• Title/Summary/Keyword: in vitro experiments

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Influence of barley grain treated with alkaline compounds or organic extracts on ex vivo site and extent of digestion of starch

  • Naseroleslami, Reza;Mesgaran, Mohsen Danesh;Tahmasbi, Abdolmansour;Vakili, Seyed Alireza;Ebrahimi, Seyed Hadi
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.2
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    • pp.230-236
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    • 2018
  • Objective: Two ex vivo experiments were conducted to verify the effect of barley grain (Nusrat cultivar) treated with alkaline compounds (AC) including alum, ammonium, and sodium hydroxide or cation-exchanged organic extracts (OE) prepared from alfalfa hay, sugar beet pulp and Ulva Fasciata, on extent and digestion of starch. Methods: In the first study, the in vitro first order disappearance kinetic parameters of dry matter (DM), crude protein (CP) and starch were estimated using a non-linear model ($D_{(t)}=D_{(i)}{\cdot}e^{(-k_d{\cdot}time)}+I$, where: $D_{(t)}$ = potentially digestible residues at any time, $D_{(i)}$ = potentially digestible fraction at any time, $k_d$ = fractional rate constant of digestion (/h), I = indigestible fraction at any time). In the second experiment, the ruminal and post-ruminal disappearance of DM, CP, and starch were determined using in situ mobile nylon bag. Results: Barley grains treated with alum and alfalfa extract had a higher constant rate of starch digestion (0.11 and 0.09/h) than others. Barley grain treated with OE had a higher constant rate of CP digestion and that of treated with AC had a higher constant rate of starch digestion (0.08 and 0.11/h) compared with those of the other treatments. The indigestible fraction of starch treated with alum and sugar beet pulp extract was higher than that of the control group (0.24 and 0.25 vs 0.21). Barley grain treated with AC and OE had significant CP disappearance in the rumen, post-rumen and total tract, and also starch disappearance for post-rumen and total tract compared with the untreated (p<0.001). Conclusion: This study demonstrated that AC and OE might have positive effects on the starch degradation of the barley grain. In addition, treating barley grain with alum and sugar beet pulp extract could change the site and extend digestion of protein and starch.

Antifungal Activity of the Extracts from Veratrum album L. var. grandiflorum Max. (백려노근경(白藜蘆根莖) 추출물(抽出物)의 항진균작용(抗眞菌作用))

  • Lee, Jong-Hwa;Kim, Won-Ja;Song, Byung-Sook;Cho, Sun-Hee
    • The Korean Journal of Pharmacology
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    • v.9 no.2
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    • pp.1-6
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    • 1973
  • Although numerous drugs are available for the treatment of superficial fungi infections of skin, the clinical effects of the majority of such drugs are not satisfactory. In the hope of searching the effective drugs for superficial fungi infections, authors studied whether Veratrum rhizoma extracts had any effect on fungi, with water extract (VRWE), ethanol extract (VREE) and methanol extract (VRME) from Veratrum album L. var. grandiflorum Max. In in vitro studies, the spores of fungi were inoculated on Sabouraud's glucose agar media which contained three extracts of Veratrun rhizoma in each concentration of $500\;{\mu}g/ml$, $1,000\;{\mu}g/ml$ and $5,000\;{\mu}g/ml$ respectively, and the growth of the fungi were observed for 3 weeks. The species of the fungi used in these experiments were Epidermophyton floccosum, Microsporum canis, Microsporum nanum, Microsporum gypseum, Microsporum cookei, Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans and Trichophyton verrucosum. The results of the studies were as follows: 1. The growth of M. canis, M. nanum, T mentagrophytes and T. tonsurans were slightly inhibited by VRWE $1,000\;{\mu}g/ml$, and with VRWE $5,000\;{\mu}g/ml$, the growth of E. floccosum, M. gypseum and T. rubrum were slightly inhibited, moderate inhibition on the growth of M. canis, M. nanum, M. cookei, T. mentagrophytes and T. tonsurans were showed by VRWE $5,000\;{\mu}g/ml$. 2. With $500\;{\mu}g/ml$ of VREE, the inhibition on growth of E. floccosum, M. nanum and M. gypseum were slight, however significant inhibition on the growth of M. canis, M. cookei, T. mentagrophytes, T. rubrum and T. tonsurans were observed. The growth of M. nanum and M. gypseum were moderately inhibited, and significant inhibition on the growth of E. floccosum, M. canis, M. cookei, T. mentagrophytes, T. rubrum and T. tonsurans were observed by VREE $1,000\;{\mu}g/ml$. By VREE $5,000\;{\mu}g/ml$, the growth of all tested fungi were significantly inhibited except T. verrucosuia being showed slight inhibition. 3. Significant inhibition on the growth of M. canis, T, mentagrophytes, T. rubrum and T. tonsurans were noted, and moderate inhibition of M. nanum, slight inhibition of E. floccosum and M. gypseum in growth were observed by VRME $500\;{\mu}g/ml$. The growth of E. floccosum, M. canis, M. nanum, M. cookei, T. mentsgrophytes, T. rubrum and T. tonsurans were significantly inhibited by VRME $1,000\;{\mu}g/ml$, and that of M. gypseum was moderate. With $5,000\;{\mu}g/ml$ of VRME, significant inhibition on the growth of E. floccosum, M. canis, M. nanum, M. gypseum, M. cookei, T mentagrophytes, T. rubrum and T. tonsurans were observed, and T. verrucosum was showed only slight inhibition. From the above results, it was found that the extracts of organic solvents from Veratrum rhizoma (VREE & VRME) exerted significant antifungal activity, and their effects were probably derived from the pharmacological action of steroidal alkaloids.

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Melatonin inhibits the Migration of Colon Cancer RKO cells by Down-regulating Myosin Light Chain Kinase Expression through Cross-talk with p38 MAPK

  • Zou, Duo-Bing;Wei, Xiao;Hu, Ruo-Lei;Yang, Xiao-Ping;Zuo, Li;Zhang, Su-Mei;Zhu, Hua-Qing;Zhou, Qing;Gui, Shu-Yu;Wang, Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5835-5842
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    • 2015
  • Background: Melatonin, which is mainly produced by the pineal gland, has a good inhibitory effect on cell growth of multiple cancer types. However, the underlying molecular mechanisms of anti-tumor activity for colon cancer have not been fully elucidated. In this study, we investigated the effects of melatonin on migration in human colon cancer RKO cells and the potential molecular mechanisms. Materials and Methods: The viability of RKO cells was investigated by MTT assay after treatment with melatonin, SB203580 (p38 inhibitor) and phorbol 12-myristate 13-acetate (PMA, MAPK activator) alone or in combination for 48h. The effects of melatonin, and ML-7, a selective inhibitor of myosin light chain kinase (MLCK), and SB203580, and PMA on the migration of RKO cells were analyzed by in vitro scratch-wound assay. The relative mRNA levels of MLCK was assessed by real-time quantitative RT-PCR. Western blotting analysis was performed to examine the expression of MLCK, phosphorylation of myosin light chain (pMLC) and p38 (pp38). Results: The proliferation and migration of human colon cancer RKO cells were inhibited significantly after treatment with melatonin. The expression levels of MLCK and phosphorylation of MLC of RKO cells were reduced, and real-time quantitative RT-PCR showed that melatonin had significant effects on suppressing the expression of MLCK. Furthermore, the phosphorylation level of p38, which showed the same trend, was also reduced when cells were treated by melatonin. In addition, ML-7 (25umol/l) could down-regulate the phosphorylation of p38. Conclusions: Melatonin could inhibit the proliferation and migration of RKO cells, and further experiments confirmed that p38 MAPK plays an important role in regulating melatonin-induced migration inhibition through down-regulating the expression and activity of MLCK.

Harmfulness of Particulate Matter in Disease Progression (미세먼지의 질병에 미치는 유해성)

  • Choi, Jong Kyu;Choi, In Soon;Cho, Kwang Keun;Lee, Seung Ho
    • Journal of Life Science
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    • v.30 no.2
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    • pp.191-201
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    • 2020
  • As society develops rapidly, environmental pollution is becoming a greater risk factor threatening human health. One of the major causes of air pollution that affects human health is particulate matter (PM), which contains a heterogeneous mixture of different particle sizes and chemical compositions. PM is classified by size into general PM (PM10; diameter below 10 ㎛) and fine PM (PM2.5; diameter below 2.5 ㎛). PM2.5 can pass through the respiratory tract into the circulatory system and thence throughout the body. PM2.5 is known to stimulate oxidative stress and inflammatory responses to cells, promoting diseases such as asthma, chronic respiratory disease, cardiovascular disease, diabetes mellitus, and immunological disorders. Although detailed molecular mechanisms for how PM stimulates disease progression still need to be elucidated, together with national efforts to reduce PM production, significant research has been conducted that demonstrates the harmfulness of PM in disease progression through in vitro and in vivo experiments. This review focuses on the harmfulness of PM in disease progression; we also introduce a biological verification method for determining the hazards of PM.

Studies on the Quality and Productivity Improvement by Mixed Sowing of Oat-Hairy Vetch (연맥-헤어리베치 혼파에 의한 사료가치 및 생산성 향상 연구)

  • Kim, J.G.;Chung, E.S.;Yoon, S.H.;Seo, S.;Seo, J.H.;Park, G.J.;Kim, C.K.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.1
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    • pp.31-36
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    • 2002
  • The yield and quality of winter forage crops such as oats could increase by mixed sowing with a winter legume such as hairy vetch(Vicia villosa Roth). The purpose of this study was to improve the forage quality and productivity using Oat-Hairy vetch mixture in winter season for three years. The results of this experiments were summarized as follows. In mix seeding of oat and hairy vetch, the dry matter(DM) content of herbage was 15∼16% in fall-harvest and 19∼20% in spring-harvest. Crude protein(CP) content of hairy vetch was the highest as 30.1% and the mixture was higher than that of oat. Acid detergent fiber(ADF) and neutral detergent fiber(NDF) content of hairy vetch was lower, but in vitro dry matter digestibility(IVDMD) and total dry matter digestibility(TDN) content was higher than that of oat. Although oat showed the low yield by 3.7∼4.4 DM t/ha, the Fall-seeded-Spring-harvest hairy vetch plot showed highly production by 9.5 DM t/ha. Crude protein and TDN yield of Fall-seeded-Spring-harvest plots were higher than those of others. According to this study, the highest dry matter yield could be obtained by Fall-seeded-Spring-harvest. It would be recommend in cropping field for oat.

Anti-oxidant Activities of Punica granatum Extracts (석류 추출물의 항산화 작용)

  • Roh, Bo-Kyung;Kim, Jee-Young;Kim, Ju-Yon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.2 s.51
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    • pp.207-212
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    • 2005
  • Pomegranate (Punica granatum L.) which is very rich in Polyphenols and tannins was recently reported its anti-oxidant activities and phytoestrogenic activities in vivo test and many clinical studies, but the effects of them on the skin have not been reported. The experiments were tarried out in vitro to determine anti-oxidant activities of pomegranate extracts on DPPH radical scavenging assay, NBT/Xanthine Oxidase-superoxide scavenging assay, silica-induced intracellular $H_2O_2$, hydroperoxide and superoxide generation assay in RAW 264.7 cells. It showed that the methanolic extract of dried pomegranate peels have the most significant anti-oxidant activities on free radical scavenging assay and inhibitory activities on silica-induced intracellular free radical generation in RAW 264.7 cells. The concentrated juice of pomegranate showed only DPPH radical scavenging activities and inhibited hyaluronidase activity. Moreover, pomegranate seed oil inhibited specially silica-induced intracellular hydroperoxide generation in RAW 264.7 cells. These results suggest that the methanolic extract of dried pomegranate peels and pomegranate seed oil have more anti-oxidant activity than concentrated juice of pomegranate. Thus the extracts of pomegranate peels and seed oil could be developed cosmetic ingredients for anti-aging.

The effects of Two Terpenoids, UA and ONA on Skin Barrier and Its Application

  • S. W. Lim;S. W. Jung;Kim, Bora;H. C. Ryoo;Lee, S. H.;S. K. Ahn
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.108-109
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    • 2003
  • Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol, prunol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ON A are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepatoprotective, anti-inflammatory, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. To clarify the effects of UA and ONA on skin barrier recovery, both flank skin of 8-12 weeks hairless mice were topically treated with samples (2mg/ml) after tape stripping, then measured recovery rate using TEWL on hairless mice. The recovery rate increased in UA and ONA treated groups at 6h more than 20% compared to vehicle treated group (p <0.05). For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/ml per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to Vehicle group from 1 week without TEWL alteration (p<0.005). EM examination using Ru04 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent (ONA$\geq$UA>Vehicle). LM finding showed that stratum corneum was slightly increased and especially epidermal thickening and flattening was observed (UA>ONA>Vehicle). Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber increasing by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory experiments were also confirmed in vivo findings. This result suggested that the effects of UA and ONA related to not only skin barrier but also collagen and elastic fibers. Taken together, UA and ONA can be relevant candidates to improve barrier function and pertinent agents for cosmetic applications.

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In vitro evaluation of nano zinc oxide (nZnO) on mitigation of gaseous emissions

  • Sarker, Niloy Chandra;Keomanivong, Faithe;Borhan, Md.;Rahman, Shafiqur;Swanson, Kendall
    • Journal of Animal Science and Technology
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    • v.60 no.11
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    • pp.27.1-27.8
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    • 2018
  • Background: Enteric methane ($CH_4$) accounts for about 70% of total $CH_4$ emissions from the ruminant animals. Researchers are exploring ways to mitigate enteric $CH_4$ emissions from ruminants. Recently, nano zinc oxide (nZnO) has shown potential in reducing $CH_4$ and hydrogen sulfide ($H_2S$) production from the liquid manure under anaerobic storage conditions. Four different levels of nZnO and two types of feed were mixed with rumen fluid to investigate the efficacy of nZnO in mitigating gaseous production. Methods: All experiments with four replicates were conducted in batches in 250 mL glass bottles paired with the ANKOM$^{RF}$ wireless gas production monitoring system. Gas production was monitored continuously for 72 h at a constant temperature of $39{\pm}1^{\circ}C$ in a water bath. Headspace gas samples were collected using gas-tight syringes from the Tedlar bags connected to the glass bottles and analyzed for greenhouse gases ($CH_4$ and carbon dioxide-$CO_2$) and $H_2S$ concentrations. $CH_4$ and $CO_2$ gas concentrations were analyzed using an SRI-8610 Gas Chromatograph and $H_2S$ concentrations were measured using a Jerome 631X meter. At the same time, substrate (i.e. mixed rumen fluid+ NP treatment+ feed composite) samples were collected from the glass bottles at the beginning and at the end of an experiment for bacterial counts, and volatile fatty acids (VFAs) analysis. Results: Compared to the control treatment the $H_2S$ and GHGs concentration reduction after 72 h of the tested nZnO levels varied between 4.89 to 53.65%. Additionally, 0.47 to 22.21% microbial population reduction was observed from the applied nZnO treatments. Application of nZnO at a rate of $1000{\mu}g\;g^{-1}$ have exhibited the highest amount of concentration reductions for all three gases and microbial population. Conclusion: Results suggest that both 500 and $1000{\mu}g\;g^{-1}$ nZnO application levels have the potential to reduce GHG and $H_2S$ concentrations.

Antimetastatic Effects of Crude Polysaccharide Isolated from Polygonati Rhizoma on 4T1 Breast Cancer Cells by Activation of Innate Immune System (황정(黃精)으로부터 유래한 조다당류의 선천면역 활성에 의한 유방암 세포주 전이 억제 효과)

  • Ji, Hae-Ri;Hwang, Deok-Sang;Lee, Chang-Hoon;Jang, Jun-Bok;Lee, Jin-Moo
    • The Journal of Korean Obstetrics and Gynecology
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    • v.32 no.4
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    • pp.1-13
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    • 2019
  • Objective: This study is aimed to investigate the anti-tumor metastasis by innate immunomodulating effects of crude polysaccharide isolated from Polygonati Rhizoma (CP-PR) on 4T1 breast cancer cells. Methods: CP-PR was isolated from Polygonati Rhizoma. Antimetastatic experiments were conducted in vivo mouse model by using 4T1 breast cancer cells. The cell viability of CP-PR was tested with normal spleen and 4T1 breast cancer cells. To observe the activation of macrophages with/without 4T1 breast cancer cells, production of tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), IL-10 and IL-12 were measured with enzyme-linked immunosorbent assay (ELISA), respectively. In addition, the lysis of YAC-1 cells and the production of granzymes were measured to observe the activation of natural killer (NK) cell. Results: Intravenous administration of CP-PR significantly inhibited metastasis of 4T1 breast cancer cells. In an in vitro cytotoxicity analysis, CP-PR affected the growth of normal spleen and 4T1 breast cancer cells above specific concentration. The production of $TNF-{\alpha}$, IL-6, IL-10 and IL-12 were significantly increased in macrophages with CP-PR. As compared with control, CP-PR showed significantly higher production of $TNF-{\alpha}$, IL-10 and IL-12 in macrophages co-cultured with 4T1 breast cancer cells. The lysis of YAC-1 cells and the production of granzymes were significantly up regulated by CP-PR. Conclusion: CP-PR appears to have considerable activity on the anti-metastasis by activation of innate immune system.

Effects of red ginseng and Baeknyeoncho complex on the arthritis improvement (홍삼·백년초 복합물이 관절염 개선에 미치는 효과)

  • Lee, Seung-Hyeon;Lee, Chang-Hyun;Kim, Hong-Jun;Kim, Young-Sik;Shin, Min-Ji;Kim, Ji-Hyun;Ma, Sang-Yong;Lee, Se-Youn;Kwon, Jin;Oh, Chan-Ho
    • Herbal Formula Science
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    • v.29 no.2
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    • pp.81-91
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    • 2021
  • Objectives : This study was conducted to evaluate the efficacy of a complex mixture of natural substances of ginseng and baeknyeoncho on the arthritic rats. Methods : In vitro experiments were conducted to ensure the stability of the complex. After setting toxicity and concentration by MTT assay, the antioxidant effect was measured through DPPH and ABTS radical scavenging activity. To confirm the anti-inflammatory effects of the complex, levels of nitric oxide (NO) and pro-inflammatory cytokines (IL-1β, TNF-α) were measured in LPS-treated macrophage cell lines (RAW264.7). We injected monosodium iodoacetate (MIA) 50 μl (60 mg/ml) into knee joints of rats to induce osteoarthritis. The rats were divided into three groups (normal (n=5), control (n=5), and OR (n=5) group). The control group consumed 2 mg/kg of physiological saline once a day for 4 weeks, and the OR group was mixed at a concentration of 416.5 mg/kg of Baengnyeoncho (O) and 208.25 mg/kg of red ginseng (R) and ingested 1 mL each 5 days a week. Results : This complex increased the DPPH and ABTS radical scavenging rate. The complex decreased NO production and pro-inflammatory cytokine production of macrophages. In the OR group, the secretion of cytokine in serum was decreased. In histopathological examination, the joint tissue of the composite showed less damage to the synovial membrane, cartilage, and fibrous tissue than the control group. Conclusions : As a result of this study, natural complexes have antioxidant, anti-inflammatory and cartilage protection effects. Therefore, we expect the complex to be effective in treating osteoarthritis.