• Title/Summary/Keyword: Concentration Gradient

Search Result 583, Processing Time 0.024 seconds

Exploring the feasibility of Salmonella Typhimurium-specific phage as a novel bio-receptor

  • Choi, In Young;Park, Do Hyeon;Chin, Brayan A.;Lee, Cheonghoon;Lee, Jinyoung;Park, Mi-Kyung
    • Journal of Animal Science and Technology
    • /
    • v.62 no.5
    • /
    • pp.668-681
    • /
    • 2020
  • The purpose of this study was aimed to isolate a Salmonella Typhimurium-specific phage (KFS-ST) from washing water in a poultry processing facility and to investigate the feasibility of the KFS-ST as a novel bio-receptor for the magnetoelastic (ME) biosensor method. KFS-ST against S. Typhimurium was isolated, propagated, and purified using a CsCl-gradient ultracentrifugation. Morphological characteristics of KFS-ST were analyzed using transmission electron microscopy (TEM). Its specificity and efficiency of plating analysis were conducted against 39 foodborne pathogens. The temperature and pH stabilities of KFS-ST were investigated by the exposure of the phage to various temperatures (-70℃-70℃) and pHs (1-12) for 1 h. A one-step growth curve analysis was performed to determine the eclipse time, latent time and burst size of phage. The storage stability of KFS-ST was studied by exposing KFS-ST to various storage temperatures (-70℃, -20℃, 4℃, and 22℃) for 12 weeks. KFS-ST was isolated and purified with a high concentration of (11.47 ± 0.25) Log PFU/mL. It had an icosahedral head (56.91 ± 2.90 nm) and a non-contractile tail (225.49 ± 2.67 nm), which was classified into the family of Siphoviridae in the order of Caudovirales. KFS-ST exhibited an excellent specificity against only S. Typhimurium and S. Enteritidis, which are considered two of the most problematic Salmonella strains in the meat and poultry. However, KFS-ST did not exhibit any specificity against six other Salmonella and 27 non-Salmonella strains. KFS-ST was stable at temperature of 4℃ to 50℃ and at pH of 4 to 12. The eclipse time, latent time, and burst size of KFS-ST were determined to be 10 min, 25 min and 26 PFU/ infected cell, respectively. KFS-ST was relatively stable during the 12-week storage period at all tested temperatures. Therefore, this study demonstrated the feasibility of KFS-ST as a novel bio-receptor for the detection of S. Typhimurium and S. Enteritidis in meat and poultry products using the ME biosensor method.

Quantitative Analysis of Various Carotenoids from Different Colored Paprika Using UPLC (UPLC를 이용한 색상별 파프리카 유래 카로티노이드의 정량적 평가)

  • Hwang, Jeong Rok;Hwang, In Kyeong;Kim, Suna
    • Korean Journal of Food Science and Technology
    • /
    • v.47 no.1
    • /
    • pp.1-5
    • /
    • 2015
  • This study aimed to simultaneously determine various carotenoids from different colored paprika using an ultra performance liquid chromatograph (UPLC) equipped with a HSS T3 column. Analysis was performed at 450 nm using gradient conditions with acetonitrile/methanol/methylene chloride (65/25/10) and distilled water. We improved the peak resolution and performed carotenoid analysis within 30 min. We qualitatively analyzed 11 carotenoids (neoxanthin, capsorubin, violaxanthin, capsanthin, zeaxanthin, lutein, ${\alpha}$-cryptoxanthin, ${\beta}$-cryptoxanthin, lycopene, ${\alpha}$-carotene, and ${\beta}$-carotene). For the validation of UPLC methods, we validated the precision and accuracy of capsanthin. Capsanthin showed good linearity ($R^2$=0.9998) in the concentration range of $1-200{\mu}g/mL$ with 2.4 and $7.2{\mu}g/mL$ of limit of detection (LOD) and limit of quantification (LOQ), respectively. The relative standard deviation (RSD) for intra- and inter-day precision was less than 3.83%. Recovery was in the range of 91.86-99.87%. We quantitatively analyzed carotenoid contents from 8 different colored paprika (red, orange, yellow, and green). The most abundant carotenoids were capsanthin in red paprika, and zeaxanthin in orange, yellow, and green paprika.

Development of the Duplex PCR Method of Identifying Trachurus japonicus and Trachurus novaezelandiae (다중 PCR 분석법을 이용한 전갱이속 어종의 신속한 종판별 분석법 개발)

  • Park, Yeon Jung;Lee, Mi Nan;Kim, Eun Mi;Noh, Eun Soo;Noh, Jae Koo;Park, Jung Youn;Kang, Jung-Ha
    • Journal of Life Science
    • /
    • v.28 no.9
    • /
    • pp.1062-1067
    • /
    • 2018
  • Reliable labeling of fish products can reassure consumers regarding the identity and quality of seafoods. Therefore, techniques that can identify adulteration or mislabeling are valuable. To rapidly identify two Trachurus species, Trachurus japonicus and Trachurus novaezelandiae, a highly efficient, rapid, duplex polymerase chain reaction (PCR) having two species-specific primers simultaneously was identified. This species-specific primer focused on a single nucleotide mismatch in the 3'-terminal base of a primer designed in the mitochondrial cytochrome c oxidase (COI) subunit I DNA. To optimize the duplex PCR condition, gradient PCR reactions were conducted to determine the primer annealing temperature and the primer concentration. The PCR's product was observed on the gel, suggesting that DNA molecules may be useful in differentiating the two species. The length of the amplification fragments were 103 bp for Trachurus japonicus and 214 bp for Trachurus novaezelandiae, which, along with the species-specific primer visualized by agarose gel electrophoresis, enabled accurate distinction of the species of the Trachurus genus. These results indicate that the duplex PCR, which has a species-specific primer based on single nucleotide polymorphism (SNP), can be useful for rapidly differentiating the two species of Trachurus. This duplex PCR analysis is simple, rapid, and reliable, and could be beneficial to protecting consumers' rights.

pH Variance Model Depending on Phosphate Ion Form (인산염 이온 형태에 의한 pH 변량 모형)

  • Soh, Jae-Woo;Soh, Soon-Yil;Nam, Sang-Yong
    • Horticultural Science & Technology
    • /
    • v.33 no.6
    • /
    • pp.854-859
    • /
    • 2015
  • This experiment was performed to develop a model for nutrition ion concentration and EC in regard to change in pH from 4.0 to 8.0 in nutrient solution. The pH changes according to the variation of $HPO_4{^{-2}}$ and $H_2PO_4{^-}$ in the nutrient solution while variation of EC increased from pH 4.0 to 5.0, stabilized from pH 5.0 to 7.0 and increased again from pH 7.0 to 8.0. For the variance of major elements in the nutrient solution, K, Ca, N and P increased while pH was higher, especially the variables for K and P were increased largely. On the other hand, variables of Mg and S were stable. Based on analysis of the ion balance model of nutrient solution, the cation increased than anion over rising the variation of pH while balance point of ion moved from a-side to d-side. In addition, the imbalance increased while it moved away from the EC centerline as variance of pH increased. It was larger than effect of EC variance to correction values of equivalence ratios of K and Ca about variation of $HPO_4{^{-2}}$ and $H_2PO_4{^-}$ while as variance of pH increased, K decreased but Ca increased. These showed the result that variance of pH about correction values of equivalence ratios of K and Ca gave a second-degree polynomial model rating of 0.97. Through this research, it was identified the pH variable model about variance among pH, ion and EC according to gradient of phosphate.

Study on Heat-Loss-Induced Self-Excitation in Laminar Lifted Jet Flames (층류제트 부상화염에서 열손실에 의한 자기진동에 관한 연구)

  • Yoon, Sung-Hwan;Park, Jeong;Kwon, Oh-Boong;Kim, Jeong-Soo;Bae, Dae-Seok;Yun, Jin-Han;Keel, San-In
    • Transactions of the Korean Society of Mechanical Engineers B
    • /
    • v.35 no.3
    • /
    • pp.309-319
    • /
    • 2011
  • We experimentally investigated lifted propane jet flames diluted with nitrogen to obtain flame-stability maps based on heat-loss-induced self-excitation. We found that heat-loss-induced self-excitations are caused by conductive heat loss from premixed flame branches to trailing diffusion flames as well as soot radiation. The conductive-heat-loss-induced self-excitation at frequencies less than 0.1 Hz is explained well by a suggested mechanism, whereas the oscillation of the soot region induces a self-excitation of lift-off height of the order of 0.1 Hz. The suggested mechanism is also verified from additive experiments in a room at constant temperature and humidity. The heat-loss-induced self-excitation is explained by the Strouhal numbers as a function of the relevant parameters.

Role of Electrode Reaction of Electrolyte in Electrokinetic-Fenton Process for Phenanthrene Removal (동전기-펜턴 공정에서 전해질의 전극반응이 처리효율에 미치는 영향)

  • Park Ji-Yeon;Kim Sang-Joon;Lee You-Jin;Yang Ji-Won
    • Journal of Soil and Groundwater Environment
    • /
    • v.11 no.1
    • /
    • pp.7-13
    • /
    • 2006
  • The effects of electrolytes were investigated on the removal efficiency when several different electrolytes were used to change the electrode reaction in an electrokinetic (EK)-Fenton process to remediate phenanthrene-contaminated soil. Electrical potential gradient decreased initially due to the ion entrance into soil and then increased due to the ion extraction from soil under the electric field. Accumulated electroosmotic flow was $NaCl>KH_2PO_4>MgSO_4$ at the same concentration because the ionic strength of $MgSO_4$ was the highest and $Mg(OH)_2$ formed near the cathode reservoir plugged up soil pore to inhibit water flow. When hydrogen peroxide was contained in electrolyte solution, removal efficiency increased by Fenton reaction. When NaCl was used as an electrolyte compound, chlorine ($Cl_2$) was generated at the anode and dissolved to form hypochlorous acid (HClO), which increased phenanthrene removal. Therefore, the electrode reaction of electrolyte in the anode reservoir as well as its transport into soil should be considered to improve removal efficiency of EK-Fenton process.

Extraction of Glycosaminoglycan from Sea Hare, Aplysia kurodai, and Its Functional Properties 1. Optimum Extraction of Polysaccharide and Purification of Glycosaminoglycan (군소(Aplysia kurodai)에 분포하는 글루코사미노글리칸의 추출과 기능특성 1. 다당류 추출의 최적화와 글루코사미노글리칸의 정제)

  • Yoon, Bo-Yeong;Choi, Byeong-Dai;Choi, Yeung-Joon
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.39 no.11
    • /
    • pp.1640-1646
    • /
    • 2010
  • The optimum condition was investigated for the extraction of glycosaminoglycan (GAG) from sea hare, Aplysia kurodai. The most effective enzyme was Flavourzyme for extraction of glycosaminoglycan. The optimum incubation temperature and time for hydrolysis were $60^{\circ}C$ and 15 hr, respectively. The yield of precipitated polysaccharide depended on Brix and ethanol volume. The most effective concentration of Brix and ethanol were sixty and 5 volume of ethanol, respectively. Most GAG was eluted between 0.5 M and 0.75 M NaCl gradient on DEAE-Sepharose column, and identified by electroconductivity. The contents of hexuronic acid from polysaccharide extract and GAG were 1.0 g/100 g and 6.0 g/100 g, respectively. Hexosamine of polysaccharide and GAG as indicator of GAG component was 5.6 g/100 g and 25.7 g/100 g, respectively. GAG was identified as heparan sulfate compared with bands of other GAG on agarose gel electrophoresis, and its molecular weight was 29.6 kDa on Superdex 200 HR column.

Isolation and Development of Quantification Method for Cyanidin-3-Glucoside and Cyanidin-3-Rutinoside in Mulberry Fruit by High-Performance Countercurrent Chromatography and High-Performance Liquid Chromatography

  • Choi, Soo-Jung;Jeon, Heejin;Lee, Chang Uk;Yoon, Shin Hee;Bae, Soo Kyung;Chin, Young-Won;Yoon, Kee Dong
    • Natural Product Sciences
    • /
    • v.21 no.1
    • /
    • pp.20-24
    • /
    • 2015
  • Cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) were isolated by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water/trifluoroacetic acid (1 : 3 : 1 : 5 : 0.01, v/v) to give pure C3G (34.1 mg) and C3R (14.3 mg) from 1.5 g crude mulberry fruit extract. Using the pure C3G and C3R, a reliable high-performance liquid chromatography (HPLC) method was developed and validated to determine the C3G and C3R contents in mulberry fruit. C3G and C3R were separated simultaneously using an Eclipse XDB-C18 column ($4.6{\times}250mm$ I.D., $5{\mu}m$) coupled with a photodiode array detector (PDA). The gradient elution of the mobile phase consisting of acetonitrile (0.5% formic acid) and water (0.5% formic acid) was applied (1.0 mL/min), and the detection wavelength was 520 nm. The calibration curves of C3G and C3R showed good linearity (both with $r^2=0.9996$) in the concentration range $15.625-500{\mu}g/mL$, and the relative standard deviations (RSD%) of intra- and inter-day variability were in the ranges 2.1 - 8.2% and 4.1 - 17.1%, respectively. The accuracies were ranged 96.5 - 102.6% for C3G and C3R, respectively. The developed HPLC method was used to determine the contents of C3G and C3R in newly harvested mulberry from eight different provinces of Korea.

Inositol 1,4,5-Trisphosphate-induced Increase in $Ca^{2+}-ATPase$ Activity in the Microsomes of Tracheal Epithelial Cells

  • Cho, Hyoung-Jin;Park, Sung-Shin;Kim, Young-Kee
    • The Korean Journal of Physiology
    • /
    • v.29 no.2
    • /
    • pp.269-277
    • /
    • 1995
  • Membrane vesicles were prepared by differential centrifugation from epithelial cells of porcine trachea. Total activity of microsomal ATPases was measured spectrophotometrically by a coupled enzyme assay. The steady-state activity of the enzyme was $329{\pm}10$ nmol/min mg protein. Thapsigargin, a specific antagonist of intracellular $Ca^{2+}-ATPase$, inhibited about 50% of the activity, leaving $178{\pm}18\;nmol/min .mg$ protein (n=6), indicating that the $Ca^{2+}-ATPase$ is one of the major microsomal ATPases. The microsomes used in this study appeared to be tight-sealed vesicles since they showed saturation in $^{45}Ca^{2+}$ uptake experiments. Inositol 1,4,5-trisphosphate $InsP_{3}, 4\;{\mu}M$, an agonist of $InsP_{3}$-sensitive $Ca^{2+}$ release channel ($InsP_{3}$, receptor), and Ca-ionophore A23187 $(10\;{\mu}M)$ induced $^{45}Ca^{2+}$ releases of 20% and 50% of stored $^{45}Ca^{2+}$, respectively. The addition of $(10\;{\mu}M\;InsP_{3}$ also increased the microsomal ATPase activity from $282{\pm}8$ nmol/min mg protein to $334{\pm}21$ nmol/min . mg protein in the intact vesicles. Similar increase in the activity was observed by making microsomes leaky (uncoupling) using the Ca-ionophore A23187. ;$InsP_{3}-induced$ effects were blocked by either thapsigargin or heparin suggesting that: 1) the $InsP_{3}-induced$ increase in ATPase activity is mediated by microsomal $Ca^{2+}-ATPase$, and 2) dissipation of $Ca^{2+}$ gradient across the microsomal membrane is responsible for the $InsP_{3}-induced$ effect. In order to test the dependence of the $Ca^{2+}-ATPase$ activity on the activity of $InsP_{3}-induced$ the activity of ATPases was monitored in various concentrations of free $Ca^{2+}$ using $EGTA-Ca^{2+}$ buffers. The $Ca^{2+}$-dependent biphasic change is the well-known character of $InsP_{3} receptor but not of microsomal $Ca^{2+}-ATPase$ in non-excitable cells; however, the activity of microsomal ATPase appeared biphasic and a maxim진 activity of $397{\pm}36nmol/min\;.mg$ protein was obtained in the solution containing 100 nM free $Ca^{2+}$. Below or above this concentration, the activity of ATPases was lower. These results strongly support a positive correlation of microsomal $Ca^{2+}-ATPase$ to the $InsP_{3}$ receptors in epithelial microsomes.

  • PDF

Multiresidue Determination of Tetracyclines in Eggs using Liquid Chromatography with Ultraviolet Detection (액체크로마토그래피를 이용한 계란 중 테트라사이클린계 항생물질의 동시분석법 개발)

  • Lee, Sang-Hee;Shim, You-Sin;Choi, Yoon-Hee;Lee, Beom-Gil;Kim, Hyun-Ju;Shin, Dong-Bin
    • Journal of Food Hygiene and Safety
    • /
    • v.22 no.4
    • /
    • pp.370-374
    • /
    • 2007
  • An analytical method for the simultaneous determination of four tetracycline (oxytetracycline, tetracycline, chlortetracycline, doxycycline) in egg samples was developed and validated using liquid chromatography with ultraviolet detection. Egg samples were extracted by the liquid-liquid extraction based on acetonitrile. The chromatographic separation was achieved on a reverse phase C8 column with gradient elution using a mobile phase of 20 mM oxalic acid (pH 1.5)/acetonitrile. The procedure was validated according to the Food Drugs Administration guideline determining accuracy, precision, and limit of detection. Mean recovery of tetracyclines from spiked egg samples (50, 100, 200, 400, and $800{\mu}g/kg$) were 78.8-109.3%. Linearity in concentration range of $50-800{\mu}g/kg$ was obtained with the correlation coefficient $(r^2)$ of 0.994-0.999. The intra- and inter-day precision (relative standard deviation; RSD) was between 0.3-12.8 and 0.2-11.7%, respectively. Limit of detection (LOD) and limit of quantification (LOQ) for the investigated tetracyclines were 30 and $50{\mu}g/kg$ depending on egg samples, respectively. This method was reliable, sensitive, economical and suitable for routine monitoring of tetracycline residues in dairy egg.