• Proceedings of the Korean Nutrition Society Conference
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• 1998.11a
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• pp.89-89
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• 1998

• Journal of Yeungnam Medical Science
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• v.3 no.1
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• pp.41-48
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• 1986

Band 3, the predominent 95,000 dalton anion transport protein, is the major intrinsic glycoprotein of the human erythrocyte membrane. This anion carrier exists as a dimer and binds the cytoskeletons such as spectrin, ankyrin and actin. And the liposomes are vesicular structures which form spontaneouly upon hydration of phospholipids. These artificial lipid vesicles have been investigated as model of the biological membranes and as a mean of improving the delivery of nucleic acids, drugs, proteins and biological substances to specific target tissues and cells. In this study, we were purified Band 3 from the human erythrocyte membrane(ghost) was prepared by hemolysis of intact human erythrocyte with weak alkali-hypotonic solution. Band 6 was removed from ghost by extracting with solution of an ionic strength of 0.15. Band 3 and Band 4 were solubilized selectively by extracting Band 6-depleted ghosts with Triton X-100 under nondenaturing conditions. Band 3 was then purified from Triton X-100 extract treated with p-chloromercuribenzoate by sucrose density gradient ultracentrifugation. This purified Band 3 was incorporated into liposomes prepared by reverse-phase evaporation. Phosphatidyl L-serine and cholesterol(1 : 1 molar ratio) were dissolved in chloroform and then chloroform was removed by rotatory evaporation under reduced pressure. Band 3 solution without Triton X-100 was introduced into a mixture of lipids and diethylether. Diethylether was subsequently removed by evaporation. This purified Band 3 and its incorporation into liposomes were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

• BMB Reports
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• v.30 no.4
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• pp.240-245
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• 1997

Interactions between ganglioside $G_{M3}$ and glucose transporter, GLUT1 were studied by measuring the effect of $G_{M3}$ on steady-state and time-resolved fluorescence of purified GLUT1 in synthetic lipids and on the 3-O-methylglucose uptake by human erythrocytes. The intrinsic tryptophan fluorescence showed a GLUT 1 emission maximum of 335 nm, and increased in the presence of $G_{M3}$ by 12% without shifting the emission maximum, The fluorescence lifetimes of intrinsic tryptophan on GLUT1 consisted of a long component of 7.8 ns and a short component of 2,3 ns and $G_{M3}$ increased both lifetime components. Lifetime components were quenched by acrylamide and KI. Acrylarnide-mduced quenching of long-lifetime components was partly recovered by $G_{M3}$ However. KI-induccd quenching of short- and long-lifetime components was not rescued by $G_{M3}$. The anisotropy of 1.6-diphenyl-1.3.5-hexatriene (DPH)-probed dimyristoylphosphatidylcholine (DMPC) model membrane was also increased with $G_{M3}$ incorporation, The transport rate of 3-O-methylglucose increased by 20% with $G_{M3}$ incorporation on the erythrocytes, Therefore, $G_{M3}$ altered the environment of lipid membrane and induced the conformational change of GLUT1.

• Advances in Traditional Medicine
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• v.3 no.4
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• pp.205-211
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• 2003

Stem bark of Cinchona sp. (Rubiaceae) is one of the well known drugs for its therapeutic values in traditional as well as modern medicine. Even though a lot of work has been carried out on quinoline alkaloids of Cinchona, its phenolic constituents received very little attention. In the present study, we evaluated antioxidant properties of C. officinalis stem bark methanolic extract and water extract containing phenolic compounds (total phenolics 21.37, 5.18% w/w respectively in the two extracts) in different in vitro and ex vivo models viz., antiradical activity by DPPH reduction, superoxide radical scavenging activity in riboflavin/light/NBT system, nitric oxide radical scavenging activity in sodium nitroprusside/Greiss reagent system and inhibition of lipid peroxidation induced by iron-ADP-ascorbate in liver homogenate and haemolysis of erythrocytes induced by phenylhydrazine in erythrocyte membrane stabilization study. Both the extracts exhibited very good antioxidant activity in all the models tested. The phenolic compounds including tannins present in the stem bark seem to offer protection from the oxidative damage.

• The Korean Journal of Physiology
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• v.17 no.2
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• pp.135-142
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• 1983

$PGE_2$ and $PGF_{2{\alpha}}$ are known to act similarly in a number of animal tissues. They both facilitate regression of corpus luteum(Poyser, 1972; Fuch et al, 1974; Coudert et at, 1974) and stimulate contraction of uterine muscle (Laudanski et al, 1977; Porter et al, 1979; Hollingsworth et al, 1980). It is, however, not known whether these two prostaglandins exert similar actions in osmotic fragility of erythrocytes (Rasmussen et al, 1975) and $PGF_{2{\alpha}}$ alters conformation of membrane proteins (Meyers aud Swislocki, 1974). The former effect may not be mediated through changes in c- AMP concentration in the cell, since the adenylate cyclase activity in human erythrocyte is extremely low (Rodan et al, 1976; Sutherland et al, 1962) and the latter effect implies that physical state (or fluidity) of the membrane is altered by $PGF_{2{\alpha}}$. The present study was undertaken to elucidate mechanisms of action of $PGE_2$ and $PGF_{2{\alpha}}$ on the human erythocyte membrane by examining their effects on osmotic fragility and $Ca^{++}$ binding to the membrane fragments. The results are summarized as follows: 1) $PGE_2$ and $PGF_{2{\alpha}}$ increased osmotic fragility at concentrations above $10^{11}\;M$, the effect being similar for both hormones. The concentration of NaCl for 100% hemolysis was $1/16{\sim}1/17\;M$ in the presence of $10^{11}\;M\;PGE_2$ or $PGF_{2{\alpha}}$ and 1/18 M in the absence of the hormone (control). 2) When erythrocytes were suspended in 1/15 M NaCl solution, $44.2{\pm}4.3%$ of cells were hemolyzed. Addition of $10^{12}\;M\;PGE_2$ or $PGF_{2{\alpha}}$ did not increase hemolysis. When the concentration of the hormones was increased to $10^{11}\;M$, however the degree of hemolysis increased markealy to about 80%. No further increase in hemolysis was observed at concentration of the hormones above $10^{11}\;M$. 3) The additional hemolysis due to $10^{11}\;M\;PGE_2$ and $PGF_{2{\alpha}}$ appeared to he identical regardless of absence or presence of $Ca^{++}\;(0.5{\sim}10\;mM)$ in the suspending medium. 4) In the absence of prostaglandin, the binding of $Ca^{++}$ to the erythrocyte membrane increased curvilinearly as the $Ca^{++}$ concentration increased up to 5 mM above which it leveled off. A similar dependence of $Ca^{++}$ binding on the $Ca^{++}$ concentration was observed in the presence of $10^{11}\;M\;PGE_2$ or $PGF_{2{\alpha}}$, however, the amount of $Ca^{++}$ bound at a given $Ca^{++}$ concentration was significantly higher than in the absence of the hormones. 5) As in the hemolysis, $PGE_2$ and $PGF_{2{\alpha}}$ did not affect the $Ca^{++}$ binding at a concentration of $10^{12}\;M$, but increased it by about 100% at concentration above $10^{11}\;M$. These result indicate that both tile osmotic fragility of erythrocyte and the $Ca^{++}$ binding to the erythrocyte membrane are similarly enhanced by $PGE_2$ and $PGF_{2{\alpha}}$, but these two effects are not causally related. It is, therefore, concluded that the prostaglandin-induced hemolysis is not directly associated with alterations of the $Ca^{++}$ content in the membrane.

• Journal of Veterinary Clinics
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• v.16 no.2
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• pp.276-280
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• 1999

It is well-known that onion and garlic of Allium family have been cultivated for major food since ancient times. Especially garlic is known for containing protective materials against cardiovascular disease, neoplasm and cancer, and decreasing cholesterol value. Hemolytic anemia was first reported by Goldsmith in onion-fed cattle in 1909. Subsequently, similar effects have been reported in cattle, horses, sheep, dogs and cats. This toxic effect has been attributed to npropyl disulfide which causes oxidant damage to erythrocyte cell membrane. There are few reports on overconsumption of garlic. Kim et al. reported in 1999 that 45% of Jindo dogs in korea have erythrocytes characterized by high Na-K-ATPase activity with high K and low Na concentrations (HK) which easily results in hemolytic anemia. The purpose of present report is to determine whether garlic-induced hemolytic anemia occurs in normal erythrocytes characterized by complete lack of Na-K-ATPase activity with low K and high Na concentrations (LK) when LK dogs are fed large amounts of garlics. We found in this experiment that Erythrocyte count, PCV and Hemoglobin concentration decreased significantly, and GSH and Met-hemoglobin concentrations increased. Consequently, we determined that overconsumption of garlic induces hemolytic anemia in LK dogs.

• Journal of Veterinary Clinics
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• v.18 no.4
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• pp.341-344
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• 2001

It is well-known that onion and garlic of Allium family have been cultivated for major food since ancient times. Especially garlic is known for containing protective materials against cardiovascular disease, neoplasm and cancer, and decreasing cholesterol value. Hemolytic anemia was first reported by Goldsmith in onion-fed cattle in 1909. Subsequently, similar effects have been reported in cattle, horses, sheep, dogs and cats. This toxic effect has been attributed to n-propyl disulfide which causes oxidant damage to erythrocyte cell membrane. There are few reports on overconsumption of garlic. Kim et al. reported in 1999 that 45% of Jindo dogs in korea have erythrocytes characterized by high Na-K-ATPase activity with high K and low Na concentrations (HK) which easily results in hemolytic anemia. The purpose of present report is to determine whether garlic-induced hemolytic anemia occurs in normal erythrocytes characterized by complete lack of Na-K-ATPase activity with low K and high Na concentrations (LK) dogs are fed large amounts of garlics. We found in this experiment that Erythrocyte count, PCV and Hemoglobin concentration decreased significantly, and GSH and Met-hemoglobin concentrations increased. Consequently we determined that overconsumption of garlic induces hemolytic anemia in LK dogs.

• Journal of Microbiology
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• v.43 no.3
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• pp.272-276
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• 2005

Bacillus subtilis ATCC 6633 was grown in BHIB medium supplemented with $Mn^{2+}$ for 96 h at $37^{\circ}C$ in a shaker incubator. After removing the microbial biomass, a lipopeptide biosurfactant was extracted from the supernatant. Its structure was established by chemical and spectroscopy methods. The structure was confirmed by physical properties, such as Hydrophile-Lipophile Balance (HLB), surface activity and erythrocyte hemolytic capacity. The critical micelle concentration (cmc) and erythrocyte hemolytic capacity of the biosurfactant were compared to those of surfactants such as SDS, BC (benzalkonium chloride), TTAB (tetradecyltrimethylammonium bromide) and HTAB (hexadecyltrimethylammonium bromide). The maximum hemolytic effect for all surfactants mentioned was observed at concentrations above cmc. The maximum hemolytic effect of synthetic surfactants was more than that of the biosurfactant produced by B. subtilis ATCC 6633. Therefore, biosurfactant would be considered a suitable surface-active agent due to low toxicity to the membrane.

• The Korean Journal of Physiology
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• v.20 no.2
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• pp.209-217
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• 1986

The functional molecular weight of band 4.5 polypeptide was measured by applying the classical target theory to radiation inactivation data of the cytochalasin B binding. Band 4.5 polypeptides purified from human erythrocyte membranes were irradiated at -45 to $-50^{\circ}C$ with an increasing dose of 1.5 MeV electron beam, and after thawing, cytochalasin B binding activities were assayed. Each activity measured was reduced as a simple exponential function of radiation dose. $D_{37}$, dose appeared to be 6.7 mega rads, from which the target size (radiation sensitive mass) of band 4.5 polypeptide was calculated to be 95,500 daltons. This result with other informations available in literature suggests that band 4.5 polypeptide may exist as a dimer in human erythrocytes.

• Proceedings of the Korean Biophysical Society Conference
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• 1996.07a
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• pp.37-37
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• 1996

Tenecin fragments are antimicrobial and antifungal peptide from Tenebrio molitor with highly positive charged amino acid residues. To elucidate their membrane selectivity and molecular mechanism, various forms of tenecin fragments were synthesized, and their interaction with acidic phospholipid, Gram (+), fungal and human erythrocyte membrane were investigated by ANTS/DPX leakage, membrane binding and fusion assay. (omitted)