• Title/Summary/Keyword: phosphate-buffered saline

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Detection of Protein Molecules by Electrical Current Response Using Two-Electrode Method

  • Lyu, Hong-Kun;Woo, Sung-Ho;Han, Yoon-Soo;Lee, Hee-Ho;Shin, Jang-Kyoo
    • Journal of Sensor Science and Technology
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    • v.20 no.2
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    • pp.90-95
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    • 2011
  • In order to protect human lives from disease, various biosensors having the potential to analyze a variety of biomolecules have been utilized. Biosensors constitute one of the most promising ways to monitor and detect various biomolecules corresponding to diseases. In this study, we demonstrate that the reaction of streptavidin molecules with biotin on a gold electrode can be detected using the twoelectrode method with a gold electrode and a platinum reference electrode. We also show the characteristics of the electrical current response. While detecting 2-${\mu}M$ streptavidin molecules dissolved in phosphate buffered saline(PBS) solution, we found that an analytical biosensor can operate on the principle of detecting an antigen-antibody reaction event of protein molecules using the two-electrode method. We think that the "potential step" method might be useful to detect the occurrence of any antigen-antibody reactions and can be combined with other devices or ICs such as BJTs, MOSFETs, and OP-amps for the detection of biomolecules of diseases.

PCR Diagnosis of Entamoeba histolytica Cysts in Stool Samples

  • Moon, Joung-Ho;Cho, Shin-Hyeong;Yu, Jae-Ran;Lee, Won-Ja;Cheun, Hyeng-Il
    • Parasites, Hosts and Diseases
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    • v.49 no.3
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    • pp.281-284
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    • 2011
  • Amebiasis is a protozoan disease caused by Entamoeba histolytica and a potential health threat in areas where sanitation and hygiene are inappropriate. Highly sensitive PCR methods for detection of E. histolytica in clinical and environmental samples are extremely useful to control amebiasis and to promote public health. The present study compared several primer sets for small subunit (SSU) rDNA and histone genes of E. histolytica cysts. A 246 bp of the SSU rDNA gene of pure cysts contained in phosphate-buffered saline (PBS) and in stool samples was successfully amplified by nested PCR, using the 1,147-246 bp primer set, of the primary PCR products which were pre-amplified using the 1,147 bp primer as the template. The detection limit of the nested PCR using the 1,147-246 primer set was 10 cysts in both groups (PBS and stool samples). The PCR to detect histone gene showed negative results. We propose that the nested PCR technique to detect SSU rDNA can be used as a highly sensitive genetic method to detect E. histolytica cysts in stool samples.

Screening of Immune-Enhancing Substance(s) from Korean Wheats (우리밀의 면역증강능 규명)

  • Choe, Myeon;Park, Jae-Bong;Kim, Hyun-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.2
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    • pp.307-311
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    • 2000
  • The purpose of this study was to identify excellent immune-enhancing substance from Korean wheats(Eunpa, Gueru, Alchan, Topdong, Suwon 267, Gobun) compared with imported ones(Australian standard white, ASW; Dark northern spring, DNS). Phagocytic activities of PBS (phosphate buffered saline, pH 7.4) and EA(ethanol-acetic acid) extracts from the wheats were determined using mouse macrophage J774 cell line. In order to set the optimal experimental condition up, the cultured cells were tested in varying experimental conditions. About two to five times higher phagocytic activity was shown in EA extract of Korean wheats compared to that of imported wheats. PBS extracts of wheats did not show increased phagocytic activity compared to control that did not add any extract. The EA extract of Gobun wheat showed the highest phagocytic activity. From the experiment we found that the optimal experimental condition was shown in two hours of reaction time and 0.05mg amout of EA extract added to J774 cells.

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Impact of pH on the response of bovine serum albumin to gold surface plasmon resonance chip (소 혈청 알부민의 금 표면 플라즈몬 공명 칩과의 반응에 대한 pH의 영향)

  • Sohn, Young-Soo
    • Journal of Sensor Science and Technology
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    • v.30 no.5
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    • pp.326-330
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    • 2021
  • Reactions between gold (Au) surface plasmon resonance (SPR) chips and bovine serum albumin (BSA) dissolved in solutions of different pH were investigated. The charge on the BSA depends on the pH of the solution in which it is dissolved. Thus, dissolving BSA in different pH solutions resulted in different charges of BSA. Among the BSA dissolved in solutions with pH 4.01, 7.4, and 10.01, the SPR response was the highest for BSA dissolved in the solution of pH 4.01. To eliminate the response variation owing to the difference in the refractive indices of the solutions, phosphate buffered saline (PBS) was injected into the system after the reaction of BSA with the Au SPR chip had happened. In this case too, the BSA dissolved in the solution with pH 4.01 exhibited the highest response. This may be attributed to the non-uniform distribution of ionic patches on the BSA, which can induce electrostatic attraction to the surface even though BSA has a positive charge at pH 4.01, and the absolute values of the net charge of BSA at pH 4.01 and 7.4 were very close.

Effects of Ionic Strength in the Medium on Sample Preconcentration Utilizing Nano-interstices between Self-Assembled Monolayers of Gold Nanoparticles

  • Nguyen, Ngoc-Viet;Wu, Jian-Sheng;Jen, Chun-Ping
    • BioChip Journal
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    • v.12 no.4
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    • pp.317-325
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    • 2018
  • This paper investigated the effects of ionic strength in the medium on a preconcentrator for a protein sample with low concentration. The preconcentration chip was designed and fabricated using a polydimethylsiloxane replica through standard lithophotography. A glass substrate is silanized prior to functionalizing the nanoparticles for self-assembly at a designed region. Due to the overlap of electrical double layers in a nanofluidic channel, a concentration polarization effect can be achieved using an electric field. A nonlinear electrokinetic flow is induced, resulting in the fast accumulation of proteins in front of the induced ionic depletion zone, so called exclusion-enrichment effect. Thus, the protein sample can be driven by electroosmotic flow and accumulated at a specific location. The chip is used to collect fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) diluted in phosphate-buffered saline (PBS) buffer solution. Different concentrations of the buffer media were studied herein. Fluorescence intensity images show that the buffer concentration of 4 mM is more appropriate than all the other ones. The sample of FITC-BSA with an initial concentration of $10{\mu}M$ in the 4 mM PBS solution increases its concentration at the desired region by up to 50 times within 30 min, demonstrating the results in this investigation.

A new in vitro method for evaluating the antimicrobial activity of toothpaste

  • Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.46 no.2
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    • pp.94-97
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    • 2021
  • The purpose of this study was to introduce a new in vitro method for evaluating the antimicrobial activity of toothpaste, reflecting the actual toothbrushing time and the dilution of toothpaste by salivation. We designed three experimental groups and one negative control group. The experimental groups were (1) 90 μL of toothpaste + 10 μL 1X phosphate-buffered saline (PBS, 9/10 dilution group), (2) 50 μL of toothpaste + 40 μL 1X PBS (1/2 dilution group), and (3) 25 μL of toothpaste + 65 μL 1X PBS (1/4 dilution group). During toothbrushing, saliva is continuously secreted into the oral cavity and the toothpaste concentration is diluted over time during toothbrushing. Therefore, the 1/2 and 1/4 dilution experimental groups were added. The negative control group was toothpaste diluted 20,000-fold with 1X PBS. Miracle Fresh Doctor toothpaste and Streptococcus mitis KCOM 1350, Prevotella intermedia KCOM 1107, Fusobacterium nucleatum subsp. polymorphum KCOM 1322, and Aggregatibacter actinomycetemcomitans KCOM 1306 were used as the toothpaste and target bacterial strains, respectively. The number of bacterial cells plated on agar plates in the negative control group was 1,000 CFU. If the number of colonies on the experimental group plate was less than one, the treatment was considered to have > 99.9% bactericidal activity. These results suggest that this new in vitro method for antimicrobial evaluation could be used as the standard method for testing the antimicrobial activity of toothpaste.

The Therapeutic Role of Nanoparticle Shape in Traumatic Brain Injury : An in vitro Comparative Study

  • Youn, Dong Hyuk;Jung, Harry;Tran, Ngoc Minh;Jeon, Jin Pyeong;Yoo, Hyojong
    • Journal of Korean Neurosurgical Society
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    • v.65 no.2
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    • pp.196-203
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    • 2022
  • Objective : To perform a comparative analysis of therapeutic effects associated with two different shapes of ceria nanoparticles, ceria nanorods (Ceria NRs) and ceria nanospheres (Ceria NSs), in an in vitro model of traumatic brain injury (TBI). Methods : In vitro TBI was induced using six-well confluent plates by manually scratching with a sterile pipette tip in a 6×6-square grid. The cells were then incubated and classified into cells with scratch injury without nanoparticles and cells with scratch injury, which were treated separately with 1.16 mM of Ceria NSs and Ceria NRs. Antioxidant activities and anti-inflammatory effects were analyzed. Results : Ceria NRs and Ceria NSs significantly reduced the level of reactive oxygen species compared with the control group of SH-SY5Y cells treated with Dulbecco's phosphate-buffered saline. The mRNA expression of superoxide dismutases was also reduced in nanoparticle-treated SH-SY5Y cells, but apparently the degree of mRNA expression decrease was not dependent on the nanoparticle shape. Exposure to ceria nanoparticles also decreased the cyclooxygenase-2 expression, especially prominent in Ceria NR-treated group than that in Ceria NS-treated group. Conclusion : Ceria nanoparticles exhibit antioxidant and anti-inflammatory effects in TBI models in vitro. Ceria NRs had better anti-inflammatory effect than Ceria NSs, but showed similar antioxidant activity.

Isolation and Characterization of Halophilic Kocuria salsicia Strains from Cheese Brine

  • Youn, Hye-Young;Seo, Kun-Ho
    • Food Science of Animal Resources
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    • v.42 no.2
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    • pp.252-265
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    • 2022
  • Kocuria salsicia can survive in extreme environments and cause infections, including catheter-related bacteremia, in humans. Here, we investigated and evaluated the characteristics of nine K. salsicia strains (KS1-KS9) isolated from cheese brine from a farmstead cheese-manufacturing plant in Korea from June to December, 2020. Staphylococcus aureus American Type Culture Collection (ATCC) 29213 was used as a positive control in the growth curve analysis and biofilm-formation assays. All K. salsicia isolates showed growth at 15% salt concentration and temperatures of 15℃, 25℃, 30℃, 37℃, and 42℃. KS6 and KS8 showed growth at 5℃, suggesting that they are potential psychrotrophs. In the biofilm-formation analysis via crystal violet staining, KS6 exhibited the highest biofilm-forming ability at various temperatures and media [phosphate buffered saline, nutrient broth (NB), and NB containing 15% sodium chloride]. At 25℃ and 30℃, KS3, KS6, and KS8 showed higher biofilm-forming ability than S. aureus ATCC 29213. The antimicrobial resistance of the isolates was evaluated using the VITEK® 2 system; most isolates were resistant to marbofloxacin and nitrofurantoin (both 9/9, 100%), followed by enrofloxacin (7/9, 77.8%). Five of the nine isolates (5/9, 55.6%) showed multidrug resistance. Our study reports the abilities of K. salsicia to grow in the presence of high salt concentrations and at relatively low temperatures, along with its multidrug resistance and tendency to form biofilms.

Development of Enzyme Linked Immunosorbent Assay for Determination of Zearalenone in Animal Feeds (가축사료 중 Zearalenone 분석을 위한 Enzyme Linked Immunosorbent Assay법의 개발)

  • 하정기;정덕화;김성영
    • Journal of Food Hygiene and Safety
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    • v.6 no.3
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    • pp.111-117
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    • 1991
  • We examined to develop the enzyme linked immunosorbent assay (ELISA) for determination of zearalenone in animal feeds. Zearalenone was first converted to 6'-(carboxymethyl) zearalenone oxime(zearalenone oxime) to get a coupling site and then conjugated to bovine serum albumin(BSA) for use as immunogen and to horseradish peroxidase(HRP) for use as enzyme marker. Antibody against zearalenone was obtained after 11 weeks of immunization of rabbit with zearalenone oxime-BSA. Cross reactivity of the antibody with ${\alpha}-zearalanol,\;{\beta}-zearalenol,\;{\alpha}-zearalanol\;and\;{\beta}-zearalanol$ were 168, 46, 26 and 20% respectiviely. A simple procedure was devised for the screening of zearalenone in feeds using ELISA. Feeds samples(5g) were extracted by blending with 25 ml of methanol-phospate butTered saline-dimethylformate(70 : 29 : 1) and the extract was filtered and aqueous filterate analyzed. It took only 1 hours to do whole procedure for the analysis of zearalenone in feeds by the direct competitive ELISA, and detectable limit was 1-100 ppb. Using this procedure, only 4 of 24 feed samples showed positive results with 3.93-7.43 ppb levels.

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Seasonal Changes in Concentrations of Proteins and Lipids in Growing Goat Oocytes

  • Sangha, G.K.;Bhatia, H.;Khera, K.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.1
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    • pp.36-40
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    • 2007
  • Proteins and lipids not only provide a source of energy to the cell, but also play vital roles in modifying the physical properties and function of the biological membranes. In the present study, we investigated the biochemical constituents, viz. proteins and lipids, in growing oocytes of goat antral follicles during summer and winter seasons. Goat genitalia in phosphate buffered saline (pH 7.4) were brought to the laboratory within one hour of slaughter under aseptic conditions at $37^{\circ}C$. Oocytes were aspirated from normal small (<3 mm in diameter) and large (>3 mm) follicles and pooled for biochemical estimations. A significant increase in the amount of protein and lipid was observed with the growth of the oocyte. The amount of protein varied non-significantly with the season, while the amount of lipid varied significantly. The amounts of phospholipid, cholesterol, free fatty acid, and triglyceride increased with the growth of the oocyte, but no significant effect of season in these constituents was observed. Lysolecithin, sphingomyelin, and sterols were the polar lipids identified in both oocytes prepared from small follicles (small oocytes) as well as large follicles (large oocytes). In addition, the small oocytes also contained phosphatidyl serine, while large oocytes contained phosphatidyl glycerol phosphate and phosphatidyl inositol. Among non-polar lipids, triglycerides and long chain alcohols appear only in small oocytes and not in large oocytes. Monoglycerides, 1,2-diglycerides, 1,3-diglycerides and o-dialkyl glycerol ethers, fatty acids, fatty acid methyl esters, and wax esters were identified in both small and large oocytes. Information on biochemical composition of growing oocytes is relevant to oocyte and embryo competence, culture and cryopreservation.