• Title, Summary, Keyword: Escape Protein

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Effect of TLR4 and B7-H1 on Immune Escape of Urothelial Bladder Cancer and its Clinical Significance

  • Wang, Yong-Hua;Cao, Yan-Wei;Yang, Xue-Cheng;Niu, Hai-Tao;Sun, Li-Jiang;Wang, Xin-Sheng;Liu, Jing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1321-1326
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    • 2014
  • Background/Aim: Toll-like receptor 4 (TLR4) and B7-H1, both normally expressed restricted to immune cells, are found to be aberrantly expressed in a majority of human tumors and may play important roles in regulation of tumor immunity. It has been shown that urothelial bladder cancer (UBC) patients can manifest tumoral immune escape which may be a potential critical factor in tumor pathogenesis and progression. However, so far, the mechanisms of UBC-related immune escape have not been clarified. The aim of this study was to investigate the effect of TLR4 and B7-H1 on immune escape of UBC. Methods: Bladder cancer T24 cells were pre-incubated with LPS and co-cultured with tumor specific CTLs. CTL cytotoxicity and apoptosis rates were measured by MTT assay and flow cytometry, respectively. The effects of an ERK inhibitor on B7-H1 expression and CTL cytotoxicity against T24 cells were also evaluated. In addition, TLR4, B7-H1 and PD-1 protein expression was analyzed by immunohistochemistry in 60 UBC specimens and 10 normal urothelia. Results: TLR4 activation protected T24 cells from CTL killing via B7-H1 overexpression. However PD98059, an inhibitor of ERK, enhanced CTL killing of T24 cells by reducing B7-H1 expression. TLR4 expression was generally decreased in UBC specimens, while B7-H1 and PD-1 were greatly overexpressed. Moreover, expression of both B7-H1 and PD-1 was significantly associated with UICC stage and WHO grade classification. Conclusions: TLR4 and B7-H1 may contribute to immune escape of UBC. Targeting B7-H1 or the ERK pathway may offer new immunotherapy strategies for bladder cancer.

Protein Folding and Diseases

  • Lee, Cheol-Ju;Yu, Myeong-Hee
    • BMB Reports
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    • v.38 no.3
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    • pp.275-280
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    • 2005
  • For most of proteins to be active, they need well-defined three-dimensional structures alone or in complex. Folding is a process through which newly synthesized proteins get to the native state. Protein folding inside cells is assisted by various chaperones and folding factors, and misfolded proteins are eliminated by the ubiquitin-proteasome degradation system to ensure high fidelity of protein expression. Under certain circumstances, misfolded proteins escape the degradation process, yielding to deposit of protein aggregates such as loop-sheet polymer and amyloid fibril. Diseases characterized by insoluble deposits of proteins have been recognized for long time and are grouped as conformational diseases. Study of protein folding mechanism is required for better understanding of the molecular pathway of such conformational diseases.

Duckweed as a Protein Source for Fine-Wool Merino Sheep: Its Edibility and Effects on Wool Yield and Characteristics

  • Damry, J.V. Nolan;Bell, R.E.;Thomson, E.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.4
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    • pp.507-514
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    • 2001
  • Two experiments were carried out to investigate whether duckweed is useful as a dietary protein source for fine-wool Merino sheep and to evaluate its effects on wool yield and characteristics. In Experiment 1, the sheep were given one of three maintenance diets consisting of oaten chaff (520-700 g/d) supplemented with 16-32 g crude protein/d in the form of fresh (1 kg/d) or sun-dried (50-100 g/d) duckweed. Each ration was estimated to provide 5.4 MJ (1.3 Mcal)/d of metabolisable energy (ME). The sheep readily ingested the fresh or dried duckweed. None of the wool measures (yield, rate of fibre elongation, fibre diameter) differed (p>0.05) between dietary treatments. In Experiment 2, oaten-chaff-based diets (800 g/d) supplying 6.5-7.2 MJ (1.6-1.7 Mcal)/d of ME were supplemented with iso-nitrogenous amounts (4-5 g N) either of urea (8 g), cottonseed meal (60 g) or dried duckweed (100 g). In this experiment, the rate of wool fibre elongation, thought to be related to intestinal amino acid absorption, was lower (p<0.05) for sheep given the oaten chaff/urea diet than for those given either oaten chaff/cottonseed meal or oaten chaff/duckweed for which the rates did not differ (p>0.05). Fibre diameter, which ranged from 16.0-16.7 mm, did not differ (p>0.05) between diets, but tended to be lower on the oaten chaff/urea diet so that volume of wool produced was also significantly lower (p<0.05) on this diet than on the diets containing duckweed or cottonseed meal. Rumen ammonia concentrations at 4.5 and 7.5 h after feeding were higher (p<0.05) for sheep given the oaten chaff/urea diet than for those given the other two diets. A comparison of the rumen ammonia concentrations, wool growth rate and predicted flows of amino acids from the rumen of sheep supplemented with duckweed rather than cottonseed meal suggested that duckweed is a valuable source of 'escape protein' for ruminants.

Analysis of antigenic sites on the VP4 of porcine rotavirus, Gottfried strain (돼지 로타바이러스(Gottfried 주)의 VP4 항원구조분석)

  • Song, Yun-kyung;Kim, Won-yong;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.41 no.3
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    • pp.343-350
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    • 2001
  • The neutralization epitopes of the outer capsid protein VP4 of a porcine rotavirus, Gottfried strain, were studied using neutralizing monocolonal antibodies(N-MAbs). Eight N-MAbs which are specific for the VP4 of Gottfried strain were used for analyzing the antigenic sites of VP4. Three different approaches were used for this analysis; i)testing the serological reactivity of each N-MAb against different G and P types of human and animal rotavirusese ii) analyzing N-MAb-resistant viral escape mutants and iii) performing nucleotide sequence analysis of the VP4 gene of each N-MAb-resistant viral escape mutant. From experimental results, at least four antigenic sites(I, II, III, and IV) were identified. Antigenic site I recognized by N-MAbs 24B9, 23G10, and 26A2 was separated from antigenic site II recognized by N-MAbs 30H5, 32B3, and 29B3. However, these antigenic sites were overlapped with antigenic site III recognized by N-MAb 21A1. The other antigenic site IV recognized by N-MAb 16D2 was separated from antigenic sites I, II, and III.

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Control of Encapsulation Efficiency and Initial Burst in Polymeric Microparticle Systems

  • Yeo, Yeon;Park, Ki-Nam
    • Archives of Pharmacal Research
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    • v.27 no.1
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    • pp.1-12
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    • 2004
  • Initial burst is one of the major challenges in protein-encapsulated microparticle systems. Since protein release during the initial stage depends mostly on the diffusional escape of the protein, major approaches to prevent the initial burst have focused on efficient encapsulation of the protein within the microparticles. For this reason, control of encapsulation efficiency and the extent of initial burst are based on common formulation parameters. The present article provides a literature review of the formulation parameters that are known to influence the two properties in the emulsion-solvent evaporation/extraction method. Physical and chemical properties of encapsulating polymers, solvent systems, polymer-drug interactions, and properties of the continuous phase are some of the influential variables. Most parameters affect encapsulation efficiency and initial burst by modifying solidification rate of the dispersed phase. In order to prevent many unfavorable events such as pore formation, drug loss, and drug migration that occur while the dispersed phase is in the semi-solid state, it is important to understand and optimize these variables.

Responses of Dairy Cows to Supplemental Highly Digestible Rumen Undegradable Protein and Rumen-protected Forms of Methionine

  • Sun, T.;Yu, X.;Li, S.L.;Dong, Y.X.;Zhang, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.5
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    • pp.659-666
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    • 2009
  • Metabolizable protein (MP) supply and amino acid balance in the intestine were manipulated through selection of highly digestible rumen-undegradable protein (RUP) sources and protected methionine (Met) supplementation. Four ruminallycannulated, multiparous Holstein cows averaging 193${\pm}$13 days in milk were used in a 4${\times}$4 Latin square design to assess N utilization and milk production responses to changes in RUP level, post-ruminal RUP digestibility and protected Met supplementation. Treatments were A) 14.0% crude protein (CP), 8.0% rumen degradable protein (RDP) and 6.0% RUP of low intestinal digestibility (HiRUP-LoDRUP); B) 14.1% CP, 8.1% RDP and 6.0% RUP of high intestinal digestibility (HiRUP-HiDRUP); C) 13.1% CP, 7.9% RDP and 5.2% RUP of high intestinal digestibility (LoRUP-HiDRUP), and D) 13.1% CP, 7.9% RDP and 5.2% RUP of high intestinal digestibility plus rumen escape sources of Met (LoRUP-HiDRUP+Met). Experimental diets were formulated to have similar concentrations of RDP, net energy of lactation ($NE_L$), neutral detergent fiber (NDF), acid detergent fiber (ADF), calcium, phosphorus and ether extract using the NRC model (2001). Results showed that dry matter intake (DMI), production of milk fat and protein were similar among treatments. Milk production was similar for diet HiRUP-LoDRUP, HiRUP-HiDRUP and LoRUP-HiDRUP+Met, and significantly higher than diet LoRUP-HiDRUP. Milk fat and protein percentage were higher for cows receiving HiDRUP treatments, with the greatest increases in the diet LoRUP-HiDRUP+Met. There was no significant change in ruminal pH, $NH_3g-N$ and volatile fatty acid (VFA) concentration among all treatments. Apparent digestibility of dry matter (DM), CP, NDF and ADF and estimated bacterial CP synthesis were similar for all treatments. Nitrogen intakes, blood and milk urea-N concentrations were significantly higher for cows receiving HiRUP diets. Urine volume and total urinary N excretion were significantly lowered by LoRUP diets. Lowering dietary RUP level while supplementing the highly digestible RUP source with rumen escape sources of Met resulted in similar milk production, maximal milk fat and protein concentration and maximum N efficiency, indicating that post-ruminal digestibility of RUP and amino acid balance in the small intestine can be more important than total RUP supplementation.

Crystallization and preliminary X-ray analysis of API5-FGF2 complex

  • Bong, Seoung Min;Lee, Byung Il
    • Biodesign
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    • v.6 no.4
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    • pp.92-95
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    • 2018
  • API5 is a unique oncogenic, non-BIR type IAP nuclear protein and is up-regulated in several cancers. It exerts several functions, such as apoptosis inhibition, cell cycle progression, cancer immune escape, and anticancer drug resistance. Although structural studies of API have revealed that API5 mediates protein-protein interactions, its detailed molecular functions remain unknown. Since FGF2 is one of API5's major interacting proteins, structural studies of the API5-FGF2 complex will provide insight into both proteins' molecular function. We overexpressed and purified API5 and FGF2 in Escherichia coli and crystallized the API-FGF2 complex using polyethylene glycol (PEG) 6000 as a precipitant. Diffraction data were collected to a $2.7{\AA}$ resolution using synchrotron X-rays. Preliminary diffraction analysis revealed that the API5-FGF2 complex crystal belongs to the space group $P2_12_12_1$ with the following unit cell parameters: a = 46.862, b = 76.523, $c=208.161{\AA}$. One asymmetric unit with 49.9% solvent contains one API5-FGF2 complex. Molecular replacement calculation, using API5 and FGF2 coordinates, provided a clear electron density map for an API5-FGF2 complex.

Influence of Caecectomy on the Bioavailability of Minerals from Vegetable Protein Supplements in Adult Roosters

  • Vasan, P.;Dutta, Narayan;Mandal, A.B.;Sharma, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.8
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    • pp.1178-1182
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    • 2008
  • The present study was designed to assess the influence of caeca on the availability of calcium, phosphorus, magnesium, manganese and copper from soybean, sunflower, rapeseed, sesame, fish and meat cum bone meal in adult roosters. The excretion of endogenous origin minerals viz., copper, magnesium, manganese and calcium was significantly (p<0.001) higher in caecectomized than in normal roosters. The difference in the endogenous excretion was 50; 60.45; 40.35 and 29.63 per cent for copper, magnesium, manganese and calcium, respectively, in caecectomized roosters. The caeca played a pivotal role in the reabsorption of endogenous origin calcium, magnesium, manganese and copper. The mechanism of phosphorus absorption by the caecal epithelium was negligible. The caecectomized roosters underestimated the bioavailability of copper in sunflower meal and manganese in almost all the test feedstuffs. The present investigation revealed that the caeca played a critical role in the absorption of minerals from vegetable protein feedstuffs which escape digestion and absorption in the small and large intestinal segments.

Effects of Micronization on the In situ and In vitro Digestion of Cereal Grains

  • McAllister, T.A.;Sultana, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.929-939
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    • 2011
  • The effects of micronization on in situ and in vitro nutrient disappearances of wheat, barley and corn were investigated in a series of experiments. In Experiment 1, chemical composition and in situ dry matter disappearance (DMD) of six varieties of wheat were determined. In addition, an in vitro study was completed using ground micronized and unmicronized wheat (var. Kansas). In Experiment 2, three varieties of wheat (Kansas, Sceptre and Laura) and in Experiment 3, three cereal grains (wheat, barley and corn) were either micronized for 1 min to attain internal kernel temperatures of 90-100$^{\circ}C$ or not (controls), and DM, protein and starch disappearances were estimated. In Experiment 2, an in vitro study was also completed using ground micronized and unmicronized wheat (var. Kansas). Wheat samples varied with respect to crude protein (10.0-21.2%), starch (61.6-73.9%), NDF (8.5-11.8%), volume weight (753-842 g/L) and kernel hardness (0.0-32.0). Rate (p = 0.003) and extent (p = 0.001) of in situ DMD differed among wheat varieties. Correlations between in situ kinetics, and chemical and physical properties of wheat varieties showed that protein content was negatively correlated with the rate of disappearance ($r^2$ = -0.77). Micronization of all grains markedly reduced (p = 0.001) the rate and extent of DM, and protein disappearances as compared to control samples. Micronization increased (p<0.05) the digestion of starch in wheat. However, release of ammonia into the incubation medium was markedly reduced (p<0.05), suggesting that micronization increased the resistance of protein to microbial digestion. Disappearances of DM, protein and starch differed (p = 0.001) among cereal grains with wheat>barley>corn. Micronization reduced the rate of DM disappearance (p = 0.011) and slowly degradable protein fractions (p = 0.03), however, increased (p = 0.004) slowly degradable starch fractions of all three cereals. Examination of in situ samples by scanning electron microscopy confirmed that microbial colonization focused on starch granules in micronized grains, and that the protein matrix exhibited resistance to microbial colonization. These results suggest that micronization may be used to increase the ruminal escape value of protein in cereal grains, but may lead to increased starch digestion if grains are finely ground.