• Asian-Australasian Journal of Animal Sciences
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• v.28 no.3
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• pp.420-427
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• 2015

It is necessary to understand the cellular uptake and cytotoxicity of food-grade delivery systems, such as ${\beta}$-lactoglobulin (${\beta}$-lg) nanoparticles, for the application of bioactive compounds to functional foods. The objectives of this study were to investigate the relationships between the physicochemical properties of ${\beta}$-lg nanoparticles, such as particle size and zeta-potential value, and their cellular uptakes and cytotoxicity in Caco-2 cells. Physicochemical properties of ${\beta}$-lg nanoparticles were evaluated using particle size analyzer. Flow cytometry and confocal laser scanning microscopy were used to investigate cellular uptake and cytotoxicity of ${\beta}$-lg nanoparticles. The ${\beta}$-lg nanoparticles with various particle sizes (98 to 192 nm) and zeta-potential values (-14.8 to -17.6 mV) were successfully formed. A decrease in heating temperature from $70^{\circ}C$ to $60^{\circ}C$ resulted in a decrease in the particle size and an increase in the zeta-potential value of ${\beta}$-lg nanoparticles. Non-cytotoxicity was observed in Caco-2 cells treated with ${\beta}$-lg nanoparticles. There was an increase in cellular uptake of ${\beta}$-lg nanoparticles with a decrease in particle size and an increase in zeta-potential value. Cellular uptake ${\beta}$-lg nanoparticles was negatively correlated with particle size and positively correlated with zeta-potential value. Therefore, these results suggest that the particle size and zeta-potential value of ${\beta}$-lg nanoparticles play an important role in the cellular uptake. The ${\beta}$-lg nanoparticles can be used as a delivery system in foods due to its high cellular uptake and non-cytotoxicity.

• Nutrition Research and Practice
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• v.17 no.4
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• pp.616-630
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• 2023

BACKGROUND/OBJECTIVES: Indole-3-propionic acid (IPA) is a tryptophan-derived microbial metabolite that has been associated with protective effects against inflammatory and metabolic diseases. However, there is a lack of knowledge regarding the effects of IPA under physiological conditions and at the intestinal level. MATERIALS/METHODS: Human intestinal epithelial Caco-2 cells were treated for 2, 24, and/or 72 h with IPA or its precursors - indole, tryptophan, and propionate - at 1, 10, 100, 250, or 500 μM to assess cell viability, integrity, differentiation, and proliferation. RESULTS: IPA induced cell proliferation and this effect was associated with a higher expression of extracellular signal-regulated kinase 2 (ERK2) and a lower expression of c-Jun. Although indole and propionate also induced cell proliferation, this involved ERK2 and c-Jun independent mechanisms. On the other hand, both tryptophan and propionate increased cell integrity and reduced the expression of claudin-1, whereas propionate decreased cell differentiation. CONCLUSIONS: In conclusion, these findings suggested that IPA and its precursors distinctly contribute to the proliferation, differentiation, and barrier function properties of human intestinal epithelial cells. Moreover, the pro-proliferative effect of IPA in intestinal epithelial cells was not explained by its precursors and is rather related to its whole chemical structure. Maintaining IPA at physiological levels, e.g., through IPA-producing commensal bacteria, may be important to preserve the integrity of the intestinal barrier and play an integral role in maintaining metabolic homeostasis.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.357-364
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• 1995

The protozoan parasite, Entcmoeba histoIWticc, is one of major causative agents of intestinal disease all over the world. In acute experimental infection, the early host response to 5. histoIHtica is characterized by an infiltration of neutrophils. However, the chemotactic signal for this response is not well known. Based on the (jading that human epithelial cells produce the potent neutrophil chemoattractant and activator, interleukin-8 (IL-8), IL-8 gene expression was examined thoroughly in human colon epithelial cells exposed to 5. histolvtica trophozoites. Cellular RNAs were extracted from HT-29 or Caco-2 human colon epithelial cells exposed to 5. histoLvtica trophozoites for 30 minutes, 1 and 3 hours. IL-8 mRNA transcripts were measured by reverse transcriptional polprnerase chain reaction (RT-PCR) using synthetic standard RNA. The number of IL-8 mRNA molecules increased from 30 minutes to 3 hours of exposure period, reaching 3.1 H 107 molecules/ug of total RNA. Expression pattern of IL-8 mRNA transcripts was parallel to the amounts of IL-8 protein measured by enzyme-linked immunosorbent assay (ELISA) . Lysates of 5. histoIVtica also induced expression of mRNA for IL-8 in colon epithelial cells. These results sugf:esc that acute inflammatory reaction by 5. histoIVticc may be initially triggered by proinflammatory cytokines such as IL-8 secreted from epithelial cells of the colon.

• Microbiology and Biotechnology Letters
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• v.40 no.3
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• pp.226-236
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• 2012

In the present work, the influence of prebiotic fructooligosaccharide (FOS) on adhesion to Caco-2 cells, viability, acid and bile tolerance, antibacterial, antioxidant, enzymatic, and metabolic activities of the probiotic starters Lactobacillus acidophilus GK20 and Lactobacillus paracasei GK74, has been explored. Experiments were conducted with fermented yoghurt over a period of 7 days at $4^{\circ}C$. When compared to control fermentations without prebiotic, the addition of FOS was seen to significantly (p<0.05) increase the viable cell counts of the probiotics, overall viscosity, and concurrently reduce the pH of the fermented yoghurts. Both Escherichia coli ATCC 11229 and Salmonella enteritidis ATCC 13076 were inhibited by the probiotics' antibacterial activities, while the synbiotic yoghurt containing mixed probiotics and FOS was noted to highly improve antagonistic action. When fermented with mixed starters, the addition of FOS (1.0%) resulted in the highest proteolytic ($1.06{\pm}0.06$ unit) and ${\beta}$-galactosidase activities ($20.14{\pm}0.31$ unit). However, FOS did not affect acid and bile tolerance, adhesion to Caco-2 cells or the antioxidant activity of the probiotics, although both L. acidophilus GK20 and L. paracasei GK74 had functionality as probiotic strains. Hence, a significant synbiotic effect was observed in fermented yoghurt after 7 days of storage at $4^{\circ}C$, and as a result, such synbiotic yoghurt can be said to possess synergistic actions which improve the gastrointestinal environment and promote of health.

• Journal of Microbiology and Biotechnology
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• v.26 no.6
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• pp.1035-1045
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• 2016

To evaluate the effects of lactic acid bacteria (LAB) on Peyer's patch cells, mice were treated with a high dose of kanamycin to disturb the gut microbial environment. The overarching goal was to explore the potential of LAB for use as a dietary probiotic that buffers the negative consequences of antibiotic treatment. In vitro, LAB stimulated the production of immunoglobulin A (IgA) from isolated Peyer's patch cells. Inflammation-related genes (TNF-α, IL-1β, and IL-8) were up-regulated in Caco-2 cells stimulated with lipopolysaccharide (LPS), while tight-junction-related genes (ZO-1 and occludin) were down-regulated; the effects of LPS on inflammatory gene and tight-junction gene expression were reversed by treatment with LAB. Mice treated with a high dose of kanamycin showed increased serum IgE levels and decreases in serum IgA and fecal IgA levels; the number of Peyer's patch cells decreased with kanamycin treatment. However, subsequent LAB treatment was effective in reducing the serum IgE level and recovering the serum IgA and fecal IgA levels, as well as the number of Peyer's patch cells. In addition, ZO-1 and occludin mRNA levels were up-regulated in the ileum tissues of mice receiving LAB treatment. Lactic acid bacteria can enhance the intestinal immune system by improving the integrity of the intestinal barrier and increasing the production of IgA in Peyer's patches. Lactic acid bacteria should be considered a potential probiotic candidate for improving intestinal immunity, particularly in mitigating the negative consequences of antibiotic use.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.04a
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• pp.103-103
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• 2003

This study was carried out to investigate of the biologically active components in germinating Mung bean(Phaseolus radiata L.) and Buckwheat (Fagopyrum esculentum Moench) seeds. During the initial germination, germination ratio of 24 hours pre-soaking Mung bean and Buckwheat seeds were higher about 2∼3% than that of non-soaking. This experiment also was peformed to observe cytotoxic effect of the germinating seeds(germination length : 2, 5, 10mm) extracts against cancer cell lines including human lung carcinoma(Calu-6), human breast adenocarcinoma(MCT-7), human great intestine carcinoma(Caco-2) and human leukemia carcinoma(AML-2/WT). The growth of the cancer cells in medium containing Mung bean and Buckwheat extracts were significantly inhibited degree in proportion to the length of germination seeds, Especially, the results show that a significant shrinkage of Calu-6 cells was observed when the cells were exposed into extract of 10mm germination seeds in germinating Mung bean and Buckwheat seeds.

• The Journal of Internal Korean Medicine
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• v.21 no.1
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• pp.13-22
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• 2000

목적 : 본(本) 연구(硏究)는 배기음(排氣飮)이 인간(人間)의 장관내(腸管內)에서 산화물(酸化物)에 의해 유발(誘發)된 세포(細胞)의 사망(死亡) 및 DNA의 손상(損傷)을 방지할수 있는지를 검증(檢證)하기 위한 실험(實驗)이다. 방법 : 배양(培養)된 인체장관(人體腸管) 세포계열(細胞系列)인 Caco-2 세포(細胞)에서 세포(細胞)의 사망(死亡)은 trypan bile의 소실정도에 의해서 평가했으며, DNA의 손상(損傷)은 double stranded DNA의 파괴정도를 측정하여 평가하였다. $H_2O_2$는 표본(標本) 산화제(酸化劑)로 사용되었다. 결과 : $H_2O_2$에 노출된 세포들의 세포사망(細胞死亡) 정도는 노출시간과 용량에 비례하여 증가하는 양상을 보였다. 배기음(排氣飮)은 $H_2O_2$에 의해 유발(誘發)되는 세포방지를 방지하였고, 0.05-1%의 농도범위에 걸쳐서는 그 효과가 용량에 비례하여 증가하는 양상을 보였다. $H_2O_2$에 의해 유발(誘發)된 세포손상(細胞損傷)은 catalase(hydrogen peroxide scavenger enzyme)와 deferoxamine(iron chelator)에 의해 억제되었다. 그러나 강력한 항산화제(抗酸化劑)인 DPPD는 $H_2O_2$에 의해 유발(誘發)되는 세포손상(細胞損傷)에는 영향을 주지 못했다. $H_2O_2$에 의해 유발(誘發)된 지질(脂質)의 과산화(過酸化)는 배기음(排氣飮)과 DPPD에 의해 억제되었다. $H_2O_2$에 의해 유발(誘發)된 DNA의 손상(損傷)은 배기음(排氣飮)에 의해 방지되었으며 용량에 의존하는 양상을 보였다. $H_2O_2$에 의해 유발(誘發)된 DNA의 손상은 catalase와 deferoxamine에 의해 억제되었지만 DPPD는 억제시키지 못했다. 배기음(排氣飮)은 $H_2O_2$에 의해 유발(誘發)된 ATP의 소실을 회복시켰다. 이러한 실험결과 $H_2O_2$에 의해 유발(誘發)된 세포(細胞)의 손상(損傷)은 지질(脂質)의 과산화(過酸化)와는 다른 독립적인 기전에 의해 일어남을 나타낸다. 결론 : 이러한 결과들로 볼 때 Caco-2 세포(細胞)에서 배기음(排氣飮)이 항산화작용(亢酸化作用)보다는 다른 기전을 통하여 Caco-2 세포안에서 산화제(酸化劑)에 의해 유발(誘發)된 세포(細胞)의 사망(死亡)와 DNA의 손상(損傷)을 방지할 수 있다는 것을 가리킨다. 따라서 본 연구(硏究)는 배기음(排氣飮)이 반응성산소기(反應性酸素基)에 의해 매개된 인체(人體) 위장관질환(胃腸管疾患)의 치료(治療)에 사용할 수 있을 가능성(可能性)이 있음을 제시하고 있다.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.584-589
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• 2001

Isopropryl 2-(1-3-dithiethane-2-ylidene)-2 [N-(4-methyl-thiazole-2-yl) carbamoyl] acetate (YH439) is currently under phase ll clinical trials by the Yuhan Research Center for use as a hepatoprotective agent. Unfortunately, the oral bioavailbility of YH439, which is sparingly soluble in water (i.e., $0.3{\;}\mu\textrm{g}/ml{\;}or{\;}0.91{$\mu}M$ at room temperature), reportedly, is negligibleregardless of the dose administered to rats in the 10-300 mg/kg range. The bioavailability of the compound increased up to 24%, when administered in the form of a micellar solution ($700{\;}\mu\textrm{g}/ml$or 2.1 mM for YH439) at a dose of 10 mg/kg, suggesting that its limited solubility is associated with its negligible bioavailability. In order to obtain additional informmation concerning the bioavailability of YH439, the mechanism(s) involved in gastrointestinal (Gl) absorption were investigated in the present study. For this purpose, the transport of YH430 across a Caco-2 cell monolayer was measured in a $Transwell^{\circledR}$. A permeability of $4.07{\times}10^{-5}{\;}cm/s$ was obtained for the absorptive (i.e., apical to basolateral direction) transport of $0.42{\mu}M$ YH439, implicating that the in vivo Cl absorption is nearly complete. The absorptive transport exhibited a slight concentration-dependency with an intrinsic clearance ($CL_{i}$) of $0.38{\mu}L/{\textrm{cm}^2}/sec$, which accounted for 28.1% of the total intrinsic clearance (i.e., $CL_i$ plus the intrinsic clearance for the linear component) of the transport. Thus, saturation of the absorption process appears to be a minor factor in limiting the bioavailability of the compound. The apparent permeability of YH439 from the basolateral to the apical direction (i.e., efflux, $6.67{\times}10^{-5}{\;}cm/s$) was comparable to that for absorptive transport, but, interestingly, a more distinct concentration-dependency was observed for this transport. However, the efflux does not appear to influence the bioavailability of the compound, as evidenced by the sufficiently high permeability in the absorption direction. Rather, a reportedly extensive first-pass hepatic metabolism appears to be a principal factor in limiting the bioavailability. In this respect, reducing the first-pass metabolism by some means would lead to a higher bioavailability of the compound. Thus, elevation of the absorption rate of YH439 becomes a necessity. From a practical point of view, increasing the concentration of YH439 in the Cl fluid appears to be a feasible way to increase the absorption rate, because the compound is primarily absorbed via a linear mechanism. In summary, the solubilization of YH439, as previously demonstrated for a micellar solution of the compound, appears to be a practical way to increase the oral bioavailability of YH439.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.1 s.32
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• pp.115-129
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• 2007

Objectives : This study was carried out to evaluate anti-oxidative, anti-microbial and anti-cancer effect for clinical application of Bibangtalmyungsan (BTS) Results : 1. The oxidative effects were measured by polyphenol, DPPH radical scavenging activity. BTS water extract was showed more effective than ethanol extract and also various solvent fractions from BTS water extract showed effective in the following order : ethyl acetate fraction > butanol fraction > hexane fraction ${\fallingdotseq}$ chloroform fraction ${\fallingdotseq}$ aqueous fraction 2. The results of anti-microbial effects were as follows. 1) Antibacterial activities of BTS extracts against Gram's negative and positive bacteria were ineffective 2) Antifungal activities of the BTS extracts against Aspergillus spp. , Trichohyton mentagrophyte KTCC 1077 were not effective. 3. The result of anti-cancer effects were as followings: 1) BTS ethanol extract was more effective than water extract against Caco-2, Calu-6, but it had cytotoxic effect against NIH3T3. 2) On the Caco-2, effective only in ethyl acetate $fraction(IC_{50}:$ 35.81 ${\mu}g/ml)$. 3) On the Calu-6, the most effective in ethyl acetate $fraction(IC_{50}:$ 189.65 ${\mu}g/ml)$ and effective In butanol $fraction(IC_{50}:$ 299.74 ${\mu}g/ml)$ and hexane $fraction(IC_{50}:$ 345.13 ${\mu}g/ml)$ 4) On the SUN-601, $IC_{50}$ value was within 80 ${\mu}g/ml$ in ethyl acetate fraction and hexane fraction. 5) On the HCT-1l6, the most effective in ethyl acetate $fraction(IC_{50}:$ 82.94 ${\mu}g/ml)$ and effective in hexane $fraction(IC_{50}:$ 374.56 ${\mu}g/ml)$. 6) On the AML-2/WT, the most effective in ethyl acetate $fraction(IC_{50}:$ 41.44 ${\mu}g/ml)$ and effective In hexane $fraction(IC_{50}:$ 303.01 ${\mu}g/ml)$. 7) On the NIH3T3, effective only in ethyl acetate $fraction(IC_{50}:$ 203.42 ${\mu}g/ml)$, but it was more ineffective than other cancer cells. Conclusion : These result suggest that BTS has antioxidative, antifungal activities and cytotoxic effects against Caco-2, Calu-6, SUN-601, HCT-116, AML-2/WT and NIH3T3, especially ethyl acetate fraction from water extract has more effective in antioxidative and anticancer effects.

• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.72-80
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• 2022

In this study, the survival capacity (acid and bile salt tolerance, and adhesion to gut epithelial cells) and probiotic properties (enzyme activity-inhibition and anti-inflammatory activities, inhibition of adipogenesis, and stress hormone level reduction) of Lactiplantibacillus plantarum LRCC5314, isolated from kimchi (Korean traditional fermented cabbage), were investigated. LRCC5314 exhibited very stable survival at ph 2.0 and in 0.2% bile acid with 89.9% adhesion to Caco-2 intestinal epithelial cells after treatment for 2 h. LRCC5314 also inhibited the activities of α-amylase and α-glucosidase, which are involved in elevating postprandial blood glucose levels, by approximately 72.9% and 51.2%, respectively. Treatment of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells with the LRCC5314 lysate decreased the levels of the inflammatory factors nitric oxide, tumor necrosis factor (TNF-α), interleukin (IL)-1β, and interferon-γ by 88.5%, 49.3%, 97.2%, and 99.8%, respectively, relative to those of the cells treated with LPS alone. LRCC5314 also inhibited adipogenesis in differentiating preadipocytes (3T3-L1 cells), showing a 14.7% decrease in lipid droplet levels and a 74.0% decrease in triglyceride levels, as well as distinct reductions in the mRNA expression levels of adiponectin, FAS, PPAR/γ, C/EBPα, TNF-α, and IL-6. Moreover, LRCC5314 reduced the level of cortisol, a hormone with important effect on stress, by approximately 35.6% in H295R cells. L. plantarum LRCC5314 is identified as a new probiotic with excellent in vitro multifunctional properties. Subsequent in vivo studies may further demonstrate its potential as a functional food or pharmabiotic.