• Title/Summary/Keyword: Human T-cell

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Preparation and Characterization of PE Liposomes Containing Antibody (항체를 포함하는 Phosphatidylethanolamine 리포좀의 제조와 그 특성)

  • 박성호;신현재양지원최태부
    • KSBB Journal
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    • v.10 no.2
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    • pp.204-211
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    • 1995
  • A target-sensitive liposome was prepared by using a dioleoyl-phosphatidylethanolamine(DOPE) and a palmitic acid coupled antibody(p-IgG). For the preparation of stable PE-liposomes, the key factors such as antibody modification method with palmitic acrid, molar ratio of p-IgG to lipid and the amount of various additives, were examined. The optimum molar ratio of p-IgG to lipid was found to be $2.5{\times}10^{-4}$ and the final concentration of deoxycholate for the stable liposome formation was about 0.09%. Two kinds of target-sensitive liposomes, containing polyclonal anti-SRBC(Sheep Red Blood Cell)-antibody and monoclonal anti-${\beta}$-HCG(Human Chorionic Gonadotropin)-antibody, were successfully prepared. The destabilization of liposomes was examined by measuring the release of calcein entrapped in the liposome vesicles. Calcein was released only when the liposomes were contacted with the specific target cells. The calcein release with non-specific target cells was negligible. From this result, it is clear that p-IgG is indispensible for the maintenance of stable PE-liposome and the calcein release is mainly due to the specific interactions between the liposomes containing antibody and the target cells containing antigen.

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Protective effect of gallic acid derivatives from the freshwater green alga Spirogyra sp. against ultraviolet B-induced apoptosis through reactive oxygen species clearance in human keratinocytes and zebrafish

  • Wang, Lei;Ryu, BoMi;Kim, Won-Suk;Kim, Gwang Hoon;Jeon, You-Jin
    • ALGAE
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    • v.32 no.4
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    • pp.379-388
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    • 2017
  • In the present study, we enhanced the phenolic content of 70% ethanol extracts of Spirogyra sp. (SPE, $260.47{\pm}5.21$ gallic acid equivalent $[GAE]mg\;g^{-1}$), 2.97 times to $774.24{\pm}2.61GAE\;mg\;g^{-1}$ in the ethyl acetate fraction of SPE (SPEE). SPEE was evaluated for its antiradical activity in online high-performance liquid chromatography-ABTS analysis, and the peaks with the highest antiradical activities were identified as gallic acid derivatives containing gallic acid, methyl gallate, and ethyl gallate. Isolation of ethyl gallate from Spirogyra sp. was performed for the first time in this study. In ultraviolet B (UVB)-irradiated keratinocytes (HaCaT cells), SPEE improved cell viability by 8.22%, and 23.33% and reduced accumulation of cells in the sub-$G_1$ phase by 20.53%, and 32.11% at the concentrations of 50 and $100{\mu}g\;mL^{-1}$, respectively. Furthermore, SPEE (50 and $100{\mu}g\;mL^{-1}$) reduced reactive oxygen species generation in UVB-irradiated zebrafish by 66.67% and 77.78%. This study suggests a protective activity of gallic acid and its derivatives from Spirogyra sp. against UVB-induced stress responses in both in vitro and in vivo models, suggesting a potential use of SPEE in photoprotection.

Effect of Lactoferrin from Korean Native Cattle on the Production of Tumor Necrosis Factor-$\alpha$ and Nitric Oxid (한우 Lactoferrin이 TNF-$\alpha$와 NO 생성에 미치는 영향)

  • 이수원;양희진;황보식
    • Food Science of Animal Resources
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    • v.21 no.4
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    • pp.374-382
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    • 2001
  • Lactoferrin(Lf) has the function of modulation in the host defense systems, including cytokine production and immune responses. We have tested the effect of Lf and Lf ydrolysates(Lf-H) on the productions of tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) and nitric oxide(NO) in macrophage cells. Lf from Korean native cattle(K-Lf) and hydrolyzed K-Lf(K-Lf-H) increased the production of TNF-${\alpha}$ in RAW264.7 cells with dose-dependency. Bovine Lf(B-Lf), human Lf(H-Lf), and its hydrolysates did not induce either TNF-${\alpha}$ production or NO production. On the other hand those didn\`t affect on the production of TNF-${\alpha}$ in lipopolysaccharide(LPS)-stimulated RAW264.7 cells. K-Lf induced the production of NO similar to its role on the TNF-${\alpha}$ production.

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Anti-aging Potential of Extracts Prepared from Fruits and Medicinal Herbs Cultivated in the Gyeongnam Area of Korea

  • Shon, Myung-Soo;Lee, Yunjeong;Song, Ji-Hye;Park, Taehyun;Lee, Jun Kyoung;Kim, Minju;Park, Eunju;Kim, Gyo-Nam
    • Preventive Nutrition and Food Science
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    • v.19 no.3
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    • pp.178-186
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    • 2014
  • Many recent studies have focused on maintaining a healthy life by preventing and/or postponing the aging process. Numerous studies have reported that continuous exposure to reactive oxygen species can stimulate skin aging and that excessive accumulation of fat can cause an impaired skin barrier and tissue structure alterations. Thus, the maintenance of antioxidant homeostasis and the suppression of adipose accumulation are important strategies for skin anti-aging. Here, we prepared three types of extracts [whole juice, acetone-perchloric acid (PCA), and ethanol] from 20 fruits and medicinal herbs native to the Gyeongnam area of Korea. The total phenolic content of each extract was analyzed, and we observed higher total phenolic contents in the medicinal herbs. Consistent with this, the results of the oxygen radical absorbance activity capacity assay indicated that the in vitro antioxidant activities of the medicinal herb extracts were stronger than those of the fruit extracts. The fruits and medicinal herbs had strong effects on cell-based systems, including $H_2O_2$-induced oxidative stress in human keratinocytes and 3T3-L1 lipid accumulation. Nishimura Wase persimmon, Taishu persimmon, wrinkled giant hyssop, sweet wormwood, Chinese cedar, red perilla, tan shen, hiyodori-jogo, and cramp bark may be natural anti-aging materials with effective antioxidant and anti-adipogenic activities. Taken together, our findings may provide scientific evidence supporting the development of functional foods and nutraceuticals from fruits and medicinal herbs.

Reversal of Multidrug Resistance in Mouse Lymphoma Cells by Extracts and Flavonoids from Pistacia integerrima

  • Rauf, Abdur;Uddin, Ghias;Raza, Muslim;Ahmad, Bashir;Jehan, Noor;Siddiqui, Bina S;Molnar, Joseph;Csonka, Akos;Szabo, Diana
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.51-55
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    • 2016
  • Phytochemical investigation of Pistacia integerrima has highlighted isolation of two known compounds naringenin (1) and dihydrokaempferol (2). A crude extract and these isolated compounds were here evaluated for their effects on reversion of multidrug resistance (MDR) mediated by P-glycoprotein (P-gp). The multidrug resistance P-glycoprotein is a target for chemotherapeutic drugs from cancer cells. In the present study rhodamine-123 exclusion screening test on human mdr1 gene transfected mouse gene transfected L5178 and L5178Y mouse T-cell lymphoma cells showed excellent MDR reversing effects in a dose dependent manner. In-silico molecular docking investigations demonstrated a common binding site for Rhodamine123, and compounds naringenin and dihydrokaempferol. Our results showed that the relative docking energies estimated by docking softwares were in satisfactory correlation with the experimental activities. Preliminary interaction profile of P-gp docked complexes were also analysed in order to understand the nature of binding modes of these compounds. Our computational investigation suggested that the compounds interactions with the hydrophobic pocket of P-gp are mainly related to the inhibitory activity. Moreover this study s a platform for the discovery of novel natural compounds from herbal origin, as inhibitor molecules against the P-glycoprotein for the treatment of cancer.

Reversal of Multidrug Resistance and Computational Studies of Pistagremic Acid Isolated from Pistacia integerrima

  • Rauf, Abdur;Uddin, Ghias;Raza, Muslim;Ahmad, Aftab;Jehan, Noor;Ahmad, Bashir;Nisar, Muhammad;Molnar, Joseph;Csonka, Akos;Szabo, Diana;Khan, Ajmal;Farooq, Umar;Noor, Mah
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2311-2314
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    • 2016
  • Pistagremic acid (PA) is a bioactive triterpenoid isolated from various parts of Pistacia integerrima plants. The aim of this research was to investigate PA for reversion of multidrug resistant (MDR) mediated by P-glycoprotein using rhodamine-123 exclusion study on a multidrug resistant human ABCB1 (ATP-binding cassette, sub-family B, member 1) gene-transfected mouse T-lymphoma cell line in vitro. Results were similar to those with verapamil as a positive control. Docking studies of PA and standard Rhodamine123 were carried out against a P-gp crystal structure which showed satisfactory results. Actually, PA cannot bind exactly where co-crystallized ligand of P-gp is already present. However, the docking study predicted that if a compound gives a lesser score then it may have some potency. The docking scores of PA and Rhodamine were similar. Therefore, we can conclude that there are certain important chemical features of PA which are responsible for the inhibiting potency of P-gp.

A Novel Mutation in the DNA Binding Domain of NFKB is Associated with Speckled Leukoplakia

  • Govindarajan, Giri Valanthan Veda;Bhanumurthy, Lokesh;Balasubramanian, Anandh;Ramanathan, Arvind
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3627-3629
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    • 2016
  • Background: Activation and inactivation of nuclear factor of kappa light chain gene enhancer in B cells (NFKB) is tightly regulated to ensure effective onset and cessation of defensive inflammatory signaling. However, mutations within NFKB, or change in activation and inactivation molecules have been reported in a few cancers. Although oral squamous cell carcinoma is one of the most prevalent forms of cancer in India, with a development associated with malignant transformation of precancerous lesions, the genetic status of NFKB and relative rates of change in oral precancerous lesions remain unknown. Hence in the present study we investigated all twenty four exons of NFKB gene in two precancerous lesions, namely oral submucous fibrosis (OSMF) and oral leukoplakia (OL) to understand its occurrence, incidence and assess its possible contribution to malignant transformation. Materials and Methods: Chromosomal DNA isolated from twenty five each of OSMF and OL tissue biopsy samples were subjected to PCR amplification with intronic primers flanking twenty four exons of the NFKB gene. The PCR amplicons were subsequently subjected to direct sequencing to elucidate the mutation status. Results: Sequence analysis identified a novel heterozygous mutation, c.419T>A causing substitution of leucine with glutamine at codon 140 (L140Q) in an OL sample. Conclusions: The identification of a substitution mutation L140Q within the DNA binding domain of NFKB in OL suggests that NFKB mutation may be relatively an early event during transformation. To the best of our knowledge, this study is the first to have identified a missense mutation in NFKB in OL.

In Vitro Studies on Phytochemical Content, Antioxidant, Anticancer, Immunomodulatory, and Antigenotoxic Activities of Lemon, Grapefruit, and Mandarin Citrus Peels

  • Diab, Kawthar AE
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3559-3567
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    • 2016
  • Background: In recent years, there has been considerable research on recycling of agro-industrial waste for production of bioactive compounds. The food processing industry produces large amounts of citrus peels that may be an inexpensive source of useful agents. Objective: The present work aimed to explore the phytochemical content, antioxidant, anticancer, antiproliferation, and antigenotxic activities of lemon, grapefruit, and mandarin peels. Materials and Methods: Peels were extracted using 98% ethanol and the three crude extracts were assessed for their total polyphenol content (TPC), total flavonoid content (TFC), and antioxidant activity using DPPH (1, 1-diphenyl-2-picrylhydrazyl). Their cytotoxic and mitogenic proliferation activities were also studied in human leukemia HL-60 cells and mouse splenocytes by CCK-8 assay. In addition, genotoxic/antigenotoxic activity was explored in mouse splenocytes using chromosomal aberrations (CAs) assay. Results: Lemon peels had the highest of TPC followed by grapefruit and mandarin. In contrast, mandarin peels contained the highest of TFC followed by lemon and grapefruit peels. Among the extracts, lemon peel possessed the strongest antioxidant activity as indicated by the highest DPPH radical scavenging, the lowest effective concentration 50% ($EC_{50}=42.97{\mu}g\;extract/mL$), and the highest Trolox equivalent antioxidant capacity (TEAC=0.157). Mandarin peel exhibited moderate cytotoxic activity ($IC_{50}=77.8{\mu}g/mL$) against HL-60 cells, whereas grapefruit and lemon peels were ineffective anti-leukemia. Further, citrus peels possessed immunostimulation activity via augmentation of proliferation of mouse splenocytes (T-lymphocytes). Citrus extracts exerted non-cytotoxic, and antigenotoxic activities through remarkable reduction of CAs induced by cisplatin in mouse splenocytes for 24 h. Conclusions: The phytochemical constituents of the citrus peels may exert biological activities including anticancer, immunostimulation and antigenotoxic potential.

Phenotypic Characterization of MPS IIIA (Sgshmps3a/ Sgshmps3a) Mouse Model

  • Park, Sung Won;Ko, Ara;Jin, Dong-kyu
    • Journal of mucopolysaccharidosis and rare diseases
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    • v.4 no.1
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    • pp.26-36
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    • 2018
  • Mucopolysaccharidosis IIIA is a heritable neurodegenerative disorder resulting from the dysfunction of the lysosomal hydrolase sulphamidase. This leads to the primary accumulation of the complex carbohydrate heparan sulphate in a wide range of tissues and CNS degeneration. Characterization of animal model is the beginning point of the therapeutic clinical trial. Mouse model has a limitation in that it is not a human and does not have all of the disease phenotypes. Therefore, delineate of the phenotypic characteristics of MPS IIIA mouse model prerequisite for the enzyme replace treatment for the diseases. We designed 6-month duration of phenotypic characterization of MPS IIIA mouse biochemically, behaviorally and histologically. We compared height and weight of MPS IIIA mouse with wild type from 4 weeks to 6 months in both male and female. At 6 months, we measured GAG storage in urine kidney, heart, liver, lung and spleen. The brain GAG storage is presented with Alcian blue staining, immunohistochemistry, and electron-microscopy. The neurologic phenotype is evaluated by brain MRI and behavioral study including open field test, fear conditioning, T-maze test and Y-maze test. Especially behavioral tests were done serially at 4month and 6month. This study will show the result of the MPS IIIA mouse model phenotypic characterization. The MPS IIIA mouse provides an excellent model for evaluating pathogenic mechanisms of disease and for testing treatment strategies, including enzyme or cell replacement and gene therapy.

Screening of Anti-Atopic Dermatitis Material by Using NC/Nga Mouse Whole Blood System (NC/Nga 마우스 전혈을 이용한 항 아토피 피부염 물질 탐색)

  • Park, Dong-Hoon;Kim, Youn-Uck
    • IMMUNE NETWORK
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    • v.8 no.3
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    • pp.98-105
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    • 2008
  • Background: Allergic inflammation was induced by activated Th2 lymphocytes, leading to IgE production and eosinophil activation. A Th2 disproportion was shown in atopic children soon after birth. During specific allergen stimulation, an increase of Th2 cells was observed in most cases. In this study, we prepared new screening "whole blood" system for searching the anti-atopic materials. Cytokine production and IgE secretion from whole blood system were assessed and we confirmed the results by using animal system. Methods: Pathological features in NC/Nga mice are similar to those observed in human atopic dermatitis. Whole blood from NC/Nga mouse was stimulated by using TNCB (Th2 activator) or candidate materials of anti-atopic dermatitis, and the production of cytokines (IL-4, IL-12, and IFN-${\gamma}$) were measured by ELISA. In order to confirm the results of whole blood system, in vivo test was done by using NC/Nga mice. Results: In whole blood system, LPS and extracts of green tea, hardy orange and onion induced the production of IL-12 and IFN-${\gamma}$ while they reduced the production of IL-4. Also, LPS and extracts of onion reduced IgE production. Though atopic dermatitis was observed from a mouse stimulated with TNCB, it was not when a mouse was co-stimulated in LPS or extracts of onion. The results are same as those observed in whole blood system. Conclusion: Whole blood system was simple and speedy methods for searching a materials compared with the conventional high-cost animal system. And the results using whole blood system was proved to be reliable in our experiments for screening anti-atopic material. We expect that the system can be applied to other experiments for searching similar materials.