Ophicephalus argus, and Misgurnus anguillicaudatus were investigated by vertical starch gel electrophoresis. The number of the hemoglobin band of normal C. cararsius on the anode was one, and that of O. argus was two; neither the number of the homoglobin bands of the fishes nor the mobility of them was affected by copper or salt. The hemoglobin mobility of O. argus was faster than those of C. carassius and M. anguillicaudatus. Normal M. anguillicaudatus had two hemoglobin bands with nearly identical mobilities, one band migrating anodically and the other cathodically. When M. anguillicaudatus was exposed to copper, the mobility of band on the anode was faster than those of the other groups of control and exposed to salt, whereas the hemoglobin band on the cathode was not affected.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.4
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• pp.697-701
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• 2011

The purpose of this study was to fine proteins, which have significantly different level in plasma between Qi-deficiency and Fire-heat group of Korean Oriental Stroke pattern identification (PI) among Korean stroke patients. Eighteen stroke patients with Qi-deficiency and forty nine patients with Fire-heat, which had critical syndrome of each PI, were participated in this study. Plasma protein pattern were analyzed by SELDI-TOF MS using Q10 strong anion exchange chip and Mass spectral data (m/z) statistically determined. The expression level of proteins, which were different between Qi-deficiency and Fire-heat in the results by SELDI-TOF MS, were confirmed by western blot. As a result of analyzing plasma protein by SELDI-TOF MS, six protein peaks were significantly higher in Fire-heat group than Qi-deficiency group. Two peaks among of them, M15003 and M15745, were respectively identified as hemoglobin alpha and beta in previous study. Expression level of plasma free hemoglobin of Fire-heat group was also confirmed higher in Fire-heat group than in Qi-deficiency group. These findings suggest that plasma free hemoglobin is a candidate for discriminating Qi-deficiency and Fire-heat group according to pattern identification (PI) of stroke.