• Title/Summary/Keyword: Bovine serum albumin

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Peroxynitrite Scavenging Mechanism of Psoralea corylifolia (보골지(補骨脂)의 Peroxynitrite 제거 기전)

  • Jang, Yong-Suk;Min, Gun-Woo;Jeong, Ji-Cheon
    • The Journal of Internal Korean Medicine
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    • v.25 no.2
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    • pp.268-276
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    • 2004
  • Objective : Peroxynitrite $(ONOO^-)$, formed from the reaction of $O_2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors. NO and $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123.. Methods : Psoralea corylifolia was crushed. The crushed Psoralea corylifolia was extracted 3 times, each time with 3 volumes of methyl alcohol at $60^{\circ}C$ for 24 h. The extract was filtered and evaporated under reduced pressured using a rotary evaporator to yield 16g. This was done to investigate scavenging activities of $ONOO^-$, NO, $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123. Results : After Psoralea corylifolia was added authentic $ONOO^-,\;{\cdot}\;O_2^-$ and NO was markedly scavenged. Also, $ONOO^-$ induced by $O_2^-$ and NO (these derived from SIN-1) was inhibited. The data showed a decrease in $ONOO^-$ mediated nitration of tyrosine through electron donation after Psoralea corylifolia was added. Data showed a dose-dependent correlation with inhibition of nitration of bovine serum albumin induced by $ONOO^-$, Furtheremore, LPS-induced ROS and RNS generation was blocked. Conclusions: These results suggest potential for use of Psoralea corylifolia as an effective $ONOO^-$ scavenger to counter the aging process and age-related diseases.

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Effects of Epoxidised Soya Bean Oil on Humoral Immune Response in Mice

  • Chung, Seung-Tae;Kim, Young-Ok;Chung, Hye-Joo;Kim, Jin-Ho;Park, Jae-Hyun;Kang, Sun-Kyung;Kim, Hyung-Soo;Cho, Dae-Hyun;Kim, Dong-Sup
    • Toxicological Research
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    • v.16 no.4
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    • pp.269-274
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    • 2000
  • Epoxidised soya bean oil (ESBO, 1000, 2000 or 4000 mg/kg) was orally administered to BALB/c mice daily for 28 consecutive days, and the control mice were exposed to vehicle (corn oil). Mice were immunized and challenged with sheep red blood cells (SRBC) or bovine serum albumin (BSA). In groups exposed to ESBO, the body weight gains and the relative lymphoid organ weights were not significantly changed as compared with control group. Secondary IgG antibody response to BSA was not significantly changed by ESBO, but plaque-forming cell (PFC) response to SRBC was significantly suppressed in mice treated with 4000 mg ESBO/kg/day. The mitogenic response of splenic B cells induced by LPS was not effected by ESBO in any of the groups. These results indicate that ESBO did not induce significant humoral immune response at a dose less than 2000 mg/kg/day in mice.

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Poly(Ethylene Glycol)-branched Polyethylenimine-poly(L-phenylalanine) Block Copolymer Synthesized by Multi-initiation Method for Formation of More Stable Polyelectrolyte Complex with Biotherapeutic Drugs

  • Park, Woo-Ram;Na, Kun
    • Journal of Pharmaceutical Investigation
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    • v.41 no.2
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    • pp.95-102
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    • 2011
  • An amphiphilic cationic branched methoxy poly (ethylene glycol)-branched polyethylenimine - poly(L-phenylalanine) (mPEG-bPEI-pPhe) block copolymer was successfully synthesized by ring-opening polymerization (ROP) of N-carboxyanhydride of L-phenylalanine (Phe-NCA) with mPEG-bPEI for the preparation of more stable polyelectrolyte complex (PEC) included a hydrophobic interaction. mPEG-bPEI was firstly prepared by the coupling of mPEG and bPEI using hexamethylene diisocyanate (HMDI). The structural properties of mPEG-bPEI-pPhe copolymers were confirmed by $^1H$ NMR. The copolymers exhibited a self-assemble behavior in water above critical aggregate concentration (CAC) in the range of 0.01-0.14 g/L. The CAC of copolymers obviously depended on the hydrophobic block content in the copolymers (the value decreased with the increase of the pPhe block content). The cationic copolymers have the ability to form multi-interaction complex (MIC) with bovine serum albumin (BSA) and plasmid DNA through multi-interaction (electrostatic and hydrophobic interaction). The physicochemical characterization of the complex was carried out by the measurement of zeta potential and particle size. Their zeta-potentials were positive (approximately +10 mV) and their sizes decreased with increasing pPhe contents in the copolymers (PPF/BSA wt% ratio = 2). The complex showed good stability at high ionic strength. Therefore, mPEG-bPEI-pPhe block copolymer was considered as a potential material to enhance the stability of complex including biotherapuetic drugs.

Phospholipase and Aspartyl Proteinase Activities of Candida Species Causing Vulvovaginal Candidiasis in Patients with Type 2 Diabetes Mellitus

  • Bassyouni, Rasha H.;Wegdan, Ahmed Ashraf;Abdelmoneim, Abdelsamie;Said, Wessam;AboElnaga, Fatma
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1734-1741
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    • 2015
  • Few research had investigated the secretion of phospholipase and aspartyl proteinase from Candida spp. causing infection in females with type 2 diabetes mellitus. This research aimed to investigate the prevalence of vulvovaginal candidiasis (VVC) in diabetic versus non-diabetic women and compare the ability of identified Candida isolates to secrete phospholipases and aspartyl proteinases with characterization of their genetic profile. The study included 80 females with type 2 diabetes mellitus and 100 non-diabetic females within the child-bearing period. Candida strains were isolated and identified by conventional microbiological methods and by API Candida. The isolates were screened for their extracellular phospholipase and proteinase activities by culturing them on egg yolk and bovine serum albumin media, respectively. Detection of aspartyl proteinase genes (SAP1 to SAP8) and phospholipase genes (PLB1, PLB2) were performed by multiplex polymerase chain reaction. Our results indicated that vaginal candidiasis was significantly higher among the diabetic group versus nondiabetic group (50% versus 20%, respectively) (p = 0.004). C. albicans was the most prevalent species followed by C. glabrata in both groups. No significant association between diabetes mellitus and phospholipase activities was detected (p = 0.262), whereas high significant proteinase activities exhibited by Candida isolated from diabetic females were found (82.5%) (p = 0.000). Non-significant associations between any of the tested proteinase or phospholipase genes and diabetes mellitus were detected (p > 0.05). In conclusion, it is noticed that the incidence of C. glabrata causing VVC is increased. The higher prevalence of vaginal candidiasis among diabetics could be related to the increased aspartyl proteinase production in this group of patients.

The optimization of ELISA for methamphetamine determination : the effect of immunogen, tracer and antibody purification method on the sensitivity

  • Choi, Jeongeun;Choi, Myung-Ja;Kim, Choonmi;Cho, Young-Shik;Chin, Jaeho;Jo, Young-Ah
    • Archives of Pharmacal Research
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    • v.20 no.1
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    • pp.46-52
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    • 1997
  • To obtain more sensitive immunoassay for methamphetamine (MA) determination, the optimum condition of enzyme-linked immunosorbent assay (ELISA) was investigated in regard to immunogens, antibody purification methods and coating tracers. Activated MA, N-(4-aminobutyl)methamphetamine (4-ABMA), was conjugated with bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) and used as immunogen. The antibodies were purified by protein G chromatography or various immunoaffinity chromatography-linked MA-protein ligands, such as MA-BSA, MA-KLH or MA-ovalbumin (OVA). Each purified antibody was characterized by means of sensitivity and cross-reactivity using the three MA-protein coating tracers, MA-BSA, MA-KLH and MA-OVA. The best sensitivity of each antibody was acquired with the MA-OVA tracer although the tracer concentration and the antibody titer level at optimum condition were varied. The antibody with high titer level did not always yield good sensitivity. At optimum condition, immunoaffinity chromatography-purified antibodies were better for sensitivity and for specificity than protein G-purified antibodies. The cross-reactivity of the purified antibodies seemed to be affected by immunogen structure and showed somewhat different patterns according to the immunoaffinity ligand utilized. These data show that the antibody purification method as well as choice of coating tracer and immunogen is essential for the sensitivity and specificity of EIA; the optimum condition for assay should be discovered using various methods and combinations.

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Synthesis and Characterization of Biodegradable Thermo- and pH-Sensitive Hydrogels Based on Pluronic F127/Poly($\varepsilon$-caprolactone) Macromer and Acrylic Acid

  • Zhao, Sanping;Cao, Mengjie;Wu, Jun;Xu, Weilin
    • Macromolecular Research
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    • v.17 no.12
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    • pp.1025-1031
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    • 2009
  • Several kinds of biodegradable hydrogels were prepared via in situ photopolymerization of Pluronic F127/poly($\varepsilon$-caprolactone) macromer and acrylic acid (AA) comonomer in aqueous medium. The swelling kinetics measurements showed that the resultant hydrogels exhibited both thermo- and pH-sensitive behaviors, and that this stimuli-responsiveness underwent a fast reversible process. With increasing pH of the local buffer solutions, the pH sensitivity of the hydrogels was increased, while the temperature sensitivity was decreased. In vitro hydrolytic degradation in the buffer solution (pH 7.4, $37^{\circ}C$), the degradation rate of the hydrogels was greatly improved due to the introduction of the AA comonomer. The in vitro release profiles of bovine serum albumin (BSA) in-situ embedded into the hydrogels were also investigated: the release mechanism of BSA based on the Peppas equation was followed Case II diffusion. Such biodegradable dual-sensitive hydrogel materials may have more advantages as a potentially interesting platform for smart drug delivery carriers and tissue engineering scaffolds.

Dye-binding Capacities of Proteins using Coomassie blue G250 in an Acidic Solution (단백질과 색소(色素) Comassie blue G250과의 결합능력)

  • Park, Ro-Dong;Lee, Yeon;Shin, Yong-Kwang
    • Applied Biological Chemistry
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    • v.25 no.4
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    • pp.248-251
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    • 1982
  • Commassie blue G250 produced metachromatic effect with some solvents. The absorptivity and molar absorptivity of the dye in ethanol were 82.4 and $70.4{\times}10^3$ at maximum absorption wavelength 610nm, respectively. The dye had a red from$({\lambda}_m=465nm)$ in ethanol-phosphoric acid-water solution and converted to a blue form$({\lambda}_m=590nm)$ after binding to protein. Absorbance at 590nm gave linear responses with respect to protein contents. The dye-binding capacities of proteins varied considerably with the content and source of proteins. Under the experimental condition the dye-binding capacities of bovine serum albumin, cytochrome C and ${\gamma}-globulin$ were 110, 103, and $88{\mu}g$ commassie blue G250 bound per $100{\mu}g$ protein, respectively.

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Release Properties of BSA from Pectin Heads for Colonic Drug Delivery (Colonic Delivery를 위한 펙틴 비드로부터 BSA의 방출 특성)

  • 최춘순;박상무;송원현;이창문;이기영;김동운;김진철
    • KSBB Journal
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    • v.18 no.2
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    • pp.161-164
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    • 2003
  • Oral drug delivery system using pectin gel was developed for colon-targeting of peptide drug. BSA(bovine serum albumin)-loaded pectin and pectin-alginate beads were prepared for drug release properties in vitro. Morphological studies by electron microscopy indicated that pectin and pectin-alginate beads were spherical in shape and approximately 1.0 mm. In order to find the suitable beads, effects of cross-linking agents (calcium chloride or zinc acetate) and drying temperature of beads were investigated. Drug release decreased with concentration of cross-linking agents and drying temperature. For colonic drug delivery from pectin and pectin-alginate beads, pectin degradable enzymes were added at 5 hrs from the beginning of drug release. After addition of enzymes, drug release was suddenly increased against free enzymes. Therefore, pectin and pectin-alginate beads can be promised as useful drug release carriers for colon-targeted delivery.

THE EFFECT OF SUPEROXIDE DISMUTASE ON EXPERIMENTAL GINGIVITIS AND ACTIVITY OF 3T3 FIBROBLAST (Superoxide Dismutase가 백서의 실험적 치은염과 3T3 섬유모 세포의 활성에 미치는 영향)

  • Kim, Yoon-Seong;Yoo, Hyung-Keun;Kang, Hyun-Ku;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.25 no.2
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    • pp.222-238
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    • 1995
  • Inflammatory cells may produce active species of oxygen in antimicrobial defense. While such species can directly damage surrounding tissue, their major secondary role may be to mediate important components of the inflammatory response. Superoxide dismutase, antioxidant, have significant anti-inflammatory properties in rheumatoid arthritis, ischemic tissue injury and gastrointestinal disease. Increased oxidative product formation diseases. And superoxide dismutase produced by Porphyromonas Gingivalis is resistant to killing by polymorphonuclear leukocyte. The purpose of this study was to investigate on the effects of superoxide dismutase in 3T3 fibroblast and in experimental gingivitis in the rats. The effect of superoxide dismutase(SOD) to cell morphology and cell activity was measured in cultured mouse 3T3 fibroblast. After experimental gingivitis were induced by lipopolysaccharide(LPb) and bovine serum albumin(BSA), injection of SOD were done. WBC count and histologic findings were observed at 1, 2, 3, and 7 days. The results were as follows; 1. There was a little difference between LPS treated groups and SOD treated groups in 3T3 fibroblast morpholoy. 2. There was no difference between only SOD treated groups (except SOD 150U at 3days) and control in 3T3 fibroblast activity. 3. LPS $0.5{\mu}g/ml$ and SOD treated groups (except 150U) had decreased 3T3 fibroblast activity and no significant difference at 3 days. 4. LPS $5.0{\mu}g/ml$ and SOD treated groups were significantly increased cell activity of 3T3 fibroblast than control group at 1 day(P<0.05). 5. In LPS induced gingivitis, the number of leukocytes in SOD treated was significantly decreased than in saline treated at 1 day(P<0.05). 6. In histopathologic findings of LPS or BSA induced gingivitis, inflammatorycell infiltration in SOD treated groups were less than in saline treated group at 1, 2 and 3 days.

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Production of Monoclonal Antibodies Against the Immunoglobulin M of Olive Flounder Paralichthys Olivaceus (넙치(Paralichthys olivaceus)의 immunoglobulin M에 대한 단클론 항체 생산)

  • Kim, Wi-Sik;Kim, Ki-Hong;Kim, Choon-sup;Oh, Myung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.2
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    • pp.169-174
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    • 2017
  • Immunoglobulin M (IgM) was purified from olive flounder Paralichthys olivaceus sera using mannan-binding protein (MBP) and protein L affinity columns (designated as MBPIgM and ProLIgM, respectively). A monoclonal antibody (MAb) against olive flounder IgM was produced. The MBPIgM and ProLIgM had apparent molecular weights of 77, 73, and 28 kDa in SDS-PAGE. Nine hybridomas secreting MAbs against olive flounder IgM were established: five MAbs for MBPIgM (1, 2, 3, 4, and 5) and four for ProLIgM (6, 7, 8, and 9). Western blotting indicated that seven MAbs recognized heavy (H; MAbs 1, 2, 3, 4, 5, 6, and 7) chains and one recognized light (L; MAb 9) chains of IgM, while MAb 8 did not recognize IgM. The results of enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the nine MAbs revealed that the optical density (OD) values of sera differed significantly between BSA- and non-immunized fish, despite some sera from non-immunized fish with slight high OD values. These results suggest that the MAbs produced in this study reacted specifically with the IgM from olive flounder.