• Korean Journal of Clinical Laboratory Science
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• v.48 no.2
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• pp.124-129
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• 2016

Endotoxin, also known as lipopolysaccharide (LPS) produced by the cell wall of gram negative bacteria can be present in any liquid or on any biomaterial. Endotoxin in blood can cause fever and inflammation. In this study, we compared bacterial endotoxin using Escherichia coli O157:H7, Klebsiella oxytoca, Salmonella Typhi, Shigella sonnei and Morganella morganii. Bacteria were cultured for use in the experiment, and diluted to $1.5{\times}10^8CFU/mL$. A check marked sensitivity confirmatory test of the Limulus amebocyte lysate (LAL) reagent was performed to examine the validity. The end point reaction to each bacteria sample was confirmed with 10 fold dilution and then the final reaction end point was confirmed by 2 fold dilution between the dilution step and the upper dilution step. According to the results, in detection of endotoxins in more than 0.015 EU/mL, E. coli O157 was 75~37.5 CFU/mL, K. oxytoca 37.5~18.75 CFU/mL, M. morganii and S. Typhi 3.75~1.875 CFU/mL, and S. sonnei 7.5~3.75 CFU/mL. The resulting value was finally ensured by a confirmation test for the inhibitory factor. Based on this study, conduct of further research on bacterial endotoxin is encouraged.

• Korean Journal of Food Science and Technology
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• v.18 no.4
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• pp.249-254
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• 1986

An experiment was performed to investigate the combined effect of preservatives and the synergistic effect of sodium chloride to them on the inhibition of bacterial growth. Escherichia coli and Salmonella typhimurium were cultured with or without shaking in liquid media (pH 6) of tryptone-glucose-yeast extract or tryptic soy broth which contained 0.1% potassium sorbate and/or 0.03% sodium benzoate, equivalent to half of the maximum permissible levels, respectively. The growth of E. coli was more inhibited with one or both of the two preservatives by shaking culture than by non-shaking culture. For S. typhimurium the single treatment of the preservatives did not show inhibitory effect whereas the combined treatment of them showed bacteriostatic effect in shaking culture and a prolongation of lag phase in non-shaking culture. Addition of 2% sodium chloride to either potassium sorbate or potassium sorbate plus sodium benzoate remarkably increased the growth inhibition of E. coli for non-shaking cultivation but no effect observed for shaking cultivation. S. typhimurium was more sensitive to the addition of sodium chloride than E. coli in both shaking and non-shaking culture to show lower viable cell counts than initial numbers.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.381-385
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• 2003

An efficient method of plant regeneration from Acanthopanax chiisanensis somatic embryos was developed. Cotyledonary somatic embryos were obtained in liquid Murashige and Skoog (MS) medium from embryogenic cell suspension cultures. They were desiccated for 0 to 72 hr and then cultured on MS medium containing NAA, BA, GA$_3$, (0-0.5mg/L). The highest multiple shoots formation (100％) was obtained from 72 hr desiccated somatic embryos on ifs medium with 0.5mg/L NAA＋0.5mg/L BA or 0.5 mg/L NAA＋0.5mg/L BA＋0.5mg/L GA$_3$ after 6 weeks culture. Plant conversion from multiple shoots was not high. The highest plant conversion from multiple shoots was obtained on 1/3MS medium with 1.0mg/L GA$_3$. Converted plantlets were transferred to ex vitro condition and the highest survival rate (70％) of the plantlets was obtained on plastic pots containing vermiculite and sand. These results indicate that micropropagation procedure can be applied for an efficient mass propagation of Acanthopanax chiisanensis.

• Korean Journal of Medicinal Crop Science
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• v.26 no.5
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• pp.391-400
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• 2018

Background: Atractylodes radix is a well-known medicinal crop having many physiological effects. This study was conducted to select useful Atractylodes japonica ${\times}$ Atractylodes macrocephala (AJM) cultivars by comparing anti-oxidative and anti-inflammatory efficacies. Methods and Results: Seven extracts from AJM cultivars were used to treat lipopolysacchride (LPS)-treated BV2 cells, and the effects on cell viability and inhibition on reactive oxygen species (ROS) and nitric oxide (NO) production were analyzed. In vitro scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and peroxynitrite ($NOO^-$) radicals were also investigated. Contents of total phenol, atractylenolide I, and atractylenolide III in the AJM extracts were measured using high performance liquid chromatography (HPLC) or spectrophotometry. The experiments show that none of the seven extracts was cytotoxic above 89.2% at $20-250{\mu}g/m{\ell}$. Extracts of Gowon, Dawon, Sangchul, and Huchul inhibited ROS generation in a dose-dependent manner, and Sangchul extract showed the highest inhibition on ROS production. All the AJM extracts showed effective inhibitory activity after on NO release in the LPS-treated BV2 cells, and Sangchul extract showed the highest activity. Sangchul extract had the most potent scavenging activities for $NOO^-$ and had some DPPH radical scavenging effect. Sangchul extract also had the highest content at total phenol and atractylenolide I content. Atractylenolide III was not detected in the AJM extracts. Conclusions: The results suggested that Sangchul was the most useful anti-oxidative and anti-inflammatory resource among the AJM cultivars.

• Journal of Applied Biological Chemistry
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• v.62 no.4
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• pp.399-406
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• 2019

The roots of Scrophularia buergeriana were extracted with 80% aqueous Methanol and the concentrates were partitioned into EtOAc, n-BuOH, and H₂O fractions. The repeated silica gel or octadecyl SiO₂column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of phenylethanoid glycosides and iridoid glycosides. The chemical structures of these compounds were determined as harpagoside (1), angoroside C (2), aucubin (3) and acetoside (4) based on spectroscopic analyses including nuclear magnetic resonance and MS. A simple and efficient HPLC with UV detection method for the simultaneous determination of the four compounds (1-4) has been developed and applied to their content determination in the S. buergeriana. The roots were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to 11.5, 7.6, 41.2, and 4.8 mg/g, respectively. Additionally, angoroside C (2) and acetoside (4) exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

• Korean Journal of Food Science and Technology
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• v.21 no.6
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• pp.795-799
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• 1989

This study was carried out to hydrolyze lactose in commercial milk by ${\beta}-galactosidase$ from Kluyveromyces fragilis and to compare some physicochemical properties of yoghurts made from control and lactase-treated commercial milks. Quantitative analysis of sugars was performed by gas liquid chromatograph (GLC) on trimethylsilyl (TMS) derivatives. In commercial milk, 94.6% of lactose was hydrolyzed after 2 hours incubation at $40^{\circ}C$ with 6.0 units/ml of ${\beta}-galactosidase$. pH, titratable acidity and viable cell number of yoghurt made from lactase-hydrolyzed (LH) commercial milk were 4.1, 1.04% and $6.5{\times}10^8/ml$ of Str. thermophilus, $8.9{\times}10^8/ml$ of L. bulgaricus after 8 hours incubation at $40^{\circ}C$, respectively, The total contents of amino acid were 2.63% in control and 2.19%. in LH yoghurt. The total contents of free amino acid were 26.95 mg% in control and 17.55mg% in LH yoghurt. Analysis of free fatty acids resulted in that the contents of short chain fatty acids in LH yoghurt were a little higher than those in control. Both in control and LH yoghurt, the palmitic acid content was highest and that was followed by oleic and myristic acid.

• Analytical Science and Technology
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• v.29 no.5
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• pp.234-241
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• 2016

The approach presented in this article refers to the bioanalytical method validation for the detection and quantitative determination of arsenic species including arsenite (As(III)), arsenate (As(V)), dimethylarsinic acid (DMA) and monomethylarsonic acid (MMA) in dog plasma by high-performance liquid chromatography inductively coupled plasma mass spectrometry (HPLC-ICP/MS). The arsenic species were separated using an agilent As speciation column by a mobile phase of 2 mM sodium phosphate monobasic, 0.2 mM ethylenediaminetetraacetic acid disodium salt dehydrate, 10 mM sodium acetate, 3 mM sodium nitrate and 1 % ethyl alcohol at pH 11 (adjusted with 1M NaOH). The method validation experiment was obtained selectivity, linearity, accuracy, precision, matrix effect, recovery, system suitability, dilution integrity and various stabilities. All calibration curves showed good linearity (R²>0.999) within test ranges. The lower limit of quantitation (LLOQ) was 5 ng/mL for As(III), As(V) and DMA, and 20 ng/mL for MMA. The system suitability and dilution values were within 6.5 % and 7.7 %. Subsequently, the developed and validated HPLC-ICP/MS method was also successfully applied to determine the arsenic speciation in dog plasma samples, and the recoveries for the spiked samples were in the range of 91.5–102.2 %. Therefore, this method could be applied to the evaluation of arsenic exposure, health effect assessment and other bio-monitoring studies in biological samples.

• Journal of Food Hygiene and Safety
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• v.33 no.5
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• pp.383-388
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• 2018

In this study, we investigated the effect of bioprocessed polysaccharides (BPPs) from liquid culture of Lentinus edodes fungal mycelia containing rice bran (BPP-RB) on a chicken-derived macrophage cell line, HD-11, when infected with Salmonella Gallinarum, an etiological agent of fowl typhoid. Experimental results demonstrated water extract of BPP-RB did not show growth inhibitory effects on S. Gallinarum 277. Protein expression profiles were also not altered by its treatment. Nonetheless, it could (i) enhance phagocytic activity of HD-11 cells, (ii) activate transcriptional expression of Th1-type cytokines such as tumor necrosis factor-${\alpha}$ and interleukin $(IL)-1{\beta}$, iNOS, as well as an immunosuppressive cytokine IL-10, and (iii) negatively regulate Th2-type cytokines such as IL-4 and IL-6. Together results suggest that BPP-RB may be applicable for preventing fowl typhoid or other Salmonella infections in poultry farms as a potential feed additive.

• Korean Journal of Food Science and Technology
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• v.27 no.6
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• pp.985-990
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• 1995

Grapefruit seed extracts(GFSE) have some unknown compounds which exhibit the antibiotic activities aganist microorganisms including bacteria and fungi. We have examined the effects of GFSE on the growth of Enterobacter pyrinus which was isolated from necrotic lesions of pear trees. During the cultivation, the growth of the bacteria was strongly inhibited at the low concentration(0.01%, w/w) of GFSE. Hydrophobic fraction extracted from GFSE by mixed solvents (chloroform : methanol : water, 1 : 2 : 0.8, v/v/v) had components which inhibited the growth of bacteria. There was, however, no inhibitory effect of GFSE on the activities of several enzymes including hexokinase, glucose 6-phosphate dehydrogenase, malate dehydrogenase and succinate dehydrogenase. $O-nitrophenyl-{\beta}-D-galactopyranoside(ONPG)$, the artificial substrate of ${\beta}-galactosidase$ was hydrolyzed in the presence of GFSE, indicating that the membrane was pertubated by the GFSE. From the results it was suggested that the antibiotic activity of GFSE is due to the change of membrane permeability of cell. GFSE was fractionated by high performance liquid chromatography equipped with $C_{18}$ reverse phase column. Among active fractions, three peaks were identified as 1-chloro-2-methyl-benzene (o-toluene), N,N-dimethyl-benzenemethaneamine, 1-[2-(2-ethylethoxy)ethoxy]-4- (1,1,3,3-tetramethyl)-bezene, respectively, while the other three remained unidentified.

• Journal of Korean Medicine Rehabilitation
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• v.21 no.2
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• pp.63-85
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• 2011

Objectives : This study was carried out to know the effects of Danggwisayeokgaohsuyusaenggang-tang(hereinafter referred to DST) on arthritis induced by collagen on DBA/1 OlaHsd mice. Methods : For this purpose, DST was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, high performance liquid chromatograph(HPLC) analysis, arthritis index, value of immunocyte in draining lymph node and paw joint, cytokine were measured in vivo. Results : 1. The cytotoxicity against human fibroblast cells(hFCs) was not measured in any concentration. 2. In HPLC analysis, There are high peak patterns at 8 minute(min), 12 min, 35 min, 45 min. 3. The arthritis index was decreased significantly. 4. The degree of arthritis induced damage of joint of DST group is slight compared with control group in histopathologic observation(Hematoxylin and eosin stain(H&E), Masson's trichrome(M-T) staining). 5. In total cell counts of draining lymph node(DLN) and paw joint, the cells in DLN decreased significantly on DST 200 mg/kg and the cells in paw joint decreased significantly on 200 mg/kg and 50 mg/kg. 6. In DLN, $CD4^+/CD25^+$, $CD3^+/CD69^+$, major histocompatibility complex(MHC), class-II/$CD11c^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg $CD3^+/CD8^+$ cells decreased significantly on DST 200 200 mg/kg, $CD4^+$, $CD3^+/CD44^+$ cells decreased. 7. In paw joints, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg. 8. In joints, levels of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, cyclo-oxygenase-2(COX-2), NOS-II were decreased on DST 200 mg/kg and DST 50 mg/kg. 9. In analysing of cytokine in CD3/CD28 activated spleen, IL-17 was decreased significantly, IL-4 was increased significantly $INF-{\gamma}$ was decreased on DST 200 mg/kg. 10. In analysing of cytokine in collagen activated spleen, IL-17 were decreased significantly, IL-4 was increased significantly. Conclusions : This results demonstrated that DST suppressed the inflammatory progression of collagen-induced arthritis(CIA) mice and supported further studies are required to survey continuously in looking for the effective substance and mechanism in the future.