• Title/Summary/Keyword: S-100 protein

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Effects of Polygalae Radix Preparata Cum Glycyrrhizae Radix on 4-HNE-induced Apoptosis in PC-12 cell (지감초자(志甘草煮)가 4-HNE로 유도된 Apoptosis에 미치는 영향)

  • Ham, Mee-Jin;Kim, Youn-Sub
    • The Korea Journal of Herbology
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    • v.31 no.1
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    • pp.77-82
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    • 2016
  • Objectives : The study was designed to observe the effect of Polygalae Radix Preparata Cum Glycyrrhizae Radix on 4-Hydroxynonenal (4-HNE)-induced apoptosis in PC-12 cell.Methods : A cytotoxic test on Polygalae Radix Preparata Cum Glycyrrhizae Radix (PG) was conducted and another MTT assay was conducted to observe the cytoprotective effect against 4-HNE that cause oxidative stress. In addition, in order to observe the expression of Bax, Bcl-2, Caspase-3 and TNF-α protein involved with apoptosis, western blot was conducted.Results : The groups treated with 25 ㎍, 50 ㎍ and 100 ㎍ of PG water extract had no toxicity for PC-12 cell. The groups treated with 25 ㎍, 50 ㎍ and 100 ㎍ of PG water extract showed a significant increase of cell survival rate in comparison with the control group injected by only 4-HNE. The groups treated with 25 ㎍ and 50 ㎍ of PG water extract showed a significant supression on increase of Bax protein expression in the control group. The group treated with 100 ㎍ of PG water extract showed a significant promotion on decrease of Bcl-2 protein expression in the control group. The group treated with 50 ㎍ of PG water extract showed a significant supression on increase of Caspase-3 protein expression in the control group. The group treated with 25 ㎍ of PG water extract showed a significant supression on increase of TNF-α protein expression in the control group.Conclusions : These results suggest that Polygalae Radix Preparata Cum Glycyrrhizae Radix is effective in reducing apoptosis by 4-HNE-dameged cell.

S100A8 and S100A9 Secreted by Allergens in Monocytes Inhibit Spontaneous Apoptosis of Normal and Asthmatic Neutrophils via the Lyn/Akt/ERK Pathway (단구에서 분비되는 S100A8과 S100A9의 Lyn/Akt/ERK 경로를 통한 정상인과 천식질환 호중구의 세포고사 억제 효과)

  • Kim, In Sik;Lee, Ji-Sook
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.2
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    • pp.128-134
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    • 2017
  • Der p 1 and Der p 2 are essential allergens of house dust mite associated with the development of asthma. In the present study, we examined whether Der p 1 and Der p 2 induce a release of S100A8 and S100A9 in monocytes, which are involved in the regulatory mechanism of neutrophil apoptosis. We found that Der p 1 and Der p 2 significantly increased the secretion of S100A8 and S100A9 in normal monocytes. Moreover, S100A8 and S100A9 strongly suppressed the spontaneous apoptosis of normal and asthmatic neutrophils. The inhibitory effect of S100A9 was stronger than that of S100A8, and asthmatic neutrophils showed a higher inhibitory effect than normal neutrophils. S100A8 and S100A9 induced activation of Lyn, Akt, and ERK in a time-dependent manner. These findings elucidate the roles of Der p 1 and Der p 2 in the interaction between monocytes and neutrophils, as well as contributing to our knowledge of the pathogenesis of allergic diseases.

Antitumor Activity of Kp, a Protein-polysaccharide from the Mycelial Culture of Phellinus linteus (Phellinus linteus 균사 배양물로부터 분리한 단백다당체 Kp의 항암활성)

  • Chung, Kyeong-Soo;Kim, Shin-Sook;Kim, Hee-Soo;Han, Man-Woo;Kim, Byong-Kak
    • YAKHAK HOEJI
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    • v.38 no.2
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    • pp.158-165
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    • 1994
  • A protein-polysaccaride fraction Kp(53.9% polysaccharide, 14.2% protein) was separated from the shake-cultured mycelia of a basidiomycetous fungus, Phellinus linteus, and its antitumor activity against sarcoma 180 in ICR mice was investigated. When administered after the tumor implantation, Kp exerted antitumor activity by inhibiting the growth of the sarcoma 180 solid tumor by 71.5% and increasing the life span of the sarcoma 180 ascitic mice by 51.5% at 100 mg/kg. In pretreatment tests, in which Kp was administered once daily for 9 days before the tumor implantation, Kp inhibited the growth of the solid and ascites form of sarcoma 180, respectively, by 35.4% and by 80.3% at 100 mg/kg. However, Kp showed no in vitro cytotoxic activity against a murine leukemia L1210 and a human gastric tumor SNU.1 upto the concentration of $200\;{\mu}g/ml$. From these results, it is clear that the antitumor activity of Kp is exerted through its immunomodulating activity on the host's immune system.

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Determination of Adequate Method for Protein Extraction from Rice Bran and the Substitution of Dried Skim Milk with Protein Concentrate from Rice Bran in Early Weaned Pigs

  • Phipek, W.;Nagasinha, C.;Vallisuth, S.;Nongyao, C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1268-1273
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    • 2011
  • The present study was conducted to determine a feasible method of protein concentrate extraction from rice bran (RBPC) and its effect as a substitution for skim milk in early weaning pig diets. An investigation to extract protein concentrate from full fat rice bran was undertaken to determine the best ratio of water and rice bran, the amount of NaOH and a HCl solvent to use in a simple paddle-type mixer with modified spinning to produce RBPC. The results stated that the best ratio for water mixing in the RBPC extraction process was 1:5 with 20 g NaOH and 30 min in a paddle-type mixer at 300 rpm. A mix of 250 ml 0.2 N HCl was optimum for neutralization and protein precipitation. After the fluid was spun out with a washing machine, the sediment was left for 12-14 hours to complete the filtration. One kilogram of rice bran could produce an average of 324.5 gram RBPC and it contained 3.40% ash, 496.48 kcal of GE/100 gram, 1.94% crude fiber, 28.20% ether extract, 7.64% moisture and 16.66% crude protein, respectively. A total of 45 crossbred piglets, weaned at 3 weeks of age were allotted into control diet (A) and dietary treatments formulated with a four different rates of RBPC substitution for skim milk at a percentage of 25 (B), 50 (C), 77 (D) and 100 (E) respectively, in a randomized complete block (RCB) design. All piglets had free access to feed and water until 8 week of age when the experiment ended. Feed intake, average daily gain, growth rate and feed efficiency were not affected by dietary treatments. Blood test parameters after completion of the growth trial indicated normal health. Even though the mean of cell hemoglobin concentration was significantly different between treatments (p<0.05) it was still within the normal range. The cost difference for BW gain of 100% RBPC substituted for skim milk in the weaning diet was approximately 35% lower than that of the control and the relative cost of production was 96.67, 92.85, 70.75 and 64.48% lower for the replacement of 25, 50, 75 and 100% of skim milk respectively. These results implied that this technology is feasible for use by small scale farmers to improve their self-reliance.

Iodine Supplementation of Leucaena leucocephala Diet for Goats. II. Effects on Blood Metabolites and Thyroid Hormones

  • Rajendran, D.;Pattanaik, A.K.;Khan, S.A.;Bedi, S.P.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.6
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    • pp.791-796
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    • 2001
  • Twelve adult male goats, comprising of six castrated and six intacts, (2.5-3 years; $24.4{\pm}0.62kg$) were randomly but evenly divided into two groups ($I_0$ and $I_{100}$) and fed conventional concentrate mixture along with Leucaena leucocephala leaf meal (100 g/head approx.), the latter to supply 50 per cent of the crude protein (CP) requirements. The $I_{100}$ group was provided with supplemental iodine as potassium iodide solution at 0.1 mg/day/animal. Wheat straw was provided ad libitum as sole source of roughage during the experimental period of 105 d. Blood samples were collected at the begining (0 d) and thereafter at 30, 60 and 90 d of experimental feeding. The study revealed that the serum glucose level was significantly higher (p<0.01) in $I_{100}$ group as compared to $I_0$. Haemoglobin, packed cell volume and serum concentrations of total protein, albumin, globulin, calcium, inorganic phosphorus and alkaline phosphatase did not show significant differences as a result of iodine supplementation. Though the serum levels of triiodothyronine ($T_3$) were comparable between the two groups, that of thyroxine ($T_4$) increased significantly (p<0.001) in the $I_{100}$ group. The $T_3:T_4$ ratio was also similar between both the groups. The study indicated that the adverse effect of Leucaena feeding on thyroid gland could possibly be alleviated by provision of extra iodine. However, this needs further confirmation using long duration studies.

Iodine Supplementation of Leucaena leucocephala Diet for Goats. I. Effects on Nutrient Utilization

  • Rajendran, D.;Pattanaik, A.K.;Khan, S.A.;Bedi, S.P.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.6
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    • pp.785-790
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    • 2001
  • Twelve indigenous male goats, comprising of six intact and six castrated (2.5-3 years; $24.4{\pm}0.62kg$) were assigned evenly into two dietary treatments, viz. $I_0$ and $I_{100}$ and were used to study the effect of supplementation of iodine on the nutrient utilization when their diet contained Leucaena leaf meal. They were offered a conventional concentrate mixture along with Leucaena leucocephala leaf meal, the latter to meet 50% of their crude protein (CP) requirements, and supplemented with either no iodine ($I_0$) or 0.1 mg of iodine ($I_{100}$)/day/animal as potassium iodide for a period of 105 days. Wheat straw given ad libitum was the sole source of roughage. A metabolism trial of 8 days duration was conducted after 90 days of experimental feeding. It was observed that the overall dry matter (DM) intake during experimental period was higher (p<0.05) in $I_{100}$ group as compared to $I_0$ group (508.6 vs. $443.7g\;d^{-1}$). The intake of CP, digestible crude protein (DCP) and metabolisable energy (ME), although non-significant, tended to be higher in the iodine supplemented group, $I_{100}$. Digestibility of dry matter, organic matter (OM), CP, ether extract and crude fiber (CF) did not differ (p>0.05) between the treatments. However, nitrogen retention was higher (p<0.01) in $I_{100}$ than $I_0$ with the values being 2.63 and $1.70g\;d^{-1}$, respectively. No difference (p>0.05) was evident in the retention of calcium and phosphorus between the two groups. The castrated animal exhibited lower DM intake concurrent with higher digestibility of DM and crude fibre (p<0.05), and organic matter and total carbohydrates (p<0.01) when compared to intact ones. It was concluded that supplementation of iodine to leucaena based ration may help in improving the DM intake and nitrogen utilization by goats.

Characterization of a Monoclonal Antibody Specific to Human Siah-1 Interacting Protein (인체 SIP 단백질에 특이적인 단일클론 항체의 특성)

  • Yoon, Sun Young;Joo, Jong Hyuck;Kim, Joo Heon;Kang, Ho Bum;Kim, Jin Sook;Lee, Younghee;Kwon, Do Hwan;Kim, Chang Nam;Choe, In Seong;Kim, Jae Wha
    • IMMUNE NETWORK
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    • v.4 no.1
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    • pp.23-30
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    • 2004
  • Background: A human orthologue of mouse S100A6-binding protein (CacyBP), Siah-1-interacting protein (SIP) had been shown to be a component of novel ubiquitinylation pathway regulating $\beta$-catenin degradation. The role of the protein seems to be important in cell proliferation and cancer evolution but the expression pattern of SIP in actively dividing cancer tissues has not been known. For the elucidation of the role of SIP protein in carcinogenesis, it is essential to produce monoclonal antibodies specific to the protein. Methods: cDNA sequence coding for ORF region of human SIP gene was amplified and cloned into an expression vector to produce His-tag fusion protein. Recombinant SIP protein and monoclonal antibody to the protein were produced. The N-terminal specificity of anti-SIP monoclonal antibody was conformed by immunoblot analysis and enzyme linked immunosorbent assay (ELISA). To study the relation between SIP and colon carcinogenesis, the presence of SIP protein in colon carcinoma tissues was visualized by immunostaining using the monoclonal antibody produced in this study. Results: His-tag-SIP (NSIP) recombinant protein was produced and purified. A monoclonal antibody (Korea patent pending; #2003-45296) to the protein was produced and employed to analyze the expression pattern of SIP in colon carcinoma tissues. Conclusion: The data suggested that anti-SIP monoclonal antibody produced here was valuable for the diagnosis of colon carcinoma and elucidation of the mechanism of colon carcinogenesis.

Purification and Characterization of Farnesyl Protein Transferase from Bovine Testis

  • Ryo, Kwon-Yul;Baik, Young-Jin;Yang, Chul-Hak
    • BMB Reports
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    • v.28 no.3
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    • pp.197-203
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    • 1995
  • Famesyl protein transferase involved in the first step of post-translational modification of $p21^{ras}$ proteins transfers the famesyl moiety from famesyl pyrophosphate to a cysteine residue in $p21^{ras}$ proteins. The enzyme was first purified 30,000-fold from bovine testis by use of 30~50% ammonium sulfate fractionation, DEAE-Sephacel ion exchange chromatography, Sephacryl S-300 gel filtration chromatography, Sephacryl S-200 gel filtration chromatography, and hexapeptide (Lys-Lys-Cys-Val-Ile-Met) affinity chromatography. The molecular weight of the purified enzyme was estimated to be ~100 kDa by gel filtration and SDS-polyacrylamide gels showed two closely spaced bands of ~50 kDa protein. These indicate that the enzyme consists of two nonidentical subunits, a and 13, which have slightly different molecular weights. The enzyme was inhibited by hexapeptide (Lys-Lys-Cys-Val-Ile-Met), which acted as an alternative substrate that competed for famesylation. Kinetic analysis by measuring initial velocities showed that famesyl protein transferase is a very slow enzyme. EDTA-treated famesyl protein transferase showed little activity with $Mg^{2+}$ or $Zn^{2+}$ alone, but required both $Mg^{2+}$ and $Zn^{2+}$ for the catalytic activity.

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Property Comparison of Polyclonal Anti-Soy Protein Antibodies Produced for ELISA (효소면역측정법을 위한 다클론 항대두단백 항체의 생산 및 특성비교)

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Yun, Sung-Seob
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1221-1226
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    • 2000
  • Specific antibodies were produced to develope the enzyme-linked immunosorbent assay for analysis of soy proteins and the properties of the antibodies were compared. Isolate soy protein(ISP), and ISP heated with SDS and urea (ISP(SU)), acidic subunits(AS) of 11S globulin were immunized to produce polyclonal antibodies. By using competitive indirect ELISA(ciELISA), the reactivities of the antibodies toward soy proteins treated with different methods were investigated and shown as $IC_{50}$. $IC_{50}'s$ of anti-ISP antibodies to ISP, ISP(SU), ISP treated with 2-ME(ISP(ME)), and crude 11S were 20, 30, 36, and $1000\;{\mu}g/mL$, respectively. And the values of anti-ISP(SU) antibodies to the same antigens were 100, 5, 4, and $220\;{\mu}g/mL$ and those of anti-AS antibodies were 20, 2, 2.5, and $200\;{\mu}g/mL$, respectively. Therefore, anti-AS antibodies showed the highest reactivities toward soy proteins among the produced antibodies as determined by ciELISA.

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Physico-chemical Properties and In Vivo Anti-cancer Effects of Potato Kimchi Prepared by adding Hot Water Extracts of Potato (생감자의 열탕 추출물을 첨가한 감자 김치의 이화학적 특성 및 In Vivo에서의 항암 효과)

  • Chang, Sang-Keun;Kim, Hee-Joo
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.3
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    • pp.302-310
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    • 2008
  • In this study, potato kimchi was prepared by applying heat to raw potatoes, and then the physico-chemical properties and anti-cancer effects of the kimchi were analyzed. The texture results indicated the potato kimchi had very good hardness and springiness attributes. During th late storage period, total vitamin C content of the kimchi slowly increased. In addition, the potato kimchi had non-volatile organic acid changes that promoted early aging; however, after the complete aging period, it was comparatively similar to other types of kimchi. Using the methanol extracts of various kimchi samples, the potato kimchi(solid 100%) showed the highest anti-carcinogenic effects in terms of anti-tumor activity in tumor bearing Balb/c mice with sarcoma-180 cells. In addition, the effects of the methanol extracts on hepatic glutathione S-transferase content were $289.76\;{\mu}mol/mg$ protein/min, $250.97\;{\mu}mol/mg$ protein/min, $251.20\;{\mu}mol/mg$ protein/min, $219.53\;{\mu}mol/mg$ protein/min, $183.79\;{\mu}mol/mg$ protein/min, for control kimchi, mul kimchi, and two potato kimchis [(solid 100%) and(solid 60%+kimchi juice 40%)], respectively. The in vivo anti-cancer effects of the potato kimchi were investigated using AGS human gastric adenocarcionoma cells and HT-29 human colon adenocarcionoma cells. Overall, an MTT assay revealed that the methanol extract of the potato kimchi showed the highest anti-carcinogenic effects.

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