• Title/Summary/Keyword: biological assays

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Evaluation of Cryptosporidiurn Disinfection by Ozone and Ultraviolet Irradiation Using Viability and Infectivity Assays (크립토스포리디움의 활성/감염성 판별법을 이용한 오존 및 자외선 소독능 평가)

  • Park Sang-Jung;Cho Min;Yoon Je-Yong;Jun Yong-Sung;Rim Yeon-Taek;Jin Ing-Nyol;Chung Hyen-Mi
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.534-539
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    • 2006
  • In the ozone disinfection unit process of a piston type batch reactor with continuous ozone analysis using a flow injection analysis (FIA) system, the CT values for 1 log inactivation of Cryptosporidium parvum by viability assays of DAPI/PI and excystation were $1.8{\sim}2.2\;mg/L{\cdot}min$ at $25^{\circ}C$ and $9.1mg/L{\cdot}min$ at $5^{\circ}C$, respectively. At the low temperature, ozone requirement rises $4{\sim}5$ times higher in order to achieve the same level of disinfection at room temperature. In a 40 L scale pilot plant with continuous flow and constant 5 minutes retention time, disinfection effects were evaluated using excystation, DAPI/PI, and cell infection method at the same time. About 0.2 log inactivation of Cryptosporidium by DAPI/PI and excystation assay, and 1.2 log inactivation by cell infectivity assay were estimated, respectively, at the CT value of about $8mg/L{\cdot}min$. The difference between DAPI/PI and excystation assay was not significant in evaluating CT values of Cryptosporidium by ozone in both experiment of the piston and the pilot reactors. However, there was significant difference between viability assay based on the intact cell wall structure and function and infectivity assay based on the developing oocysts to sporozoites and merozoites in the pilot study. The stage of development should be more sensitive to ozone oxidation than cell wall intactness of oocysts. The difference of CT values estimated by viability assay between two studies may partly come from underestimation of the residual ozone concentration due to the manual monitoring in the pilot study, or the difference of the reactor scale (50 mL vs 40 L) and types (batch vs continuous). Adequate If value to disinfect 1 and 2 log scale of Cryptosporidium in UV irradiation process was 25 $mWs/cm^2$ and 50 $mWs/cm^2$, respectively, at $25^{\circ}C$ by DAPI/PI. At $5^{\circ}C$, 40 $mWs/cm^2$ was required for disinfecting 1 log Cryptosporidium, and 80 $mWs/cm^2$ for disinfecting 2 log Cryptosporidium. It was thought that about 60% increase of If value requirement to compensate for the $20^{\circ}C$ decrease in temperature was due to the low voltage low output lamp letting weaker UV rays occur at lower temperatures.

Effect of Unripe Apple Peel Water Extracts on Tyrosinase Activity and Melanin Production in B16F10 Melanoma Cells (B16F10 melanoma 세포에서 미성숙 사과 과피 열수추출물의 tyrosinase 활성과 melanin 생 성에 미치는 영향)

  • Jang, Young-Ah;Lee, Jin-Tae
    • Journal of Life Science
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    • v.28 no.8
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    • pp.900-907
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    • 2018
  • This study was carried out to evaluate the possibility of unripe apple peel water extracts as cosmetic materials and to evaluate the biological activities of the antioxidant and whitening effects of the samples. The antioxidative properties of the samples were confirmed by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) cation radical scavenging ability. To evaluate the whitening effect of the samples, several analytical techniques were used, including toxicity evaluations of the samples by MTT assays. Measurements of the inhibition rates of cellular tyrosinase, melanin synthesis rates, and expression rates of whitening-related proteins and genes were confirmed using melanoma (B16F10 cell). At equivalent unripe apple peel water concentrations ($1,000{\mu}g/ml$), the DPPH radical scavenging and the ABTS cation radical scavenging activities were 77.3% and 93.1%, respectively. The whitening activity evaluation showed that tyrosinase activity and melanin synthesis were inhibited by 19.8% and 17.3%, respectively, at unripe apple peel water extract concentrations of $50{\mu}g/ml$. In B16F10 cells induced by ${\alpha}$-MSH, the expression of tyrosinase, TRP-1, and TRP-2 decreased. Also, the activity of the transcription factor MITF was inhibited. In real-time PCR experiments, the expression of related genes at the upstream signal level was also found to be progressively lowered as the concentration of unripe apple peel water extracts increased. From these results, it was confirmed that the unripe apple peel water extracts showed excellent whitening efficacy and could be used as safe, natural, raw cosmetic material in the future.

SAMD4B, a novel SAM-containing protein, inhibits AP-1-, p53- and p21-mediated transcriptional activity

  • Luo, Na;Li, Guan;Li, Yongqing;Fan, Xiongwei;Wang, Yuequn;Ye, Xiangli;Mo, Xiaoyan;Zhou, Junmei;Yuan, Wuzhou;Tan, Ming;Xie, Huaping;Ocorr, Karen;Bodmer, Rolf;Deng, Yun;Wu, Xiushan
    • BMB Reports
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    • v.43 no.5
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    • pp.355-362
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    • 2010
  • The sterile alpha motif (SAM) is a putative protein interaction domain involved in a wide variety of biological processes. Here we report the identification and characterization of a novel gene, SAMD4B, which encodes a putative protein of 694 amino acids with a SAM domain. Northern blot and RT-PCR analysis showed that SAMD4B is widely expressed in human embryonic and adult tissues. Transcriptional activity assays show SAMD4B suppresses transcriptional activity of L8G5-luciferase. Over-expression of SAMD4B in mammalian cells inhibited the transcriptional activities of activator protein-1 (AP-1), p53 and p21, and the inhibitory effects can be relieved by siRNA. Deletion analysis indicates that the SAM domain is the main region for transcriptional suppression. The results suggest that SAMD4B is a widely expressed gene involved in AP-1-, p53- and p21-mediated transcriptional signaling activity.

MicroRNA-301b promotes cell proliferation and apoptosis resistance in triple-negative breast cancer by targeting CYLD

  • Song, Hongming;Li, Dengfeng;Wu, Tianqi;Xie, Dan;Hua, Kaiyao;Hu, Jiashu;Deng, Xiaochong;Ji, Changle;Deng, Yijun;Fang, Lin
    • BMB Reports
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    • v.51 no.11
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    • pp.602-607
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    • 2018
  • Aberrant expression of microRNAs (miRNAs) plays important roles in carcinogenesis and tumor progression. However, the expression and biological role of miR-301b in triple-negative breast cancer (TNBC) remains unclear. Here we aimed to evaluate the roles and mechanisms of miR-301b in TNBC cells. miR-301b expression was assessed in TNBC specimens and cell lines by quantitative Real-Time PCR (qRT-PCR). TNBC cells were transfected with miR-301b mimics, inhibitors or Cylindromatosis (CYLD) small interfering RNA (siRNA) using Lipofectamine 2000. The functional roles of miR-301b were determined by cell proliferation, colony formation, and apoptosis assays. Western blots and qRT-PCR were used to measure the expression of mRNAs and proteins in the cells. We found that miR-301b was upregulated in TNBC specimens and cell lines. Overexpression of miR-301b promoted cell proliferation in TNBC cells, while inhibited the apoptosis induced by 5-FU. CYLD was downregulated by miR-301b at both mRNA and protein levels in TNBC cells. Dual-luciferase report assay confirmed that miR-301b downregulated CYLD by direct interaction with the 3'-untranslated region(3'-UTR) of CYLD mRNA. $NF-{\kappa}B$ activation was mechanistically associated with miR-301b-mediated downregulation of CYLD. However, inhibition of miR-301b reversed all the effects of miR-301b. In conclusion, miR-301b plays an oncogenic role in TNBC possibly by downregulating CYLD and subsequently activating $NF-{\kappa}B$ p65, and this may provide a novel therapeutic approach for TNBC.

Nutritional Characteristics and Screening of Biological Activity of Agarum cribrosum (구멍쇠미역의 식품영양학적 접근 및 생리활성 분석)

  • Park, Sung-Jin;Min, Kyung-Jin;Park, Tae-Gil
    • The Korean Journal of Food And Nutrition
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    • v.25 no.4
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    • pp.842-849
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    • 2012
  • The purpose of this study is to determine the possibility of using Agarum cribrosum as a natural health food source. To accomplish this purpose, the contents of the general and antioxidant activities of Agarum cribrosum were measured. The contents of carbohydrate, crude protein, crude lipid and ash were 45.4%, 15.0%, 2.3% and 33.1%. The calories of Agarum cribrosum were 262.3 kcal and total dietary fiber of Agarum cribrosum was 34.0%. The protein contained a total of 18 different kinds of amino acids. The content of amino acids was 12,402.42 mg/100 g. The K was the largest mineral followed by Ca, Na and Mg, implying that Agarum cribrosum is an alkali material. The antioxidant activity of Agarum cribrosum was assessed by various radical scavenging assays using DPPH (2,2-diphenyl-1-picrylhydrazyl), FRAP (ferric ion reducing antioxidant power), reducing power, and ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)). All antioxidant activity of Agarum cribrosum extract increased the concentration of the dependents. Total phenolic contents of Agarum cribrosum extract were $34.1{\pm}2.56mg/g$, and total flavonoids contents were estimated at $4.9{\pm}0.26mg/g$. General nutrients and other antioxidant bioactive materials in Agarum cribrosum were also potential materials for good health food. It is expected that a follow-up study of Agarum cribrosum through developing processed food and evaluation of their functional properties would provide useful information or sources of functional foods.

Repellent Activity of the Extracts of Acorus Gramineus against Mosquito (Culex pipinens pallens) (석창포 추출물의 모기에 대한 기피활성)

  • Lee, Seung-Ho;Jung, Jin-Kwan;Park, Sang-Hyun;Kim, Mun-Sik;Han, Yeon-Soo;Seo, Jung-Mi;Kim, In-Seon
    • Korean Journal of Environmental Agriculture
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    • v.30 no.1
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    • pp.76-81
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    • 2011
  • BACKGROUND: Acorus gramineus is of medicinal plants that exhibit variable biological activities for human health and against insect pests. The extracts of A. gramineus was examined in an attempt to develop a natural repellent against human disease-mediating Culex pipiens. METHODS AND RESULTS: The roots of A. gramineus dried under dark conditions were homogenized and extracted with ethanol. The extracts were subjected to repellent activity assays against C. pipiens in a hand-made acrylamide box with three accessible rooms. Significantly low number of mosquitos was found in the room previously fumigated with the extracts at 50 mg/L on the filter paper, exhibiting less than 20% of mosquitos tested. More than 50% of mosquitos tested was found in the room without the extracts, but less than 30% was found in the room that released mosquitos. GC/MS analysis detected ${\beta}$-asarone as a main component of the extracts. The commercial asarones (${\alpha}$ and ${\beta}$) showed a repellent activity at 50 mg/L on the filter paper similar to the extracts. CONCLUSION(S): A. gramineus has potential for use as a mosquito repellent since ${\beta}$-asarone, a main component of the plant, exhibited a strong repellent activity against C. pipiens.

Comparative study on stability and efficacy of Banhasasim-tang decoction depending on the preservation temperature and periods (반하사심탕 전탕액의 보관 온도 및 기간에 따른 안정성 및 유효성 연구)

  • Jin, Seong Eun;Kim, Ohn Soon;Seo, Chang-Seob;Shin, Hyeun-Kyoo;Jeong, Soo-Jin
    • The Journal of Korean Medicine
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    • v.37 no.1
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    • pp.21-33
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    • 2016
  • This study aimed to investigate the stability and biological activities of BHSST decoction depending on the preservation temperature and periods. Methods: BHSST decoction was preserved at room temperatures (R/T, $23{\pm}1^{\circ}C$) or refrigeration ($4^{\circ}C$) for 0, 30, 60 and 90 days. To evaluate the stability of BHSST decoction, pH and sugar content were estimated. In addition, high-performance liquid chromatography (HPLC) analysis was performed to determine marker compounds of BHSST decoction. To evaluate anti-inflammatory effect, nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) productions were measured in LPS-stimulated RAW 264.7 macrophages. Antioxidant activity was examined using the assays for 3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities. Results: There was no change in pH and sugar content depending on the preservation temperature and periods of BHSST decoction. Among the major components of BHSST, contents of liquiritin, baicalein and wogonin was reduced time-dependently both at R/T and $4^{\circ}C$. Inhibitory effects of BHSST decoction on NO and PGE2 productions were slightly decreased in a time-dependent manner by 90 days of preservation. In addition, BHSST decoction maintained ABTS and DPPH radical scavenging activities by 60 days while significantly reducing the activities in 90 days of preservation at R/T. By contrast, BHSST decoction had no significant change of ABTS and DPPH radical scavenging activities by 90 days at $4^{\circ}C$. Conclusions: Our results suggest that the stability and efficacy of BHSST decoction are maintained for 60 days at $4^{\circ}C$ rather than R/T.

Different Potential of Hematopoietic Differentiation in Two Distinct Mouse Embryonic Stem Cells (두 개의 다른 마우스 배아줄기세포의 차별적인 조혈세포 분화능)

  • Kim, Jin-Sook;Kang, Ho-Bum;Song, Jee-Yeon;Oh, Goo-Taeg;Nam, Ki-Hoan;Lee, Young-Hee
    • Development and Reproduction
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    • v.9 no.2
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    • pp.105-114
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    • 2005
  • Embryonic stem(ES) cells have tremendous potential as a cell source for cell-based therapies. Realization of that potential will depend on our ability to understand and manipulate the factors that influence cell fate decision and to develop methods for getting enough cell numbers for clinical applications. Hematopoiesis has been widely studied, and hematopoietic differentiation from ES cells is a good model to study lineage commitment. In this study, we investigated stemness and compared the efficiency of hematopoietic differentiation using two different mouse embryonic stem cell lines TC-1 and B6-1. Although the two cell lines showed known stem cell properties with minor differences, the embryoid body formation efficiency in methylcellulose was much higher in TC-1 than B6-1. When measured potentials of hematopoietic differentiation using functional(colony-forming cell) and phenotypic(specific marker expression) assays, we found that TC-1 can differentiate into hematopoietic cells in methylcellulose culture but B6-1 cannot. These results imply that we can improve the efficiency of hematopoietic cell differentiation by selection of proper cell lines and this may be also applied in the differentiation of human embryonic stem cells.

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The Adjuvant Effect of Sophy ${\beta}$-Glucan to the Antibody Response in Poultry Immunized by the Avian Influenza A H5N1 and H5N2 Vaccines

  • Le, Thanh Hoa;Le, Tran Binh;Doan, Thanh Huong Thi;Quyen, Dong Van;Le, Kim XuyenThi;Pham, Viet Cuong;Nagataki, Mitsuru;Nomura, Haruka;Ikeue, Yasunori;Watanabe, Yoshiya;Agatsuma, Takeshi
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.405-411
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    • 2011
  • Avian influenza virus vaccines produced in oil-emulsified inactivated form with antigen content of at least 160 hemagglutinin units (HAU) induced immunity in birds. However, in addition to enhancing the effect of the adjuvant(s), other additional supplemented biological compounds included in inactivated vaccines could produce higher levels of antibody. We examined in chickens, Vietnamese ducks, and muscovy ducks the adjuvant effect of Sophy ${\beta}$-glucan (SBG), a ${\beta}$-1,3-1,6 glucan produced by the black yeast Aureobasidium pollulans strain AF0-202, when administered with an avian influenza H5 subtype vaccine. In Experiment 1, 40 chickens (ISA Brown hybrid), allocated to four groups of ten each, were immunized with Oil-H5N1(VN), Oil-H5N1(CN), Oil-H5N2(CN), and saline (control group), respectively. In Experiment 2, chickens (ISA Brown hybrid), muscovy ducks (French hybrid), and Vietnamese ducks (indigenous Vietnamese) were used to further assess the effect of SBG on immunogenicity of the Oil-H5N1(VN) Vietnamese vaccine. ELISA and hemagglutination inhibition (HI) assays were used to assess the antibody response. The H5 subtype vaccines initiated significantly higher immune responses in the animals dosed with SBG, with 1.0-1.5 $log_2$ higher HI titers and 10-20% ELISA seroconversion, compared with those not dosed with ${\beta}$-glucan. Notably, some of the animals dosed with SBG induced HI titers higher than 9.0 $log_2$ following boosting immunization. Taken together, our serial studies indicated that SBG is a potential effector, such as enhancing the immune response to the H5 vaccines tested.

Evaluation of Pregnancy and Thyroid Function (임신과 갑상선 기능의 평가)

  • Park, Chang-Eun
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.1
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    • pp.1-10
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    • 2018
  • During early pregnancy, before the development of a functioning thyroid gland, thyroid stimulating hormone (TSH) is a very sensitive marker of thyroid dysfunction during pregnancy. Normal values have been modified during gestation with a downward shift. The fetus is influenced by the TSH supplied by the mother. TSH and free thyroxine (FT4) concentrations vary during pregnancy and conventional units can vary between laboratories. A downward shift of the TSH reference range occurs during pregnancy, with a decrease in both the lower and upper limits of maternal TSH, relative to the typical non-pregnant TSH reference range. Each laboratory produces its own reference TSH and FT4 concentrations because there are many different assays that yield different results in pregnancy. Therefore, automated immunoassays used for serum FT4 analysis are still used widely, but the important considerations discussed above must be noted. The use of population-based, trimester-specific reference ranges remains the best way to handle this issue The slight downward shift in the upper reference range of TSH occurring in the latter first trimester (7~12 weeks) of pregnancy, typically not observed prior to 7 weeks. Their use indicates high or low levels in a quantitative manner independent of the reference ranges. These data highlight the importance of calculating population-based pregnancy-specific thyroid parameter reference intervals. A precision medicine initiative in this area will require the collection and analysis of a large number of genetic, biological, psychosocial, and environmental variables in large cohorts of individuals. Large prospective randomized controlled trials will be needed to resolve these controversies.