• Title/Summary/Keyword: in vitro and in vivo protein quality

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Ginsenoside Rg5 overcomes chemotherapeutic multidrug resistance mediated by ABCB1 transporter: in vitro and in vivo study

  • Feng, Sen-Ling;Luo, Hai-Bin;Cai, Liang;Zhang, Jie;Wang, Dan;Chen, Ying-Jiang;Zhan, Huan-Xing;Jiang, Zhi-Hong;Xie, Ying
    • Journal of Ginseng Research
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    • v.44 no.2
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    • pp.247-257
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    • 2020
  • Background: Multidrug resistance (MDR) to chemotherapy drugs remains a major challenge in clinical cancer treatment. Here we investigated whether and how ginsenoside Rg5 overcomes the MDR mediated by ABCB1 transporter in vitro and in vivo. Methods: Cytotoxicity and colon formation as well as the intracellular accumulation of ABCB1 substrates were carried out in MDR cancer cells A2780/T and A549/T for evaluating the reversal effects of Rg5. The expressions of ABCB1 and Nrf2/AKT pathway were determined by Western blotting. An A549/T cell xenograft model was established to investigate the MDR reversal activity of Rg5 in vivo. Results: Rg5 significantly reversed ABCB1-mediated MDR by increasing the intracellular accumulation of ABCB1 substrates without altering protein expression of ABCB1. Moreover, Rg5 activated ABCB1 ATPase and reduced verapamil-stimulated ATPase activity, suggesting a high affinity of Rg5 to ABCB1 binding site which was further demonstrated by molecular docking analysis. In addition, co-treatment of Rg5 and docetaxel (TXT) suppressed the expression of Nrf2 and phosphorylation of AKT, indicating that sensitizing effect of Rg5 associated with AKT/Nrf2 pathway. In nude mice bearing A549/T tumor, Rg5 and TXT treatment significantly suppressed the growth of drug-resistant tumors without increase in toxicity when compared to TXT given alone at same dose. Conclusion: Therefore, combination therapy of Rg5 and chemotherapy drugs is a strategy for the adjuvant chemotherapy, which encourages further pharmacokinetic and clinical studies.

Structural Identification of Modified Amino Acids on the Interface between EPO and Its Receptor from EPO BRP, Human Recombinant Erythropoietin by LC/MS Analysis

  • Song, Kwang-Eun;Byeon, Jaehee;Moon, Dae-Bong;Kim, Hyong-Ha;Choi, Yoo-Joo;Suh, Jung-Keun
    • Molecules and Cells
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    • v.37 no.11
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    • pp.819-826
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    • 2014
  • Protein modifications of recombinant pharmaceuticals have been observed both in vitro and in vivo. These modifications may result in lower efficacy, as well as bioavailability changes and antigenicity among the protein pharmaceuticals. Therefore, the contents of modification should be monitored for the quality and efficacy of protein pharmaceuticals. The interface of EPO and its receptor was visualized, and potential amino acids interacting on the interface were also listed. Two different types of modifications on the interface were identified in the preparation of rHu-EPO BRP. A UPLC/Q-TOF MS method was used to evaluate the modification at those variants. The modification of the oxidized variant was localized on the Met54 and the deamidated variants were localized on the Asn47 and Asn147. The extent of oxidation at Met54 was 3.0% and those of deamidation at Asn47 and Asn147 were 2.9% and 4.8%, respectively.

In vitro and in vivo evaluation of kenaf (Hibiscus cannabinus L.) as a roughage source for beef cattle

  • Oh, Seongjin;Mbiriri, David Tinotenda;Ryu, Chaehwa;Lee, Kangheon;Cho, Sangbuem;Choi, Nag-Jin
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.10
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    • pp.1598-1603
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    • 2018
  • Objective: The goal of this study was to evaluate kenaf as a roughage source in vitro and its effects on meat quality of Hanwoo (Korean native) cattle. Methods: Three roughage materials, rice straw silage, ryegrass silage, and kenaf silage, were tested in a batch culture and feeding trial. Rumen fermentation parameters, including gas, pH, volatile fatty acid (VFA), and ammonia were analyzed. In the feeding trial, Hanwoo steers ($373.5{\pm}5.1kg$, n = 36, 11 month of age) were divided into three feeding groups (n = 12 each). Animals were fed with each silage and concentrate until the fattening stage. Results: Crude protein, ether extract, and non-structural carbohydrates were greater in kenaf silage. Total gas production was higher in ryegrass silage, followed by kenaf silage and rice straw silage (p<0.05). Total VFA and individual VFA (acetate, propionate, and n-butyrate) were greater in kenaf silage than rice straw silage (p<0.05). In vitro dry matter digestibility showed a similar trend to that of total gas and VFA production; it was higher in ryegrass silage and lower in rice straw (p<0.05). Throughout the feeding trial, the rice straw silage group showed significantly greater average daily gain than did the others (p<0.05). The feed conversion ratio in the group fed kenaf silage was significantly greater than that of others (p<0.05). No significant differences were observed in yield or quality traits, including carcass weight, ribeye area, backfat thickness, and scores for marbling, meat color, and fat color (p>0.05). Conclusion: The results indicated that no negative effects on growth performance and carcass characteristics occurred across treatments. Therefore, kenaf could be substituted for rice straw, which is most widely used as a roughage source in Korea.

Broken rice in a fermented total mixed ration improves carcass and marbling quality in fattened beef cattle

  • Kotupan, Salisa;Sommart, Kritapon
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1331-1341
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    • 2021
  • Objective: This study aimed to determine the effects of replacing cassava chips with broken rice in a fermented total mixed ration diet on silage quality, feed intake, ruminal fermentation, growth performance, and carcass characteristics in the final phase of fattening beef cattle. Methods: Eighteen Charolais-Thai native crossbred steers (average initial body weight: 609.4±46 kg; average age 31.6 mo) were subjected to three ad libitum dietary regimes and were maintained in individual pens for 90 d before slaughter. The experimental design was a randomized complete block design by initial age and body weight with six replicates. The dietary regimens used different proportions of broken rice (0%, 16%, and 32% [w/w] of dry matter [DM]) instead of cassava chips in a fermented total mixed ration. All dietary treatments were evaluated for in vitro gas production and tested in in vivo feeding trials. Results: The in vitro experiments indicated that organic matter from broken rice was significantly more digestible than that from a cassava-based diet (p<0.05). Silage quality, nutrient intake, ruminal fermentation characteristics, carcass fat thickness, and marbling score substantially differed among treatments. The ruminal total volatile fatty acids, propionate concentration, dietary protein intake, and digestibility increased linearly (p<0.05) with broken rice, whereas acetate concentration and the acetate:propionate ratio decreased linearly (p<0.05) with broken rice (added up to 32 g/kg DM). Broken rice did not influence plasma metabolite levels or growth performance (p>0.05). However, the marbling score increased, and the carcass characteristics improved with broken rice. Conclusion: Substitution of cassava chips with broken rice in beef cattle diets may improve fattened beef carcass quality because broken rice increases rumen fermentation, fatty acid biosynthesis, and metabolic energy supply.

The antioxidant icariin protects porcine oocytes from age-related damage in vitro

  • Yoon, Jae-Wook;Lee, Seung-Eun;Park, Yun-Gwi;Kim, Won-Jae;Park, Hyo-Jin;Park, Chan-Oh;Kim, So-Hee;Oh, Seung-Hwan;Lee, Do-Geon;Pyeon, Da-Bin;Kim, Eun-Young;Park, Se-Pill
    • Animal Bioscience
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    • v.34 no.4
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    • pp.546-557
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    • 2021
  • Objective: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. Methods: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. Results: Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. Conclusion: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.

Potential to mitigate ammonia emission from slurry by increasing dietary fermentable fiber through inclusion of tropical byproducts in practical diets for growing pigs

  • Nguyen, Quan Hai;Le, Phung Dinh;Chim, Channy;Le, Ngoan Duc;Fievez, Veerle
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.4
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    • pp.574-584
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    • 2019
  • Objective: Research was conducted to test the effect of including fiber-rich feedstuffs in practical pig diets on nutrient digestibility, nitrogen balance and ammonia emissions from slurry. Methods: Three Vietnamese fiber sources were screened, namely cassava leaf meal (CL), cassava root residue (CR), and tofu by-product (TF). Accordingly, a control diet (Con) with 10% of dietary non-starch polysaccharides (NSP) and three test diets including one of the three fiber-rich feedstuffs to reach 15% of NSP were formulated. All formulated diets had the same level of crude protein (CP), in vitro ileal protein digestible and metabolisable energy, whereas the in vitro hindgut volatile fatty acid (VFA) production of the test diets was 12% to 20% higher than the control diet. Forty growing barrows with initial body weight at $28.6{\pm}1.93kg$ ($mean{\pm}standard$ deviation) were allocated to the four treatments. When pigs reached about 50 kg of body weight, four pigs from each treatment were used for a nitrogen balance trial and ammonia emission assessment, the remaining six pigs continued the second period of the feeding trial. Results: The TF treatment increased fecal VFA by 33% as compared with the control treatment (p = 0.07), suggesting stimulation of the hindgut fermentation. However, urinary N was not significantly reduced or shifted to fecal N, nor was slurry pH decreased. Accordingly, ammonia emissions were not mitigated. CR and CL treatments failed to enhance in vivo hindgut fermentation, as assessed by fecal VFA and purine bases. On the contrary, the reduction of CP digestibility in the CL treatment enhanced ammonia emissions from slurry. Conclusion: Dietary inclusion of cassava and tofu byproducts through an increase of dietary NSP from 10% to 15% might stimulate fecal VFA excretion but this does not guarantee a reduction in ammonia emissions from slurry, while its interaction with protein digestibility even might enhance enhanced ammonia emission.

Study on the Anti-tumor Effect of Gekko (천룡(天龍)의 항암효과에 대한 고찰)

  • Ahn, Tae-Kyu;Son, Chang-Gue;Jeong, Tae-Yong;Yoo, Hwa-Seung;Cho, Jung-Hyo
    • Journal of Korean Traditional Oncology
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    • v.14 no.1
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    • pp.75-84
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    • 2009
  • Gekko has been used for several diseases including cancer in Oriental medicine and fork herbalogy. Nevertheless, its origin as herbal medicine and its efficacy and mechanism as anti-tumor drug have not yet been thoroughly reported in Korea. This study aimed to investigate anti-tumor effect of Gekko through selected articles from cqvip database in China. In vitro and In vivo, Gekko could obviously inhibit tumor growth, induce tumor cells apoptosis, reduce micro-vessel density in tumor tissue through down regulating VEGF & bFGF protein expression, promote cytotoxicity of lymphocyte. Gekko could improve survival rate, relive clinical symptoms, improve quality of life, and relieve anti-tumor treatment reaction, suggesting that Gekko might be a effective anti-tumor drug.

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Evaluation of nutritive value of chestnut hull for ruminant animals using in vitro rumen fermentation (밤 가공 부산물의 반추가축용 사료 가치 평가: in vitro 반추위 배양)

  • Jeong, Sin-Yong;Jo, Hyeon-Seon;Park, Gi-Su;Kang, Gil-Nam;Jo, Nam-Chul;Seo, Seongwon
    • Korean Journal of Agricultural Science
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    • v.39 no.3
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    • pp.335-340
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    • 2012
  • During the manufacturing process of chestnut, 50% of biomass is produced as chestnut shell (CS) or chestnut hull (CH), a forestry by-product. Due to its high fiber content and economic benefit, there is a possibility of using chestnut hull as a supplement for a ruminant diet. Few studies, however, have been conducted on evaluating nutritive value of chestnut hull for ruminant animals. The objective of this study were thus to analyze chemical composition of CS, a by-product after the first processing of chestnut, and CH, a by-product after the second processing, and access in vitro rumen fermentation characteristics of them. For the in vitro fermentation using strained rumen fluid obtained from a fistulated Hanwoo steer, commercial total mixed ration (TMR) for dairy goat was used as a basal diet and was replaced with different proportions of chestnut shell and hull. A total number of 13 treatments were carried out in this study: 100% TMR, 100% CS, 100% CH, a mix with 50% CS and 50% of CH (MIX), TMR replaced with 5%, 10%, or 15% of CS, CH, or MIX, respectively. For each treatment, in vitro dry matter digestibility (IVDMD) and pH after 48 hours of rumen fermentation were measured. Gas production at 6, 12, 24, 48 hours of incubation was also analyzed. Compared to CH, CS contains higher level of fiber (NDF, ADF, lignin) and consequently has a lower amount of non-fiber carbohydrate, but no difference was observed in the other nutrients (i.e. crude protein, crude fat, and ash). IVDMD was significantly (p<0.05) the highest in 100% CH (71.97%) and the lowest in 100% CS (42.80%). Addition of CH by replacing TMR did not affect IVDMD, while an increase in the proportion of CS tended to decrease IVDMD. The total gas production after 48 hours of incubation and the rate of gas production were also the highest in 100% CH and the lowest in 100% CS (P<0.05). Likewise, the pH after 48 hours of fermentation was significantly (p<0.05) the lowest in 100% CH (6.33) and the highest in 100% CS (6.50), and no significant difference in gas production was observed when TMR was replaced with CS or CH up to 15% (P>0.05). In conclusion, CH may successfully be used for a supplement in a ruminant diet. The nutritive value of CS is relative low, but can replace, if not 100%, low quality forage. This study provides valuable information about the nutritive value of CS and CH. An in vivo trials, however, is needed for conclusively accessing the nutritive value of CS and CH.

Recent Progress in Research on Anticancer Activities of Ginsenoside-Rg3 (Ginsenoside Rg3의 항암효능 연구의 진보)

  • Nam, Ki Yeul;Choi, Jae Eul;Hong, Se Chul;Pyo, Mi Kyung;Park, Jong Dae
    • Korean Journal of Pharmacognosy
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    • v.45 no.1
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    • pp.1-10
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    • 2014
  • Ginsenoside Rg3 (G-Rg3) is one of protopanaxadiol ginsenosides characteristic of red ginseng, steamed and dried ginseng (Panax ginseng), which has recently attracted much attention for its antitumor properties in vitro and in vivo animal models. Experimental studies have demonstrated that it could promote cancer cell apoptosis, inhibit cancer cell growth, the apoptosis of cancer cells, adhesion, invasion and metastasis, and also prevent an angiogenetic formation in prostate, breast, ovarian, colorectal, gastric, liver and lung cancer etc. It has shown the antitumor activities by modulation of diverse signaling pathways, including regulation of cell proliferation mediators (CDKs and cyclins), growth factors (vascular endothelial growth factor), tumor suppressors (p53 and p21), cell death mediators (caspases, Bcl-2, Bax), inflammatory response molecules ($NF-{\kappa}B$ and COX-2), protein kinases (JNK, Akt, and AMP-activated protein kinase) and Wnt/${\beta}$-catenin signaling. In addition, the combination of Rg3 and chemotherapeutic agents have synergistically enhanced therapeutic efficacy and reduced antagonistically side effects. Furthermore, it can reverse the multidrug resistance of cancer cells, prolong the survival duration and improve life quality of cancer patients. Taken together, accumulating evidences could provide the potential of G-Rg3 in the treatment of cancers and the feasibility of further randomized placebo controlled clinical trials.

Effects of Dietary Herbaceous Peat on In Vitro Fermentation and Milk Production in Dairy Cows (허브부식토의 사료내 첨가에 따른 In Vitro 발효특성과 젖소의 유생산성에 미치는 영향)

  • Kim, Hyeon-Shup;Park, Joong-Kook;Kim, Hong-Yun;Kim, Sang-Bum;Yang, Seung-Hak;Kim, Chang-Hyun;Ahn, Jong-Ho
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.2
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    • pp.177-190
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    • 2011
  • This study was conducted to determine effects of dietary herbaceous peat on in vitro fermentation and milk production in dairy cows. Ruminal pH, gas production, VFA (volatile fatty acid), Ammonia-N, and rumen degradability were examined by the addition of three times over 0, 1, and 5% herbaceous peat with substrate of timothy hay, and the change of rumen fermentation characteristics were evaluated. In 0, 3, 12 and 24 hours cultivation, all treatments did not show a significant difference but the control at 6 hours appeared significantly lower pH compared to 1 and 5% treatments (p<0.05). The gas production of the treatments significantly increased until 12 hours of cultivation compared to control (p<0.05), the rumen ammonia concentration showed a tendency to increase until 24 hours in all treatment groups, and there was no significant difference between treatments. About the rumen degradability, 5% treatment showed higher rumen degradability in all hours than control and 1% treatment (p<0.05). Meanwhile, for in vivo trial, 16 heads of Holstein lactation dairy cows were selected for experiment for four weeks in order to research the change of milk yield, milk compositions and change of somatic cell counts of lactation dairy cows by herbaceous peat feeding. The milk yield of vitamin C and herbaceous peat treatments (T3) was 25.0 kg but the control was 23.2 kg, herbaceous peat treatment (T1) was 23.1 kg, and vitamin C treatment (T2) was 23.4 kg, so there was linear increase effect of milk yield by T3. The partial significance of the milk (fat, milk protein, lactose, MUN and SNF) and change of somatic cell count before and after experiment by the control and treatments about change of milk and somatic cell counts (p<0.05) were recognized. About change of milk in the first half (1~2 weeks) and latter half (3~4 weeks) during four weeks of experiments period, the herbaceous peat supplement treatments showed a tendency of significant decrease of quality of milk protein and SNF. The control and treatments did not show significant change of blood nutrients (total protein, cholesterol, NEFA, BUN), liver function component (AST, GGT) and minerals (Ca, P, Mg) before and after experiment. In summary, it is judged that herbaceous peat feeding for lactation dairy cows would be recommendable based on the results of milk, somatic cell count physiologically.