• Title/Summary/Keyword: phosphate-buffered saline

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Antibacterial Effect of the Surface-Modified Biomedical Polyurethane against Staphylococcus aureus and Staphylococcus epidermidis

  • Jeon, Sung-Min;Kim, Hyun-Jung;Lee, Kyu-Back;Kim, Jong-Won;Kim, Mal-Nam
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.259-265
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    • 2001
  • Staphylococal infection still remains to be one of the most serious infections, having various complications in the clinical use of indwelling polymeric medical devices. However, there are a few promising systems showing a high antibacterial effect without causing any demage of polymer backbone under biological environments such as blood or body fluid. In order to resolve this problem, we have designed a new antibiotic releasing system via a hydrolysis mechanism. The surface of biomedical polyurethane (PU) was modified by using 1,6-diisocyanatohexane (HMDI) to immobilize the rifampicon. Also, the immobilized rifampicin was designed to be released by a selective cleavage of the unstable carbamate linkage that exists on the rifampicin-immobilized polyurethane (PHR). The immobilization of rifampicin on the surface of polyurethane was confirmed by the disappearance of the characteristics IR absorbance peak of the isocyanate (-NCO) group at $2,267\;cm^{-1}$. The PHR showed a continuous rifampicin release profile under an aqueous environment of 10 mM of PBS (phosphate-buffered saline) for ove 6 days. The rifampicin molecules, which are released from PHR under an optimal bacterial infection environment, had a higher antibacterial activity against both S. aureus and S. epidermidis than rifampicin-incorporated polyurethane (RIP). In addition, the PHR maintained a stable antibacterial effect under a blood-mimic aqueous environment such as bovine calf serum.

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Cloning and Expression of the Duck Leptin Gene and the Effect of Leptin on Food Intake and Fatty Deposition in Mice

  • Dai, Han Chuan;Long, Liang Qi;Zhang, Xiao Wei;Zhang, Wei Min;Wu, Xiao Xiong
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.850-855
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    • 2007
  • Leptin is the adipocyte-specific product of the obese gene and plays a major role in food intake and energy metabolism. Leptin research was mainly focused on mammalian species, but understanding of leptin and its function in poultry is very poor. In this study, the duck leptin gene was amplified using the reverse transcription-polymerase chain reaction (RT-PCR) from duck liver RNA. The cDNA fragment was inserted into the pET-28a expression vector, and the resulting plasmid was expressed in Escherichia coli BL21 (DE3). Experimental mice were given an intraperitoneal injection of 10 mg/kg leptin dissolved in phosphate buffered saline (PBS), while the control mice were injected with PBS. The effect of leptin on food intake, body weight and fatty deposition in mice was detected. Sequence analysis revealed that duck leptin had a length of 438 nucleotides which encoded a peptide with 146 amino acid residues. The sequence shares highly homology to other animals. The coding sequence of duck leptin was 84 and 86% identical to human and pig leptin nucleotides sequence. Highest identity was with the rat coding sequence (95%). The identity of the amino acid sequence was 84, 82 and 96% respectively compared to that of the human, pig and rat. Results of SDS-PAGE analysis indicated that a fusion protein was specifically expressed in E. coli BL21 (DE3). The purified product was found to be biologically active during tests. Continuous administration of recombinant duck leptin inhibited food intake. Despite the decrease of food intake, leptin significantly induced body weight and fatty deposition. These changes were accompanied by a significant down-secretion of plasma glucose, cholesterol, triglyceride and insulin levels in mice. The observations provide evidence for an inhibitory effect of leptin in the regulation of food intake and for a potential role of duck leptin in the regulation of lipogenesis.

Drug Release and Skin Irritancy of Poloxamer Gel Containing Kojic Acid (코지산을 함유한 폴록사머 겔 제제의 약물방출 및 피부자극성)

  • Park, Eun-Woo;Cho, Seong-Wan;Kim, Dong-Sup;Choi, Ki-Hwan;Choi, Young-Wook
    • Journal of Pharmaceutical Investigation
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    • v.28 no.3
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    • pp.177-183
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    • 1998
  • Low toxicity, reverse thermal gelation and high drug loading capabilities suggest that poloxamer 407 gels have great potential as a topical drug delivery system. Kojic acid (KA) is an antimelanogenic agent which has been widely used in cosmetics to whiten the skin color. However, it has the drawbacks of skin irritancy due to its acidic pH. Poloxamer gels of different polymer contents were formulated to overcome the problem and compared to the cream type formulations of either w/o/w multiple emulsion cream or o/w type emulsion cream. Using Franz diffusion cells mounted with a synthetic cellulose membrane (MWCO 12,000), drug release characteristics of the formulations were evaluated by the HPLC assay of KA concentration in the receptor compartment of pH 7.4 phosphate buffered saline solutions. Drug release from w/o/w multiple emulsion cream was controlled by oil membrane, showing the apparent zero order release kinetics. The KA release from the poloxamer gels was also controlled by the gel matrix, showing that drug release increased linearly as KA contents increase, but decreased exponentially as the polymer contents increase. In the skin irritancy test, the primary irritancy index(PII) of poloxamer gel base was lower than those of multiple emulsion cream base and o/w cream. Depending on KA contents or polymer contents in the gel. PH values in poloxamer gels were ranged from 1.3 to 2.0, which are interpreted as low or negligible irritation on skin. There was a good correlation between the log value of flux in drug release and PII value in skin irritation. It was possible to conclude that the poloxamer gels containing KA might be a good candidate for an antimelanogenic topical delivery system by virtue of the controlled release of the drug and the reduced skin irritancy.

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Enterobacter aerogenes ZDY01 Attenuates Choline-Induced Trimethylamine N-Oxide Levels by Remodeling Gut Microbiota in Mice

  • Qiu, Liang;Yang, Dong;Tao, Xueying;Yu, Jun;Xiong, Hua;Wei, Hua
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1491-1499
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    • 2017
  • Trimethylamine N-oxide (TMAO), which is transformed from trimethylamine (TMA) through hepatic flavin-containing monooxygenases, can promote atherosclerosis. TMA is produced from dietary carnitine, phosphatidylcholine, and choline via the gut microbes. Previous works have shown that some small molecules, such as allicin, resveratrol, and 3,3-dimethyl-1-butanol, are used to reduce circulating TMAO levels. However, the use of bacteria as an effective therapy to reduce TMAO levels has not been reported. In the present study, 82 isolates were screened from healthy Chinese fecal samples on a basal salt medium supplemented with TMA as the sole carbon source. The isolates belonged to the family Enterobacteriaceae, particularly to genera Klebsiella, Escherichia, Cronobacter, and Enterobacter. Serum TMAO and cecal TMA levels were significantly decreased in choline-fed mice treated with Enterobacter aerogenes ZDY01 compared with those in choline-fed mice treated with phosphate-buffered saline. The proportions of Bacteroidales family S24-7 were significantly increased, whereas the proportions of Helicobacteraceae and Prevotellaceae were significantly decreased through the administration of E. aerogenes ZDY01. Results indicated that the use of probiotics to act directly on the TMA in the gut might be an alternative approach to reduce serum TMAO levels and to prevent the development of atherosclerosis and "fish odor syndrome" through the effect of TMA on the gut microbiota.

Anticandidal Activity of the Protein Substance from Coptidis Rhizoma (황련에서 분리된 단백질성분의 항진균효과)

  • Kim Hyunkyung;Lee Jue-Hee;Shim Jin Kie;Han Yongmoon
    • YAKHAK HOEJI
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    • v.49 no.4
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    • pp.323-329
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    • 2005
  • Antimicrobial peptides are evolutionary ancient weapons for animal and plant species to depend themselves against infectious microbes. In the present study, we investigated if an antimicrobial peptide was produced from Coptidis Rhizoma. For the determination, protein substance from the medicinal plant was isolated by various preparations. Among the preparations, the protein portion dissolved in phosphate-buffered saline solution (CRP-DS) that contained the most amount of protein $(90\%)$ resulted in maximal inhibition of Candida albicans which causes local and systemic infections. Analyses by gel-electrophoresis and gel-permeation chromatography showed the CRP-DS formed a single band of approximately 11.8 KDa as molecular size. Antifungal activity of the CRP-DS was almost equivalent to antifungal activity by fluconazole, resulting in MIC (minimal inhibitory concentration) of approximately $50{\mu}g/ml$. The antifungal activity was a dose-dependent. The antifungal activity appeared to be inactivated by heat-treatment and ionic strength, respectively. In a murine model, the CRP-DS enhanced resistance of mice against disseminated candidiasis. The HPLC analysis demonstrated maximum $4\%$ of berberine as residual content in the CRP-DS preparation resulted in no influence on the antifungal activity. In addition, protein portion isolated from Phellodendri Cortex producing the alkaloid component like Coptidis Rhizoma had no such anticandidal effect. These results indicate that the protein substance from Coptidis Rhizoma was responsible for the antifungal activity.

Preparation and Release Profile of N8f-loaded Polylactide Scaffolds for Tissue Engineered Nerve Regeneration (조직공학적 신경재생을 위한 NGF를 함유한 PLA 담체의 제조 및 방출)

  • 전은경;황혜진;강길선;이일우;이종문
    • Polymer(Korea)
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    • v.25 no.6
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    • pp.893-901
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    • 2001
  • We developed the nerve growth factor (NGF) loaded poly (L - lactide) (PLA) scaffolds by means of emulsion freeze drying method to the possibility for the application of the nerve regeneration of spinal cord disease and the degeneration in Alzheimer's disease. The release amount of NGF from NGF loaded PLA scaffold were analyzed over a 4 week period in vitro at phosphate buffered saline (PBS), pH 7.4, at $37^{\circ}C$. It can be observed the open cell pore structure of porous scaffolds and can be easily controlled the pore structure by the controlling of formulation factors resulting in the controlling of the release rate and the release period. The stability of NGF during the preparation of PLA scaffold was evaluated by comparing the released amounts of total NGF, assayed NGF enzyme - linked immunosorbent assay (ELISA). Released NGF has been found to enhance the neurite sprouting and outgrowth from pheochromocytoma (PC-12) cells. These results suggest that the released NGF from NGF loaded PLA scaffold such as conduit type can be very useful for the nerve regeneration in the neural tissue engineering area.

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Effect of Kinematic Motion on Changes in Coefficients of Friction of Porcine Knee Joint Cartilage (기구학적 운동이 돼지 무릎 관절연골의 마찰계수 변화에 미치는 영향)

  • Kim, Hwan;Kim, ChoongYeon;Lee, KwonYong;Kim, DaeJoon;Kim, DoHyung
    • Tribology and Lubricants
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    • v.29 no.1
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    • pp.46-50
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    • 2013
  • In this study, the frictional behaviors of articular cartilage against a Co-Cr alloy in two types of kinematic motions were compared. Cartilage pins were punched from the femoral condyles of porcine knee joints, and Co-Cr alloy disks were machined from orthopedic-grade rods and polished to a surface roughness ($R_a$) of 0.002. Friction tests were conducted by using a pin-on-disk-type tribotester in phosphate buffered saline (PBS) under pressures of 0.5, 1, and 2 MPa. All tests were performed in the repeat pass rotational (ROT) and the linear reciprocal (RCP) sliding motions with the same sliding distance and speed of 50 mm/s. The coefficients of friction of the cartilage against the Co-Cr alloy increased with the sliding time in both kinematic motions for all contact pressures. The maximum coefficients of friction in RCP motion were 1.08, 2.82, and 1.96 times those in ROT motion for contact pressures of 0.5, 1, and 2 MPa, respectively. As the contact pressure increased, the coefficients of friction gradually increased in RCP motion, whereas they decrease and then increased in ROT motion. The interaction between the directional change of the shear stress and the orientation of collagen fiber in the superficial layer of the cartilage could affect the change in the frictional behaviors of the cartilage. A large difference in the coefficients of friction between the two kinematic motions could be interpreted as differences in the directional change of shear stress at the contact surface.

The Modified Electrode by PEDOP with MWCNTs-Palladium Nanoparticles for the Determination of hydroquinone and Catechol

  • Naranchimeg, Orogzodmaa;Kim, Seul-Ki;Jeon, Seung-Won
    • Bulletin of the Korean Chemical Society
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    • v.32 no.8
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    • pp.2771-2775
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    • 2011
  • Poly-ethylenedioxypyrrole (PEDOP) coated thiolated multiwall carbon nanotubes palladium nanoparticles (MWCNTs-Pd) modified glassy carbon electrode (GCE) [PEDOP/MWCNTs-Pd/GCE] for the determination of hydroquinone (HQ) and it’s isomer catechol (CA) were synthesized and compared with bare GCE and thiolated multiwall carbon nanotubes (MWCNTs-SH/GCE). The modification could be made by simple processes on a GCE with MWCNTs-Pd covered by PEDOP in a 0.05 M tetrabutylammonium perchlorate (TBAP)/MeCN solution system. A well-defined peak potential evaluation of the oxidation of hydroquinone to quinone at 0.05 V (vs. Ag/AgCl), and electrochemical reduction back to hydroquinone were found by cyclic voltammetry (CV) in phosphate buffered saline (PBS) at pH 7.4. Peak current values increased linearly with increasing hydroquinone contents. The peak separation between the anodic and cathodic peaks at the PEDOP/MWCNTs-Pd/GCE was ${\Delta}Ep$ = 40 mV for HQ and ${\Delta}Ep$ = 70 mV for CA, resulting in a higher electron transfer rate. Moreover, good reproducibility, excellent storage stability, a wide linear range (0.1 ${\mu}M$ - 5 mM for HQ and 0.01 ${\mu}M$ - 6 mM for CA), and low detection limits ($2.9{\times}10^{-8}$ M for HQ and $2.6{\times}10^{-8}$ M for CA; S/N = 3) were determined using differential pulse voltammetry (DPV) and amperometric responses; this makes it a promising candidate as a sensor for determination of HQ and CA.

Anti-inflammatory Activities of Coumarins Isolated from Angelica gigas Nakai on LPS-stimulated RAW 264.7 Cells

  • Ma, Yong-Fen;Jung, Jae-Yeon;Jung, Yu-Jung;Choi, Ji-Hye;Jeong, Woo-Sik;Song, Young-Sun;Kang, Jae-Seon;Bi, Kaishun;Kim, Myo-Jeong
    • Preventive Nutrition and Food Science
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    • v.14 no.3
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    • pp.179-187
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    • 2009
  • Five kinds of coumarin compounds were successfully purified from Angelica gigas Nakai by using recyclingpreparative HPLC and identified as decursin (1), decursinol angelate (2), 7-demethylsuberosine (3), marmesin (4), and decursinol (5) by NMR analyses. None of the purified compounds in ethanol showed DPPH radical scavenging activity, while the A. gigas extract (AGNEX) displayed a significant level of activity. Interestingly, compounds 3 in phosphate buffered saline (PBS) showed good $ABTS^+$ radical scavenging activity ($IC_{50}=8.1{\mu}g$/mL) as did compounds 4 and 5. The anti-inflammatory activities of the purified compounds were evaluated and compared using the NO concentration assay and western blot analysis on LPS-stimulated RAW 264.7 cells. NO production was significantly suppressed by all the compounds in a dose-dependent manner among which compounds 1, 2, and 3 showed very good activities with $IC_{50}$ values of 7.4, 6.5, and $7.6{\mu}g$/mL, respectively. Treatment with compounds 1-5 effectively suppressed the expression levels of iNOS, IL-1$\beta$, and COX-2, which are responsible for promoting the inflammatory process. Thus, the ethanol extract and coumarin compounds of A. gigas Nakai hold promise for use as potential anti-inflammatory agents.

Self-organized Pullulan/Deoxycholic Acid Nanogels: Physicochemical Characterization and Anti-cancer Drug-releasing Behavior

  • Na, Kun;Park, Kyong-Mi;Jo, Eun-Ae;Lee, Kwan-Shik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.262-267
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    • 2006
  • The objective of this study was to develop new self-organized nanogels as a means of drug delivery in patients with cancer. Pullulan (PUL) and deoxycholic acid (DOCA) were conjugated through an ester linkage between the hydroxyl group in PUL and the carboxyl group in DOCA. Three types of PUL/DOCA conjugates were obtained, differing in the number of DOCA substitutions (DS; 5, 8, or 11) per 100 PUL anhydroglucose units. The physicochemical properties of the resulting nanogels were characterized by dynamic light scattering, transmission electron microscopy, and fluorescence spectroscopy. The mean diameter of DS 11 was the smallest (approx. 100 nm), and the size distribution was unimodal. To determine the organizing behavior of these conjugates, we calculated their critical aggregation concentrations (CACs) in a 0.01-M phosphate buffered saline solution. They were $10.5{\times}10^{-4}mg/mL,\;7.2{\times}10^{-4} mg/mL,\;and\;5.6{\times}10^{-4} mg/mL$ for DS 5, 8, and 11, respectively. This indicates that DOCA can serve as a hydrophobic moiety to create self-organized nanogels. To monitor the drug-releasing behavior of these nanogels, we loaded doxorubicin (DOX) onto the conjugates. The DOX-loading efficiency increased with the degree of DOCA substitution. The release rates of DOX from PUL/DOCA nanogels varied inversely with the DS. We concluded that the PUL/DOCA nanogel has some potential for use as an anticancer drug carrier because of its low CAC and satisfactory drug-loading capacity.