• Korean Journal of Microbiology
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• v.39 no.1
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• pp.27-35
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• 2003

No currently available methods to monitor pathogenic protozoa, Cryptosporidium and Giardia in environmental water come close to acceptable sensitivity, specificity and reproducibility, and so it has to be accompanied by thorough quality control and performance evaluation to credibly predict the distribution of them. We collected surface water samples from the Han River and spiked our prepared (oo)cysts, determined Matrix Spike recoveries using USEPA Method 1623 and considered what factors influence MS recovery and validity. As a result, average 46% of spiked oocysts and 60% of spike cysts were recovered, but repetitive sampling and statistical approach seemed to be necessary to determine the environmental pollution level of two protozoa as their variation coefficients was so much as 35oio and 26%. And MS recoveries with two acid dissociations during immunomagnetic separation were improved more 10% than that with one dissociations and the use of spiked suspension enumerated by flow cytometry instead of manual preparation enhanced the validity and reliability in spiking tests. Because fluorescence characteristics of (oo)cysts stained on well slides with FITC-labeled monoclonal antibodies and DAPI was not always same, well Elides from spiked field samples were helpful to evaluate the performance of staining. We found many (oo)cyst-like objects with typical fluorescence, not (co)cysts, from the Han River water samples, and then it was concluded that nuclei staining by DAPI (4',6-diamidino-2-phenylindole) and examination by Differential Interference Contrast Microscope should be critical for valid identification.

• The korean journal of orthodontics
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• v.28 no.6 s.71
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• pp.915-926
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• 1998

The cytoskeleton has been shown to form a network, connecting the extracelluar matrix via integrin with the nucleus and the cytoplasmic constituents of the cell. It is therefore assumed that the cytoskeleton may mediate signals generated by perturbations originating in the matrix. The purpose of this study is to examine the effect of cytoskeletal change on osteoblastic cell activities. The author cultured osteoblastic cells obtained from neonatal mouse calvaria. The cells were teated with cytochalasin B(CB) or colchicine (COL) at four concentrations for 3 hours and after another 24 hours the conditioned media was collected and assayed for prostaglandin $E_2\;(PGE_2)$, interleukin-6(IL-6), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and matrix metalloproteinase-1(MMP-1). In addition, the cytoskeletal protein actin were observed by immuno-fluorescence. The results were as follows: 1. The production of $PGE_2$ showed the tendency to be increased in CB-treated group. $PGE_2$ was increased in COL-treated group dose-dependantly, 2. IL-6 production, in CB-treated group, was increased, except at 1.0 ${\mu}g/ml$. IL-6 was induced in COL-treated group. 3. TNF-$\alpha$ production was increased in CB-treated group, except at 1.0 ${\mu}g/ml$, and in COL-treated group, that was increased. 4. The MMP-1 production was decreased in CB-treated soup and was not changed in COL-treated group, which could be selectively visualized by immunoblotting with monospecific antibody. 5. The cytoskeletal actin stress fibers were disappeared and the cells showed to be rounded in CB-treated group. These results indicated that there are a relationship between the cytoskeletal rearrangements and osteoblastic cell activities, especially in release of paracrine/autocrine factors, such as $PGE_2$, IL-6, and TNF-$\alpha$.

• Journal of Bio-Environment Control
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• v.27 no.4
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• pp.371-380
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• 2018

Aim of this study was to investigate the effects of different nutrient solutions and various light qualities generated by LED on the growth and glucosinolates contents of watercress (Nasturtium officinale) grown under hydroponics for 3 weeks. The seeds of watercress were sown on crushed rockwool media and raised them for two weeks. They were transplanted in a semi-DFT (deep flow technique) hydroponics system. A controlled-environment room was maintained at $20{\pm}1^{\circ}C$ and $16{\pm}1^{\circ}C$ temperatures and $65{\pm}10%$ and $75{\pm}10%$ relative humidity (day and night, respectively), with a provided photosynthetic photon flux density (PPFD) of $180{\pm}10{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ and a photoperiod of 16/8h. To find out the best kinds of nutrient solutions for growing watercress, Otsuka House 1A (OTS), Horticultural Experiment Station in Korea (HES), and Netherland's Proefstaion voor Bloemisterij en Gasgroente (PBG) were adapted with initial EC of $1.0-1.3dS{\cdot}m^{-1}$ and pH of 6.2, irradiating PPFD with fluorescent lamps (Ex-1). Either monochromatic (W10 and R10) or mixed LEDs (R5B1, R3B1, R2B1G1, and W2B1G1) were irradiated with a differing ratio of each LED's PPFD to understanding light quality on the growth and glucosinolates contents of watercress (Ex-2). Although significant difference in the shoot growth of watercress was not found among three nutrient solutions treatments, but the root fresh weight increased by 13.7% and 55.1% in PBG and OTS compared to HES, respectively. OTS increased the gluconasturtiin content by 96% and 65% compared to PBG and HES. Compared with the white light (W10), the red light (R10) showed a 101.3% increase in the shoot length of watercress. Increasing blue light portion positively affected plant growth. The content of total glucosinolates in watercress was increased by 144.5% and 70% per unit dry weight in R3B1 treatment compared with R2B1G1 and W10 treatments, respectively. The growth and total glucosinolates contents of the watercress were highest under R3B1 among six light qualities.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.205-214
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• 2002

There are many methods to introduce exogenous DNA into embryo to produce transgenic animals. Exogenous gene can be integrated into oocyte by sperm vector. In this study, sperm was used as a vector for a transgene, which is encoding enhanced green fluorescent protein (EGFP). The objective of this study was to investigate the expression of exogenous gene in bovine embryos after injection of spermatozoa cocultured with EGFP DNA fragment. Spermatozoa were plunged into liquid nitrogen and thawed several times or shook in 0.2% Triton X-100 to remove sperm membrane followed by DTT treatment. The injected oocytes were co-cultured with vero cells in CR1aa, and expression of EGFP gene was observed under fluorescent microscope. Blastocyst formation rates of oocytes injected with sperm treated with DTT, DTT-freezing or DTT-Triton X-100 were 34.7, 39.4 and 31.9%, respectively. The rates of EGFP expression in oocytes injected with 54 ng DNA after DTT-treated, DTT-freezing and DTT-Triton X-100-treated sperm were 0, 19.1 and 13.9%. On the other hands, expression rate of oocytes injected with sperm cocultured with 13.5, 27 and 63.5 ng of EFGP DNA were 6.7, 9.0 and 5.1%, respectively. When intact sperm was mixed with 63.5 ng/${mu}ell$ EGFP DNA fragment, and then electroporated before injection, the expression rate of injected oocyte was 2%. Unexpectedly, electro-poration could not increase the expression rate. These results suggest that sperm can be used as a transgene vector, even if the efficiency was low (19.1%).

• Journal of Digital Convergence
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• v.12 no.6
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• pp.407-414
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• 2014

The aim of this study was to evaluate the photodynamic therapy effects against staphylococci using Photofrin and Radachlorin with Light emitting diode(LED). Experimental methods, The bacterial suspensions containing Staphylococcus aureus and Staphylococcus epidermidis at $1{\times}10^5$ were prepared and diluted to different concentrations of photosensitizer, Photofrin or Radachlorin, on 1.25, 2. 5,5 and $10{\mu}g/ml$. The bacterial suspensions were exposed to 630 and 670 nm LED light at the energy density of 14.4 and $19.8J/cm^2$, respectively. The CFU results of S. aureus and S. epidermidis were showed 33 and 50 colony forming at $5{\mu}g/ml$ of Photofrin, respectively and both of them perfectely were dead at $5{\mu}g/ml$ of Radachlorin. The fluorescent intensity by flow cytometry was showed the increase in the dead cells than the normal cells. In the TEM photograph, the damage of bacterial membrane and the distortion of cell morphology were observed. These results suggest that photodynamic therapy combine with Photofrin and Radachlorin can be applied a new modality for antibacterial therapy.

• Journal of fish pathology
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• v.9 no.1
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• pp.65-77
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• 1996

Tachyplesin I is an antimicrobial peptide isolated from horseshoe crab. To investigate the mechanism of action of tachyplesin I for phospholipid bilayers, tachyplesin I and five analogs have been synthesized by the solution method. The synthesized five analogs are [$Phe^2$]-tachyplesin I, [$Phe^{8,13}$]-tachyplesin I, [$Cys(Acm)^{3,7,12,16}$]-tachyplesin I with no disulfide bonds, 7(Acm) and 10 (Acm) which denote the fragments [$Cys(Acm)^{3,7,12,16}$]-tachyplesin I. Circular dichroism spectra showed that tachyplesin I took an antiparallel $\beta$-structure in buffer solution and a less ordered structure in acidic liposomes. The carboxyfluorescein leakage experiment indicated that tachyplesin I interacted strongly with neutral and acidic phospholipid bilayers. In fluorescence experiment, the hydrophobic part of the peptide was shown to be embedded in lipid bilayers. All the peptides except for 7(Acm) and 10(Acm) were almost equally active in lipopolysaccharide binding. Therefore, the present study suggested that phospholipid bilayers induced a conformational change of tachyplesin I from the stable $\beta$-structure to a less ordered one.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.5
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• pp.589-594
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• 2007

The effect of synergists having chelating ability on inhibiting browning were studied with a giucose-glutamic acid model for doenjang containing citric acid as the anti-browning agent and iron ion. The model solutions were prepared by dissolving 0.1 M glucose, 0.1 M glutamic acid, 50 mM citric acid, 0.2 mM $FeCl_2$ and synergist in 1 M phosphate buffer (pH 7.0), heating at $50^{\circ}C$ for 24 hr and storing at $30^{\circ}C\;or\;40^{\circ}C$ for four weeks. Synergists were chitosan, gallic acid, methyl benzoate, pyrophosphate and tannic acid; they were used at the following concentrations: gallic acid, pyrophosphate and tannic acid at 0.015% and 0.15%; chitosan and methyl benzoate at 0.0075% and 0.015%. Anti-browning capacities had a tendency to decrease greatly after three weeks in the case of storage at $30^{\circ}C$, whereas they decreased with storage time during storage at $40^{\circ}C$. However, anti-browning capacities of samples containing 0.015% tannic acid and 0.15% pyrophosphate were higher than that of sample without synergist by 32% after storage at $30^{\circ}C$ for four weeks. Gallic acid, tannic acid and pyrophsphate also inhibited the formation of Maillard reaction intermediates such as fluorescent compound and 3-deoxyglucosone due to the high chelating ability with iron ion after four weeks of storage at $30^{\circ}C$. The effect of these compounds on the inhibition of formation of Maillard reaction intermediates was higher at 0.15% than at 0.015%. Moreover, gallic acid increased the browning by forming colored complexes, and tannic acid generated black precipitates. Therefore, pyrophosphate of food additives was found to be the most useful synergist of citric acid, the anti-browning agent for doenjang.

• Journal of Food Hygiene and Safety
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• v.23 no.4
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• pp.324-329
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• 2008

An analytical method for the simultaneous determination of nine quinolones (QNs) namely, marbofloxacin, norfloxacin(IS), ciprofloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, oxolinic acid, and flumequine in flatfish and egg was developed and validated using liquid chromatography with fluorescence detection (LC-FD). The samples were extracted using a traditional liquid-liquid extraction process; deproteinization was accomplished by the addition of trichloroacetic acid and acetonitrile (ACN), and defatting was performed with hexane. Chromatographic separation was achieved on a reverse phase C8 column with gradient elution using a mobile phase of 200 mM ammonium acetate buffer (pH 4.5) and ACN. The proposed method was validated according to the CODEX guideline. Mean recoveries of QNs from flatfish and egg were 89.6-106.5% with relative standard deviations (RSDs) below 15% at three different concentrations of 50, 100 and $500{\mu}g/kg$. Linearity was obtained with a correlation coefficient ($r^2$) of 0.9989-1.0000. The LOD for the investigated QNs was $1-16{\mu}g/kg$ depending on flatfish and egg. The present method can be applied simultaneously to determine QNs in muscle of flatfish and egg.

• Development and Reproduction
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• v.5 no.1
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• pp.17-21
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• 2001

This study was conducted to investigate the expression pattern of Trypsin-like enzyme and the effect of a trypsin inhibitor(benzimidine) on hatching process during in-vitro culture of mouse preimplantation embryos. The Trypsin-like enzyme was identified by rhodamine-conjugated Trypsin substrate probe. The expression of trypsin-like enzyme was firstly detected at the late morula stage, and the enzyme was uniformly localized in the trophectoderm of late blastocysts. Especially, intense fluorescence was observed in the blebbing area of hatching blastocysts. Bisbenzamidine, contained in culture media, did not alter embryonic development from 4-cell stage to the expanded blastocyst but decrease the hatching rate in ImM concentration (15.8% vs 89.7%, p<0.02). In the treatment of bisbenzimidine (5mM) for 12 hours according to the embryonic stage of mouse, the hatching rate of control (83.0%) and treatment in late blastocysts (8.7%) were significantly (p<0.01) different. From these results, we suggested that the hatching enzyme having trypsin-like activity was localized from the late morula stage, and the hatching process by this enzyme was activated in the late blastocyst stage of mouse embryos.

• Journal of Life Science
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• v.23 no.9
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• pp.1104-1108
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• 2013

Ethylene glycol (EG) is the most commonly used for automotive antifreeze, and it's easily misuseful for human. EG poisoning occurs in suicide attempts and infrequently, either intentionally through misuse or accidentally because of sweet taste. Though EG itself is mild toxic to the human body, it becomes higher toxic organic acids by in vivo broken down that are responsible for extensive cellular damage in various tissues caused principally by the metabolites. It is already well known that various cellular stresses induce gene expression of endoplasmic reticulum (ER) chaperones and ER stress sensors. This study demonstrated that regulation of gene expression of ER chaperones and ER stress sensors was induced by EG in rat tissues, and in tissues histological changes are also detected by both staining H&E and immunofluorescent.