• Title/Summary/Keyword: Lipid membrane

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Effects of protein concentration and detergent on endotoxin reduction by ultrafiltration

  • Jang, Hyun;Kim, Hyo-Seung;Moon, Seung-Cheol;Lee, Young-Rae;Yu, Kang-Yeoul;Lee, Byeong-Kil;Youn, Hyun-Zo;Jeong, Young-Ju;Kim, Byeong-Soo;Lee, Sung-Ho;Kim, Jong-Suk
    • BMB Reports
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    • v.42 no.7
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    • pp.462-466
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    • 2009
  • Lipopolysaccharide (LPS), found in the outer membrane of Gram negative bacteria, only exerts its toxic effects when in free form. LPS has three major parts, lipid A, the toxic component, along with a core polysaccharide and O-specific polysaccharide. LPS monomers are known to have molecular masses between 10 to 30 kDa. Under physiological conditions, LPS exists in equilibrium between monomer and vesicle forms. LPS removal by 100 kDa ultrafiltration was more efficient (99.6% of LPS removed) with a low concentration of protein (2.0 mg/ml) compared to a high concentration (20.1 mg/ml). In the presence of different detergents (0.5% Tween 20, 1.0% taurodeoxycholate and 1.0% Triton X-100), LPS removal was more efficient at low protein concentrations (2.0 mg/ml) compared to high protein concentrations (20.1 mg/ml).

Morphological Studies on the Epithelium of the Midgut of the Nymph and the adult in the Grasshopper, Oxya sinuosa (우리벼메뚜기 유충과 성충의 중장 점막 상피에 관한 형태학적 연구)

  • Lee, Hyeung-Sik;Lee, Jae-Hyun;Park, Sang-Ock
    • Applied Microscopy
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    • v.20 no.1
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    • pp.105-119
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    • 1990
  • In order to know the morphological characteristics and changes according to the developmental stage, the comparative observations have been studied by light and electron microscope on the midgut(gastric caecum, stomach) of the grasshopper(Oxya sinuosa Mistshenko). The results obtained are summarized as follow : In light microscopic level, the cuboidal shaped cells of the gastric caecum in the 5th instar nymph are differentiated the columnar epithelium in the adult. A number of pigment granules are appeared in the gastric caecum of the 5th instar nymph, however the pigment granules were absent in the adult. Indistinct or undifferentiated folds of the epithelial layer were appeared in the 5th instar nymph, whereas the well-developed folds were appeared the gastric caecum in the adult. The well-developed muscular layers are seen in the 5th instar numph, however in the adult the muscular layers are appeared thin or a few layers. In electron microscopic level, in the midgut epithelium, a number of well-developed rER, a few lipid droplets, multi vesicular bodies, small vesicles and glycogen granules were found. Two types of the pigment granules in the gastric caecum of the 5th instar nymph were appeared, whereas the whorl-membrane, a few secretory granules and one type of the gastro-entero-endocrine cell were found in the adult midgut. The light and dark cells could be distinguished in the stomach epithelim of the 5th instar nymph.

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Effect of Salviae Radix on renal tubular reabsorption in rabbits with mercury-induced acute renal failure (수은으로 유발된 토끼의 신장 기능 손상에 대한 단삼(丹參)의 효과)

  • Hwang, Young-Geun;Jeong, Ji-Cheon
    • The Journal of Internal Korean Medicine
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    • v.21 no.2
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    • pp.227-234
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    • 2000
  • 독성약물에 의한 급성신부전시 세뇨관세포의 물질 재흡수 장애에 대한 단삼(丹參) 추출액의 효과를 조사하였다. 토끼에 수은(HgCl2)을 10 mg/kg되게 피하 주사하여 급성신부전을 유발하였고, 단삼(丹參) 추출액의 효과는 수은을 주사하기 전 7일 동안 0.05% 액(液) 0.3 g/kg 용량을 경구 투여하여 관찰하였다. 수은을 주사하기 전 24시간 동안 요와 혈액을 채취하여 신장기능을 측정하여 대조기간(basal period)의 값으로 하였고, 수은을 주사한 후 24시간 동안 요와 혈액을 얻어 수은에 의한 신장기능 변화를 평가하였다. 수은을 처리한 후 사구체여과율이 대조값에 비해 감소하였고, 혈청내 creatinine 농도가 증가하였다. 이러한 결과들은 수은이 급성신부전을 유발하였음을 가리킨다. 수은을 처리한 동물에서 포도당 및 인산의 배설분율이 증가하였고, 이러한 변화는 brush-border membrane에서 물질의 이동장애와 Na-pump 활성의 감소에 기인하였다. 수은을 주사한 동물의 신장피질 절편에서 유기이온인 PAH와 TEA 이동이 억제되었다. 토끼의 신장조직에서 지질의 과산화가 수은을 주사한 후 증가하였다. 단삼(丹參) 추출액을 전 처리한 후 수은을 주사한 경우 수은에 의해 유발된 사구체여과율의 감소와 혈청내 creatinine 농도 증가 현상이 유의하게 완화되었다. 수은에 의한 세뇨관에서 물질의 재흡수 장애가 단삼(丹參) 추출액의 전처리에 의해 방지되었다. 단삼(丹參) 추출액은 수은에 의한 지질의 과산화를 억제하였다. 수은에 의한 급성신부전은 항산화제로 잘 알려진 DPPD에 의해 방지되었다. 이상의 결과를 종합하면 생체실험결과 수은에 의한 급성신부전의 유발과정에 지질의 과산화가 중요한 역할을 할 가능성을 보이고 있고, 단삼(丹參) 추출액은 수은에 의한 급성신부전을 방지하는 효과를 가지고 있으며, 그 효과는 단삼(丹參)의 항산화작용에 기인(起因)할 가능성이 많다.

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Anti-Skin Aging Effect of Syriacusins from Hibiscus Syriacus on Ultraviolet-Irradiated Human Dermal Fibroblast Cells

  • Ryoo, In-Ja;Moon, Eun-Yi;Kim, Young-Hee;Lee, Ik-Soo;Choo, Soo-Jin;Bae, Ki-Hwan;Yoo, Ick-Dong
    • Biomolecules & Therapeutics
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    • v.18 no.3
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    • pp.300-307
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    • 2010
  • Photosensitized peroxidation of membrane lipids has been implicated in skin pathologies such as phototoxicity and premature aging. We have previously reported that syriacusin compounds isolated from Hibiscus Syriacus inhibited lipid peroxidation. Here, we investigated whether syriacusins could be effective inhibitor to skin aging using ultraviolet-irradiated human dermal fibroblast cells (HDFCs). Syriacusins A, B, and C inhibit the activity of human neutrophil elastase (HNE), a serine protease to degrade extracellular matrix (ECM) proteins including elastin, with $IC_{50}s$ of 8.0, 5.2, and $6.1\;{\mu}M$, respectively. No changes in cell viability were detected by syriacusins A and B in UV-B ($10\;mJ/cm^2$) irradiated HDFCs. Matrix metallo-proteinase (MMP)-1 expression in HDFCs was increased by UV-B irradiation. MMP-1 expression in UV-B irradiated HDFCs was decreased by $10\;{\mu}M$ and $20\;{\mu}M$ syriacusin A to 50% and 20% of untreated control, respectively. Syriacusin B treated with $20\;{\mu}M$ reduced MMP-1 expression in UV-B irradiated HDFCs to 60% of untreated control. Syriacusin A also inhibited MMP-2 expression accompanying the increase of type-I pro-collagen in UV-B irradiated HDFCs. These results demonstrate that syriacusin A could be a more effective compound to inhibit skin aging caused by UV irradiation. It suggests that syriacusins A and B might be developed as possible agents to treat or prevent skin aging.

Biochemical Characteristics and Function of a Fucosyltransferase Encoded by ste7 in Ebosin Biosynthesis of Streptomyces sp. 139

  • Chang, Ming;Bai, Li-Ping;Shan, Jung-Jie;Jiang, Rong;Zhang, Yang;Guo, Lian-Hong;Zhang, Ren;Li, Yuan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1092-1097
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    • 2009
  • A novel exopolysaccharide named Ebosin was produced by Streptomyces sp. 139, with medicinal activity. Its biosynthesis gene cluster (ste) has been previously identified. For the functional study of the ste7 gene in Ebosin biosynthesis, it was disrupted with a double crossover via homologous recombination. The monosaccharide composition of EPS-7m produced by the mutant strain Streptomyces sp. 139 ($ste7^-$) was found altered from that of Ebosin, with fucose decreasing remarkably. For biochemical characterization of Ste7, the ste7 gene was cloned and expressed in Escherichia coli BL21. With a continuous coupled spectrophotometric assay, Ste7 was demonstrated to have the ability of catalyzing the transfer of fucose specifically from GDP-$\beta$-L-fucose to a fucose acceptor, the lipid carrier located in the cytoplasmic membrane of Streptomyces sp. 139 ($ste7^-$). Therefore, the ste7 gene has been identified to code for a fucosyltransferase, which plays an essential role in the formation of repeating sugars units during Ebosin biosynthesis.

Application of Enzymatic method to Determine Choline Concentration in Bovine Blood and Muscle (소의 혈액 및 근육 중 choline 농도 분석을 위한 효소측정법의 적용기법의 개발)

  • Kim, Young-Il;Jung, Won-Chul;Shon, Ho-Yeong;Kim, Suk;Hur, Yoen;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.23 no.3
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    • pp.271-275
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    • 2008
  • Choline is important an organic compound for normal membrane function, acetylcholine synthesis, lipid transport, and methyl metabolism. In biological tissues and foods, there are multiple choline compounds that contribute to choline content. There are so many analytical methods for choline determination, such as radioisotopic, high-performance liquid chromatography, and gas chromatography/mass spectrometry. However, these existing methods are expensive, unmanageable, and time-consuming. In this study, we modified enzymatic method, which is applicable for the determination of choline in milk and infant formulas, and applied to bovine serum and muscle. The calibration curves were linear with higher correlation coefficients than 0.994. Recoveries obtained by calibration curves from the spiked bovine serum and muscle samples varied between 70.6 and 85.2%. The method may be suitable for use as a routine method in the determination of choline for biological tissue and food samples.

Effects of Reactive Oxygen Species on the Function of Porcine Spermatozoa (활성산소계가 돼지 정자의 기능에 미치는 영향)

  • Kim, Byung-Gak;Kim, Ki-Jung;Lee, Yong-An;Kim, Bang-Jin;Kim, Yong-Hee;Ryu, Buom-Yong
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.49-54
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    • 2009
  • The current study was designed to evaluate the effects of the reactive oxygen species (ROS) generated with a xanthine (X) and xanthine oxidase system (XO) on sperm function and DNA fragmentation in porcine spermatozoa. ROS were produced by a combination of $1,000{\mu}M$ X and 50 mU/ml XO. The ROS scavengers such as superoxide dismutase (SOD) (200 U/ml) and catalase (CAT) (500 U/ml) were also tested. Spermatozoa were incubated for 2 hours in BWW medium with a combination of X-XO supplemented with or without antioxidants at $37^{\circ}C$ under 5% $CO_2$ incubator. Ca-ionophore-induced acrosome reaction, the proportion of swollen spermatozoa under hypo-osmotic condition, malondialdehyde formation for the analysis of lipid peroxidation, and the proportion of DNA fragmentation were determined after 2 hours incubation. The action of ROS on porcine spermatozoa resulted in decreased Ca-ionophore-induced acrosome reaction and membrane integrity, increased the formation of malondialdehyde, and the proportion of sperm with DNA fragmentation(p<0.05). The toxic effects caused by ROS were completely alleviated by CAT in terms of sperm function and characteristics, however SOD did not serve the same scavenger effect as CAT. To conclude, the ROS can cause significant damage to porcine sperm functions and characteristics, which can be minimized by the use of antioxidants.

Physiologically Active Fatty Acids their Metabolism and Function (생리활성지방산;그 대사와 기능)

  • Mitsu, Kayama
    • Journal of the Korean Applied Science and Technology
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    • v.13 no.3
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    • pp.15-24
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    • 1996
  • Essentiality was proposed in the field of lipid by Burr and Burr in 1929. When rats were raised on the fat-free diet, their growth retarded and their skin and tails showed the characteristic deficient symptoms, which were relieved by the addition of ${\omega}6(n-6)$ polyunsaturated fatty acids as linoleic(LA) and arachidonic(AA) acids to the basal diet. LA is dehydrogenated to ${\gamma}-linolenic$ acid(GLNA) by ${\Delta}6$ desaturase, then GLNA is 2 carbon chain elongated by elongase to $dihomo-{\gamma}-linolenic$ acid(DGLNA), which is desaturated by ${\Delta}5$ desaturase to AA. These acids are called LA family or ${\omega}6(n-6)$ polyunsaturated fatty acids(PUFA). ${\alpha}-Linolenic$ acid(ALNA) is converted through the series of desaturation and elongation steps to docosahexaenic acid(DHA) via eicosapentaenoic acid(EPA). These acids belong to ALNA family or ${\omega}3(n-3)$PUFA. Human who consume large amounts of EPA and DHA, which are present in fatty fish and fish oils, have increased levels of these two fatty acids in their plasma and tissue lipids at the expense of LA and AA. Alternately, vegetarians, whose intake of LA in high, have more elevated levels of LA and AA and lower levels of EPA and DHA in plasma lipids and in cell membranes than omnivores. AA and EPA are metabolized to substances called eicosanoids. Those derived form AA are known as prostanocids(prostaglandins and prostacyclins) of the 2-types and leukotrienes of the 4-series, whereas those derived from EPA are known as prostanoids of the 3-types and leukotrienes of the 5-series. DGLNA is a precursor of the 1-types of prostaglandins. The metabolites of AA and EPA have competitive functions. Ingestion of EPA from fish or fish oil replaces AA from membrane phospholipids in practically all cells. So this leads to a more physiological state characterized by the production of proatanoids and leukotrienes that have antithrombic, antichemotactic, antivasoconstrictive and antiinflammatory properties. It is evident that ${\omega}3$ fatty acids can affect a number of chronic diseases through eicosanoids alone.

Enhancement of Arsenic Trioxide ($As_2O_3$)-Mediated Apoptosis Using Berberine in Human Neuroblastoma SH-SY5Y Cells

  • Kim, Dae-Won;Ahan, Song-Ho;Kim, Tae-Young
    • Journal of Korean Neurosurgical Society
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    • v.42 no.5
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    • pp.392-399
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    • 2007
  • Objective : Arsenic trioxide ($As_2O_3$) has been used as an anticancer agent in traditional Chinese medicine for thousand years and berberine is an isoquinoline alkaloid present that has indicated significant antimicrobial activity. We have examined the combined anticancer effects of $As_2O_3$ and berberine against the human neuroblastoma (HNB) SH-SY5Y cells in vitro, and to elucidate underlying molecular mechanism. Methods : HNB SH-SY5Y cells were treated with $2\;{\mu}M\;As_2O_3$ and $75\;{\mu}g/ml$ berberine, and their survival, cell death mechanism as well as synergistic cytotoxic effects were estimated by using MTT assay, DAPI staining, agarose gel electrophoresis, flow cytometric analysis, and western blot analysis. Results : The combined treatment of two drugs also markedly decreased cell viability. The cytotoxic effects of two drugs were revealed as apoptosis characterized by chromatin condensation, DNA fragmentation, and the loss of mitochondrial membrane potential. The apoptotic cytotoxicity was accompanied by activation of caspase-3 protease as well as decreased the expression of Bcl-2, Bid, and Bcl-x/L. In addition, the cells treated with combination of two drugs also showed significantly increased intracellular reactive oxygen species levels and lipid peroxidation compared to cells $As_2O_3$or berberine only. Conclusion : Combined treatment of $As_2O_3$ with berberine induced activation of apoptotic signaling pathways in HNB SH-SY5Y cells. These results suggest that the possibility of the combined treatment of two chemotherapeutic agents with low concentration improving cytotoxic effect for cancer cells with minimal side effects.

Cloning and Initial Analysis of Porcine MPDU1 Gene

  • Yang, J.;Yu, M.;Liu, B.;Fan, B.;Zhu, M.;Xiong, T.;Li, Kui
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.9
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    • pp.1237-1241
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    • 2005
  • Mannose-P-dolichol utilization defect 1 (MPDU1) gene is required for utilization of the mannose donor MPD in synthesis of both lipid-linked oligosaccharides (LLOs) and glycosylphosphatidylinositols (GPI) which are important for functions such as protein folding and membrane anchoring. The full length cDNA of the porcine MPDU1 was determined by in silico cloning and rapid amplification of cDNA ends (RACE). The deduced amino acid showed 91% identity to the corresponding human sequence with five predicted transmembrane regions. RT-PCR was performed to detect its expression pattern in five tissues and results showed that it is expressed ubiquitously among the tissues checked. A single nucleotide substitution resulting in the amino acid change (137 Tyr-137 His) was detected within exon 5. Allele frequencies in six pig breeds showed distinctive differences between those Chinese indigenous pigs breeds and European pigs. Using the pig/rodent somatic cell hybrid panel (SCHP), we mapped the porcine MPDU1 gene to SSC12, which is consistent with the comparative mapping result as conservative syntenic groups presented between human chromosome 17 and pig chromosome 12.