WorldWideScience

Sample records for silage protein degradability

  1. Postruminal degradation of crude protein, neutral detergent fibre and starch of maize and grass silages in dairy cows

    DEFF Research Database (Denmark)

    Ali, M.; Weisbjerg, Martin Riis; Cone, J.W.

    2012-01-01

    samples of grass silage were used. The samples were selected to represent a broad range in digestibility and chemical composition. Prior to the intestinal incubations, samples were incubated in the rumen for 6 h (starch), 12 h (CP) or 24 h (aNDFom) using the rumen nylon bag technique. Residues from....... The objective of this study was to develop a unique dataset on the ruminal degradability and the postruminal digestibility of CP, NDF (aNDFom, amylase neutral detergent fibre organic matter basis) and starch in maize and grass silages, using the mobile nylon bag technique. Twenty samples of maize silage and 20...... in the maize and grass silages. The results proved the assumption of the Dutch feed evaluation system that the rumen undegraded starch is completely digested in the small intestine of dairy cows. Regression showed that the rumen degradability, the intestinal digestibility and the total tract undigested...

  2. Ruminal degradability and carbohydrates and proteins fractioning of triticale silages in singular culture or in mixtures with oat and/or legumes

    OpenAIRE

    Bumbieris Junior, Valter Harry; Jobim, Cloves Cabreira; Emile, Jean Claude; Rossi, Robson; Calixto Junior, Moyses; Branco, Antonio Ferriani

    2011-01-01

    It was aimed to evaluate the ruminal degradability, and the fractioning of carbohydrates, as well as of the nitrogen fractions of triticale silages in singular culture or in mixtures with oats and/or legumes. The treatments had been: triticale silage (X. Triticosecale Wittimack) (ST); triticale silage + forage pea (Pisum arvense) (STE); triticale silage + oats (Avena strigosa Scheb) + forage pea + vetch (Vicia sativa) (STAE). Three castrated bovine Prim’Holstein males had been used, with aver...

  3. Pea and pea-grain mixtures as whole crop protein silage for dairy cows

    OpenAIRE

    Rondahl, Tomas

    2004-01-01

    In this review the use of pea and pea/grain mixtures as whole crop protein silage for dairy cows is discussed. An introductory discussion concerns the ensilage process and protein degradation and effects of different silage additives. To minimise protein loss, prewilting time should be kept short. An acid additive will reduce respiration and thereby reduce protein degradation. The main part of the review discusses nutritional and botanical changes during development as well as results from bo...

  4. Degradability of dry matter and crude protein of dry grains and wet grain silages from different processing corn hybrids (Zea mays

    Directory of Open Access Journals (Sweden)

    Wagner dos Reis

    2013-03-01

    Full Text Available The objective of this work was to evaluate the effect of processing two corn hybrids conserved, dry and humid grains, the dry matter (DM and crude protein (CP degradability in situ. The particle size was determined and difference was verified in MGD (Medium Geometric Diameter of processed ingredients. Three sheep were used with rumen canulated, in a completely randomized design, using a factorial outline 2 x 2 x 3, being two corn hybrid, two conservation methods and three processing forms (whole, coarsely and finely ground, with five times of incubation (3, 6, 12, 24 and 48 hours. The fraction A in SDC (silage of dent corn of DM was superior to GDC (grain of dent corn in all of the particles size. The ensiling process increased the DM solubility, reducing the fraction B in comparison to dry grain. The values regarding the fractions DP and DE the 5% per hour of the protein, were larger for SDC and GDC, it presents a decreasing when the incubation time advances. The fermentation rate was superior for SDC and GDC. The ensiling process has positive effect in the decreasing of DM and CP in comparison to GDC.

  5. Effect of enzyme addition to forage at ensiling on silage chemical composition and NDF degradation characteristics

    DEFF Research Database (Denmark)

    Dehghani, Mohammad Reza; Weisbjerg, Martin Riis; Hvelplund, Torben

    2012-01-01

    , and two varieties of maize stover, lucerne and grass clover were used to study NDF degradation characteristics in experiment 2. Forages were treated with enzymes (500 mg crude protein of the enzyme products/kg DM) and ensiled for 60 days in vacuum-sealed bags. Samples of forage (before ensiling......) and silage were analysed for chemical composition and silages were analysed for pH and fermentation products. The in vitro NDF degradation characteristics of four forages treated with selected enzymes were measured by incubation for up to 96 h with rumen fluid. Enzymes with glucanase, β......-glucanase and pectinase activity increased lactic acid and decreased butyric acid, ammonia and pH compared with control silage, and increased glucose concentration in lucerne silage. NDF concentration generally decreased due to enzyme treatment with glucanase, β-glucanase and xylanase activity and in vitro organic matter...

  6. Influence of stage of maturity of grass silages on protein digestion and microbial protein synthesis in the rumen.

    NARCIS (Netherlands)

    Bosch, M.W.; Bruchem, van J.; Bongers, L.J.G.M.; Tamminga, S.

    1994-01-01

    In four change-over experiments, wilted grass silages, differing in growth stage at harvesting and as a consequence in cell wall content, were fed ad libitum to dairy cows in early and late lactation. Ruminal degradation rate of the crude protein fraction of the silages was investigated using nylon

  7. Aerobic stability, chemical composition and ruminal degradability of sugarcane silage with glycerin from biodiesel

    Directory of Open Access Journals (Sweden)

    Marco Antonio Bensimon Gomes

    2015-06-01

    Full Text Available The experiment was performed with the objective of studying the ensiled sugarcane silage with 0, 5, 10, 15 and 20% of glycerin in experimental PVC silos. The aerobic stability was assessed by measuring the pH and the temperature of the silage at 0, 24, 48, 72, 96 and 120h. The chemical composition, the levels of non-fiber carbohydrates (NFC and the total digestible nutrients (TDN were evaluated. The in vitro digestibility of dry matter (IVDDM and the in vitro digestibility of the cell wall (IVDCW in the silages were evaluated. In three fistulated cattle the in situ degradability of dry matter (DM and the disappearance percentage of the neutral detergent fiber (NDF in samples incubated at 0, 2, 6, 12, 24, 48, 72 and 96h were analyzed. The experimental design was completely randomized and the statistical analyzes were done using Bayesian inference. Increases were observed in DM, TDN, mineral matter, NFC and reductions in NDF, acid detergent fiber, crude protein and ether extract as the inclusion of glycerin was higher. IVDDM increased (P <0.05 in silage with 15 and 20% of glycerin in relation to those with 0, 5 and 10%. The IVDCW at levels of 10, 15 and 20% of glycerin was higher (P <0.05 compared to the other treatments. Increases were observed in the soluble portion (a, a reduction in the insoluble fraction (b, and an increase in the degradability fraction constant (c of the silages with 5, 10, 15 and 20% of glycerin (P <0.05 compared to the control. Glycerin improved aerobic stability while maintaining a low pH and temperature during the observation period at levels of 15 and 20% of glycerin against the silage with 0, 5 and 10%. These results indicate glycerin as a promising additive for sugarcane silage, being able to enhance energy density and improve the aerobic stability of the ensiled matter when its inclusion is from 10 to 20%.

  8. Evaluation of anaerobic degradation, biogas and digestate production of cereal silages using nylon-bags.

    Science.gov (United States)

    Negri, Marco; Bacenetti, Jacopo; Fiala, Marco; Bocchi, Stefano

    2016-06-01

    In this study, the degradation efficiency and the biogas and digestate production during anaerobic digestion were evaluated for the cereal silages most used to feed biogas plants. To this purpose, silages of: maize from the whole plant, maize from the ear, triticale and wheat were digested, inside of nylon bags, in laboratory scale digesters, for 75days. Overall, the test involved 288 nylon bags. After 75days of digestion, the maize ear silage shows the highest degradation efficiency (about 98%) while wheat silage the lowest (about 83%). The biogas production ranges from 438 to 852Nm(3)/t of dry matter for wheat and ear maize silage, respectively. For all the cereal silages, the degradation as well as the biogas production are faster at the beginning of the digestion time. Digestate mass, expressed as percentage of the fresh matter, ranges from 38% to 84% for wheat and maize ear silage, respectively. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Impact of NDF degradability of corn silage on the milk yield potential of dairy cows

    Directory of Open Access Journals (Sweden)

    Mauro Spanghero

    2010-01-01

    Full Text Available The neutral detergent fibre (NDF degradability of corn silage samples, measured in vitro (ivNDFd by a filter bag system, was used to examine (i the relationship between the ivNDFd and that calculated from acid detergent lignin (L content (NDFd and (ii the impact of ivNDFd variations on the predicted milk yield (MY of dairy cows fed corn silage based diets. A total of 173 samples of corn silage were collected during a period of three years (2001-03 in different dairy farms of the Po Valley (Northern Italy. Each sample was analysed for chemical composition and was also tested in triplicate for the ivNDFd using the DaisyII incubator (Ankom, Tech. Co., Fairport, NY, USA with incubation time of 48hs. Moreover, the NDFd of samples was calculated from the L contents, while the measured ivNDFd values were used to estimate the NEl, the potential dry matter intakes (DMI and to predict the MY of cows. Corn silage samples of the three years were similar for NDF and starch contents (44.2 and 30.7% DM, on average, respectively while samples from 2003, in comparison with 2001 and 2002, had lower crude protein (6.9 vs 8.3-8.4% DM, P<0.01 and L contents (3.3 vs 3.6-3.9% DM, P<0.01 and higher ivNDFd values (53.3 vs 45.6-47.8%, P<0.01. The relationship between ivNDFd and NDFd was weak (R2=0.09, not significant. The MY predicted from the NEl content and DMI of corn silage (5.5 MJ/kg DM and 8.9 kg/d minus the maintenance energy costs, was 11.5 kg/d on average (coefficient of variation 20%. Our simulations indicate that a variation of ivNDFd by +1.0% changes the NEl of corn silage to have an expected variation in milk yield of +0.15 kg/d. If the ivNDFd is also used to predict the corn silage DMI then a +1.0% variation in ivNDFd of corn silage produces an overall +0.23 kg/d MY variation. The present results indicate that ivNDFd is highly variable in corn silage populations and differences in this nutritional parameter have an appreciable impact on the predicted milk

  10. Effect of nitrogen fertilization on the protein quality of timothy grass and silage

    Directory of Open Access Journals (Sweden)

    Liisa Syrjälä-Qvist

    1984-09-01

    Full Text Available Timothy grass given N fertilizer at the rates of 40, 80 and 120 kg N/ha was preserved in 3 glass-fibre silos of 0.4 m3. The crude protein content of DM in the grass increased with the increase of N fertilization as follows: N40 14.8 %, N80 18.4 % and N120 22.1 %, but the proportion of true protein in crude protein decreased: N40 82 %, N80 78 % and N120 76 %. The proportion of watersoluble N in the total N in the grass was: N40 27 %, N80 30 % and N120 33 %. The higher was the N fertilization level, the more rapidly was the protein of the grass degraded in the rumen. The amino acid profile of the protein was similar at all the N fertilization levels. The quality of all the silages was good. The NH3-N fraction of total N was 2.8—3.9 % and the proportion of water-soluble N in total N was 51—55 %, In silage the decrease during ensiling in the proportion of true protein in crude protein and the increase in the proportion of water-soluble N in total N were smaller than in the other silages. The rumen degradability of protein during the first two hours was also lowest in this silage.

  11. Silage fermentation and ruminal degradation of stylo prepared with lactic acid bacteria and cellulase.

    Science.gov (United States)

    Li, Mao; Zhou, Hanlin; Zi, Xuejuan; Cai, Yimin

    2017-10-01

    In order to improve the silage fermentation of stylo (Stylosanthes guianensis) in tropical areas, stylo silages were prepared with commercial additives Lactobacillus plantarum Chikuso-1 (CH1), L. rhamnasus Snow Lact L (SN), Acremonium cellulase (CE) and their combination as SN+CE or CH1 + CE, and the fermentation quality, chemical composition and ruminal degradation of these silages were studied. Stylo silages treated with lactic acid bacteria (LAB) or cellulase, the pH value and NH 3 -N ⁄ total-N were significantly (P fermentation and ruminal degradation than SN+CE treatment. The results confirmed that LAB or LAB plus cellulase treatment could improve the fermentation quality, chemical composition and ruminal degradation of stylo silage. Moreover, the combined treatment with LAB and cellulase may have beneficial synergistic effects on ruminal degradation. © 2017 Japanese Society of Animal Science.

  12. The effect of partial replacement of corn silage on rumen degradability, milk production and composition in lactating primiparous dairy cows

    Directory of Open Access Journals (Sweden)

    Hakan Biricik

    2010-01-01

    Full Text Available The objective of this experiment was to evaluate the effects of partial replacement of corn silage with long alfalfa hay and/or coarse chopped wheat straw on neutral detergent fibre (NDF rumen degradability, milk yield and composition in late lactating dairy cows fed diets with 50% forage on dry matter basis. Twelve late lactating Holstein primiparous cows including four cows equipped with a rumen cannula, averaging 210 ± 20 d in milk and weighing 575 ± 50 kg were randomly assigned in a 4x4 Latin square design. During each of four 21-d periods, cows were fed 4 total mixed diets that were varied in the forage sources: 1 50% corn silage (CS, 2 35% corn silage + 15% wheat straw (CSW, 3 35% corn silage + 15% alfalfa hay (CSA, 4 25% corn silage + 10% wheat straw + 15% alfalfa hay (CSWA. The production of milk averaged 18.55, 20.41 and 20.06 kg/d for unadjusted milk production, 4% fat corrected milk and solid corrected milk, respectively, and was not affected by treatments. Likewise, milk composition or production of milk components was not affected by diets and averaged 4.69% fat, 3.66% protein, 4.51% lactose, 866 g/d fat, 665 g/d protein, 824 g/d lactose. Treatments had no effect on in situ NDF soluble, degradable and potential degradability of all diets, whereas the effective degradability (ED of NDF was greater for cows fed CS diet than for cows fed CSW, CSA and CSWA diets (P<0.05. These values suggested that the partial replacement of corn silage with alfalfa hay and/or wheat straw has no unfavourable effect on the productive parameters.

  13. Degradability of dry matter and crude protein of dry grains and wet grain silages from different processing corn hybrids (Zea mays)

    OpenAIRE

    Wagner dos Reis; Ciniro Costa; Paulo Roberto de Lima Meirelles; Marina Gabriela Berchiol da Silva; Marco Aurélio Factori; Janaína Conte Hadlich; Kátia de Oliveira; Erikelly Aline Ribeiro de Santana; Cristiano Magalhães Pariz; Josineudson Augusto II de Vasconcelos Silva

    2013-01-01

    The objective of this work was to evaluate the effect of processing two corn hybrids conserved, dry and humid grains, the dry matter (DM) and crude protein (CP) degradability in situ. The particle size was determined and difference was verified in MGD (Medium Geometric Diameter) of processed ingredients. Three sheep were used with rumen canulated, in a completely randomized design, using a factorial outline 2 x 2 x 3, being two corn hybrid, two conservation methods and three processing forms ...

  14. Ruminal degradability and carbohydrates and proteins fractioning of triticale silages in singular culture or in mixtures with oat and/or legumesDegradabilidade ruminal e fracionamento de carboidratos e proteínas em silagens de triticale em cultivo singular ou em misturas com aveia e/ou leguminosas

    Directory of Open Access Journals (Sweden)

    Moysés Calixto Junior

    2011-07-01

    Full Text Available It was aimed to evaluate the ruminal degradability, and the fractioning of carbohydrates, as well as of the nitrogen fractions of triticale silages in singular culture or in mixtures with oats and/or legumes. The treatments had been: triticale silage (X. Triticosecale Wittimack (ST; triticale silage + forage pea (Pisum arvense (STE; triticale silage + oats (Avena strigosa Scheb + forage pea + vetch (Vicia sativa (STAE. Three castrated bovine Prim’Holstein males had been used, with average live weight of 300 kg, having ruminal cannulas. The incubation times had duration of 0, 6, 12, 24, 48, 72 and 96 hours. Fractions, a, c and ED of the DM of STAE silage was higher (30,33; 0,026 and 45,45% than others silages. Fraction b of the DM of ST silage was higher (58,45% than STE silages (45,36% and STAE (44,37%. In CP the ST silage presented higher fraction (72.12%. For the potentially degradability fraction (b of CP it was not observed difference among treatments. The degradation rate (c of the CP was higher for STE silage (0.063% than ST ensilage (0,012%, however this was similar to the STAE (0,045%. ED of CP was better for STE silage (77,71%. ST silage presented fraction a and b for NDF highest (8,62 and 81,99% than others silages. The degradation rate (c of NDF was higher for STAE (0,027%. ED for NDF did not present difference among treatments. ST silage presented the best values for the total carbohydrates (83,97% than STE (79,87% and STAE silages (76,77%. STE and STAE silages presented better DM degradability and suggest to be a potential source of non degradable protein in the rumen. The exclusive triticale silage revealed superior with regard to the degradability of fiber fraction, also presenting higher amount of total carbohydrates potentially degraded.O objetivo desse trabalho foi avaliar a degradabilidade ruminal, e o fracionamento de carboidratos, assim como das frações nitrogenadas das silagens de triticale em plantio singular ou em

  15. Effects of microbial enzymes on starch and hemicellulose degradation in total mixed ration silages

    Directory of Open Access Journals (Sweden)

    Tingting Ning

    2017-02-01

    Full Text Available Objective This study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR silage. Methods The TMRs were prepared with soybean curd residue, alfalfa hay (ATMR or Leymus chinensis hay (LTMR, corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties. Results Both ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens, B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively. Conclusion The microbial amylase contributes to starch hydrolysis during the

  16. Ensiling Characteristics and the In situ Nutrient Degradability of a By-product Feed-based Silage.

    Science.gov (United States)

    Kim, Y I; Oh, Y K; Park, K K; Kwak, W S

    2014-02-01

    This study was conducted to evaluate the ensiling characteristics and the in situ degradability of a by-product feed (BF)-based silage. Before ensilation, the BF-based mixture was composed of 50% spent mushroom substrate, 21% recycled poultry bedding, 15% ryegrass straw, 10.8% rice bran, 2% molasses, 0.6% bentonite, and 0.6% microbial inoculant on a wet basis and ensiled for up to 4 weeks. The BF-based silage contained on average 39.3% moisture, 13.4% crude protein (CP), and 52.2% neutral detergent fiber (NDF), 49% total digestible nutrient, and 37.8% physically effective NDF1.18 on a dry matter (DM) basis. Ensiling the BF-based silage for up to 4 weeks affected (p<0.01) the chemical composition to a small extent, increased (p<0.05) the lactic acid and NH3-N content, and decreased (p<0.05) both the total bacterial and lactic acid bacterial counts from 10(9) to 10(8) cfu/g when compared to that before ensiling. These parameters indicated that the silage was fermented and stored well during the 4-week ensiling period. Compared with rice or ryegrass straws, the BF-based silage had a higher (p<0.05) water-soluble and filterable fraction, a lower insoluble degradable DM and CP fraction (p<0.05), a lower digestible NDF (p<0.05) fraction, a higher (p<0.05) DM and CP disappearance and degradability rate, and a lower (p<0.05) NDF disappearance and degradability rate. These results indicated that cheap, good-quality BF-based roughage could be produced by ensiling SMS, RPB, rice bran, and a minimal amount of straw.

  17. Comparison of Chemical and Degradability Characteristics of Green Forage and Silage of Sorghums Varieties with Corn Using In vitro

    Directory of Open Access Journals (Sweden)

    A. Hedayatipour

    2012-10-01

    Full Text Available The chemical and fermentative parameters of three fresh forages and silages of sorghum including Sweet, Pegah and Speedfeed varieties were compared with corn using in vitro method, also degradability coefficients of forages and silages were determined by in situ method. Forages were planted in the same condition and harvested in soft dough stage, then ensilaged in four replicates for each time of 30, 60 and 90 days of preservation in mini silos. Buffering capacity in green Sweet sorghum was lower than corn and Speedfeed, and acid detergent fiber and water soluble carbohydrates respectively were significantly highest and lowest in fresh forage of Speedfeed sorghum. In time of 60 days, percent of acid detergent lignin of corn silage was lower than Sweet and Speedfeed sorghum silages; similarly, residual water soluble carbohydrate was lowest in corn silage. The lactate Concentration in corn and Pegah sorghums was higher than Sweet and Speedfeed silages. In corn and Sweet sorghum silages, Contents of acetic acid and ammonium nitrogen were highest and lowest, respectively. In nylon bag experiment, Degradation rate of corn and Pegah sorghum forages were significantly higher than Sweet and Speedfeed sorghums that cause to more effective degradability with passage rate of 0.08 in this forages. Also, the slowly degradation coefficient of corn silage was higher than sorghums silages. In conclusion, Speedfeed sorghum forage is not suitable for making silage in comparison others, and corn silage had more potential of degradability.

  18. In situ degradability of dry matter and fibrous fraction of sorghum silage

    Directory of Open Access Journals (Sweden)

    Renê Ferreira Costa

    2016-05-01

    Full Text Available This study aimed to evaluate in situ degradability and degradation kinetics of DM, NDF and ADF of silage, with or without tannin in the grains. Two isogenic lines of grain sorghum (CMS-XS 114 with tannin and CMS-XS 165 without tannin and two sorghum hybrids (BR-700 dual purpose with tannin and BR-601 forage without tannin were ensiled; dried and ground silage samples were placed in nylon bags and introduced through the fistulas. After incubation for 6, 12, 24, 48, 72 and 96 hours, bags were taken for subsequent analysis of fibrous fractions. The experimental design was completely randomized with 4 replicates and 4 treatments and means compared by Tukey’s test at 5% probability. As for the DM degradation rate, silage of CMSXS165without tannin was superior. Silages of genotypes BR700 and CMSXS 114 with tannin showed the highest values of indigestible ADF (59.54 and 43.09%. Regarding the NDF, the potential degradation of silage of CMSXS165 line without tannin was superior. Tannin can reduce ruminal degradability of the dry matter and fibrous fractions.

  19. Utilization of protein in red clover and alfalfa silages by lactating dairy cows and growing lambs.

    Science.gov (United States)

    Broderick, Glen A

    2018-02-01

    Feeding trials were conducted with lactating cows and growing lambs to quantify effects of replacing dietary alfalfa silage (AS) with red clover silage (RCS) on nutrient utilization. The lactation trial had a 2 × 4 arrangement of treatments: AS or RCS fed with no supplement, rumen-protected Met (RPM), rumen-protected Lys (RPL), or RPM plus RPL. Grass silage was fed at 13% of dry matter (DM) with AS to equalize dietary neutral detergent fiber (NDF) and crude protein contents. All diets contained (DM basis) 5% corn silage and 16% crude protein. Thirty-two multiparous (4 ruminally cannulated) plus 16 primiparous Holstein cows were blocked by parity and days in milk and fed diets as total mixed rations in an incomplete 8 × 8 Latin square trial with four 28-d periods. Production data (over the last 14 d of each period) and digestibility and excretion data (at the end of each period) were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Although DM intake was 1.2 kg/d greater on AS than RCS, milk yield and body weight gain were not different. However, yields of fat and energy-corrected milk as well as milk content of fat, true protein, and solids-not-fat were greater on AS. Relative to AS, feeding RCS increased milk and energy-corrected milk yield per unit of DM intake, milk lactose content, and apparent N efficiency and reduced milk urea. Relative to AS, apparent digestibility of DM, organic matter, NDF, and acid detergent fiber were greater on RCS, whereas apparent and estimated true N digestibility were lower. Urinary N excretion and ruminal concentrations of ammonia, total AA, and branched-chain volatile fatty acids were reduced on RCS, indicating reduced ruminal protein degradation. Supplementation of RPM increased intake, milk true protein, and solids-not-fat content and tended to increase milk fat content. There were no silage × RPM interactions, suggesting that RPM was equally limiting on both AS and RCS. Supplementation of RPL did not

  20. Metabolisable protein supply to lactating dairy cows increased with increasing dry matter concentration in grass-clover silage

    DEFF Research Database (Denmark)

    Johansen, Marianne; Hellwing, Anne Louise Frydendahl; Lund, Peter

    2017-01-01

    The aim of this experiment was to study the effect of increased dry matter (DM) concentration in grass-clover silage, obtained by extending the pre-wilting period before ensiling, on the amount of metabolisable protein (MP) supplied to lactating dairy cows. Spring growth and first regrowth of grass...... and faeces, respectively, were collected over 94 h to cover the diurnal variation, pooled, and subsequently analysed. Rumen fluid was collected in same sampling procedure. To estimate the duodenal flow of microbial protein, microbes were isolated from the rumen and analysed for amino acids (AA) and purines...... flow of AA. The higher duodenal flow of AA derived from a lower rumen degradation of feed protein and a tendency towards a higher microbial synthesis in the rumen. Fibre digestibility and CH4 production were not affected by silage DM concentration. In conclusion, MP concentration in grass-clover silage...

  1. Effect of monensin on in vitro fermentation of silages and microbial protein synthesis.

    Science.gov (United States)

    Wischer, Gerald; Boguhn, Jeannette; Steingaß, Herbert; Schollenberger, Margit; Hartung, Karin; Rodehutscord, Markus

    2013-06-01

    The objective of the study was to investigate the effects of monensin on silage fermentation and microbial net protein synthesis. In Experiment 1, monensin (0.5, 1, 2, 4, 6, or 10 µg) was added to syringes that contained 120 mg of grass silage (GS), grass silage and concentrate (GS + C), or maize silage (MS), resulting in concentrations of 4.2, 8.3, 16.7, 33.3, 50.0 and 83.3 mg monensin/kg feed. Samples were incubated for 24 h to determine the monensin concentration that resulted in the maximum reduction in methane production without effects on the total gas production. In Experiment 2, GS and GS + C were incubated in a rumen simulation technique (Rusitec) to assess the monensin effects (133 and 266 mg/kg feed) on the production of total gas, methane and volatile fatty acids (VFA), degradation of nutrients and microbial net protein synthesis. In Experiment 1, methane production was reduced without significant effects on the total gas production; the reductions were 17% (GS), 10% (GS + C) and 13% (MS) with 16.7 (GS), 50.0 (GS + C) and 33.3 (MS) mg monensin/kg feed. Monensin reduced the total gas and methane production in GS and GS + C in Experiment 2. Propionate production was enhanced by monensin, accompanied by a decrease in acetate production. Along with a reduction in crude protein (CP) degradation, monensin reduced the ammonia nitrogen concentration in the effluent of both treatments. While the protein produced by liquid-associated microbes increased with monensin, protein production by solid-associated microbes was reduced. Total microbial net protein synthesis increased in the presence of monensin. Monensin influenced the production of total gas, methane and VFA from the silages without an effect on the degradation of organic matter (OM). Different microbial fractions were affected differently by monensin supplementation. If monensin is used as a tool to reduce methane emission, the supplementation level must be carefully chosen to avoid negative effects on

  2. Estimating grass and grass silage degradation characteristics by in situ and in vitro gas production methods

    Directory of Open Access Journals (Sweden)

    Danijel Karolyi

    2010-01-01

    Full Text Available Fermentation characteristics of grass and grass silage at different maturities were studied using in situ and in vitro gas production methods. In situ data determined difference between grass and silage. Degradable fraction decreased as grass matured while the undegradable fraction increased. Rate of degradation (kd was slower for silage than fresh grass. Gas production method (GP data showed that fermentation of degradable fraction was different between stage of maturity in both grass and silage. Other data did not show any difference with the exception for the rate of GP of soluble and undegradable fraction. The in situ degradation characteristics were estimated from GP characteristics. The degradable and undegradable fractions could be estimated by multiple relationships. Using the three-phases model for gas production kd and fermentable organic matter could be estimated from the same parameters. The only in situ parameter that could not be estimated with GP parameters was the soluble fraction. The GP method and the three phases model provided to be an alternative to the in situ method for animal feed evaluations.

  3. Variation between individual cows in in situ rumen degradation characteristics of maize and grass silages

    NARCIS (Netherlands)

    Ali, M.; Cone, J. W.; van Duinkerken, G.; Klop, A.; Blok, M. C.; Bruinenberg, M.; Khan, N. A.; Hendriks, W. H.

    2016-01-01

    Different numbers of animals have been used in different studies to cover the variation between individual animals in in situ rumen degradation characteristics of maize and grass silages. The objective of this study was to determine whether three cows are sufficient or not to cover the variation

  4. Effects of Lactobacillus plantarum and hydrolytic enzymes on fermentation and ruminal degradability of orange pulp silage

    DEFF Research Database (Denmark)

    Taghizadeh, Akbar; Paya, Hamid; Lashkari, Saman

    2015-01-01

    The current study was carried out to examine the effect of inoculants, enzymes and mixtures of them on the fermentation, degradability and nutrient value of orange pulp silage. Orange pulp was treated with water (control), inoculant (Lactobacillus plantarum), enzymes (multiple enzyme) or inoculants...

  5. Effects of moisture content or particle size on the in situ degradability of maize silage and alfalfa haylage in lactating dairy cows

    OpenAIRE

    Yang Zou; Shuangzhao Dong; Yun Du; Shengli Li; Yajing Wang; Zhijun Cao

    2016-01-01

    A study using four Holstein cows with ruminal cannulas was conducted to evaluate the degradability of different moisture content or particle size of maize silage and alfalfa haylage. The maize silage (MS; 20-mm length) and alfalfa haylage (AH; 40-mm length) samples were wet (wet maize silage, MSW; wet alfalfa haylage, AHW), dried (dried maize silage, MSD; dried alfalfa haylage, AHD), or ground to pass through a 2.5-mm screen (dried ground maize silage, MSG; dried ground alfalfa haylage, AHG)....

  6. Ruminal degradation kinetic of Brachiaria decumbens silages with different nitrogen additives

    Directory of Open Access Journals (Sweden)

    Odimári Pricila Pires do Prado

    2014-02-01

    Full Text Available This study aimed to assess levels of nitrogen additive Silogen® pasto on ruminal in vitro degradability gas production in Brachiaria decumbens silages. The nitrogen additive contained bacterial strains (Bacillus subtilis, Lactobacillus curvatus, Lactobacillus plantarum and Pediococcus acidilactici and minimum nitrogen to 360 g/kg. The levels were: 0.0; 1.0; 1.5; 2.0 and 2.5%. In fractionation were calculated total carbohydrates (TC, non-fiber carbohydrates (NFC, soluble and rapidly degradable (A + B1, potentially degradable (B2 and degradable (C. The kinetic parameters of FC and NFC were estimated using the technique of in vitro gas production. Data were subjected to analysis of variance at 5%. The level additive did not influence the TC and fraction A + B1. The levels of nitrogen additive influenced the NFC, fractions B2 and C. For NFC content 2.0% showed the lowest (22.0 %. The lower levels of additive nitrogen (0, 1.0 and 1.5 % had higher fractions of B2 (average 40.2 % and lower values for the fraction C (average 20.0 % . There was no difference in the volume of gas CNF (average 86.73 mL and final volume of gas produced (average 195.79 mL. Was no influence of nitrogen additives for the time of colonization, the lowest time of 3.89 h to 1.0%. The volume of gas of FC was influenced by levels of nitrogen additives with higher values to 0 % from 1.0 %, and 114.74 and 115.09 mL, respectively. Degradation rates of FC and FNC were also affected by the concentrations of nitrogenous additives, which presented higher rates to the levels of 2.0 and 2.5%. It follows that the lower levels of additives to 1.5 % promoted the reduction of the C fraction and increased B2, and greater production of gas volume of the fiber in these silages, showing better nutritional value in these silages.

  7. Effects of Feeding Corn-lablab Bean Mixture Silages on Nutrient Apparent Digestibility and Performance of Dairy Cows

    Directory of Open Access Journals (Sweden)

    Yongli Qu

    2013-04-01

    Full Text Available This study estimated the fermentation characteristics and nutrient value of corn-lablab bean mixture silages relative to corn silages. The effects of feeding corn-lablab bean mixture silages on nutrient apparent digestibility and milk production of dairy cows in northern China were also investigated. Three ruminally cannulated Holstein cows were used to determine the ruminal digestion kinetics and ruminal nutrient degradability of corn silage and corn-lablab bean mixture silages. Sixty lactating Holstein cows were randomly divided into two groups of 30 cows each. Two diets were formulated with a 59:41 forage: concentrate ratio. Corn silage and corn-lablab bean mixture silages constituted 39.3% of the forage in each diet, with Chinese wildrye hay constituting the remaining 60.7%. Corn-lablab bean mixture silages had higher lactic acid, acetic acid, dry matter (DM, crude protein (CP, ash, Ca, ether extract concentrations and ruminal nutrient degradability than monoculture corn silage (p<0.05. Neutral detergent fiber (NDF and acid detergent fiber (ADF concentrations of corn-lablab bean mixture silages were lower than those of corn silage (p<0.05. The digestibility of DM, CP, NDF, and ADF for cows fed corn-lablab bean mixture silages was higher than for those fed corn silage (p<0.05. Feeding corn-lablab bean mixture silages increased milk yield and milk protein of dairy cows when compared with feeding corn silage (p<0.05. The economic benefit for cow fed corn-lablab bean mixture silages was 8.43 yuan/day/cow higher than that for that fed corn silage. In conclusion, corn-lablab bean mixture improved the fermentation characteristics and nutrient value of silage compared with monoculture corn. In this study, feeding corn-lablab bean mixture silages increased milk yield, milk protein and nutrient apparent digestibility of dairy cows compared with corn silage in northern China.

  8. Nutrient composition, ruminal degradability and whole tract digestibility of whole crop maize silage from nine current varieties.

    Science.gov (United States)

    Gruber, Leonhard; Terler, Georg; Knaus, Wilhelm

    2018-04-01

    Since maize silage is an important forage in cattle nutrition, it is important to know its nutritive value. Much effort is put into breeding maize, and several new varieties are introduced on the market every year. This requires periodical analyses of the nutritive value of current maize varieties for the formulation of cattle rations. The aim of this study was to examine the nutritive value of whole crop maize silage (WCMS) from nine maize varieties in 3 consecutive years. For the analysis of nutrient composition and ruminal degradability of organic matter (OM), crude protein (CP), neutral detergent fibre (aNDFom) and non-fibre carbohydrates (NFC), varieties were harvested at three harvest dates (50%, 55% and 60% dry matter content in ear). Due to capacity limitations, the digestibility of WCMS was tested only for the middle harvest date. The CP and acid detergent fibre (ADFom) content was affected (p content was not influenced by variety. With advancing maturity, CP, aNDFom and ADFom content declined while NFC content increased. Variety influenced effective ruminal degradability (ED) of nutrients, except for CP. The ED of all examined nutrients decreased as maturity advanced from first to third harvest date. Digestibility of OM, ADFom and NFC was significantly and digestibility of aNDFom was tendentially (p = 0.064) influenced by variety. Additionally, an effect of year and a harvest date × year interaction was found for almost all examined parameters. In conclusion, variety, harvest date and year influence the nutritive value of WCMS. A comparison with earlier studies shows that current varieties have a higher fibre digestibility and a slower-ripening stover compared to older varieties.

  9. Parâmetros de degradabilidade potencial da matéria seca e da proteína bruta das silagens de seis genótipos de sorgo (Sorghum bicolor (L. Moench, com e sem tanino no grão, avaliados pela técnica in situ Potential degradability parameters of the dry matter and crude protein of six sorghum silage genotypes (Sorghum bicolor (L. Moench, with or without tannin on grain, evaluated by in situ technique

    Directory of Open Access Journals (Sweden)

    Lívio Ribeiro Molina

    2003-02-01

    Full Text Available Para testar o efeito da presença de tanino no grão sobre os parâmetros de degradabilidade potencial da matéria seca e da proteína bruta de seis genótipos de sorgo para silagem, foi conduzido um experimento utilizando a técnica da degradabilidade in situ. Quatro novilhos canulados no rúmen, alimentados individualmente, duas vezes ao dia ad libitum com feno de Tifton 85, foram utilizados para a incubação de silagens de sorgo no estádio de grão leitoso, pertencentes a seis genótipos: BR 303, BR 304, BR 601e AG 2006 (sem tanino no grão; BR 700 e BR 701 (com tanino. Foram incubadas 5 g de amostra nos tempos de incubação: 6, 12, 24, 48, 72 e 96 horas. O tempo zero (t0 foi utilizado para cálculo da fração solúvel. A presença do tanino reduziu a extensão de degradação da matéria seca e da proteína bruta nas silagens de sorgo BR 700 e BR 701. As degradabilidades potenciais destes nutrientes foram influenciada pelo tanino. Não foi observado efeito depressivo do tanino sobre as taxas de degradação para nenhum nutriente, em nenhuma silagem testada.The aim of this experiment was to evaluate the effect of tannin on grain over potential degradability parameters of the dry matter and crude protein of six sorghum silage genotypes by in situ technique. It was used four crossbred steers, canulated in rumen, fed ad libitum twice at day with a diet comprised by Cynodon dactylon hay. Six sorghum silage genotypes, harvested at milk stage, were incubated in the rumen: BR 303, BR 304, BR 601, and AG 2006 (without tannin on grain; BR 700, and BR 701 (with tannin on grain. Five grams of samples were incubated at times: 6, 12, 24, 48, 72, and 96 hours. The time zero (t0 was used to calculate the soluble fraction. Tannin decrease the extent of degradation of dry matter and crude protein of BR 700 and BR 701 sorghum silages. The potential degradabilities of these nutrients were reduced by tannin presence on grain. There was not effect of tannin

  10. Determination of microbial protein in perennial ryegrass silage

    NARCIS (Netherlands)

    Driehuis, F.; Wikselaar, van P.G.

    2001-01-01

    The microbial matter fraction was determined in perennial ryegrass silages of different dry-matter (DM) contents, ensiled with or without Lactobacillus plantarum. 15N-Leucine and the bacterial cell wall constituent diaminopimelic acid (DAPA) were used as markers for microbial-N. Perennial ryegrass

  11. Using brown midrib 6 dwarf forage sorghum silage and fall-grown oat silage in lactating dairy cow rations.

    Science.gov (United States)

    Harper, M T; Oh, J; Giallongo, F; Lopes, J C; Roth, G W; Hristov, A N

    2017-07-01

    Double cropping and increasing crop diversity could improve dairy farm economic and environmental sustainability. In this experiment, corn silage was partially replaced with 2 alternative forages, brown midrib-6 brachytic dwarf forage sorghum (Sorghum bicolor) or fall-grown oat (Avena sativa) silage, in the diet of lactating dairy cows. We investigated the effect on dry matter (DM) intake, milk yield (MY), milk components and fatty acid profile, apparent total-tract nutrient digestibility, N utilization, enteric methane emissions, and income over feed cost. We analyzed the in situ DM and neutral detergent fiber disappearance of the alternative forages versus corn silage and alfalfa haylage. Sorghum was grown in the summer and harvested in the milk stage. Oats were grown in the fall and harvested in the boot stage. Compared with corn silage, neutral detergent fiber and acid detergent fiber concentrations were higher in the alternative forages. Lignin content was highest for sorghum silage and similar for corn silage and oat silage. The alternative forages had less than 1% starch compared with the approximately 35% starch in the corn silage. Ruminal in situ DM effective degradability was similar, although statistically different, for corn silage and oat silage, but lower for sorghum silage. Diets with the alternative forages were fed in a replicated 3 × 3 Latin square design experiment with three 28-d periods and 12 Holstein cows. The control diet contained 44% (DM basis) corn silage. In the other 2 diets, sorghum or oat silages were included at 10% of dietary DM, replacing corn silage. Sorghum silage inclusion decreased DM intake, MY, and milk protein content but increased milk fat and maintained energy-corrected MY similar to the control. Oat silage had no effect on DM intake, MY, or milk components compared to the control. The oat silage diet increased apparent total-tract digestibility of dietary nutrients, except starch, whereas the sorghum diet slightly

  12. Particle passage kinetics and neutral detergent fiber degradability of silage of pineapple waste (aerial parts under different packing densities

    Directory of Open Access Journals (Sweden)

    Graciele Araújo de Oliveira Caetano

    2014-01-01

    Full Text Available The objective of this study was to determine the kinetics of in situ degradability parameters of the dry matter (DM and neutral detergent fiber (NDF and the passage of materials originating from the ensilage of the waste from pineapple cultivation (aerial parts. The four treatments utilized were silage of pineapple waste compacted at 600, 700, 900 and 1000 kg/m³. After ensiling the material from the pineapple cultivation, the particle-transit and rumen-degradation kinetics were analyzed. For the analysis of particle transit, chromium was utilized as a marker to mark the fiber. Passage rates were determined by retrieving the markers in the feces of the animals. In the degradation assay, samples were incubated in nylon bags for 0, 6, 18, 48 and 96 hours. The behavior observed in the regression curves of the variables analyzed describes high correlation between them, i.e., the time during which the silage is retained in the rumen influences its digestibility and its degradation rate. Although the silage compacted at 900 kg/m³ shows a larger potentially digestible fraction, it is recommended that it be ensiled at a compaction density of approximately 750 kg/m³ due to the lower cost and shorter mean retention time in the rumen-reticulum and rumen fill, thereby increasing the ruminal degradation and passage dynamics.

  13. Wilting and biological additive effect on in situ degradability and chemical composition of Arachis pintoi cv Belomonte silage

    OpenAIRE

    Rosana Aparecida Possenti; Evaldo Ferrari Júnior; Valdinei Tadeu Paulino; Ivani Pozar Otsuk; Patrícia Brás

    2010-01-01

    The purpose of this work was to evaluate the effect of wilting and biological additive amendment on chemical composition, fermentation and ruminal degradability of Arachis pintoi cv Belmonte silage. The following treatments were analysed: T1- Arachis pintoi cv Belmonte fresh forage; T2 - Arachis pintoi cv Belmonte fresh forage plus bacterial additive added to the forage prior to the ensilage; T3- Arachis pintoi cv Belmonte wilted by the sun for 4 hours; T4- Arachis pintoi cv Belmonte wilted b...

  14. Microbial protein synthesis, digestion and lactation responses of cows to grass or grass-red clover silage diet supplemented with barley or oats

    Directory of Open Access Journals (Sweden)

    A. VANHATALO

    2008-12-01

    Full Text Available The study was conducted to evaluate effects of silage type (grass-red clover vs. pure grass and grain supplement (oats vs. barley on rumen fermentation, post-ruminal nutrient flows, diet digestion and milk production. Four primiparous Finnish Ayrshire cows fitted with cannulae in the rumen and duodenum were used in a 4 × 4 Latin square experiment with four 28-d experimental periods and 2 × 2 factorial arrangements of treatments. Using red clover-containing (40% silage rather than pure grass silage had minor effects on rumen fermentation or diet digestion but increased non-ammonia nitrogen (N flow in terms of increased flows of microbial and dietary N entering to the small intestine. This was reflected as a reduced ruminal N degradability on grass-red clover diets. Furthermore, grass-red clover diets in comparison to grass silage diets increased milk lactose concentration and yields of milk, protein and lactose. Feeding oats in replacement for barley had minor effects on rumen fermentation or post-ruminal non-ammonia N flows but reduced digestibility of organic matter and neutral detergent fibre in the diet. Using oats rather than barley increased yields of milk and lactose but reduced milk protein concentration. Oats also increased proportions of C18:0 and C18:1 in milk fat and reduced those of C10:0 to C16:0. It is concluded that inclusion of red clover and replacement of barley with oats in grass silage based diets have beneficial effects in dairy cow production.;

  15. Trichoderma longibrachiatum acetyl xylan esterase 1 enhances hemicellulolytic preparations to degrade corn silage polysaccharides

    NARCIS (Netherlands)

    Neumüller, K.G.; Streekstra, H.; Gruppen, H.; Schols, H.A.

    2014-01-01

    Supplementation of a Trichoderma longibrachiatum preparation to an industrial Aspergillus niger/Talaromyces emersonii enzyme mixture demonstrated synergy for the saccharification of corn silage water-unextractable solids (WUS). Sub-fractions of the crude T. longibrachiatum preparation obtained after

  16. The Growth Rate and Efficiency of Rumen Microbial Protein Digestion of Red Clover Silage (Trifolium pratense cv. Sabatron)

    International Nuclear Information System (INIS)

    Asih Kurniawati

    2004-01-01

    (Trifolium pratense cv. Sabatron). Red clover silage supplemented with different level of carbohydrates has been examined using the in-vitro gas production technique. Cumulative gas production, hydro.gen sulfite production, and ammonia was followed and used as indicators of microbial growth rate and extent of protein degradation. Microbial nitrogen production, VFA, and efficiency microbial production was used as indicator of nitrogen use efficiency. 15 N was used as a microbial marker to estimate the amount of nitrogen incorporation into microbial protein. Supplementation of Red clover with increasing 5 levels; 0 g; 0.625 g; 0.15 g; 0.225 g and 0.3 g of maize starch led to graded increase in microbial growth and protein degradation. This was reflected in the increasing gas production and the accumulation of hydrogen sulfite. Diurnal change in ammonia production reflected the microbial utilization of ammonia for protein synthesis. Protein microbe (P<0.001) as VFA (P<0.001) increased due to carbohydrate addition as well as utilization of nitrogen (P<0.001). There was also the efficiency of nitrogen utilization which increased significantly. This result suggested that energy supply can increased efficiency of nitrogen use in the rumen and may reduce nitrogen losses into the environment. (author)

  17. Effects of moisture content or particle size on the in situ degradability of maize silage and alfalfa haylage in lactating dairy cows.

    Science.gov (United States)

    Zou, Yang; Dong, Shuangzhao; Du, Yun; Li, Shengli; Wang, Yajing; Cao, Zhijun

    2016-09-01

    A study using four Holstein cows with ruminal cannulas was conducted to evaluate the degradability of different moisture content or particle size of maize silage and alfalfa haylage. The maize silage (MS; 20-mm length) and alfalfa haylage (AH; 40-mm length) samples were wet (wet maize silage, MSW; wet alfalfa haylage, AHW), dried (dried maize silage, MSD; dried alfalfa haylage, AHD), or ground to pass through a 2.5-mm screen (dried ground maize silage, MSG; dried ground alfalfa haylage, AHG). Samples were incubated in the rumen for 2, 6, 12, 24, 36, 48, and 72 h. Cows were fed ad libitum and allowed free access to water. High moisture content treatment of MSW expressed a lower rinsing NDF and ADF degradability at 2 h ( P  < 0.05) compared with dried samples (MSD and MSG). Different moisture content and particle size had a significant impact ( P  < 0.05) on the NDF degradability at 72 h, ADF degradability at 36, 48, and 72 h, and ruminally degradable ADF. All of the highest values were observed in small particle size and low moisture content AHG treatment. Based on this study, sample processing, such as drying and grinding, should be considered when evaluating nutritive values of forages.

  18. Effects of moisture content or particle size on the in situ degradability of maize silage and alfalfa haylage in lactating dairy cows

    Directory of Open Access Journals (Sweden)

    Yang Zou

    2016-09-01

    Full Text Available A study using four Holstein cows with ruminal cannulas was conducted to evaluate the degradability of different moisture content or particle size of maize silage and alfalfa haylage. The maize silage (MS; 20-mm length and alfalfa haylage (AH; 40-mm length samples were wet (wet maize silage, MSW; wet alfalfa haylage, AHW, dried (dried maize silage, MSD; dried alfalfa haylage, AHD, or ground to pass through a 2.5-mm screen (dried ground maize silage, MSG; dried ground alfalfa haylage, AHG. Samples were incubated in the rumen for 2, 6, 12, 24, 36, 48, and 72 h. Cows were fed ad libitum and allowed free access to water. High moisture content treatment of MSW expressed a lower rinsing NDF and ADF degradability at 2 h (P < 0.05 compared with dried samples (MSD and MSG. Different moisture content and particle size had a significant impact (P < 0.05 on the NDF degradability at 72 h, ADF degradability at 36, 48, and 72 h, and ruminally degradable ADF. All of the highest values were observed in small particle size and low moisture content AHG treatment. Based on this study, sample processing, such as drying and grinding, should be considered when evaluating nutritive values of forages.

  19. Lactobacillus diolivorans sp nov., a 1,2-propanediol-degrading bacterium isolated from aerobically stable maize silage

    NARCIS (Netherlands)

    Krooneman, J; Faber, F; Alderkamp, AC; Elferink, SJHWO; Driehuis, F; Cleenwerck, [No Value; Swings, J; Gottschal, JC; Vancanneyt, M

    Inoculation of maize silage with Lactobacillus buchneri (5 x 10(5) c.f.u. g(-1) of maize silage) prior to ensiling results in the formation of aerobically stable silage. After 9 months, lactic acid bacterium counts are approximately 10(10) c.f.u. g(-1) in these treated silages. An important

  20. Kinetics of transit and degradation of the fiber from guinea grass silages enriched with waste from soybean pre-cleaning

    Directory of Open Access Journals (Sweden)

    Filipe Ton Fialho

    2015-06-01

    Full Text Available The objective was to study the kinetics of transit and degradation of the fiber from guinea grass and the waste from soybean pre-cleaning (WSPC, ensiled with different proportions of mass (0, 100, 150, and 200 g WSPC/kg total mass. Four crossbred (Gyr × Holstein, fistulated cattle with an average body mass of 400±50 kg were organized in a 4 × 4 Latin square experimental design. The fiber utilized in the study of the transit kinetics was stained with chromium mordant, whereas the in situ technique was adopted for the degradation kinetics. The level of inclusion of WSPC only affected the true digestibility and the mean retention time. The addition of waste from soybean pre-cleaning to the silage of guinea grass is beneficial, in terms of kinetics of digestion and passage, at up to levels close to 100 g/kg, because after this quantity the fiber digestion and passage in and through the reticulo-rumen are impaired and there may be alterations in the ruminal environment that will affect the use of silage by animals.

  1. Performance and metabolite profile of dairy cows fed tropical grasses and concentrates containing crude protein with low or high degradability

    Directory of Open Access Journals (Sweden)

    Raphael dos Santos Gomes

    Full Text Available ABSTRACT Ten Holstein-Zebu crossbred cows distributed into two simultaneous Latin squares (5 × 5 as a 2 × 2 factorial arrangement formed by chopped sugarcane or elephant grass silage, both with high or low protein degradability supplements and a corn silage as a control treatment, were compared using orthogonal contrasts. The studied variables were the performance, plasma concentrations of urea-N, glucose, and creatinine, urine-N and milk urea-N, and the nychthemeral variation in NH3-N in the rumen fluid of dairy cows. Nutrient intake, milk production, and milk composition were affected by the treatments. The total mixed ration containing elephant grass silage combined with rumen undegradable protein (RUP provided balanced amounts of carbon and nitrogen in the rumen. This effect may explain the 18% increase in milk yield compared with the other treatments. The diurnal pattern of ruminal NH3-N was interpreted with a sinusoid model. In general, cows fed elephant grass silage exhibited higher concentrations of blood plasma and milk urea-N than animals fed sugarcane. The cows that consumed elephant grass silage with rumen degradable protein concentrate showed a higher milk urea-N compared with animals that consumed elephant grass silage with the RUP concentrate. The use of diets based on corn silage leads to a better use of nitrogen compounds because these diets resulted in lower levels of urea-N in the plasma, urine, and milk at the same level of milk production compared with diets containing elephant grass silage or chopped sugarcane as roughages. In sugarcane-based diets, even greater nitrogen losses in the urine are observed, despite the presence of readily fermentable carbohydrates in the diet.

  2. Rumen degradation characteristics of ryegrass herbage and ryegrass silage are affected by interactions between stage of maturity and nitrogen fertilisation rate

    NARCIS (Netherlands)

    Heeren, J.A.H.; Podesta, S.C.; Hatew, B.; Klop, G.; Laar, van H.; Bannink, A.; Warner, D.; Jonge, de L.H.; Dijkstra, J.

    2014-01-01

    The objective of this experiment was to evaluate interaction effects between stage of maturity and N fertilization rate on rumen degradation characteristics determined with nylon bag incubations of ryegrass herbages and ryegrass silage. Grass herbage (n = 4) was cut after 3 or 5 weeks of regrowth

  3. Amino acid profile of metabolisable protein in lactating dairy cows is affected by dry matter concentration in grass-clover silage

    DEFF Research Database (Denmark)

    Johansen, Marianne; Lund, Peter; Weisbjerg, Martin Riis

    2018-01-01

    Our previous study showed that supply of metabolisable protein (MP) to lactating dairy cows increased with increasing dry matter (DM) concentration in grass-clover silage. The aim of this study was to examine how amino acid (AA) profile of MP was affected by silage DM concentration. Eight grass-c...

  4. Short communication. Effects of adding different protein and carbohydrates sources on chemical composition and in vitro gas production of corn stover silage

    Directory of Open Access Journals (Sweden)

    L. A. Mejía-Uribe

    2013-05-01

    Full Text Available The use of protein-rich by-products based in swine manure (SM, poultry waste (PW or chemicals compounds as urea (U, as well as energy products like molasses (M and bakery by-product (BB, is a viable method to produce good quality silage. In addition, the use of a bacterial additive can improve the fermentation characteristics of silage. The objective of this study was to determine chemical composition, in vitro gas production (GP and dry matter disappearance (DMd, using different sources of protein and energy in silage. The silages were made using SM, PW or U as protein sources and M or BB as energy source, with corn stover and with or without a bacterial additive. The organic matter (OM content was higher (p < 0.001 in silages with UBB, UM and SMBB compared with the rest of the treatments; meanwhile crude protein content was higher (p < 0.001 in silages with U. The addition of a bacterial additive increased (p < 0.05 OM content and decreased (p < 0.05 fiber content. Total GP was higher (p < 0.05 in silages containing BB, but DMd was higher (p < 0.05 in silages with U and SMBB. The inclusion of a bacterial additive decreased (p < 0.05 GP and DMd. The use of alternative sources of protein such as poultry and swine manure or urea, and of by-products of sugar industry and bakery is an alternative for silages based on corn stover. The results show that when properly formulated, the silages can provide more than 16% of crude protein and have DMd values above 60%.

  5. Rumen escape protein in grass and grass silage deterimened with a nylon bag and an enzymatic technique

    NARCIS (Netherlands)

    Cone, J.W.; Gelder, van A.H.; Mathijssen-Kamman, A.A.; Hindle, V.A.

    2004-01-01

    Rumen escape protein (REP) was determined for six grasses and 16 grass silages using a nylon bag technique and an in vitro technique using a proteolytic enzyme preparation of Streptomyces griseus. In vitro, the samples were incubated for 0, 1, 6 and 24 h. The highest correlation observed between

  6. and the quality of silage

    Directory of Open Access Journals (Sweden)

    Liisa Syrjälä-Qvist

    1984-09-01

    Full Text Available Five silages having different proportions of red clover and timothy (100/0, 75/25, 50/50, 25/75 and 0/100 were preserved in five glass-fibre silos of 0.4 m 3. Chopped red clover and timothy were mixed during ensiling and preserved with AIV II solution (80 % formic acid and 2 % orthophosphoric acid, applied at the rate of 6 l/1000kg fresh material. The timothy sward was fertilized with 100kg N/ha and the red clover with 15kg N/ha. The dry matter content in red clover was 14.5 % and in timothy 18.6%, and the crude protein content of DM 22.9 % and 11.5 %, respectively. True protein formed 75—76 % of crude protein in both herbages and the proportion of watersoluble N in total N was 32 % in red clover and 36 % in timothy. The rumen degradability of protein during the first two hours was under 10 % in both herbages; after 18 and 24h ours it was 76 % and 87 % in red clover, and 65 % and 70 % in timothy. In red clover the contents of lysine, methionine and cysteine were 5.7 g, 0.4 g and 0.9 g/16 g N, respectively, the corresponding values for timothy being 4.8 g, 0.7 g and 1.4g/16 g N. In red clover the content of water-soluble carbohydrates was 10.6 % of DM and in timothy 16.5 %; the contents of plant acids were 6.7 % and 3.5 % of DM, respectively. Red clover contained 10.9g Ca/kg DM and timothy 3.1 g Ca. The total amount of inorganic constituents was also higher in red clover than in timothy. Although the chemical composition of red clover was less suitable for ensiling than the composition of timothy, the quality of all the silages was good. During ensiling the decrease in the proportion of true protein in crude protein and the increase in the proportion of watersoluble N in total N were smaller in red clover than in timothy silage. These changes were reflected in the ruminal protein degradation, which seemed to be slower when they were small. The amino acid profile of the protein did not alter during ensiling. The ensiling losses were higher in

  7. Chemical composition and standardized ileal digestibility of protein and amino acids from blue mussel, starfish, and fish silage in pigs

    DEFF Research Database (Denmark)

    Nørgaard, Jens Vinther; Petersen, Jens Kjerulf; Tørring, Ditte Bruunshøj

    2015-01-01

    –162 g CP/kg and 5 g chromicoxide/kg. Endogenous losses of protein and AA were estimated by feeding an N-free diet.On a dry matter (DM) basis, mussel meal contained 605 g, mussel silage 575 g, starfish meal700 g, starfish juice 393 g, and fish silage 776 g CP/kg. The ratio of AA to CP ranged from0......Mussels cultured on lines for nine months and harvested in March were boiled to removeshells and processed into a dry meal or a silage acidified by formic acid. Starfish mealwas prepared from starfish caught in May, and a starfish juice fraction was obtained bypressing fresh starfish. Commercial...... fish silage from farmed salmon was also included in theexperiment. The standardized ileal digestibility (SID) of crude protein (CP) and amino acids(AA) was evaluated in a Latin square design with pigs (initial weight 39.3 kg) fitted with asimple T-cannula in the terminal ileum. Diets contained 131...

  8. Performance and metabolite profile of dairy cows fed tropical grasses and concentrates containing crude protein with low or high degradability

    OpenAIRE

    Gomes, Raphael dos Santos; Oliveira, Tadeu Silva de; Pereira, José Carlos; Vieira, Ricardo Augusto Mendonça; Henrique, Douglas Sampaio; Fernandes, Alberto Magno; Leonel, Fernando de Paula

    2016-01-01

    ABSTRACT Ten Holstein-Zebu crossbred cows distributed into two simultaneous Latin squares (5 × 5) as a 2 × 2 factorial arrangement formed by chopped sugarcane or elephant grass silage, both with high or low protein degradability supplements and a corn silage as a control treatment, were compared using orthogonal contrasts. The studied variables were the performance, plasma concentrations of urea-N, glucose, and creatinine, urine-N and milk urea-N, and the nychthemeral variation in NH3-N in th...

  9. Radiation degradation of silk protein

    International Nuclear Information System (INIS)

    Pewlong, W.; Sudatis, B.; Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu

    2000-01-01

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated using an electron beam accelerator to investigate the application of the radiation degradation technique as a means to solubilize fibroin. The irradiation caused a significant degradation of the fiber. The tensile strength of fibroin fiber irradiated up to 2500 kGy decreased rapidly with increasing dose. The presence of oxygen in the irradiation atmosphere enhanced degradation of the tensile strength. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: a calcium chloride solution(CaCl 2 /C 2 H 5 OH/H 2 O=1:2:8 in mole ratio), a hydrochloric acid (0.5 N) and a distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water soluble proteins was extracted by a distilled water. (author)

  10. Radiation degradation of silk protein

    Energy Technology Data Exchange (ETDEWEB)

    Pewlong, W; Sudatis, B [Office of Atomic Energy for Peace, Bangkok (Thailand); Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated using an electron beam accelerator to investigate the application of the radiation degradation technique as a means to solubilize fibroin. The irradiation caused a significant degradation of the fiber. The tensile strength of fibroin fiber irradiated up to 2500 kGy decreased rapidly with increasing dose. The presence of oxygen in the irradiation atmosphere enhanced degradation of the tensile strength. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: a calcium chloride solution(CaCl{sub 2}/C{sub 2}H{sub 5}OH/H{sub 2}O=1:2:8 in mole ratio), a hydrochloric acid (0.5 N) and a distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water soluble proteins was extracted by a distilled water. (author)

  11. Efeito do acipin sobre a degradabilidade e taxa de passagem de silagens de capim-elefante e de milho, em bovinos Holandês × Zebu Effect of acipin on the degradability and rate of passage of elephant-grass and corn silages in Holstein × Zebu cattle

    Directory of Open Access Journals (Sweden)

    L.T. Henriques

    2004-12-01

    Full Text Available Avaliou-se o efeito da inclusão de acipin nas silagens de capim-elefante e de milho sobre a taxa de passagem das fases sólida e líquida da digesta ruminal e sobre a degradabilidade da matéria seca (MS, proteína bruta (PB e fibra em detergente neutro (FDN, em quatro bovinos. Os animais, com média de peso de 550kg, foram confinados em baias individuais por 90 dias. O experimento foi conduzido em quatro períodos experimentais, seguindo um esquema de parcelas subdivididas, tendo nas parcelas um esquema fatorial 2×2 (duas silagens combinadas com ausência e presença de acipin e nas subparcelas o tempo de coleta de líquido ruminal, em delineamento inteiramente ao acaso com quatro repetições. A degradabilidade efetiva da MS, da PB e da FDN da silagem de capim-elefante sem adição de acipin foi menor do que a das silagens de capim-elefante com acipin, de milho sem acipin e de milho com acipin; estas foram semelhantes entre si. A taxa de passagem de sólidos ruminais foi maior nos animais que consumiram silagem de milho. A taxa de passagem de líquidos foi maior nos animais que consumiram silagem de capim-elefante. A inclusão de acipin melhorou a degradabilidade da MS, da PB e da FDN da silagem de capim-elefante.The effects of adding acipin to elephantgrass and corn silages on the passage rates of the solid and liquid phases of the ruminal digesta, and on degradabilities of dry matter (DM, crude protein (CP and neutral detergent fiber (NDF were evaluated. The animals with average live weight of 550kg, were kept in individual stalls for 90 days. The experiment was carried out in four experimental periods, according to a split plot arrangement, with 2×2 (two silages combined with absence or presence of acipin factorial treatment combination in the plot and the time of collection of ruminal liquid in the split plot, in a completely randomized design with four replicates. The degradabilities of DM, CP and NDF in the elephantgrass silage

  12. Radiation degradation of silk protein

    Energy Technology Data Exchange (ETDEWEB)

    Wachiraporn Pewlong; Boonya Sudatis [Office of Atomic Energy for Peace, Bangkok (Thailand); Takeshita, Hidefumi; Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-09-01

    Silk fibroin fiber from the domesticated silkworm Bombyx mori was irradiated in the dose range up to 2500 kGy using an electron beam accelerator to apply the radiation degradation technique as a means to solubilize fibroin. The tensile strength of irradiated fibroin fiber decreased with increasing dose and the presence of oxygen in the irradiation atmosphere enhanced the degradation. The solubilization of irradiated fibroin fiber was evaluated using the following three kinds of solutions: calcium chloride solution (CaCl{sub 2}/C{sub 2}H{sub 5}OH/H{sub 2}O = 1 : 2 : 8 in mole ratio), hydrochloric acid (0.5N) and distilled water. Dissolution of fibroin fiber into these solutions was significantly enhanced by irradiation. Especially, an appreciable amount of water-soluble protein was extracted by distilled water. (author)

  13. Degradação ruminal de silagem de capim-elefante com adição de vagem de algaroba triturada Ruminal degradation of elephant grass silage with mesquite pods

    Directory of Open Access Journals (Sweden)

    Aníbal Coutinho do Rêgo

    2011-03-01

    Full Text Available Esta pesquisa foi realizada visando-se avaliar a degradação ruminal da matéria seca (MS, proteína bruta (PB e fibra em detergente neutro (FDN de silagens de capim-elefante colhido aos 70; 90 e 110 dias após rebrota, com inclusão de 0; 5; 10 e 15% de vagem de algaroba triturada, com base na matéria natural, em delineamento inteiramente casualizado arranjado em parcelas subdivididas. Amostras de cada silagem foram incubadas no rúmen de duas vacas Jersey por 3; 6; 12; 24; 48; 72 e 96 h, sendo os saquinhos referentes ao tempo zero apenas lavados em água para determinação da fração solúvel. Não houve interação (P > 0,05 tempo de incubação x inclusão de vagem de algaroba x idade de corte para degradabilidade da MS, embora tenha ocorrido interação destes fatores para degradabilidade da PB e FDN. A maior degradabilidade efetiva (DE da MS (42,54% foi observada para 15% de inclusão de vagem de algaroba. A DE da PB foi maior (69,04% para silagem de capim-elefante com 70 dias de idade com 15% de vagem de algaroba. A inclusão de vagem de algaroba triturada à silagem de capim-elefante melhora a degradabilidade da MS, PB e FDN, enquanto o avanço da idade após rebrota resulta em redução destes parâmetros.This research was carried out to evaluate the ruminal degradation of dry matter (DM, crude protein (CP and neutral detergent fiber (NDF of silages of elephant grass (Pennisetum purpureum cutting in 70; 90 and 110 days after regrowth with inclusion of 0; 5; 10 and 15% of mesquite (Prosopis juliflora meal, based on natural matter in a completely randomized design, in split plot arrangement. Samples of silages were incubated in the rumen of two Jersey cows for 3; 6; 12; 24; 48; 72 and 96 h, and the bags at time "zero" were only washed with water to determine the soluble fraction. There was not interaction (P > 0.05 incubation time × inclusion of mesquite pods × cutting age of the grass for DM degradability, there was only

  14. Effects of different corn silage: Alfalfa silage ratios and full fat ...

    African Journals Online (AJOL)

    Full fat extruded soybeans addition increased the ether extract in diets and content of conjugated linoleic acids in milk, but it had no effect on fat, protein and lactose content in milk. Milk fat and lactose were not affected by replacing corn silage with alfalfa silage in diets. Increasing alfalfa silage content in the diets increased ...

  15. Toxic pyrrolizidinalkaloids as undesired contaminants in food and feed: degradation of the PAs from Senecio jacobaea in silage.

    Science.gov (United States)

    Becerra-Jiminez, J; Kuschak, M; Roeder, E; Wiedenfeld, H

    2013-07-01

    Pyrrolizidine alkaloids (PAs) can show a hazardous potential for men and animals. They can act as cancerogenic, mutagenic, teratogenic and fetotoxic agents. One pathway of a human intoxication is its occurence as contaminants in food and feed. Here, the contamination of cereals already led to severe and fatal intoxication episodes. Besides this, milk is of special concern as it is the main food for children which show a very high susceptibility for a PA intoxication. Milk can contain PAs in case the milk producing animals have access to contaminated feed. In this context it is of special interest whether the PA content of contaminated silage remains stable during the ensiling procedure or show a more or less high level of decomposition. We could show that ensiling will not lead to PA-free silage.

  16. Degradabilidade in situ da silagem de quatro genótipos de sorgo com e sem tanino: I - Matéria seca e proteína bruta In situ degradability of four sorgum silages with or without tannin: I - Dry mater and crude protein

    Directory of Open Access Journals (Sweden)

    W.E. Campos

    2003-04-01

    Full Text Available Avaliaram-se a degradabilidade in situ da matéria seca (MS e da proteína bruta (PB da silagem de quatro genótipos de sorgo, dois com tanino (CMS XS 210 e BR 701 e dois sem tanino (CMS XS 214 e BR007 em um delineamento de blocos inteiramente ao acaso, com três repetições (animais, em arranjo de parcelas subdivididas. Os genótipos constituíram as parcelas e os tempos de digestão as subparcelas. O genótipo CMS XS 210 apresentou menor degradabilidade da MS em relação aos demais e os genótipos que continham tanino (CMS XS 210 e BR 701 apresentaram menores degradabilidades da PB em relação aos que não o continham.The in situ procedure was used to evaluate the disappearance of dry matter (DM and crude protein (CP of four sorghum genotypes with (CMS XS 210 and BR 701 or without (CMS XS 214 and BR 007 tannin in a completely randomized block design experiment with three replicates (animals, in a split plot arrangement. The four genotypes were allotted to the plots and the time of disappearance to the split plot. The DM of CMS XS210 was less degraded when compared to the others and sorghums with tannin showed lower CP degradability when compared to the sorghums without tannin.

  17. Rumen Degradation Characteristics and Intestinal Digestibility of Whole Corn Silages from Different Areas%不同产地全株玉米青贮的瘤胃降解特性与小肠消化率的研究

    Institute of Scientific and Technical Information of China (English)

    李洋; 李春雷; 赵洪波; 张永根

    2015-01-01

    本试验旨在研究不同产地全株玉米青贮的营养价值、瘤胃降解特性以及小肠消化率的差异. 选取产自于甘肃( A)、山东( B)、四川( C)、陕西( D)和沈阳( E) 5个地区的全株玉米青贮样品,进行常规营养成分和氨基酸组成分析. 选用3头体重为550 kg左右、安装有永久性瘤胃瘘管的荷斯坦奶牛,采用尼龙袋法测定5 个产地的全株玉米青贮的干物质( DM )、粗蛋白质( CP)、中性洗涤纤维( NDF)、酸性洗涤纤维( ADF)的瘤胃降解参数以及 DM、CP 和氨基酸( AA)的小肠消化率. 结果表明:不同产地的全株玉米青贮常规营养成分存在差异. 5 个产地的全株玉米青贮的DM、CP、NDF、ADF瘤胃有效降解率之间存在差异,其高低顺序分别为B>E>A>D>C、C>A>D>B>E、B>A>D>C>E、D>A>C>B>E. 5个产地的全株玉米青贮的DM、CP和各AA小肠消化率之间存在差异,其中DM、CP和总氨基酸( TAA)小肠消化率的高低顺序分别为C>A>D>B>E、C>A>B>D>E、A>D>B>C>E. 由此得出,不同产地的全株玉米青贮的营养成分含量、瘤胃降解参数以及小肠消化率存在较大差异,在全株玉米青贮的使用中应快速准确地测量其中各种营养成分的含量,以便更有效地利用饲料资源,避免浪费.%The objective of this study was to research the differences in nutritional value, rumen degradation characteristics and intestinal digestibility of whole corn silages from different areas. Five whole corn silage sam?ples were collected from Gansu ( A) , Shandong ( B) , Sichuan ( C) , Shaanxi ( D) and Shenyang ( E) , and the common nutritional components and amino acid composition of feed samples were analysed. Three perma?nent ruminal cannulated Holstein cows with around 550 kg of body weight were used to evaluate rumen degra?dation characteristics of dry matter ( DM) , crude protein ( CP) , neutral detergent fiber ( NDF) and acid deter

  18. SILAGE QUALITY OF CORN AND SORGHUM ADDED WITH FORAGE PEANUTS

    Directory of Open Access Journals (Sweden)

    WALKÍRIA GUIMARÃES CARVALHO

    2016-01-01

    Full Text Available Corn and sorghum are standard silage crops because of their fermentative characteristics. While corn and sorghum silages have lower crude protein (CP contents than other crops, intercropping with legumes can increase CP content. Furthermore, one way to increase CP content is the addition of legumes to silage. Consequently, the research objective was to evaluate the fermentative and bromatological characteristics of corn (Zea mays and sorghum (Sorghum bicolor silages added with forage peanuts (Arachis pintoi. The experimental design was completely randomized with four replicates. The treatments consisted of corn silage, sorghum silage, forage peanut silage, corn silage with 30% forage peanut, and sorghum silage with 30% forage peanut. The results showed that the corn and sorghum added with peanut helped to improve the silage fermentative and bromatological characteristics, proving to be an efficient technique for silage quality. The forage peanut silage had lower fermentative characteristics than the corn and sorghum silages. However, the forage peanut silage had a greater CP content, which increased the protein contents of the corn and sorghum silages when intercropped with forage peanuts.

  19. Health status of cows fed maize silage covered with oxo-biodegradable foil

    Directory of Open Access Journals (Sweden)

    Piotr SZTERK

    2017-11-01

    Full Text Available In agricultural practice, silage production uses pure, low density polyethylene foil. This foil, after use, becomes farm waste, having a negative impact on the environment. Instead of conventional foil, an environmentally safe biodegradable foil can be used, made from naturally occurring polymers or from synthetic multiparticulates, easily degradable by microorganisms. Silage covered with this type of foil should be safe for animal health. The purpose of the study was to determine whether oxo-biodegradable film could be used instead of conventional film in agricultural practice, to produce silage that is safe for the cows' health. Dairy cows were fed a partly mixed ratio (PMR, the component of which was silage made of whole maize plants, covered with oxo-biodegradable foil. The cow blood serum was marked for content of: glucose, total protein, cholesterol, triacylglycerols and enzyme activity: aspartic and alanine aminotransferase, γ-glutamyl transferase, alkaline phosphatase and amylase. The total protein concentration in the serum of cows analyzed at the end of the experiment was higher than the commonly accepted normal values. The content of glucose, cholesterol, triacylglycerols and the activity of aspartate and alanine aminotransferase, γ-glutamyl transferase, alkaline phosphatase and amylase was within reference limits. Feeding of silage from whole maize plants covered withoxo-biodegradable foil did not negatively affect the biochemical indicators of the cows' blood serum. The silage proved to be safe for the cows' health.

  20. AAA-ATPases in Protein Degradation

    Directory of Open Access Journals (Sweden)

    Ravikiran S. Yedidi

    2017-06-01

    Full Text Available Proteolytic machineries containing multisubunit protease complexes and AAA-ATPases play a key role in protein quality control and the regulation of protein homeostasis. In these protein degradation machineries, the proteolytically active sites are formed by either threonines or serines which are buried inside interior cavities of cylinder-shaped complexes. In eukaryotic cells, the proteasome is the most prominent protease complex harboring AAA-ATPases. To degrade protein substrates, the gates of the axial entry ports of the protease need to be open. Gate opening is accomplished by AAA-ATPases, which form a hexameric ring flanking the entry ports of the protease. Protein substrates with unstructured domains can loop into the entry ports without the assistance of AAA-ATPases. However, folded proteins require the action of AAA-ATPases to unveil an unstructured terminus or domain. Cycles of ATP binding/hydrolysis fuel the unfolding of protein substrates which are gripped by loops lining up the central pore of the AAA-ATPase ring. The AAA-ATPases pull on the unfolded polypeptide chain for translocation into the proteolytic cavity of the protease. Conformational changes within the AAA-ATPase ring and the adjacent protease chamber create a peristaltic movement for substrate degradation. The review focuses on new technologies toward the understanding of the function and structure of AAA-ATPases to achieve substrate recognition, unfolding and translocation into proteasomes in yeast and mammalian cells and into proteasome-equivalent proteases in bacteria and archaea.

  1. AAA-ATPases in Protein Degradation.

    Science.gov (United States)

    Yedidi, Ravikiran S; Wendler, Petra; Enenkel, Cordula

    2017-01-01

    Proteolytic machineries containing multisubunit protease complexes and AAA-ATPases play a key role in protein quality control and the regulation of protein homeostasis. In these protein degradation machineries, the proteolytically active sites are formed by either threonines or serines which are buried inside interior cavities of cylinder-shaped complexes. In eukaryotic cells, the proteasome is the most prominent protease complex harboring AAA-ATPases. To degrade protein substrates, the gates of the axial entry ports of the protease need to be open. Gate opening is accomplished by AAA-ATPases, which form a hexameric ring flanking the entry ports of the protease. Protein substrates with unstructured domains can loop into the entry ports without the assistance of AAA-ATPases. However, folded proteins require the action of AAA-ATPases to unveil an unstructured terminus or domain. Cycles of ATP binding/hydrolysis fuel the unfolding of protein substrates which are gripped by loops lining up the central pore of the AAA-ATPase ring. The AAA-ATPases pull on the unfolded polypeptide chain for translocation into the proteolytic cavity of the protease. Conformational changes within the AAA-ATPase ring and the adjacent protease chamber create a peristaltic movement for substrate degradation. The review focuses on new technologies toward the understanding of the function and structure of AAA-ATPases to achieve substrate recognition, unfolding and translocation into proteasomes in yeast and mammalian cells and into proteasome-equivalent proteases in bacteria and archaea.

  2. Uptake and degradation of circulating proteins by the liver.

    NARCIS (Netherlands)

    Buys, Carolus Henricus Cornelis Maria

    1976-01-01

    Circulating proteins, like all proteins in a living animal, are subject to continual replacement or turnover. This process implies both synthesis and degradation, This thesis deals with the degradative part of turnover of circulating proteins. ... Summary

  3. Silage review: Recent advances and future uses of silage additives.

    Science.gov (United States)

    Muck, R E; Nadeau, E M G; McAllister, T A; Contreras-Govea, F E; Santos, M C; Kung, L

    2018-05-01

    to apply a direct-fed microbial strain at ensiling, have it survive ensiling, and multiply during feed out. Chemical additives traditionally have fallen in 2 groups. Formic acid causes direct acidification, suppressing clostridia and other undesired bacteria and improving protein preservation during ensiling. On the other hand, sorbic, benzoic, propionic, and acetic acids improve silage aerobic stability at feed out through direct inhibition of yeasts and molds. Current research has focused on various combinations of these chemicals to improve both aerobic stability and animal productivity. Enzyme additives have been added to forage primarily to breakdown plant cell walls at ensiling to improve silage fermentation by providing sugars for the LAB and to enhance the nutritive value of silage by increasing the digestibility of cell walls. Cellulase or hemicellulase mixtures have been more successful at the former than the latter. A new approach focused on Lb. buchneri producing ferulic acid esterase has also had mixed success in improving the efficiency of silage digestion. Another new enzyme approach is the application of proteases to corn silage to improve starch digestibility, but more research is needed to determine the feasibility. Future silage additives are expected to directly inhibit clostridia and other detrimental microorganisms, mitigate high mycotoxin levels on harvested forages during ensiling, enhance aerobic stability, improve cell wall digestibility, increase the efficiency of utilization of silage nitrogen by cattle, and increase the availability of starch to cattle. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  4. Microbial degradation of dissolved proteins in seawater

    International Nuclear Information System (INIS)

    Hollibaugh, J.T.; Azam, F.

    1983-01-01

    An experimental protocol using radiolabeled proteins was developed to investigate the rates and mechanisms whereby dissolved proteins are degraded in natural marine plankton communities. The results of field observations and laboratory experiments indicate that proteins are degraded by a particle-bound, thermolabile system, presumably bacteria-associated enzymes, with an apparent half-saturation constant of ca. 25 μg bovine serum albumin (BSA) per liter. Gel permeation chromatography indicated that peptides of chain length intermediate between BSA and the final products of degradation (MW<700) do not accumulate in the medium. Competition experiments indicate that the system is relatively nonspecific. Turnover rates for the protein pool in samples collected in the Southern California Bight were of the same order of magnitude as the turnover rate of the L-leucine pool and were correlated with primary productivity, chlorophyll a concentrations, bacterial abundance and biomass, and L-leucine turnover rate. These data suggest that amino acids derived from proteins are utilized preferentially and do not completely mix with the amino acids in the bulk phase

  5. Fracionamento dos carboidratos e proteínas de silagens de milho, sorgo e girassol Fractionation of carbohydrate and protein of corn, sorghum and sunflower silages

    Directory of Open Access Journals (Sweden)

    Renius Mello

    2004-10-01

    Full Text Available O trabalho caracteriza e quantifica as frações dos carboidratos e proteínas de silagens de milho, sorgo e girassol, com a finalidade de disponibilizar dados bromatológicos que possibilitem maximizar o aproveitamento desses alimentos e otimizar o desempenho animal. Foram avaliados dois híbridos de milho (Zea mays, DKB-215 e DKB-344, dois híbridos de sorgo (Sorghum bicolor, Ambar e AG-2005 e dois híbridos de girassol (Helianthus annuus, Rumbosol e M-734. A silagem de girassol apresentou menor valor de carboidratos totais (CT e B2 (celulose e hemicelulose e maior de C (lignina e fibra associada à lignina. A silagem de milho apresentou maior valor de carboidratos não-fibrosos (CNF, A+B1 (açúcares solúveis + amido e pectina e de CT juntamente com a silagem de sorgo, enquanto a de sorgo apresentou maior valor de B2 em função da maior contribuição de colmo. Houve diferença entre híbridos dentro da cultura do girassol para CT, sendo que o Rumbosol obteve maior valor que o M-734, em razão da aptidão dos mesmos, forrageiro e granífero respectivamente. A silagem de girassol apresentou maior valor de proteína bruta (PB e de suas frações. Não foi observada diferença entre híbridos nos valores de PB e de suas frações.This work evaluates and characterizes the carbohydrates and proteins fractions of corn, sorghum and sunflower silages. The purpose was to supply composition data that make it possible to maximize use of foods and optimize animal performance. Two corn (Zea mays hybrids, DKB-215 and DKB-344; sorghum (Sorghum bicolor hybrids, Ambar and AG-2005; and sunflower (Helianthus annuus hybrids, Rumbosol and M-734; were evaluated. Sunflower silage showed the lowest total carbohydrates (TC and B2 (cellulose and hemicelluloses values and the highest content of C (lignin and fiber associated lignin. Corn silages showed higher non-fiber carbohydrates (NFC, A+B1 (soluble sugars + starch and pectin and TC together sorghum silage while

  6. Exploring the Feasibility of Using Silage-Based Feed with Alternative Sources of Protein in Organic Pig Rations

    Directory of Open Access Journals (Sweden)

    Ruth C. Clements

    2013-12-01

    Full Text Available Current regulations for organic pig and poultry production systems permit feed ingredients of non-organic origin at an inclusion rate of up to 5 per cent. This is primarily due to concerns that there is an insufficient supply of organic protein on the European Union market, in terms of quality and quantity, to meet the nutritional requirements of pigs and poultry raised on organic farms. However, 100 per cent organic diets for monogastric livestock will become compulsory in the EU from 1 January 2018, and there is therefore a need to develop sustainable feeding strategies based on organic feeds. This feed trial conducted in the UK explores the feasibility of using a silage-based feeding system for Gloucester Old Spot pigs, and compares the inclusion of soya, beans and peas as protein sources in terms of pig growth performance. No significant difference in the pen mean daily live weight gain was observed during the grower phase (pen mean age of 11-14 weeks between the diet groups. However, during the finisher phase (pen mean age of 15-22 weeks, pigs on the soya and pea rations had significantly faster growth rates than pigs fed the bean ration. It is speculated that the slight shortfall in growth rate observed in the pigs fed the bean ration may be offset by the lower cost of production of beans in the UK. This feasibility trial demonstrates that a 100 per cent organic diet for pigs using alternative, locally-grown sources of protein as part of a forage-based ration can provide a viable alternative to a soya-based diet.

  7. Protein degradation and protection against misfolded or damaged proteins

    Science.gov (United States)

    Goldberg, Alfred L.

    2003-12-01

    The ultimate mechanism that cells use to ensure the quality of intracellular proteins is the selective destruction of misfolded or damaged polypeptides. In eukaryotic cells, the large ATP-dependent proteolytic machine, the 26S proteasome, prevents the accumulation of non-functional, potentially toxic proteins. This process is of particular importance in protecting cells against harsh conditions (for example, heat shock or oxidative stress) and in a variety of diseases (for example, cystic fibrosis and the major neurodegenerative diseases). A full understanding of the pathogenesis of the protein-folding diseases will require greater knowledge of how misfolded proteins are recognized and selectively degraded.

  8. Evaluation of continuous silage of sorghum varieties samurai 2 containing probiotic of BIOS K2 in rumen fluid of buffalo

    International Nuclear Information System (INIS)

    Shafa Imanda; Yunus Effendi; Irawan Sugoro; Sihono; Irawan Sugoro

    2016-01-01

    Ruminant forage needs could be enhanced by making silage. One of develop techniques was a continuous silage that is a modification technology of making silage by reducing the fermentation time through the addition of seeds inoculum at the beginning of the forages production. The quality of silage could be improved by adding a probiotic supplement such as BIOS K2. Forage that has a potency as silage is sorghum var. Samurai 2. The aim of this study was to evaluate the continuous silage of sorghum var. Samurai 2 that contained probiotics BIOS K2. Evaluation of sample used in vitro Hoheinheim gas test method by using buffalo rumen fluid which was incubated for 24 hours at 39 °C. Treatments were A (silage sorghum 21 days), B (continuous sorghum silage 3 days), and C (continuous silage sorghum 7 days). The parameters tested were degradation of organic matter (% DOM), ammonia, volatile fatty acids (VFA), the levels of carbon dioxide (CO_2) and methane (CH_4), as well as protein synthesis microbes (bacteria and protozoa). The analysis indicated there was a significant effect in the feed treatment to ammonia concentration, the concentration of VFA, microbial protein synthesis and % DOM, while there were no effect in levels of CO_2 and CH_4. Treatment of feed C produced ammonia, VFA and bacterial protein synthesis respectively of 0.44/ml; 0.89 mg/ml; and 5.18 mg.ml"-"1 hour"-"1, higher than treatment A and B. The treatment A had the highest DOM percentage i.e. 47.30 %, while B and C i.e. 25.18 and 37.15 % respectively. Treatment A and B had higher activity of protozoa microbial protein synthesis, respectively of 2.62 mg.ml"-"1hour"-"1 than treatment C of 2.55 mg.ml"-"1hour"-"1. It could be concluded that a continuous feed silage sorghum var. Samurai 2 with time of incubation for 7 days (C) has a better quality than sorghum silage with 21 days incubation (A). (author)

  9. Non-degradative Ubiquitination of Protein Kinases.

    Directory of Open Access Journals (Sweden)

    K Aurelia Ball

    2016-06-01

    Full Text Available Growing evidence supports other regulatory roles for protein ubiquitination in addition to serving as a tag for proteasomal degradation. In contrast to other common post-translational modifications, such as phosphorylation, little is known about how non-degradative ubiquitination modulates protein structure, dynamics, and function. Due to the wealth of knowledge concerning protein kinase structure and regulation, we examined kinase ubiquitination using ubiquitin remnant immunoaffinity enrichment and quantitative mass spectrometry to identify ubiquitinated kinases and the sites of ubiquitination in Jurkat and HEK293 cells. We find that, unlike phosphorylation, ubiquitination most commonly occurs in structured domains, and on the kinase domain, ubiquitination is concentrated in regions known to be important for regulating activity. We hypothesized that ubiquitination, like other post-translational modifications, may alter the conformational equilibrium of the modified protein. We chose one human kinase, ZAP-70, to simulate using molecular dynamics with and without a monoubiquitin modification. In Jurkat cells, ZAP-70 is ubiquitinated at several sites that are not sensitive to proteasome inhibition and thus may have other regulatory roles. Our simulations show that ubiquitination influences the conformational ensemble of ZAP-70 in a site-dependent manner. When monoubiquitinated at K377, near the C-helix, the active conformation of the ZAP-70 C-helix is disrupted. In contrast, when monoubiquitinated at K476, near the kinase hinge region, an active-like ZAP-70 C-helix conformation is stabilized. These results lead to testable hypotheses that ubiquitination directly modulates kinase activity, and that ubiquitination is likely to alter structure, dynamics, and function in other protein classes as well.

  10. Post-ruminal digestibility of crude protein from grass and grass silages in cows

    NARCIS (Netherlands)

    Cone, J.W.; Gelder, van A.H.; Mathijssen-Kamman, A.A.; Hindle, V.A.

    2006-01-01

    Grass samples were grown on a clay or sandy soil, fertilised with 150 or 300 kg N/ha per year, and harvested on different days during two consecutive growing seasons. The grass samples were stored frozen or ensiled after wilting to approximately 250 or 450 g DM/kg. The recoveries of crude protein

  11. Effect of inclusion of different levels of silage on rumen microbial population and microbial protein synthesis in dairy steers fed on rice straw

    Directory of Open Access Journals (Sweden)

    Thien Truong Giang Nguyen

    2017-02-01

    Full Text Available Objective Leucaena leucocephala (Leucaena is a perennial tropical legume that can be directly grazed or harvested and offered to ruminants as hay, silage, or fresh. However, Leucaena contain phenolic compounds, which are considered anti-nutritional factors as these may reduce intake, digestibility and thus animal performance. Therefore, the objective of this experiment was to determine effects of Leucaena silage (LS feeding levels on rumen microbial populations, N-balance and microbial protein synthesis in dairy steers. Methods Four, rumen fistulated dairy steers with initial weight of 167±12 kg were randomly assigned to receive dietary treatments according to a 4×4 Latin square design. Treatments were as followings: T1 = untreated rice straw (RS; Control, T2 = 70% RS+30% LS, T3 = 40% RS+60% LS, and T4 = 100% LS. Dairy steers were fed rice straw and LS ad libitum and supplemented with concentrate at 0.2% of body weight/d. Results Results revealed that the rumen microbial population, especially cellulolytic, proteolytic bacteria and fungal zoospores were enhanced in steers that received 60% of LS (p0.05. Protozoal population was linearly decreased with increasing level of LS (p<0.05. Moreover, N-balance and microbial protein synthesis were enhanced by LS feeding (p<0.05 and were the highest in 60% LS group. Conclusion Based on this study, it could be concluded that replacement of RS with 60% LS significantly improved microbial population and microbial protein synthesis in diary steers.

  12. Effect of the ensiling time of hydrated ground corn on silage composition and in situ starch degradability

    Directory of Open Access Journals (Sweden)

    Marcos André Arcari

    2016-04-01

    Full Text Available O objetivo deste estudo foi avaliar o efeito do tempo de ensilagem sobre a composição química e a degradabilidade in situ do amido do milho moído hidratado (MMH em grãos de média vitreosidade. Os grãos de milho foram colhidos com 83% de matéria seca (MS e vitreosidade de 67% ± 3, e foram secos até atingirem 87% de MS. Os grãos foram moídos a dois milímetros, sendo posteriormente reconstituídos, 67% MS, e ensilados (densidade de 880 kg / m³ para até 330 dias. Uma amostra MMH foi coletado mensalmente para a determinação da composição, produtos finais da fermentação e para degradabilidade in situ do amido de milho. O tempo de ensilagem não afetou o teor de MS e proteína bruta (PB. No entanto, a concentração de amido foi reduzido em 2,4 pontos percentuais em comparação de 3 com 330 dias de ensilagem. Foram observados o aumento das concentrações de N-NH3 (8,5 vezes, ácidos láctico (3,45 vezes, acético (4,1 vezes, propionico (1,7 vezes, butírico (2,8 vezes e álcool (2,4 vezes, durante o período de ensilagem. A fracção rapidamente degradável (fração A e a taxa de degradação da fracção lentamente degradável (fração C do amido do MMH foram aumentadas 3,51 e 2,21 vezes, respectivamente, durante o período de ensilagem. Por outro lado, a fração lentamente degradável (fração B do amido do MMH foi diminuída em 1,93 vezes durante o período de ensilagem. A degradabilidade efetiva do amido do MMH foi aumentado para as taxas de passagem de 0,02 / h (79,9% vs. 94,5%; 0,05 / h (65,9% vs 90,01% e de 0,08/h (56,98% vs. 86,52% quando foi comparada o período de 3 vs 330 dias de ensilagem, respectivamente. Em conclusão, o tempo de ensilagem afetou a composição química e aumentou a degradabilidade ruminal do amido do MMH de grãos com média vitreosidade.

  13. Chemical composition and ruminal degradability of the sugar cane silage treated with chemical and bacterial additivesComposição química e degradabilidade ruminal de silagens da cana-de-açúcar tratada com aditivos químicos e bacteriano

    Directory of Open Access Journals (Sweden)

    Luiz Eduardo dos Santos

    2012-02-01

    Full Text Available The objective of this trial was to evaluate the effect of the addition of chemical and bacterial additive in the ensiling of sugar cane (Saccharum officinarum L. on chemical composition, pH, kinectic fraction and in situ degradation of nutritions components of silages. Five rumen-cannulated ½ Simental + ½ Zebu steers were allotted to a completely randomized design. The steers were placed in individual cages and they were fed with diets with 76% forage (%DM. Five silages were evaluated: control – sugar cane, no additives; urea – sugar cane + 0.5% of urea (wet basis; inoculant – sugar cane inoculated with LactoSilo® (390 g/40 t forage; NaOH – sugar cane + 1.0% of sodium hydroxide (wet basis; CaOH – sugar cane + 0.6% of calcium hydroxide (wet basis. The silage additives with sodium hydroxide showed the highest pH values before (11.20 and after (4.87 for silage. No differences were observed among the silages for dry matter (26.85, crude protein (5.25 and acid detergent fiber (57.21. Fractionation of dry matter and organic matter of silages showed similar behavior, with higher values of the soluble fraction (fraction A for silages with sodium hydroxide (45.86 and 30.95% and calcium hydroxide (29.47 and 26.13%. The use of sodium hydroxide allowed obtaining higher values for the degradation of cell wall components of silages from cane sugar. The potencial and effective degradability with 3, 5 and 8%/h of passage rate were respectively 88.44, 64.45, 56.73 and 49.83% for NDF and 82.57, 55.51, 46.72 and 38.83% for ADF, indicating that the use of sodium hydroxide as chemical additives can improve the nutritive value of cane sugar silage.Objetivou-se avaliar o efeito da inclusão de aditivos químicos e bacterianos na ensilagem de cana-de-açúcar sobre a composição, pH, fracionamento e cinética de degradação in situ dos componentes nutritivos. Utilizaram-se cinco bovinos ½ Simental + ½ Zebu providos de cânula ruminal, alocados em

  14. The effect of a mixture of Lactobacillus strains on silage quality and nutritive value of grass harvested at four growth stages and ensiled for two periods

    Directory of Open Access Journals (Sweden)

    Johan De Boever

    2013-03-01

    Full Text Available   The effect of adding an inoculant containing Lactobacillus buchneri, L. plantarum and L. casei to wilted perennial ryegrass, harvested at four growth stages and ensiled for either 60 or 150 d on silage fermentation quality, chemical composition, rumen degradability of neutral detergent fibre (NDF and organic matter (OM and in vitro OM digestibility (OMd was studied. Compared to the control silage, more sugars were fermented to lactic and acetic acid with the inoculant, resulting in a lower pH, less dry matter losses and protein degradation and a better aerobic stability. The effects of the additive on fermentation quality were more pronounced after 150 than after 60 d of ensiling, because the quality of the control silage was worse after long ensiling period. The treatment lowered NDF content of grass harvested at the first two growth stages by degrading cell walls to complex sugars, but had no effect on NDF degradability of the silage. The inoculant had no effect on rumen OM degradability nor on OMd after the short ensiling period, but increased the rumen OM degradability for the first two growth stages and OMd for all growth stages after long ensiling period.

  15. Faecal particle-size distribution from ewes fed grass silages harvested at different stages of maturity

    DEFF Research Database (Denmark)

    Jalali, Alireza; Nørgaard, Peder; Nadeau, E.

    2008-01-01

    The aim of this experiment was to study the effect of maturity stage of grass at harvest on particle size in faeces from ewes fed grass silage ad libitum. Eighteen pregnant Swedish ewes bearing two foetuses were given one of three treatments as their only feed. The treatments were early (ECS......), medium (MCS) or late (LCS) cut grass silage. The ECS, MCS and LCS silages contained 449, 578, and 634 g NDF kg-1 and 166, 111 and 81 g crude protein kg-1 DM, respectively. The in situ rumen indigestible NDF (INDF) was 77, 164 and 268 g kg-1 of NDF; the degradation rate of digestible NDF (kdDNDF) was 64......, 47 and 44 g kg-1 h-1 and DM intake was 2.5, 2.1 and 1.5 kg d-1 for ECS, MCS and LCS silage, respectively. Faeces samples were collected during four days, washed in nylon bags, freeze dried and sieved into six sieving fractions; bottom bowl (B), 0.106 (C), 0.212 (D), 0.5 (S), 1.0 (M) and 2.36 (O) mm...

  16. Natural lactic acid bacteria population of tropical grasses and their fermentation factor analysis of silage prepared with cellulase and inoculant.

    Science.gov (United States)

    Khota, Waroon; Pholsen, Suradej; Higgs, David; Cai, Yimin

    2016-12-01

    lower neutral detergent fiber and acid detergent fiber contents. The results confirmed that cellulase could improve tropical silage quality, inhibiting protein degradation and promoting fiber degradation. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  17. The establishment of a protein degradability data base for dairy ...

    African Journals Online (AJOL)

    The degradability values ranged from 96,2; 91,9 and 88,7% for ... McDonald, 1979; Kristensen, et ai., 1982; Lindberg,. 1983 ... mass of 570 kg, and average production of 5000 kg ... Dicalcium phosphate'. 0,5. Commercial vitamin and trace mineral premix. 0,3 .... effects on the degradation of most protein supplements.

  18. Comparison of Maize Silage-based Diets for Dairy Cows Containing Extruded Rapeseed Cake or Extruded Full-fat Soybean as Major Protein Sources

    Directory of Open Access Journals (Sweden)

    Jiří Třináctý

    2016-01-01

    Full Text Available The trial was carried out on four Holstein cows with initial milk yield of 27.3 ± 1.7 kg.day−1. Cows were divided into two groups – the first was fed a diet based on extruded rapeseed cake (D-ERC, the second one was fed a diet based on extruded full-fat soybean (D-EFFS, both diets contained maize silage and meadow hay. The experiment was divided into 4 periods of 42 days. Intake of dry matter, crude protein and NEL was not affected by the treatment (P > 0.05 while the intake of PDIA, PDIN and PDIE was lower in D-ERC than in D-EFFS (P < 0.05. Milk yield in D-ERC (22.6 kg.d−1 was lower than in D-EFFS (24.7 kg.d−1, P < 0.001 while concentration of milk fat and protein were reverse (P < 0.05. Smaller portion of essential AADI in crude protein intake (CPI in D-ERC resulted in lower efficiency of CPI utilization for milk protein synthesis in comparison to D-EFFS being 313 and 327 g.kg−1, respectively (P < 0.01. Concentration of AA in blood plasma was not affected by the type of diet except of His and Ile that were higher in D-EFFS (P < 0.01.

  19. Cellular proteostasis: degradation of misfolded proteins by lysosomes

    Science.gov (United States)

    Jackson, Matthew P.

    2016-01-01

    Proteostasis refers to the regulation of the cellular concentration, folding, interactions and localization of each of the proteins that comprise the proteome. One essential element of proteostasis is the disposal of misfolded proteins by the cellular pathways of protein degradation. Lysosomes are an important site for the degradation of misfolded proteins, which are trafficked to this organelle by the pathways of macroautophagy, chaperone-mediated autophagy and endocytosis. Conversely, amyloid diseases represent a failure in proteostasis, in which proteins misfold, forming amyloid deposits that are not degraded effectively by cells. Amyloid may then exacerbate this failure by disrupting autophagy and lysosomal proteolysis. However, targeting the pathways that regulate autophagy and the biogenesis of lysosomes may present approaches that can rescue cells from the deleterious effects of amyloidogenic proteins. PMID:27744333

  20. Dissection of membrane protein degradation mechanisms by reversible inhibitors

    International Nuclear Information System (INIS)

    Hare, J.F.

    1988-01-01

    The degradation of slowly turning over 125I-lactoperoxidase-labeled plasma membrane polypeptides in response to reversible temperature and lysosomotropic inhibitors was studied in rat hepatoma cultures. Cells were radiolabeled and left for 24 h to allow the removal of rapidly degraded proteins. Remaining trichloroacetic acid-precipitable protein was degraded (t 1/2 = 40-68 h) by an apparent first order process 60-86% sensitive to 10 mM NH4Cl or 5 mM methylamine and greater than 95% inhibited by temperature reduction to 18 degrees C. Thus, membrane proteins are selected for degradation in a time-dependent manner by a system which is sensitive to both 18 degrees C and to lysosomotropic amines. When inhibitory conditions were removed after 40-48 h, degradation of 125I-labeled protein resumed at the same rate as that seen in their absence. Since membrane proteins do not exhibit accelerated degradation after removal of inhibitory conditions, there can be no marking or sorting of those proteins destined for degradation during the 40-h exposure to inhibitory conditions. Exposure to amines or 18 degrees C did not affect the position of two-dimensionally resolved labeled polypeptides. Fractionation of labeled cells on Percoll gradients after 40 h of exposure to low temperature or amines showed that labeled protein remained in the plasma membrane fractions of the gradient although shifted to a slightly lower buoyant density in the presence of amines. These results support the notion that selection of plasma membrane proteins for degradation requires their internalization into acidic vesicles. Lysosomotropic amines and reduced temperature interfere with the selection process by preventing membrane fusion events

  1. Proteasomal and Lysosomal Protein Degradation and Heart Disease

    OpenAIRE

    Wang, Xuejun; Robbins, Jeffrey

    2013-01-01

    In the cell, the proteasome and lysosomes represent the most important proteolytic machineries, responsible for the protein degradation in the ubiquitin-proteasome system (UPS) and autophagy, respectively. Both the UPS and autophagy are essential to protein quality and quantity control. Alterations in cardiac proteasomal and lysosomal degradation are remarkably associated with most heart disease in humans and are implicated in the pathogenesis of congestive heart failure. Studies carried out ...

  2. Knowns and unknowns of plasma membrane protein degradation in plants.

    Science.gov (United States)

    Liu, Chuanliang; Shen, Wenjin; Yang, Chao; Zeng, Lizhang; Gao, Caiji

    2018-07-01

    Plasma membrane (PM) not only creates a physical barrier to enclose the intracellular compartments but also mediates the direct communication between plants and the ever-changing environment. A tight control of PM protein homeostasis by selective degradation is thus crucial for proper plant development and plant-environment interactions. Accumulated evidences have shown that a number of plant PM proteins undergo clathrin-dependent or membrane microdomain-associated endocytic routes to vacuole for degradation in a cargo-ubiquitination dependent or independent manner. Besides, several trans-acting determinants involved in the regulation of endocytosis, recycling and multivesicular body-mediated vacuolar sorting have been identified in plants. More interestingly, recent findings have uncovered the participation of selective autophagy in PM protein turnover in plants. Although great progresses have been made to identify the PM proteins that undergo dynamic changes in subcellular localizations and to explore the factors that control the membrane protein trafficking, several questions remain to be answered regarding the molecular mechanisms of PM protein degradation in plants. In this short review article, we briefly summarize recent progress in our understanding of the internalization, sorting and degradation of plant PM proteins. More specifically, we focus on discussing the elusive aspects underlying the pathways of PM protein degradation in plants. Copyright © 2018 Elsevier B.V. All rights reserved.

  3. Protein degradation pathways in Parkinson's disease: curse or blessing.

    Science.gov (United States)

    Ebrahimi-Fakhari, Darius; Wahlster, Lara; McLean, Pamela J

    2012-08-01

    Protein misfolding, aggregation and deposition are common disease mechanisms in many neurodegenerative diseases including Parkinson's disease (PD). Accumulation of damaged or abnormally modified proteins may lead to perturbed cellular function and eventually to cell death. Thus, neurons rely on elaborated pathways of protein quality control and removal to maintain intracellular protein homeostasis. Molecular chaperones, the ubiquitin-proteasome system (UPS) and the autophagy-lysosomal pathway (ALP) are critical pathways that mediate the refolding or removal of abnormal proteins. The successive failure of these protein degradation pathways, as a cause or consequence of early pathological alterations in vulnerable neurons at risk, may present a key step in the pathological cascade that leads to spreading neurodegeneration. A growing number of studies in disease models and patients have implicated dysfunction of the UPS and ALP in the pathogenesis of Parkinson's disease and related disorders. Deciphering the exact mechanism by which the different proteolytic systems contribute to the elimination of pathogenic proteins, like α-synuclein, is therefore of paramount importance. We herein review the role of protein degradation pathways in Parkinson's disease and elaborate on the different contributions of the UPS and the ALP to the clearance of altered proteins. We examine the interplay between different degradation pathways and provide a model for the role of the UPS and ALP in the evolution and progression of α-synuclein pathology. With regards to exciting recent studies we also discuss the putative potential of using protein degradation pathways as novel therapeutic targets in Parkinson's disease.

  4. Intake, digestibility, ruminal parameters, and microbial protein synthesis in crossbred steers fed diets based on Brachiaria grass silage and sorghum silage Consumo, digestibilidade, parâmetros ruminais e síntese de proteína microbiana em novilhos cruzados alimentados com dietas a base de silagem de braquiária e silagem de sorgo

    Directory of Open Access Journals (Sweden)

    F.H.M. Chizzotti

    2009-12-01

    Full Text Available A trial was carried out with four Holstein x Nellore crossbred steers (225±22kg of BW fitted with ruminal and abomasal cannulaes in a 4 x 4 Latin Square design to evaluate the intake and the total and partial apparent digestibilities of nutrients, ruminal parameters, and microbial synthesis. Diets consisted of 60% silage and 40% concentrate formulated to be isonitrogenous (12.5% of crude protein, dry matter basis. Treatments consisted of different proportions of Brachiaria brizantha grass silage and sorghum silage: 100:0; 67:33; 33:67, and 0:100%, respectively, on dry matter basis. The intake of dry matter, organic matter, crude protein, ether extract, non-fiber carbohydrates, and total digestible nutrients linearly increased (P0.05 by the treatments. The use of 67% of sorghum silage and 33% of grass silage increased intake and digestibility of nutrients without affecting ruminal pH, ruminal NH3-N, and microbial efficiency.Realizou-se um estudo com quatro novilhos cruzados Holandês x Nelore (225±22kg de peso vivo, canulados no rúmen e abomaso, distribuídos em quadrado latino 4 x 4 para avaliar o efeito de diferentes proporções de silagem de Braquiária brizantha e silagem de sorgo sobre o consumo e a digestibilidade dos nutrientes no trato digestório total e parcial, sobre os parâmetros ruminais e sobre a eficiência microbiana. As dietas continham 60% de volumoso e 40% de concentrado e foram formuladas para serem isonitrogenadas (12,5% de proteína bruta na matéria seca. Os tratamentos consistiram em diferentes proporções de silagens de Brachiaria brizantha e de sorgo: 100:0; 67:33; 33:67 e 0:100%, respectivamente (% da matéria seca. O consumo de matéria seca, matéria orgânica, proteína bruta, extrato etéreo, carboidratos não-fibrosos e nutrientes digestíveis totais aumentou linearmente (P0,05. O uso de 67% de silagem de sorgo com 33% de silagem de braquiária aumentou o consumo e a digestibilidade dos nutrientes sem afetar

  5. Herp enhances ER-associated protein degradation by recruiting ubiquilins

    International Nuclear Information System (INIS)

    Kim, Tae-Yeon; Kim, Eunmin; Yoon, Sungjoo Kim; Yoon, Jong-Bok

    2008-01-01

    ER-associated protein degradation (ERAD) is a protein quality control system of ER, which eliminates misfolded proteins by proteasome-dependent degradation and ensures export of only properly folded proteins from ER. Herp, an ER membrane protein upregulated by ER stress, is implicated in regulation of ERAD. In the present study, we show that Herp interacts with members of the ubiquilin family, which function as a shuttle factor to deliver ubiquitinated substrates to the proteasome for degradation. Knockdown of ubiquilin expression by small interfering RNA stabilized the ERAD substrate CD3δ, whereas it did not alter or increased degradation of non-ERAD substrates tested. CD3δ was stabilized by overexpressed Herp mutants which were capable of binding to ubiquilins but were impaired in ER membrane targeting by deletion of the transmembrane domain. Our data suggest that Herp binding to ubiquilin proteins plays an important role in the ERAD pathway and that ubiquilins are specifically involved in degradation of only a subset of ubiquitinated targets, including Herp-dependent ERAD substrates

  6. Protein oxidation and degradation caused by particulate matter

    Science.gov (United States)

    Lai, Ching-Huang; Lee, Chun-Nin; Bai, Kuan-Jen; Yang, You-Lan; Chuang, Kai-Jen; Wu, Sheng-Ming; Chuang, Hsiao-Chi

    2016-09-01

    Particulate matter (PM) modulates the expression of autophagy; however, the role of selective autophagy by PM remains unclear. The objective of this study was to determine the underlying mechanisms in protein oxidation and degradation caused by PM. Human epithelial A549 cells were exposed to diesel exhaust particles (DEPs), urban dust (UD), and carbon black (CB; control particles). Cell survival and proliferation were significantly reduced by DEPs and UD in A549 cells. First, benzo(a)pyrene diolepoxide (BPDE) protein adduct was caused by DEPs at 150 μg/ml. Methionine oxidation (MetO) of human albumin proteins was induced by DEPs, UD, and CB; however, the protein repair mechanism that converts MetO back to methionine by methionine sulfoxide reductases A (MSRA) and B3 (MSRB3) was activated by DEPs and inhibited by UD, suggesting that oxidized protein was accumulating in cells. As to the degradation of oxidized proteins, proteasome and autophagy activation was induced by CB with ubiquitin accumulation, whereas proteasome and autophagy activation was induced by DEPs without ubiquitin accumulation. The results suggest that CB-induced protein degradation may be via an ubiquitin-dependent autophagy pathway, whereas DEP-induced protein degradation may be via an ubiquitin-independent autophagy pathway. A distinct proteotoxic effect may depend on the physicochemistry of PM.

  7. The establishment of a protein degradability data base for dairy ...

    African Journals Online (AJOL)

    The establishment of a protein degradability data base for dairy cattle using the nylon bag technique. 1. Protein sources. LJ Erasmus, J Prinsloo, HH Meissner. Abstract. No Abstract. Full Text: EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT · AJOL African Journals ...

  8. Human sperm degradation of zona pellucida proteins contributes to fertilization.

    Science.gov (United States)

    Saldívar-Hernández, Analilia; González-González, María E; Sánchez-Tusié, Ana; Maldonado-Rosas, Israel; López, Pablo; Treviño, Claudia L; Larrea, Fernando; Chirinos, Mayel

    2015-09-02

    The mammalian oocyte extracellular matrix known as the zona pellucida (ZP) acts as a barrier to accomplish sperm fusion with the female gamete. Although penetration of the ZP is a limiting event to achieve fertilization, this is one of the least comprehended stages of gamete interaction. Even though previous studies suggest that proteases of sperm origin contribute to facilitate the passage of sperm through the ZP, in human this process is not yet fully understood. The aim of this study was to determine the ability of human sperm to degrade recombinant human ZP (rhZPs) proteins and to characterize the proteases involved in this process. Purified rhZP2, rhZP3 and rhZP4 proteins were incubated with capacitated sperm and the proteolytic activity was determined by Western blot analysis. To further characterize the proteases involved, parallel incubations were performed in the presence of the protease inhibitors o-phenanthroline, benzamidine and MG-132 meant to block the activity of metalloproteases, serine proteases and the proteasome, respectively. Additionally, protease inhibitors effect on sperm-ZP binding was evaluated by hemizona assay. The results showed that rhZPs were hydrolyzed in the presence of capacitated sperm. O-phenanthroline inhibited the degradation of rhZP3, MG-132 inhibited the degradation of rhZP4 and benzamidine inhibited the degradation of the three proteins under investigation. Moreover, hemizona assays demonstrated that sperm proteasome inhibition impairs sperm interaction with human native ZP. This study suggests that sperm proteasomes could participate in the degradation of ZP, particularly of the ZP4 protein. Besides, metalloproteases may be involved in specific degradation of ZP3 while serine proteases may contribute to unspecific degradation of the ZP. These findings suggest that localized degradation of ZP proteins by sperm is probably involved in ZP penetration and may be of help in understanding the mechanisms of fertilization in humans.

  9. Fish silage as feed ingredient for fish and livestock

    NARCIS (Netherlands)

    Rurangwa, E.; Vuuren, van A.M.; Poelman, M.

    2014-01-01

    The present report analyses through a literature review the potential of fish silage to valorise fish processing by-products into economically relevant protein sources for fish and livestock feed production in East Africa.

  10. Investigations on the effect of forage source, grinding, and urea supplementation on ruminal fermentation and microbial protein flow in a semi-continuous rumen simulation system.

    Science.gov (United States)

    Hildebrand, Bastian; Boguhn, Jeannette; Rodehutscord, Markus

    2011-10-01

    The objective of the present study was to compare the effect of maize silage and grass silage on microbial fermentation and protein flow in a semi-continuous rumen simulation system (Rusitec) when milling screen size (MSS) during grinding was varied. Oven-dried silages were milled through screens of 1, 4 or 9 mm pore size and incubated for 48 h in a Rusitec system. Furthermore, the effect of N supplementation to maize silage (MSS: 4 mm) was investigated and single dose vs. continuous infusion of urea-N were compared. Degradation of organic matter (OM), crude protein (CP), fibre fractions and non-structural carbohydrates (NSC) as well as short-chain fatty acid production differed significantly between forage sources. Urea-N supplementation improved the degradation of NSC, but not that of fibre fractions in maize silage. The way of urea supply had only marginal effects on fermentation characteristics. An increase in MSS, and consequently in mean feed particle size, led to an improvement in the degradation of OM, CP and NSC, but efficiency of microbial net protein synthesis (EMPS; mg microbial N flow/g degraded OM) and the microbial amino acid profile were less affected. EMPS was higher in grass silage than in maize silage and was improved by urea-N supplementation in maize silage. This study indicates that fermentation of NSC as well as EMPS during incubation of maize silage was limited by availability of NH3-N. Furthermore, an increase in MSS above 1 mm seems to improve fermentation of silages in the Rusitec system.

  11. Ethanol emission from loose corn silage and exposed silage particles

    Science.gov (United States)

    Hafner, Sasha D.; Montes, Felipe; Rotz, C. Alan; Mitloehner, Frank

    2010-11-01

    Silage on dairy farms has been identified as a major source of volatile organic compound (VOC) emissions. However, rates of VOC emission from silage are not accurately known. In this work, we measured ethanol (a dominant silage VOC) emission from loose corn silage and exposed corn silage particles using wind tunnel systems. Flux of ethanol was highest immediately after exposing loose silage samples to moving air (as high as 220 g m -2 h -1) and declined by as much as 76-fold over 12 h as ethanol was depleted from samples. Emission rate and cumulative 12 h emission increased with temperature, silage permeability, exposed surface area, and air velocity over silage samples. These responses suggest that VOC emission from silage on farms is sensitive to climate and management practices. Ethanol emission rates from loose silage were generally higher than previous estimates of total VOC emission rates from silage and mixed feed. For 15 cm deep loose samples, mean cumulative emission was as high as 170 g m -2 (80% of initial ethanol mass) after 12 h of exposure to an air velocity of 5 m s -1. Emission rates measured with an emission isolation flux chamber were lower than rates measured in a wind tunnel and in an open setting. Results show that the US EPA emission isolation flux chamber method is not appropriate for estimating VOC emission rates from silage in the field.

  12. Protein degradation: recognition of ubiquitinylated substrates

    DEFF Research Database (Denmark)

    Hartmann-Petersen, Rasmus; Gordon, Colin

    2004-01-01

    A cell-free system has been developed in budding yeast that provides direct evidence that the Dsk2/Dph1, Rad23/Rhp23 and Rpn10/Pus1 multi-ubiquitin-binding proteins, long implicated in substrate recognition and presentation to the 26S proteasome, actually fulfil such a role.......A cell-free system has been developed in budding yeast that provides direct evidence that the Dsk2/Dph1, Rad23/Rhp23 and Rpn10/Pus1 multi-ubiquitin-binding proteins, long implicated in substrate recognition and presentation to the 26S proteasome, actually fulfil such a role....

  13. In vitro ruminal fermentation characteristics and utilisable CP supply of sainfoin and birdsfoot trefoil silages and their mixtures with other legumes.

    Science.gov (United States)

    Grosse Brinkhaus, A; Wyss, U; Arrigo, Y; Girard, M; Bee, G; Zeitz, J O; Kreuzer, M; Dohme-Meier, F

    2017-04-01

    The extensive protein degradation occurring during ensiling decreases the nutritive value of silages, but this might be counteracted by tannins. Therefore, silages from two legume species containing condensed tannins (CT) - sainfoin (SF) and birdsfoot trefoil (two cultivars: birdsfoot trefoil, cv. Bull (BTB) and birdsfoot trefoil, cv. Polom) - were compared for their in vitro ruminal fermentation characteristics. The effect of combining them with two CT-free legume silages (lucerne (LU) and red clover (RC)) was also determined. The supply of duodenally utilisable CP (uCP) in the forages was emphasised. The legumes were each harvested from three field sites. After 24 h of wilting on the field, the legumes were ensiled in laboratory silos for 86 days. Proximate constituents, silage fermentation characteristics, CT content and CP fractions were determined. Subsequently, silage samples and 1 : 1 mixtures of the CT-containing and CT-free silages were incubated for 24 h in batch cultures using ruminal fluid and buffer (1 : 2, v/v). Each treatment was replicated six times in six runs. The effects on pH, ammonia and volatile fatty acid concentrations, protozoal counts, and total gas and methane production were determined. uCP content was calculated by considering the CP in the silage and the ammonia in the incubation fluid from treatments and blanks. Statistical evaluation compared data from single plants alone and together with that from the mixtures. Among treatments, SF silage contained the least CP and the most CT. The non-protein nitrogen content was lower, favouring neutral detergent soluble and insoluble protein fractions, in the SF and RC silages. Absolute uCP content was lowest in SF and SF mixtures, although the ratio to total CP was the highest. In comparison with LU, the ammonia concentration of the incubation fluid was lower for SF, RC and BTB and for the mixture of SF with LU. The total gas and methane production was similar among the treatments, and the

  14. Selectivity in protein degradation during sporulation of Bacillus subtilis

    International Nuclear Information System (INIS)

    Mitani, Takahiko; Kadota, Hajime

    1976-01-01

    The breakdown of cellular protein was investigated in Bacillus subtilis ATCC 6051 labeled with glycine-2- 3 H or L-phenylalanine-U- 14 C at the different stages of vegetative growth and sporulation. The growth of the culture was determined by measuring optical density at 660 nm. The heat-resistant spores were scored by plating after heating at 80 deg C for 10 minutes. A question whether the turnover of glycine-labeled protein is similar to that of phenylalanine-labeled protein was experimentally studied. The patterns obtained with the glycine-labeled protein were different from those of phenylalanine-labeled protein. This was not multiple turnover. The cellular protein which was labeled with glycine at an early stage of sporulation showed rapid degradation, but the degradation of the protein labeled with glycine at later stages did not occur at all. Another question whether the labeled glycine incorporated into cells at the different stages of growth and sporulation was present in the spore coat fraction of matured spores was studied. Experiment demonstrated that the glycine incorporated into cells at the late sporulation stage was mainly utilized for the biosynthesis of the spore coat protein. These data suggest that the spore coat protein which contains relatively large amount of glycine is rarely subject to further degradation. (Iwakiri, K.)

  15. Lignocellulose degradation, enzyme production and protein ...

    African Journals Online (AJOL)

    Microbial conversion of corn stover by white rot fungi has the potential to increase its ligninolysis and nutritional value, thereby transforming it into protein-enriched animal feed. Response surface methodology was applied to optimize conditions for the production of lignocellulolytic enzymes by Trametes versicolor during ...

  16. In vitro microbial protein synthesis, ruminal degradation and post-ruminal digestibility of crude protein of dairy rations containing Quebracho tannin extract.

    Science.gov (United States)

    Castro-Montoya, J; Westreicher-Kristen, E; Henke, A; Diaby, M; Susenbeth, A; Dickhoefer, U

    2018-02-01

    This study evaluated the effects of Quebracho tannin extract (QTE) on in vitro ruminal fermentation, chemical composition of rumen microbes, ruminal degradation and intestinal digestibility of crude protein (iCPd). Three treatments were tested, the control (basal diet without QTE), the basal diet with 15 g QTE/kg dry matter (DM) and the basal diet with 30 g QTE/kg DM. The basal diet contained (g/kg DM): 339 grass silage, 317 maize silage and 344 concentrate. In vitro gas production kinetic was determined using the Hohenheim gas test (Experiment 1). The Ankom RF technique, a batch system with automatic gas pressure recordings, was used to determine in vitro production of short-chain fatty acids (SCFA) and ammonia-nitrogen concentration (NH 3 -N), as well as nitrogen and purine bases content in liquid-associated microbes (LAM) and in a residue of undegraded feed and solid-associated microbes (Feed+SAM) (Experiment 2). Ruminal degradation and iCPd were determined using the nylon bag technique and the mobile nylon bag technique, respectively (Experiment 3). Gas production (Experiment 1), total SCFA and NH 3 -N (Experiment 2) decreased with increasing QTE levels. Microbial mass and composition of LAM were not affected by QTE, but total mass of Feed+SAM linearly increased, likely due to decreased substrate degradation with increasing QTE levels. The total amount of N in microbial mass and undegraded feed after the in vitro incubation increased with increasing QTE levels, suggesting a potential greater N flow from the rumen to the duodenum. In contrast to in vivo studies with the same QTE, no effects were detected on ruminal effective degradability and iCPd, when using the nylon bag techniques. Based on the in vitro procedures, QTE increased the supply of N post-rumen; however, some evidence of a decreased fibre degradation were also observed. Therefore, the benefit of adding QTE to diets of cattle is still questionable. © 2017 Blackwell Verlag GmbH.

  17. Moisture Concentration Variation of Silages Produced on Commercial Farms in the South-Central USA

    Directory of Open Access Journals (Sweden)

    K. J. Han

    2014-10-01

    Full Text Available Preservation of forage crops as silage offers opportunity to avoid the high risk of rain-damaged hay in the humid south-central USA. Recent developments with baled silage or baleage make silage a less expensive option than typical chopped silage. Silage has been important in the region primarily for dairy production, but baleage has become an option for the more extensive beef cattle industry in the region. Silage samples submitted to the Louisiana State University Agricultural Center Forage Quality Lab from 2006 through 2013 were assessed for dry matter (DM and forage nutritive characteristics of chopped silage and baleage of the different forage types from commercial farms primarily in Louisiana and Mississippi. Of the 1,308 silage samples submitted, 1,065 were annual ryegrass (AR with small grains (SG, the warm-season annual (WA grasses, sorghums and pearl millet, and the warm-season perennial (WP grasses, bermudagrass and bahiagrass, providing the remaining samples. Concentration of DM was used to indicate an effective ensiling opportunity, and AR silage was more frequently within the target DM range than was the WA forage group. The AR samples also indicated a high-quality forage with average crude protein (CP of 130 g/kg and total digestible nutrient (TDN near 600 g/kg. The cooler winter weather at harvest apparently complicated harvest of SG silage with chopped SG silage lower in both CP and TDN (104 and 553 g/kg, respectively than either AR silage or baleage of SG (137 and 624 g/kg for CP and TDN, respectively. The hot, humid summer weather along with large stems and large forage quantities of the WA grasses and the inherently higher fiber concentration of WP grasses at harvest stage indicate that preservation of these forage types as silage will be challenging, although successful commercial silage samples of each forage type and preservation approach were included among samples of silages produced in the region.

  18. Prion protein degradation by lichens of the genus Cladonia

    Science.gov (United States)

    Bennett, James P.; Rodriguez, Cynthia M.; Johnson, Christopher J.

    2012-01-01

    It has recently been discovered that lichens contain a serine protease capable of degrading the pathogenic prion protein, the etiological agent of prion diseases such as sheep scrapie and cervid chronic wasting disease. Limited methods are available to degrade or inactivate prion disease agents, especially in the environment, and lichens or their serine protease could prove important for management of these diseases. Scant information is available regarding the presence or absence of the protease responsible for degrading prion protein (PrP) in lichen species and, in this study, we tested the hypothesis that PrP degradation activity in lichens is phylogenetically-based by testing 44 species of Cladonia lichens, a genus for which a significant portion of the phylogeny is well established. We categorized PrP degradation activity among the 44 species (high, moderate, low or none) and found that activity in Cladonia species did not correspond with phylogenetic position of the species. Degradation of PrP did correspond, however, with three classical taxonomic characters within the genus: species with brown apothecia, no usnic acid, and the presence of a cortex. Of the 44 species studied, 18 (41%) had either high or moderate PrP degradation activity, suggesting the protease may be frequent in this genus of lichens.

  19. Protein degradation during reconsolidation as a mechanism for memory reorganization

    Directory of Open Access Journals (Sweden)

    Bong-Kiun Kaang

    2011-02-01

    Full Text Available Memory is a reference formed from a past experience that is used to respond to present situations. However, the world is dynamic and situations change, so it is important to update the memory with new information each time it is reactivated in order to adjust the response in the future. Recent researches indicate that memory may undergo a dynamic process that could work as an updating mechanism. This process which is called reconsolidation involves destabilization of the memory after it is reactivated, followed by restabilization. Recently, it has been demonstrated that the initial destabilization process of reconsolidation requires protein degradation. Using protein degradation inhibition as a method to block reconsolidation, recent researches suggest that reconsolidation, especially the protein degradation-dependent destabilization process is necessary for memory reorganization.

  20. Proteolysis targeting peptide (PROTAP) strategy for protein ubiquitination and degradation.

    Science.gov (United States)

    Zheng, Jing; Tan, Chunyan; Xue, Pengcheng; Cao, Jiakun; Liu, Feng; Tan, Ying; Jiang, Yuyang

    2016-02-19

    Ubiquitination proteasome pathway (UPP) is the most important and selective way to degrade proteins in vivo. Here, a novel proteolysis targeting peptide (PROTAP) strategy, composed of a target protein binding peptide, a linker and a ubiquitin E3 ligase recognition peptide, was designed to recruit both target protein and E3 ligase and then induce polyubiquitination and degradation of the target protein through UPP. In our study, the PROTAP strategy was proved to be a general method with high specificity using Bcl-xL protein as model target in vitro and in cells, which indicates that the strategy has great potential for in vivo application. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Are animal models predictive for human postmortem muscle protein degradation?

    Science.gov (United States)

    Ehrenfellner, Bianca; Zissler, Angela; Steinbacher, Peter; Monticelli, Fabio C; Pittner, Stefan

    2017-11-01

    A most precise determination of the postmortem interval (PMI) is a crucial aspect in forensic casework. Although there are diverse approaches available to date, the high heterogeneity of cases together with the respective postmortal changes often limit the validity and sufficiency of many methods. Recently, a novel approach for time since death estimation by the analysis of postmortal changes of muscle proteins was proposed. It is however necessary to improve the reliability and accuracy, especially by analysis of possible influencing factors on protein degradation. This is ideally investigated on standardized animal models that, however, require legitimization by a comparison of human and animal tissue, and in this specific case of protein degradation profiles. Only if protein degradation events occur in comparable fashion within different species, respective findings can sufficiently be transferred from the animal model to application in humans. Therefor samples from two frequently used animal models (mouse and pig), as well as forensic cases with representative protein profiles of highly differing PMIs were analyzed. Despite physical and physiological differences between species, western blot analysis revealed similar patterns in most of the investigated proteins. Even most degradation events occurred in comparable fashion. In some other aspects, however, human and animal profiles depicted distinct differences. The results of this experimental series clearly indicate the huge importance of comparative studies, whenever animal models are considered. Although animal models could be shown to reflect the basic principles of protein degradation processes in humans, we also gained insight in the difficulties and limitations of the applicability of the developed methodology in different mammalian species regarding protein specificity and methodic functionality.

  2. Nutritional evaluation of elephant-grass silages with different levels of by-products from the cashew juice industry

    Directory of Open Access Journals (Sweden)

    Ana Cristina Holanda Ferreira

    2015-12-01

    Full Text Available ABSTRACT This study was conducted to evaluate the intake, apparent digestibility (AD, and degradability in situ of elephant grass (Pennisetum purpureum Schum silages containing 0, 35.0, 70.0, 105.0, and 140.0 g kg-1 by-product from dried cashew apple (DCBP (as fed basis. A completely randomized design with four replicates was adopted. For the study of degradability in situ, one adult male cattle was used in a completely randomized design with split plots. Intake and AD of dry matter (DM, crude protein (CP, neutral detergent fiber (NDF, acid detergent fiber (ADF, hemicellulose, and cellulose were evaluated, and the digestible energy (DE and nitrogen balance (NB of the silages were determined. The degradability in situ of DM, CP, and NDF was also determined. Addition of DCBP provided an increase in the intakes of DM, CP, NDF, and ADF. No effects of the levels of addition of DCBP were observed on the coefficients of AD of the silages. Regarding NB, positive values were only detected in the treatment with 105.0 g kg-1 DCBP. In the analysis of the degradability in situ, the incubation periods increased the rates of disappearance of DM, CP, and NDF. However, no effect of the levels of DCBP were observed on the effective degradability of DM. The by-product from dried cashew apple can be included at up to 140.0 g kg-1 in silages of elephant grass, but the high contents of acid detergent insoluble nitrogen may compromise the use and availability of nitrogen to the animals.

  3. Protein degradation and protein synthesis in long-term memory formation

    Directory of Open Access Journals (Sweden)

    Timothy J Jarome

    2014-06-01

    Full Text Available Long-term memory (LTM formation requires transient changes in the activity of intracellular signaling cascades that are thought to regulate new gene transcription and de novo protein synthesis in the brain. Consistent with this, protein synthesis inhibitors impair LTM for a variety of behavioral tasks when infused into the brain around the time of training or following memory retrieval, suggesting that protein synthesis is a critical step in LTM storage in the brain. However, evidence suggests that protein degradation mediated by the ubiquitin-proteasome system may also be a critical regulator of LTM formation and stability following retrieval. This requirement for increased protein degradation has been shown in the same brain regions in which protein synthesis is required for LTM storage. Additionally, increases in the phosphorylation of proteins involved in translational control parallel increases in protein polyubiquitination and the increased demand for protein degradation is regulated by intracellular signaling molecules thought to regulate protein synthesis during LTM formation. In some cases inhibiting proteasome activity can rescue memory impairments that result from pharmacological blockade of protein synthesis, suggesting that protein degradation may control the requirement for protein synthesis during the memory storage process. Results such as these suggest that protein degradation and synthesis are both critical for LTM formation and may interact to properly consolidate and store memories in the brain. Here, we review the evidence implicating protein synthesis and degradation in LTM storage and highlight the areas of overlap between these two opposing processes. We also discuss evidence suggesting these two processes may interact to properly form and store memories. LTM storage likely requires a coordinated regulation between protein degradation and synthesis at multiple sites in the mammalian brain.

  4. Whole linted cottonseed meal (Gossypium hirsutum L. protein and fiber degradability in the rumen

    Directory of Open Access Journals (Sweden)

    Deborah Clea Ruy

    1996-12-01

    3 x 3 change-over design to evaluate the following treatments: A = 0% WLC; B = 6.6% WLC; and C = 15.0% WLC. Sorghum silage contributed with 70% in all three treatments. DM degradability at 48h incubation time was statistically different (p < 0.05 (A = 54.4%; B = 54.2% and C = 58.7%, as well as PB degradability at 12h (A = 40.3%; B = 47.7% and C = 53.1% and ADF degradability at 48h (A = 40.3%; B = 41.2% and C = 45.6%. Ruminal volume, turn overtime and ruminal pH weren’t affected by the experimental diets. Substitution of WLC for cottonseed meal up to 15% diet increased degradability of DM, CP and ADF of WLC.

  5. Degradation of Akt using protein-catalyzed capture agents.

    Science.gov (United States)

    Henning, Ryan K; Varghese, Joseph O; Das, Samir; Nag, Arundhati; Tang, Grace; Tang, Kevin; Sutherland, Alexander M; Heath, James R

    2016-04-01

    Abnormal signaling of the protein kinase Akt has been shown to contribute to human diseases such as diabetes and cancer, but Akt has proven to be a challenging target for drugging. Using iterative in situ click chemistry, we recently developed multiple protein-catalyzed capture (PCC) agents that allosterically modulate Akt enzymatic activity in a protein-based assay. Here, we utilize similar PCCs to exploit endogenous protein degradation pathways. We use the modularity of the anti-Akt PCCs to prepare proteolysis targeting chimeric molecules that are shown to promote the rapid degradation of Akt in live cancer cells. These novel proteolysis targeting chimeric molecules demonstrate that the epitope targeting selectivity of PCCs can be coupled with non-traditional drugging moieties to inhibit challenging targets. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd.

  6. Total 'shrink' losses, and where they occur, in commercially sized silage piles constructed from immature and mature cereal crops.

    Science.gov (United States)

    Robinson, P H; Swanepoel, N; Heguy, J M; Price, P; Meyer, D M

    2016-07-15

    Silage 'shrink' (i.e., fresh chop crop lost between ensiling and feedout) represents losses of potential animal nutrients which degrade air quality as volatile carbon compounds. Regulatory efforts have, in some cases, resulted in semi-mandatory mitigations (i.e., dairy farmers select a minimum number of mitigations from a list) to reduce silage shrink, mitigations often based on limited data of questionable relevance to large commercial silage piles where silage shrink may or may not be a problem of a magnitude equal to that assumed. Silage 'shrink' is generally ill defined, but can be expressed as losses of wet weight (WW), oven dry matter (oDM), and oDM corrected for volatiles lost during oven drying (vcoDM). As no research has documented shrink in large cereal silage piles, 6 piles ranging from 1456 to 6297tonnes (as built) were used. Three used cereal cut at an immature stage and three at a mature stage. Physiologically immature silages had generally higher (Plosses (vcoDM) of large well managed cereal silage piles were relatively low, and a lower potential contributor to aerosol emissions of volatile carbon compounds than has often been assumed. Losses from the silage mass and the exposed silage face were approximately equal contributors to vcoDM shrink. Mitigations to reduce these relatively low emission levels of volatile organic compounds from cereal silage piles should focus on the ensiled mass and the exposed silage face. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Impact of protein uptake and degradation on recombinant protein secretion in yeast

    DEFF Research Database (Denmark)

    Tyo, Keith E. J.; Liu, Zihe; Magnusson, Ylva

    2014-01-01

    Protein titers, a key bioprocessing metric, depend both on the synthesis of protein and the degradation of protein. Secreted recombinant protein production in Saccharomyces cerevisiae is an attractive platform as minimal media can be used for cultivation, thus reducing fermentation costs...... and transcriptomics, we identify metabolic and regulatory markers that are consistent with uptake of whole proteins by endocytosis, followed by intracellular degradation and catabolism of substituent amino acids. Uptake and degradation of recombinant protein products may be common in S. cerevisiae protein secretion...... and simplifying downstream purification, compared to other systems that require complex media. As such, engineering S. cerevisiae to improve titers has been then the subject of significant attention, but the majority of previous efforts have been focused on improving protein synthesis. Here, we characterize...

  8. Effect of silage maize hybrid (dry down vs. stay green) on dairy cow performance

    NARCIS (Netherlands)

    Zom, R.L.G.; Schooten, van H.A.; Laar, van H.

    2008-01-01

    A randomized block design experiment was conducted to evaluate the effects of two contrasting silage maize hybrids (DD: dry down vs. SG: stay green) harvested at 33% dry matter (DM) on in situ degradation and dairy cow performance. Thirty-eight Red-HF cows were assigned to two silage treatments and

  9. In situ ruminal degradability of protein feeds with distinct physical ...

    African Journals Online (AJOL)

    Journal Home > Vol 47, No 1 (2017) > ... of protein feeds with different physical forms (meal versus grain) through a meta-analysis study. ... grains with the meal forms of soybean, peanut, sunflower, cottonseed, and corn gluten. The degradation parameters of dry matter did not differ significantly between meals and grains.

  10. Rate and extent of ruminal degradation of crude protein from ...

    African Journals Online (AJOL)

    Predicted crude protein degradation was calculated at rate constants for outflow of 0.04 and 0.06/h respect- ively. ... as buffers, an ionophore and an antibiotic according to general .... the non-bird resistant ('sweet') varieties. Ruminal .... have been affected by both the particle type and the math- ematical model we used.

  11. Endocytosis and intracellular protein degradation in cystic fibrosis fibroblasts

    International Nuclear Information System (INIS)

    Jessup, W.; Dean, R.T.

    1983-01-01

    Normal rates of pinocytosis of [ 3 H]sucrose were measured in cystic fibrosis fibroblasts, and were not affected by the addition of cystic fibrosis serum. Bulk protein degradation (a significant proportion of which occurs intralysosomally following autophagy) and its regulation by growth state were apparently identical in normal and cystic fibrosis cultures. (Auth.)

  12. Proteasomal and lysosomal protein degradation and heart disease.

    Science.gov (United States)

    Wang, Xuejun; Robbins, Jeffrey

    2014-06-01

    In the cell, the proteasome and lysosomes represent the most important proteolytic machineries, responsible for the protein degradation in the ubiquitin-proteasome system (UPS) and autophagy, respectively. Both the UPS and autophagy are essential to protein quality and quantity control. Alterations in cardiac proteasomal and lysosomal degradation are remarkably associated with most heart disease in humans and are implicated in the pathogenesis of congestive heart failure. Studies carried out in animal models and in cell culture have begun to establish both sufficiency and, in some cases, the necessity of proteasomal functional insufficiency or lysosomal insufficiency as a major pathogenic factor in the heart. This review article highlights some recent advances in the research into proteasome and lysosome protein degradation in relation to cardiac pathology and examines the emerging evidence for enhancing degradative capacities of the proteasome and/or lysosome as a new therapeutic strategy for heart disease. This article is part of a Special Issue entitled "Protein Quality Control, the Ubiquitin Proteasome System, and Autophagy". Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Establishment of a ruminal protein degradation data base for dairy ...

    African Journals Online (AJOL)

    Establishment of a ruminal protein degradation data base for dairy cattle using the in situ polyester bag technique. 2. Energy sources. LJ Erasmus, J Prinsloo, PM Botha, HH Meissner. Abstract. No Abstract. Full Text: EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT.

  14. Establishment of a ruminal protein degradation data base for dairy ...

    African Journals Online (AJOL)

    Establishment of a ruminal protein degradation data base for dairy cattle using the in situ polyester bag technique. 3. Roughages. LJ Erasmus, J Prinsloo, PM Botha, HH Meissner. Abstract. No Abstract. Full Text: EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT.

  15. degradable protein sources on performance of high-producing dairy ...

    African Journals Online (AJOL)

    with high-quality, low·degradable protein sources prOViding47% UDP is advocated for ... saliva and through the rumen wall (Waldo, 1968). Based on this type of ... of the feed industry, but is based on very little solid evidence. (Huber, 1984). Chalupa ...... of rumen fermentation in relation to ammonia concentration. Br. J. Nutr.

  16. Degradation of Crude Protein in Groundnut Cake, Guinea Grass ...

    African Journals Online (AJOL)

    Three West African dwarf rams fitted with rumen cannula, were used in a completely randomized design for degradation of crude protein (CP) of groundnut cake (GNC), Panicum maximum, rumen epithelial scraping (RES), and diets containing increasing levels of RES. Concentrate diets were formulated such that 0% (A), ...

  17. Degradabilidade in situ de silagens de milho confeccionadas com inoculantes bacteriano e/ou enzimático - DOI: 10.4025/actascianimsci.v28i1.658 In situ degradability of corn silages prepared with bacterial and/or enzymatic inoculants - DOI: 10.4025/actascianimsci.v28i1.658

    Directory of Open Access Journals (Sweden)

    Elzânia Sales Pereira

    2006-01-01

    Full Text Available O objetivo deste trabalho foi avaliar os efeitos de inoculantes bacterianos e/ou enzimáticos sobre a degradabilidade ruminal da silagem de milho. Foi utilizada a técnica in situ, em quatro bovinos adultos, distribuídos em quadrado latino 4x4. Os tratamentos avaliados foram: SC (silagem controle, SIB (silagem com inoculante bacteriano, SIBE (silagem com inoculante bacteriano e enzimático e SIE (silagem com inoculante enzimático. Não houve diferença entre tratamentos nas frações solúvel (a, potencialmente degradável (b, taxa de degradação da fração b (c, degradabilidade potencial (DP e degradabilidade efetiva (DE da MS e MO. A DE da PB foi maior para o tratamento SIE (63,13% e menor para o tratamento SIBE (53,69%. A fração b da FDN apresentou maior valor para SIBE (74,13% e menor para SIB (64,07%. O resíduo indigerido (I da FDN não diferiu entre os tratamentos. As frações b e I e a taxa c da FDA não diferiram entre os tratamentos. Palavras-chave: degradação, fibra, matéria orgânica, matéria seca, proteína bruta.The objective of this work was to evaluate the effects of the bacterial and/or enzymatic inoculants on corn silage degradation. The in situ technique was used in four adult steers in a 4x4 latin square design. The evaluated treatments were: CS (control silage, SBI (silage with bacterial inoculant, SBEI (silage with bacterial and enzymatic inoculant and SEI (silage with enzymatic inoculant. There was no difference among treatments in soluble fraction (a, potential degradable fraction (b, fraction b rate of degradation (c, potential degradability (PD and effective degradability (ED of DM and OM. The ED of CP was higher in SEI treatment (63.13% and lower in SBEI treatment (53.69%. The b fraction of NDF was higher for SBEI (74.13% and lower for SBI (64.07%. The NDF indigestible residue (I did not show any difference among treatments. The ADF b and I fraction and the c rate values did not show any difference among

  18. Light-induced protein degradation in human-derived cells.

    Science.gov (United States)

    Sun, Wansheng; Zhang, Wenyao; Zhang, Chao; Mao, Miaowei; Zhao, Yuzheng; Chen, Xianjun; Yang, Yi

    2017-05-27

    Controlling protein degradation can be a valuable tool for posttranslational regulation of protein abundance to study complex biological systems. In the present study, we designed a light-switchable degron consisting of a light oxygen voltage (LOV) domain of Avena sativa phototropin 1 (AsLOV2) and a C-terminal degron. Our results showed that the light-switchable degron could be used for rapid and specific induction of protein degradation in HEK293 cells by light in a proteasome-dependent manner. Further studies showed that the light-switchable degron could also be utilized to mediate the degradation of secreted Gaussia princeps luciferase (GLuc), demonstrating the adaptability of the light-switchable degron in different types of protein. We suggest that the light-switchable degron offers a robust tool to control protein levels and may serves as a new and significant method for gene- and cell-based therapies. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Fracionamento de carboidratos e proteínas de silagens de capim-elefante com casca de café, farelo de cacau ou farelo de mandioca Fractioning of carbohydrates and protein of elephant grass silages with coffee hulls, cocoa meal and cassava meal

    Directory of Open Access Journals (Sweden)

    Aureliano José Vieira Pires

    2009-03-01

    Full Text Available Este experimento foi conduzido para avaliar as frações que compõem os carboidratos e as proteínas da silagem de capim-elefante com 15% casca de café, farelo de cacau ou farelo de mandioca. A adição dos co-produtos no momento da ensilagem foi realizada na base da matéria natural (peso/peso, com dez repetições por tipo de silagem. O maior teor de carboidratos totais foi observado na silagem com farelo de mandioca e o menor, na silagem com farelo de cacau, seguida das silagens controle e com casca de café. Maiores valores de carboidratos não-fibrosos (A+B1 também foram verificados para as silagens com farelo de mandioca. Os menores valores de fração indigestível (C, %CT foram observados para a silagem com farelo de mandioca, enquanto as silagens com casca de café e farelo de cacau apresentaram os maiores valores desta fração. A silagem com farelo de cacau apresentou os maiores valores de proteína bruta e foi seguida das silagens com casca de café, controle e com farelo de mandioca. A presença dos aditivos influenciou as frações protéicas da silagem de capim-elefante: os maiores valores de fração A (%PB foram determinados nas silagens controle e com farelo de mandioca. Apesar do maior valor protéico, as silagens com farelo de cacau e casca de café apresentaram os maiores valores de fração indigestível de proteína (C, %PB. A adição de farelo de cacau e casca de café ao capim-elefante no momento da ensilagem aumenta as frações indigestíveis de carboidratos e de proteína da silagem. O farelo de mandioca ensilado com capim-elefante é um bom aditivo para a produção de silagem.This experiment was carried out to evaluate the fractions that compose carbohydrates and protein from elephant grass silage with 15% of coffee hulls, cocoa meal or cassava meal. The addition of residues at the ensilage moment was performed in natural matter basis (weight/weight, with ten replicates per treatment. The addition of residues at

  20. Fermentation Quality and Additives: A Case of Rice Straw Silage

    Directory of Open Access Journals (Sweden)

    Yusuff Oladosu

    2016-01-01

    Full Text Available Rice cultivation generates large amount of crop residues of which only 20% are utilized for industrial and domestic purposes. In most developing countries especially southeast Asia, rice straw is used as part of feeding ingredients for the ruminants. However, due to its low protein content and high level of lignin and silica, there is limitation to its digestibility and nutritional value. To utilize this crop residue judiciously, there is a need for improvement of its nutritive value to promote its utilization through ensiling. Understanding the fundamental principle of ensiling is a prerequisite for successful silage product. Prominent factors influencing quality of silage product include water soluble carbohydrates, natural microbial population, and harvesting conditions of the forage. Additives are used to control the fermentation processes to enhance nutrient recovery and improve silage stability. This review emphasizes some practical aspects of silage processing and the use of additives for improvement of fermentation quality of rice straw.

  1. Fermentation Quality and Additives: A Case of Rice Straw Silage.

    Science.gov (United States)

    Oladosu, Yusuff; Rafii, Mohd Y; Abdullah, Norhani; Magaji, Usman; Hussin, Ghazali; Ramli, Asfaliza; Miah, Gous

    2016-01-01

    Rice cultivation generates large amount of crop residues of which only 20% are utilized for industrial and domestic purposes. In most developing countries especially southeast Asia, rice straw is used as part of feeding ingredients for the ruminants. However, due to its low protein content and high level of lignin and silica, there is limitation to its digestibility and nutritional value. To utilize this crop residue judiciously, there is a need for improvement of its nutritive value to promote its utilization through ensiling. Understanding the fundamental principle of ensiling is a prerequisite for successful silage product. Prominent factors influencing quality of silage product include water soluble carbohydrates, natural microbial population, and harvesting conditions of the forage. Additives are used to control the fermentation processes to enhance nutrient recovery and improve silage stability. This review emphasizes some practical aspects of silage processing and the use of additives for improvement of fermentation quality of rice straw.

  2. FRACIONAMENTO DE PROTEÍNAS DE SILAGEM DE CAPIM-ELEFANTE EMURCHECIDO OU COM FARELO DE CACAU PROTEIN FRACTIONING OF SILAGE OF ELEPHANTGRASS WILTED OR WITH COCOA MEAL

    Directory of Open Access Journals (Sweden)

    Gleidson Giordano Pinto de Carvalho

    2008-10-01

    Full Text Available

    Desenvolveu-se o experimento para determinar as frações que compõem as proteínas da silagem de capim-elefante (Pennisetum purpureum Schum. cv. Camaroon submetido ao emurchecimento ou à adição de diferentes níveis de farelo de cacau. O capim-elefante utilizado foi colhido aos 50 dias de rebrota após o corte de uniformização e submetido aos seguintes tratamentos: capim-elefante emurchecido ao sol por oito horas, e capim-elefante sem emurchecimento com níveis de 0 %, 7 %, 14%, 21 % e 28 % de farelo de cacau (FC (% da matéria natural. Acondicionou-se o material em silos de PVC com capacidade para 5,3 litros, que foram abertos após 45 dias. Para todas as frações de proteínas estimadas, o tratamento emurchecido apresentou valores semelhantes (P>0,05 ao do tratamento sem emurchecimento. As frações protéicas foram influenciadas pelas adições de FC, verificando-se redução dos teores das frações A e B1+B2 e aumentos das frações B3 e C, para os níveis crescentes de FC.

    PALAVRAS-CHAVES: Conservação de forragens, forrageira, Pennisetum purpureum Schum. cv. Cameroon, subproduto, Theobroma cacao L.

    The experiment was conducted to determine the fractions that compose the protein of silage on the submitted elephant grass forage to wilting under the sun light for eight hours. Other treatments involved the same elephant grass without exposing to sun light but with addition of 0, 7, 14, 21, and 28% of cocoa meal (CM at the ensilage processing. The PVC silos used in the experiment were 5.3 liters in capacity, and were opened in 45 days. To all protein-estimated fractions, the wilted treatment showed similar values (P>.05 to the treatment without wilting. The protein fractions were influenced by CM addictions, verifying reduction in contents of A and B1+B2 fractions and increase in B3 and C fractions, with CM increasing levels

  3. Metabolic and productive response to ruminal protein degradability in early lactation cows fed untreated or xylose-treated soybean meal-based diets.

    Science.gov (United States)

    Jahani-Moghadam, M; Amanlou, H; Nikkhah, A

    2009-12-01

    Effects of different dietary rumen undegradable (RUP) to degradable (RDP) protein ratios on ruminal nutrient degradation, feed intake, blood metabolites and milk production were determined in early lactation cows. Four multiparous (43 ± 5 days in milk) and four primiparous (40 ± 6 days in milk) tie-stall-housed Holstein cows were used in a duplicated 4 × 4 Latin square design with four 21-day periods. Each period had 14-day of adaptation and 7-day of sampling. Diets contained on a dry matter (DM) basis, 23.3% alfalfa hay, 20% corn silage and 56.7% concentrate. Cows were first offered alfalfa hay at 7:00, 15:00 and 23:00 hours, and 30 min after each alfalfa hay delivery were offered a mixture of corn silage and concentrate. Treatments were diets with RUP:RDP ratios of (i) 5.2:11.6 (control), (ii) 6.1:10.6, (iii) 7.1:9.5 and (iv) 8.1:8.5, on a dietary DM% basis. Different RUP:RDP ratios were obtained by partial and total replacement of untreated soybean meal (SBM) with xylose-treated SBM (XSBM). In situ study using three rumen-cannulated non-lactating cows showed that DM and crude protein (CP) of SBM had greater rapidly degradable fractions. The potentially degradable fractions were degraded more slowly in XSBM. Treatment cows produced greater milk, protein, lactose, solids-non-fat and total solids than control cows. Increasing RUP:RDP reduced blood urea linearly. Feed costs dropped at RUP:RDP ratios of 6.1:10.6 and 7.1:9.5, but not at 8.1:8.5, compared with the 5.2:11.6 ratio. Intake of DM and CP, rumen pH, blood glucose, albumin and total protein, faecal and urine pH, changes in body weight and body condition score, and milk lactose and solids-non-fat percentages did not differ among treatments. Results provide evidence that increasing dietary RUP:RDP ratio from 5.2:11.6 to 7.1:9.5 optimizes nitrogen metabolism and milk production and reduces feed costs in early lactation cows. Reduced blood urea suggests reprodutive benefits.

  4. Effect of protein degradability on milk production of dairy ewes.

    Science.gov (United States)

    Mikolayunas-Sandrock, C; Armentano, L E; Thomas, D L; Berger, Y M

    2009-09-01

    The objective of this experiment was to determine the effect of protein degradability of dairy sheep diets on milk yield and protein utilization across 2 levels of milk production. Three diets were formulated to provide similar energy concentrations and varying concentrations of rumen-degradable protein (RDP) and rumen-undegradable protein (RUP): 12% RDP and 4% RUP (12-4) included basal levels of RDP and RUP, 12% RDP and 6% RUP (12-6) included additional RUP, and 14% RDP and 4% RUP (14-4) included additional RDP. Diets were composed of alfalfa-timothy cubes, whole and ground corn, whole oats, dehulled soybean meal, and expeller soybean meal (SoyPlus, West Central, Ralston, IA). Estimates of RDP and RUP were based on the Small Ruminant Nutrition System model (2008) and feed and orts were analyzed for Cornell N fractions. Eighteen multiparous dairy ewes in midlactation were divided by milk yield (low and high) into 2 blocks of 9 ewes each and were randomly assigned within block (low and high) to 3 pens of 3 ewes each. Dietary treatments were arranged in a 3 x 3 Latin square within each block and applied to pens for 14-d periods. We hypothesized that pens consuming high-RUP diets (12-6) would produce more milk and milk protein than the basal diet (12-4) and pens consuming high-RDP diets (14-4) would not produce more milk than the basal diet (12-4). Ewes in the high-milk-yield square consumed more dry matter and produced more milk, milk fat, and milk protein than ewes in the low-milk-yield square. There was no effect of dietary treatment on dry matter intake. Across both levels of milk production, the 12-6 diet increased milk yield by 14%, increased milk fat yield by 14%, and increased milk protein yield by 13% compared with the 14-4 and 12-4 diets. Gross N efficiency (milk protein N/intake protein N) was 11 and 15% greater in the 12-6 and 12-4 diets, respectively, compared with the 14-4 diet. Milk urea N concentration was greater in the 12-6 diet and tended to be

  5. ‘Shrink’ losses in commercially sized corn silage piles: Quantifying total losses and where they occur

    Energy Technology Data Exchange (ETDEWEB)

    Robinson, P.H.; Swanepoel, N.; Heguy, J.M.; Price, T.; Meyer, D.M., E-mail: phrobinson@ucdavis.edu [Department of Animal Science, UCCE Stanislaus, San Joaquin & Merced Counties, University of California, Davis, CA 95616 (United States)

    2016-01-15

    Silage ‘shrink’ (i.e., loss of fresh chopped crop between ensiling and feedout) represents a nutrient loss which can degrade air quality as volatile carbon compounds, degrade surface waterways due to seepage, or degrade aquifers due to seepage. Virtually no research has documented shrink in large silage piles. The term ‘shrink’ is often ill defined, but can be expressed as losses of wet weight (WW), oven dry matter (oDM), and oDM corrected for volatiles lost in the drying oven (vcoDM). Corn silage piles (4 wedge, 2 rollover/wedge, 1 bunker) from 950 to 12,204 tonnes as built, on concrete (4), soil (2) and a combination (1) in California's San Joaquin Valley, using a bacterial inoculant, covered within 24 h with an oxygen barrier inner film and black/white outer plastic, fed out using large front end loaders through an electronic feed tracking system, and from the 2013 crop year, were used. Shrink as WW, oDM and vcoDM were 90 ± 17, 68 ± 18 and 28 ± 21 g/kg, suggesting that much WW shrink is water and much oDM shrink is volatiles lost during analytical oven drying. Most shrink occurred in the silage mass with losses from exposed silage faces, as well as between exposed face silage removal and the total mixed ration mixer, being low. Silage bulk density, exposed silage face management and face use rate did not have obvious impacts on any shrink measure, but age of the silage pile during silage feedout impacted shrink losses (‘older’ silage piles being higher), but most strongly for WW shrink. Real shrink losses (i.e., vcoDM) of large well managed corn silage piles are low, the exposed silage face is a small portion of losses, and many proposed shrink mitigations appeared ineffective, possibly because shrink was low overall and they are largely directed at the exposed silage face. - Highlights: • Corn silage piles were used to measure ‘shrink’ from construction to feedout • Shrink was wet weight, dry weight (oDM) and oDM volatiles corrected

  6. Protein aggregation and degradation during iodine labeling and its consequences for protein adsorption to biomaterials

    DEFF Research Database (Denmark)

    Holmberg, Maria; Jensen, Karin Bagger Stibius; Ndoni, Sokol

    2007-01-01

    Protein adsorption on modified and unmodified polymer surfaces investigated through radiolabeling experiments showed a tendency for higher than expected albumin and immunoglobulin G (IgG) adsorption. Possible enhanced protein aggregation and degradation caused by the iodine labeling method used w...

  7. Direct ubiquitin independent recognition and degradation of a folded protein by the eukaryotic proteasomes-origin of intrinsic degradation signals.

    Directory of Open Access Journals (Sweden)

    Amit Kumar Singh Gautam

    Full Text Available Eukaryotic 26S proteasomes are structurally organized to recognize, unfold and degrade globular proteins. However, all existing model substrates of the 26S proteasome in addition to ubiquitin or adaptor proteins require unstructured regions in the form of fusion tags for efficient degradation. We report for the first time that purified 26S proteasome can directly recognize and degrade apomyoglobin, a globular protein, in the absence of ubiquitin, extrinsic degradation tags or adaptor proteins. Despite a high affinity interaction, absence of a ligand and presence of only helices/loops that follow the degradation signal, apomyoglobin is degraded slowly by the proteasome. A short floppy F-helix exposed upon ligand removal and in conformational equilibrium with a disordered structure is mandatory for recognition and initiation of degradation. Holomyoglobin, in which the helix is buried, is neither recognized nor degraded. Exposure of the floppy F-helix seems to sensitize the proteasome and primes the substrate for degradation. Using peptide panning and competition experiments we speculate that initial encounters through the floppy helix and additional strong interactions with N-terminal helices anchors apomyoglobin to the proteasome. Stabilizing helical structure in the floppy F-helix slows down degradation. Destabilization of adjacent helices accelerates degradation. Unfolding seems to follow the mechanism of helix unraveling rather than global unfolding. Our findings while confirming the requirement for unstructured regions in degradation offers the following new insights: a origin and identification of an intrinsic degradation signal in the substrate, b identification of sequences in the native substrate that are likely to be responsible for direct interactions with the proteasome, and c identification of critical rate limiting steps like exposure of the intrinsic degron and destabilization of an unfolding intermediate that are presumably

  8. Effects of restriction of silage fermentation with formic acid on milk production

    Directory of Open Access Journals (Sweden)

    S. JAAKKOLA

    2008-12-01

    Full Text Available The study was conducted to evaluate the effects of silage fermentation quality and type of supplementation on milk production. Thirty two Finnish Ayrshire dairy cows were used in a cyclic change-over experiment with four 21-day experimental periods and 4 × 2 × 2 factorial arrangement of treatments. Silage fermentation was modified with formic acid (FA, which was applied at the rates equivalent to 0 (FA0, 2 (FA2, 4 (FA4 or 6 (FA6 litres t-1 grass of pure formic acid (as 100% FA. Dietary treatments consisted of four silages, a protein supplementation (no supplement or rapeseed meal 1.8 kg d-1 and a glucogenic substrate (no supplement or propylene glycol 225 g d-1. Increasing the application rate of FA restricted silage fermentation curvilinearly, as evidenced by higher concentrations of ammonia N and butyric acid in FA4 than FA2 silage. Similarly the use of FA resulted in curvilinear changes in the silage dry matter intake and milk yield. The highest milk and protein yields were achieved with FA6, while the milk yield with FA2 was higher than with FA4. Interactions were observed between silage type and supplementation. Rapeseed meal increased milk yield irrespective of the extent of silage fermentation, but the magnitude of response was variable. Propylene glycol was most beneficial with restrictively fermented silages FA4 and FA6. In conclusion, restriction of silage fermentation with a high rate of formic acid is beneficial in milk production. Interactions between silage composition and concentrate types suggest that the responses to supplementary feeding depend on silage fermentation characteristics.;

  9. Model for Stress-induced Protein Degradation in Lemna minor1

    Science.gov (United States)

    Cooke, Robert J.; Roberts, Keith; Davies, David D.

    1980-01-01

    Transfer of Lemna minor fronds to adverse or stress conditions produces a large increase in the rate of protein degradation. Cycloheximide partially inhibits stress-induced protein degradation and also partially inhibits the protein degradation which occurs in the absence of stress. The increased protein degradation does not appear to be due to an increase in activity of soluble proteolytic enzymes. Biochemical evidence indicates that stress, perhaps acting via hormones, affects the permeability of certain membranes, particularly the tonoplast. A general model for stress-induced protein degradation is presented in which changes in membrane properties allow vacuolar proteolytic enzymes increased access to cytoplasmic proteins. PMID:16661588

  10. Silage review: Foodborne pathogens in silage and their mitigation by silage additives.

    Science.gov (United States)

    Queiroz, O C M; Ogunade, I M; Weinberg, Z; Adesogan, A T

    2018-05-01

    Silage is one of the main ingredients in dairy cattle diets and it is an important source of nutrients, particularly energy and digestible fiber. Unlike properly made and managed silage, poorly made or contaminated silage can also be a source of pathogenic bacteria that may decrease dairy cow performance, reduce the safety and quality dairy products, and compromise animal and human health. Some of the pathogenic bacteria that are frequently or occasionally associated with silage are enterobacteria, Listeria, Bacillus spp., Clostridium spp., and Salmonella. The symptoms caused by these bacteria in dairy cows vary from mild diarrhea and reduced feed intake by Clostridium spp. to death and abortion by Listeria. Contamination of food products with pathogenic bacteria can cause losses of millions of dollars due to recalls of unsafe foods and decreases in the shelf life of dairy products. The presence of pathogenic bacteria in silage is usually due to contamination or poor management during the fermentation, aerobic exposure, or feed-out stages. Silage additives and inoculants can improve the safety of silage as well as the fermentation, nutrient recovery, quality, and shelf life. This review summarizes the literature on the main foodborne pathogens that occasionally infest silage and how additives can improve silage safety. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  11. The relationship between protein synthesis and protein degradation in object recognition memory.

    Science.gov (United States)

    Furini, Cristiane R G; Myskiw, Jociane de C; Schmidt, Bianca E; Zinn, Carolina G; Peixoto, Patricia B; Pereira, Luiza D; Izquierdo, Ivan

    2015-11-01

    For decades there has been a consensus that de novo protein synthesis is necessary for long-term memory. A second round of protein synthesis has been described for both extinction and reconsolidation following an unreinforced test session. Recently, it was shown that consolidation and reconsolidation depend not only on protein synthesis but also on protein degradation by the ubiquitin-proteasome system (UPS), a major mechanism responsible for protein turnover. However, the involvement of UPS on consolidation and reconsolidation of object recognition memory remains unknown. Here we investigate in the CA1 region of the dorsal hippocampus the involvement of UPS-mediated protein degradation in consolidation and reconsolidation of object recognition memory. Animals with infusion cannulae stereotaxically implanted in the CA1 region of the dorsal hippocampus, were exposed to an object recognition task. The UPS inhibitor β-Lactacystin did not affect the consolidation and the reconsolidation of object recognition memory at doses known to affect other forms of memory (inhibitory avoidance, spatial learning in a water maze) while the protein synthesis inhibitor anisomycin impaired the consolidation and the reconsolidation of the object recognition memory. However, β-Lactacystin was able to reverse the impairment caused by anisomycin on the reconsolidation process in the CA1 region of the hippocampus. Therefore, it is possible to postulate a direct link between protein degradation and protein synthesis during the reconsolidation of the object recognition memory. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. The use of urea by lactating dairy cows when fed maize silage

    African Journals Online (AJOL)

    The use of urea by lactating dairy cows when fed maize silage. T.J. Dugmere, A.M. Breden and K.P. Walsh. Department of Agriculture and Water Supply, Natal Region, Pietermaritzburg. Maize silage is relatively high in energy but low in protein for the normal lactational requirements of dairy cows. Three trials were ...

  13. 'Shrink' losses in commercially sized corn silage piles: Quantifying total losses and where they occur.

    Science.gov (United States)

    Robinson, P H; Swanepoel, N; Heguy, J M; Price, T; Meyer, D M

    2016-01-15

    Silage 'shrink' (i.e., loss of fresh chopped crop between ensiling and feedout) represents a nutrient loss which can degrade air quality as volatile carbon compounds, degrade surface waterways due to seepage, or degrade aquifers due to seepage. Virtually no research has documented shrink in large silage piles. The term 'shrink' is often ill defined, but can be expressed as losses of wet weight (WW), oven dry matter (oDM), and oDM corrected for volatiles lost in the drying oven (vcoDM). Corn silage piles (4 wedge, 2 rollover/wedge, 1 bunker) from 950 to 12,204 tonnes as built, on concrete (4), soil (2) and a combination (1) in California's San Joaquin Valley, using a bacterial inoculant, covered within 24 h with an oxygen barrier inner film and black/white outer plastic, fed out using large front end loaders through an electronic feed tracking system, and from the 2013 crop year, were used. Shrink as WW, oDM and vcoDM were 90±17, 68±18 and 28±21 g/kg, suggesting that much WW shrink is water and much oDM shrink is volatiles lost during analytical oven drying. Most shrink occurred in the silage mass with losses from exposed silage faces, as well as between exposed face silage removal and the total mixed ration mixer, being low. Silage bulk density, exposed silage face management and face use rate did not have obvious impacts on any shrink measure, but age of the silage pile during silage feedout impacted shrink losses ('older' silage piles being higher), but most strongly for WW shrink. Real shrink losses (i.e., vcoDM) of large well managed corn silage piles are low, the exposed silage face is a small portion of losses, and many proposed shrink mitigations appeared ineffective, possibly because shrink was low overall and they are largely directed at the exposed silage face. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Effect of supplementation of concentrates or selenium on production and reproduction in cows grazing pastures of high protein degradability

    Energy Technology Data Exchange (ETDEWEB)

    Mongiardino, M E; Humaran, M; Corbellini, C N; Baldan, A M; Cuneo, M; Balbiani, G [Instituto Nacional de Tecnologia Agropecuaria, Moron, Provincia de Buenos Aires (Argentina). Inst. de Patobiologia

    1996-05-01

    Two experiments were carried out to determine whether the deleterious effects of high amounts of degradable protein on reproduction and production of dairy cows could be minimized by a supplemental source of undergradable protein, or grain supplementation while grazing; and to study the effect of selenium supplementation before calving on the incidence of stillbirths, mastitis, puerperal and metabolic disorders. In a first experiment, 24 Holstein cows fed on red and white clover pasture, paired by previous milk production, calving data and body condition, were supplemented with corn silage and one of two concentrates differing only in the proportion of degradable protein (Group H: 71.5% and Group L: 51.5%). The degradable protein intake from pasture supplied 93% of the requirements in both groups. The addition of undergradable protein in the concentrate of Group L did not improve reproductive performance nor milk, butterfat or solids non-fat production. In a second experiment, 132 Holstein cows and heifers were paired likewise. Both animals in each pair were fed similar forage resources, but each one was supplemented with 2 kg/cow corn grain four times a day (Herd 1) or 4 kg/cow tow times a day (Herd 2). One animal in each pair was randomly assigned to receive a barium selenate injection before calving. Rumen ammonia was higher in Herd 1 in both sampling dates (17 vs 4.2 mg/100 ml and 12 vs 9 mg/100 ml), as well as serum urea up to 50 days post-partum (26 vs 19 mg/100 ml, P<0.02). Body condition scores were similar at calving but significantly lower in Herd 1 during the lactation period (P<0.05). Total milk and butterfat production were higher in Herd 2 (6406.2 vs 6893.8 kg and 190.4 vs 203.5 kg, respectively). Selenium improved pregnancy rate to first artificial insemination in Herd 2 (71 vs 50%), and decreased the frequency of downer cows in both herds (5 vs 0%), but had not effect regarding the Wisconsin Mastitis Test results. (author). 36 refs, 4 figs, 3 tabs.

  15. Cinética ruminal da degradação de nutrientes da silagem de milho em ambiente ruminal inoculado com diferentes aditivos Ruminal degradation kinetics of corn silage in bulls inoculated with different additives in the rumen

    Directory of Open Access Journals (Sweden)

    Pedro Andrade Katsuki

    2006-12-01

    Full Text Available Objetivou-se avaliar a cinética ruminal da degradação de MS, PB e FDN da silagem de milho em ambiente ruminal inoculado com diferentes aditivos. Utilizou-se um delineamento em quadrado latino 4 x 4, com quatro bovinos holandeses e quatro períodos de incubação, em ambiente ruminal adaptado ou não com diferentes aditivos alimentares. Foram testados os seguintes tratamentos: SCL - silagem de milho em ambiente ruminal sem inoculação de aditivo; SBL - silagem de milho em ambiente ruminal inoculado com 5 g de produto comercial contendo bactérias ruminais e intestinais liofilizadas (Ruminobacter amylophilum: 3,0 x 10(11 ufc/kg; Fibrobacter succinogenes: 3,0 x 10(11 ufc/kg; Succinovibrio dextrinsolvens: 4,4 x 10(11 ufc/kg; Bacillus cereus: 3,5 x 10(11 ufc/kg; Lactobacillus acidophilus: 3,5 x 10(11 ufc/kg e Streptococcus faecium: 3,5 x 10(11 ufc/kg; SEC - silagem de milho em ambiente ruminal inoculado com 15 g de produto comercial contendo enzimas celulolíticas (xilanase 10%; e SMS - silagem de milho em ambiente ruminal inoculado com 3 g de produto comercial contendo monensina sódica. Os tratamentos SBL e SEC não afetaram a fração potencialmente degradável (b dos nutrientes avaliados da silagem de milho. A monensina sódica reduziu a fração (b da MS (51,01% e a degradabilidade potencial da silagem de milho (72,33%. Entre os aditivos estudados, a monensina sódica proporcionou a maior fração não-degradável da FDN (45,57%, reduzindo o desaparecimento desta fração a partir de 48 horas de incubação intra-ruminal. Os diferentes aditivos, nas concentrações estudadas, não proporcionaram melhora na degradabilidade efetiva da MS, PB e FDN da silagem de milho.Four bulls fitted with ruminal cannula were used in a 4 x 4 Latin square design to evaluate the effects of different ruminally inoculated additives on the degradation kinetics of DM, CP, and NDF of corn silage (CS. The treatments were: control CS incubated in rumen with no

  16. Accelerated degradation of the D2 protein of photosystem II under ultraviolet radiation

    International Nuclear Information System (INIS)

    Jansen, M.A.K.; Edelman, M.; Greenberg, B.M.; Gaba, V.

    1996-01-01

    The D2 protein of photosystem II is relatively stable in vivo under photosynthetic active radiation, but its degradation accelerates under UVB radiation. Little is known about accelerated D2 protein degradation. We characterized wavelength dependence and sensitivity toward photosystem II inhibitors. The in vivo D2 degradation spectrum resembles the pattern for the rapidly turning over D1 protein of photosystem II, with rates being maximal in the UVB region. We propose that D2 degradation, like D1 degradation, is activated by distinct photosensitizers in the UVB and visible regions of the spectrum. In both wavelength regions, photosystem II inhibitors that are known to be targeted to the D1 protein affect D2 degradation. This suggests that degradation of the two proteins is coupled, D2 degradation being influenced by events occurring at the Q B niche on the D1 protein. (Author)

  17. Secretion of intact proteins and peptide fragments by lysosomal pathways of protein degradation

    International Nuclear Information System (INIS)

    Isenman, L.D.; Dice, J.F.

    1989-01-01

    We report that degradation of proteins microinjected into human fibroblasts is accompanied by release into the culture medium of peptide fragments and intact proteins as well as single amino acids. For the nine proteins and polypeptides microinjected, acid-precipitable radioactivity, i.e. peptide fragments and/or intact proteins, ranged from 10 to 67% of the total released radioactivity. Peptide fragments and/or intact protein accounted for 60% of the radioactivity released into the medium by cells microinjected with ribonuclease A. Two major radiolabeled peptide fragments were found, and one was of an appropriate size to function as an antigen in antigen-presenting cells. The peptides released from microinjected ribonuclease A were derived from lysosomal pathways of proteolysis based on several lines of evidence. Previous studies have shown that microinjected ribonuclease A is degraded to single amino acids entirely within lysosomes. We show that release of free amino acids and peptide fragments and/or intact protein was equivalently stimulated by serum deprivation and equivalently inhibited by NH4Cl. We also show that lysosomal degradation of endocytosed [3H]ribonuclease A was accompanied by the release of two peptide fragments similar in size and charge to those from microinjected [ 3 H]ribonuclease A. These findings demonstrate that degradation within lysosomes occurs in a manner that spares specific peptides; they also suggest a previously unsuspected pathway by which cells can secrete cytosol-derived polypeptides

  18. Additives effect on chemical composition and quality of sisal co-product silage

    Directory of Open Access Journals (Sweden)

    Luiz Gustavo Neves Brandão

    2013-12-01

    Full Text Available Fermentation profile and nutritional value of sisal co-product silage (SC subjected to seven treatments (additives, were evaluated. The SC was ensiled in natura and added with: soy meal, urea, wheat meal, palm kernel cake, A. sisalana dust, licuri cake and cottonseed cake. Experimental silos with capacity for approximately 15 kg of silage, were used. The silos were opened 60 days after ensilage process. It was used a completely randomized design with three replications. The SC in natura present low values of dry mater (DM 12.3% and the additives increased dry matter silages, exception for urea. The SC silage additivated with soybean meal (pH 4.9 and palm kernel cake (butyric acid = 0.07% DM differed, respectively, for pH and butyric acid, compared with in natura SC silage (pH = 4.1 and butyric acid = 0.03% DM. The addition of soybean meal, urea, cottonseed meal, wheat bran and palm kernel, increased crude protein (CP of in natura SC silage. The NDF in silage increased with addition of cottonseed meal or palm kernel cake (60.1 and 66.2% DM in relation in natura SC silage (42.9% DM. The in natura and additivated silages of SC were considered as good or excellent quality.

  19. Nutritional potential of post extraction residues and silage from ...

    African Journals Online (AJOL)

    Nutritional potential of post extraction residues and silage from leaves of five cassava varieties as feed for ruminants. ... Results indicated that processing reduced (P < 0.05) the inherent nutrients of cassava leaf residues produced after the extraction of protein concentrate from cassava leaves with crude protein (CP) content ...

  20. Reticulophagy and Ribophagy: Regulated Degradation of Protein Production Factories

    Directory of Open Access Journals (Sweden)

    Eduardo Cebollero

    2012-01-01

    Full Text Available During autophagy, cytosol, protein aggregates, and organelles are sequestered into double-membrane vesicles called autophagosomes and delivered to the lysosome/vacuole for breakdown and recycling of their basic components. In all eukaryotes this pathway is important for adaptation to stress conditions such as nutrient deprivation, as well as to regulate intracellular homeostasis by adjusting organelle number and clearing damaged structures. For a long time, starvation-induced autophagy has been viewed as a nonselective transport pathway; however, recent studies have revealed that autophagy is able to selectively engulf specific structures, ranging from proteins to entire organelles. In this paper, we discuss recent findings on the mechanisms and physiological implications of two selective types of autophagy: ribophagy, the specific degradation of ribosomes, and reticulophagy, the selective elimination of portions of the ER.

  1. Protein Degradation in Normal and Beige (Chediak-Higashi) Mice

    Science.gov (United States)

    Lyons, Robert T.; Pitot, Henry C.

    1978-01-01

    The beige mouse, C57BL/6 (bg/bg), is an animal model for the Chediak-Higashi syndrome in man, a disease characterized morphologically by giant lysosomes in most cell types. Half-lives for the turnover of [14C]bicarbonate-labeled total soluble liver protein were determined in normal and beige mice. No significant differences were observed between the normal and mutant strain for both rapidly and slowly turning-over classes of proteins. Glucagon treatment during the time-course of protein degradation had similar effects on both normal and mutant strains and led to the conclusion that the rate of turnover of endogenous intracellular protein in the beige mouse liver does not differ from normal. The rates of uptake and degradation of an exogenous protein were determined in normal and beige mice by intravenously injecting 125I-bovine serum albumin and following, in peripheral blood, the loss with time of phosphotungstic acid-insoluble bovine serum albumin and the parallel appearance of phosphotungstic acid-soluble (degraded) material. No significant differences were observed between beige and normal mice in the uptake by liver lysosomes of 125I-bovine serum albumin (t½ = 3.9 and 2.8 h, respectively). However, it was found that lysosomes from livers of beige mice released phosphotungstic acid-soluble radioactivity at a rate significantly slower than normal (t½ = 6.8 and 3.1 h, respectively). This defect in beige mice could be corrected by chronic administration of carbamyl choline (t½ = 3.5 h), a cholinergic agonist which raises intracellular cyclic GMP levels. However, no significant differences between normal and beige mice were observed either in the ability of soluble extracts of liver and kidney to bind [3H]cyclic GMP in vitro or in the basal levels of cyclic AMP in both tissues. The relevance of these observations to the presumed biochemical defect underlying the Chediak-Higashi syndrome is discussed. PMID:202611

  2. The effects of forage proportion and rapidly degradable dry matter from concentrate on ruminal digestion in dairy cows fed corn silage-based diets with fixed neutral detergent fiber and starch contents.

    Science.gov (United States)

    Lechartier, C; Peyraud, J-L

    2010-02-01

    This study investigated the effects of the forage-to-concentrate (F:C) ratio and the rate of ruminal degradation of carbohydrates from the concentrate on digestion in dairy cows fed corn silage-based diets. Six cows with ruminal cannulas were assigned to 6 treatments in a 6x6 Latin square. Treatments were arranged in a 3x2 factorial design. Three proportions of neutral detergent fiber from forage [FNDF; 7.6, 13.2, and 18.9% of dry matter (DM)] were obtained by modifying F:C (20:80, 35:65, and 50:50). These F:C were combined with concentrates with either high or low content of rapidly degradable carbohydrates. The dietary content of rapidly degradable carbohydrates from the concentrate was estimated from the DM disappearance of concentrate after 4h of in sacco incubation (CRDM). Thus, 2 proportions of CRDM were tested (20 and 30% of DM). Wheat and corn grain were used as rapidly and slowly degradable starch sources, respectively. Soybean hulls and citrus pulp were used as slowly and rapidly degradable fiber sources, respectively. Concentrate composition was adjusted to maintain dietary starch and neutral detergent fiber contents at 35.9 and 28.9% of DM, respectively. There was no effect of the interaction between F:C and CRDM on DM intake (DMI), ruminal fermentation, chewing activity, and fibrolytic activity. When F:C decreased, DMI increased, the mean ruminal pH linearly decreased, and the pH range linearly increased from 0.95 to 1.27 pH unit. At the same time, the acetate-to-propionate ratio decreased linearly. Decreasing F:C linearly decreased the average time spent chewing per kilogram of DMI from 35.2 to 19.5min/kg of DMI and decreased ruminal liquid outflow from 11.6 to 9.2L/kg of DMI, suggesting a decrease in the salivary flow. Increasing CRDM decreased DMI and increased the time during which pH was below 6.0 (3.1 vs. 4.8h), the pH range (0.90 vs. 1.33), and the initial rate of pH drop. It also increased the volatile fatty acid range (35 vs. 59mM), thus

  3. Physiochemical Characteristics and Molecular Structures for Digestible Carbohydrates of Silages.

    Science.gov (United States)

    Refat, Basim; Prates, Luciana L; Khan, Nazir A; Lei, Yaogeng; Christensen, David A; McKinnon, John J; Yu, Peiqiang

    2017-10-18

    The main objectives of this study were (1) to assess the magnitude of differences among new barley silage varieties (BS) selected for varying rates of in vitro neutral detergent fiber (NDF) digestibility (ivNDFD; Cowboy BS with higher ivNDFD, Copeland BS with intermediate ivNDFD, and Xena BS with lower ivNDFD) with regard to their carbohydrate (CHO) molecular makeup, CHO chemical fractions, and rumen degradability in dairy cows in comparison with a new corn silage hybrid (Pioneer 7213R) and (2) to quantify the strength and pattern of association between the molecular structures and digestibility of carbohydrates. The carbohydrate-related molecular structure spectral data was measured using advanced vibrational molecular spectroscopy (FT/IR). In comparison to BS, corn silage showed a significantly (P carbohydrates were significantly (P carbohydrate content of the silages. In conclusion, the univariate approach with only one-factor consideration (ivNDFD) might not be a satisfactory method for evaluating and ranking BS quality. FT/IR molecular spectroscopy can be used to evaluate silage quality rapidly, particularly the digestible fiber content.

  4. Mechanical dewatering of silage; Mechanische Entwaesserung von Silage

    Energy Technology Data Exchange (ETDEWEB)

    Scholz, Volkhard; Daries, Werner [Leibniz-Institut fuer Agrartechnik Potsdam-Bornim e.V. (ATB), Potsdam (Germany). Abteilung Technik der Aufbereitung, Lagerung und Konservierung; Rinder, Ralf [Anhydro GmbH, Kassel (Germany)

    2009-07-01

    The use of screw presses for dewatering of silage is energetically more efficient than thermal drying. Tests with two types of screw presses show that on dry matter flow rates of more than 1.0 tDM/h the specific energy consumption is less than 30 kWh/tDM. The water content of the silage is decreased by 5 to 20 % and the share of undesired ingredients by 2 to 30 %. Besides press design and plant species, in particular moisture content, chip size and density of silage are essentially for the dewatering success. (orig.)

  5. SILAGE CANE SUGAR ADDED WITH DRIED BREWER

    Directory of Open Access Journals (Sweden)

    W. J. R. Castro

    2015-02-01

    Full Text Available The objective of this experiment was to evaluate the fermentative parameters and chemical composition of silage cane sugar added with residue dried brewery. The experimental design was completely randomized with four treatments and four replications: 100% cane sugar; 90% of cane sugar + 10% residue dried brewer; 80% of cane sugar + 20% residue dried brewer and 70% cane sugar + 30% dried brewer based on natural matter, composed silages. The sugar cane was chopped in a stationary machine with forage particle size of approximately 2 cm, and homogenized manually with the additives. For storage chopped fresh weight were used in experimental silos capacity of about 4 liters. The results showed that the contents of dry matter and crude protein showed positive linear (P0.05 with mean value of 3.81, while for ether extract and ash results were positive linear (P0.05 for N ammonia presented average value of 4.18. It is concluded that the addition of brewer dehydrated improves the fermentation process of silage cane sugar, in addition to improving their nutritional characteristics.

  6. Mass spectrometry analysis of proteome-wide proteolytic post-translational degradation of proteins

    OpenAIRE

    Shen, Yufeng; Hixson, Kim K.; Tolić, Nikola; Camp, David G.; Purvine, Samuel O.; Moore, Ronald J.; Smith, Richard D.

    2008-01-01

    Protein proteolytic degradation is an essential component to proper cell function and its life cycle. Here, we study the protein degradation in yeast Saccharomyces cerevisiae cells on a proteome-wide scale by detection of the intermediate peptides produced from the intracellular degradation of proteins using sequencing-based tandem mass spectrometry. By tracing the detected ~1,100 peptides and their ~200 protein substrate origins we obtain evidence for new insights into the proteome-wide prot...

  7. Bromatological characteristics and ruminal digestibility of grain corn hybrids with different vitreousness in silage maturity

    Directory of Open Access Journals (Sweden)

    Evandrei Santos Rossi

    2016-06-01

    Full Text Available The aim of this research was to evaluate vitreousness in commercial corn hybrids with flint and dent grains and to study the effect of this characteristic on agronomic behavior, nutritional value, and ruminal degradability of grains harvested in silage maturity. Twelve commercial corn hybrids were evaluated. They were divided in two groups (six with flint grains and six with dent grains. The experiments were conducted in two municipalities represented by Guarapuava and Laranjeiras do Sul, both in Paraná State. The harvest for the grain quality analysis was performed at ¾ of the milk line stage in the grain. The following characteristics were evaluated: grain yield, grain yield in silage maturity, vitreousness, ruminal digestibility of grain, neutral detergent fiber, acid detergent fiber and crude protein. The grains in the group of dent hybrids exhibited better degradability associated with lower vitreousness than the group of flint hybrids. The group of dent hybrids exhibited better ruminal digestibility of the grains associated with lower vitreousness versus the group of flint hybrids. There is a negative correlation between vitreousness and the digestibility of the grain; thus, vitreousness can be a criterion for selecting genotypes for forage production.

  8. Induced oligomerization targets Golgi proteins for degradation in lysosomes.

    Science.gov (United States)

    Tewari, Ritika; Bachert, Collin; Linstedt, Adam D

    2015-12-01

    Manganese protects cells against forms of Shiga toxin by down-regulating the cycling Golgi protein GPP130. Down-regulation occurs when Mn binding causes GPP130 to oligomerize and traffic to lysosomes. To determine how GPP130 is redirected to lysosomes, we tested the role of GGA1 and clathrin, which mediate sorting in the canonical Golgi-to-lysosome pathway. GPP130 oligomerization was induced using either Mn or a self-interacting version of the FKBP domain. Inhibition of GGA1 or clathrin specifically blocked GPP130 redistribution, suggesting recognition of the aggregated GPP130 by the GGA1/clathrin-sorting complex. Unexpectedly, however, GPP130's cytoplasmic domain was not required, and redistribution also occurred after removal of GPP130 sequences needed for its normal cycling. Therefore, to test whether aggregate recognition might be a general phenomenon rather than one involving a specific GPP130 determinant, we induced homo-oligomerization of two unrelated Golgi-targeted constructs using the FKBP strategy. These were targeted to the cis- and trans-Golgi, respectively, using domains from mannosidase-1 and galactosyltransferase. Significantly, upon oligomerization, each redistributed to peripheral punctae and was degraded. This occurred in the absence of detectable UPR activation. These findings suggest the unexpected presence of quality control in the Golgi that recognizes aggregated Golgi proteins and targets them for degradation in lysosomes. © 2015 Tewari et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  9. Health status of cows fed maize silage covered with oxo-biodegradable foil

    OpenAIRE

    Piotr SZTERK; Piotr DORSZEWSKI; Małgorzata GRABOWICZ; Lucyna PODKÓWKA

    2017-01-01

    In agricultural practice, silage production uses pure, low density polyethylene foil. This foil, after use, becomes farm waste, having a negative impact on the environment. Instead of conventional foil, an environmentally safe biodegradable foil can be used, made from naturally occurring polymers or from synthetic multiparticulates, easily degradable by microorganisms. Silage covered with this type of foil should be safe for animal health. The purpose of the study was to determine whether oxo...

  10. Rapid degradation of abnormal proteins in vacuoles from Acer pseudoplatanus L. cells

    International Nuclear Information System (INIS)

    Canut, H.; Alibert, G.; Carrasco, A.; Boudet, A.M.

    1986-01-01

    In Acer pseudoplatanus cells, the proteins synthesized in the presence of an amino acid analog ([ 14 C]p-fluorophenylalanine), were degraded more rapidly than normal ones ([ 14 C]phenylalanine as precursor). The degradation of an important part of these abnormal proteins occurred inside the vacuoles. The degradation process was not apparently associated to a specific proteolytic system but was related to a preferential transfer of these aberrant proteins from the cytoplasm to the vacuole

  11. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants

    OpenAIRE

    Baudisch, Bianca; Pfort, Ingrid; Sorge, Eberhard; Conrad, Udo

    2018-01-01

    Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  12. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants

    Directory of Open Access Journals (Sweden)

    Bianca Baudisch

    2018-02-01

    Full Text Available Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  13. Nanobody-Directed Specific Degradation of Proteins by the 26S-Proteasome in Plants.

    Science.gov (United States)

    Baudisch, Bianca; Pfort, Ingrid; Sorge, Eberhard; Conrad, Udo

    2018-01-01

    Here, we present data showing the directed degradation of target proteins recognized by a specific nanobody in transgenic plants. Green fluorescent protein was depleted by a chimeric nanobody fused to a distinct F-box domain, which enables protein degradation via the ubiquitin proteasome pathway. This technique could thus be used to knock out other proteins of interest in planta using specific, high-affinity binding proteins.

  14. Influence of anabolic agents on protein synthesis and degradation in muscle cells grown in culture

    Energy Technology Data Exchange (ETDEWEB)

    Roeder, R.A.; Thorpe, S.D.; Byers, F.M.; Schelling, G.T.; Gunn, J.M.

    Muscle cell culture (L/sub 6/) studies were conducted to determine whether anabolic agents have a direct effect on the muscle cell. The effect of zeranol, testosterone propionate, estradiol benzoate, progesterone, dexamethasone and anabolic agent-dexamethasone combinations on protein synthesis and degradation were measured. Myoblast and myotube cultures were pretreated with 1 ..mu..M compounds for 12, 24 and 48 h before a 6-h synthesis or degradation measuring period. Protein synthesis was determined as cpm of (/sup 3/H) leucine incorporated per mg cell protein. Protein degradation was measured by a pulse-chase procedure using (/sup 3/H) leucine and expressed as the percentage labeled protein degraded in 6 h. Progesterone slightly increased protein synthesis in myoblast cultures. Testosterone propionate had no effect on synthesis. Protein synthesis was decreased by estradiol benzoate in myotube cultures. Protein degradation was not altered appreciably by anabolic agents. Protein synthesis was initially inhibited in myotubes by dexamethasone, but increased in myoblasts and myotubes in the extended incubation time. Dexamethasone also consistently increased protein degradation, but this required several hours to be expressed. Anabolic agents did not interfere with dexamethasone-induced increases in protein synthesis and degradation. The magnitude of response and sensitivity were similar for both the myoblast and the more fully differentiated myotube for all compounds tested. These results indicate that anabolic agents at the 1 ..mu..M level do not have a direct anabolic effect on muscle or alter glucocorticoid-induced catabolic response in muscle.

  15. Influence of anabolic agents on protein synthesis and degradation in muscle cells grown in culture

    International Nuclear Information System (INIS)

    Roeder, R.A.; Thorpe, S.D.; Byers, F.M.; Schelling, G.T.; Gunn, J.M.

    1986-01-01

    Muscle cell culture (L 6 ) studies were conducted to determine whether anabolic agents have a direct effect on the muscle cell. The effect of zeranol, testosterone propionate, estradiol benzoate, progesterone, dexamethasone and anabolic agent-dexamethasone combinations on protein synthesis and degradation were measured. Myoblast and myotube cultures were pretreated with 1 μM compounds for 12, 24 and 48 h before a 6-h synthesis or degradation measuring period. Protein synthesis was determined as cpm of [ 3 H] leucine incorporated per mg cell protein. Protein degradation was measured by a pulse-chase procedure using [ 3 H] leucine and expressed as the percentage labeled protein degraded in 6 h. Progesterone slightly increased protein synthesis in myoblast cultures. Testosterone propionate had no effect on synthesis. Protein synthesis was decreased by estradiol benzoate in myotube cultures. Protein degradation was not altered appreciably by anabolic agents. Protein synthesis was initially inhibited in myotubes by dexamethasone, but increased in myoblasts and myotubes in the extended incubation time. Dexamethasone also consistently increased protein degradation, but this required several hours to be expressed. Anabolic agents did not interfere with dexamethasone-induced increases in protein synthesis and degradation. The magnitude of response and sensitivity were similar for both the myoblast and the more fully differentiated myotube for all compounds tested. These results indicate that anabolic agents at the 1 μM level do not have a direct anabolic effect on muscle or alter glucocorticoid-induced catabolic response in muscle

  16. Biochemistry of cellulose degradation and cellulose utilization for feeds and for protein

    Energy Technology Data Exchange (ETDEWEB)

    Sadara, J C; Lachke, A H; Shewale, J G

    1979-01-01

    A review discussing production of single-cell protein, fuel, and glucose from cellulose decomposition; surface or solid fermentations of single-cell protein; production of cellulases; and the biochemistry of cellulose degradation was presented.

  17. The different roles of selective autophagic protein degradation in mammalian cells.

    Science.gov (United States)

    Wang, Da-wei; Peng, Zhen-ju; Ren, Guang-fang; Wang, Guang-xin

    2015-11-10

    Autophagy is an intracellular pathway for bulk protein degradation and the removal of damaged organelles by lysosomes. Autophagy was previously thought to be unselective; however, studies have increasingly confirmed that autophagy-mediated protein degradation is highly regulated. Abnormal autophagic protein degradation has been associated with multiple human diseases such as cancer, neurological disability and cardiovascular disease; therefore, further elucidation of protein degradation by autophagy may be beneficial for protein-based clinical therapies. Macroautophagy and chaperone-mediated autophagy (CMA) can both participate in selective protein degradation in mammalian cells, but the process is quite different in each case. Here, we summarize the various types of macroautophagy and CMA involved in determining protein degradation. For this summary, we divide the autophagic protein degradation pathways into four categories: the post-translational modification dependent and independent CMA pathways and the ubiquitin dependent and independent macroautophagy pathways, and describe how some non-canonical pathways and modifications such as phosphorylation, acetylation and arginylation can influence protein degradation by the autophagy lysosome system (ALS). Finally, we comment on why autophagy can serve as either diagnostics or therapeutic targets in different human diseases.

  18. Degradation of misfolded proteins in neurodegenerative diseases: therapeutic targets and strategies.

    Science.gov (United States)

    Ciechanover, Aaron; Kwon, Yong Tae

    2015-03-13

    Mammalian cells remove misfolded proteins using various proteolytic systems, including the ubiquitin (Ub)-proteasome system (UPS), chaperone mediated autophagy (CMA) and macroautophagy. The majority of misfolded proteins are degraded by the UPS, in which Ub-conjugated substrates are deubiquitinated, unfolded and cleaved into small peptides when passing through the narrow chamber of the proteasome. The substrates that expose a specific degradation signal, the KFERQ sequence motif, can be delivered to and degraded in lysosomes via the CMA. Aggregation-prone substrates resistant to both the UPS and the CMA can be degraded by macroautophagy, in which cargoes are segregated into autophagosomes before degradation by lysosomal hydrolases. Although most misfolded and aggregated proteins in the human proteome can be degraded by cellular protein quality control, some native and mutant proteins prone to aggregation into β-sheet-enriched oligomers are resistant to all known proteolytic pathways and can thus grow into inclusion bodies or extracellular plaques. The accumulation of protease-resistant misfolded and aggregated proteins is a common mechanism underlying protein misfolding disorders, including neurodegenerative diseases such as Huntington's disease (HD), Alzheimer's disease (AD), Parkinson's disease (PD), prion diseases and Amyotrophic Lateral Sclerosis (ALS). In this review, we provide an overview of the proteolytic pathways in neurons, with an emphasis on the UPS, CMA and macroautophagy, and discuss the role of protein quality control in the degradation of pathogenic proteins in neurodegenerative diseases. Additionally, we examine existing putative therapeutic strategies to efficiently remove cytotoxic proteins from degenerating neurons.

  19. A genetic bistable switch utilizing nonlinear protein degradation.

    Science.gov (United States)

    Huang, Daniel; Holtz, William J; Maharbiz, Michel M

    2012-07-09

    Bistability is a fundamental property in engineered and natural systems, conferring the ability to switch and retain states. Synthetic bistable switches in prokaryotes have mainly utilized transcriptional components in their construction. Using both transcriptional and enzymatic components, creating a hybrid system, allows for wider bistable parameter ranges in a circuit. In this paper, we demonstrate a tunable family of hybrid bistable switches in E. coli using both transcriptional components and an enzymatic component. The design contains two linked positive feedback loops. The first loop utilizes the lambda repressor, CI, and the second positive feedback loop incorporates the Lon protease found in Mesoplasma florum (mf-Lon). We experimentally tested for bistable behavior in exponential growth phase, and found that our hybrid bistable switch was able to retain its state in the absence of an input signal throughout 40 cycles of cell division. We also tested the transient behavior of our switch and found that switching speeds can be tuned by changing the expression rate of mf-Lon. To our knowledge, this work demonstrates the first use of dynamic expression of an orthogonal and heterologous protease to tune a nonlinear protein degradation circuit. The hybrid switch is potentially a more robust and tunable topology for use in prokaryotic systems.

  20. Effects of gamma irradiation on chemical composition and ruminal protein degradation of canola meal

    Science.gov (United States)

    Shawrang, P.; Nikkhah, A.; Zare-Shahneh, A.; Sadeghi, A. A.; Raisali, G.; Moradi-Shahrebabak, M.

    2008-07-01

    Gamma irradiation of canola meal (at doses of 25, 50 and 75 kGy) could alter its ruminal protein degradation characteristics by cross-linking of the polypeptide chains. This processing resulted in decrease (linear effect, Pruminal protein degradation and increase (linear effect, Pruminant nutrition.

  1. Ruminal silage degradability and productivity of forage and grain-type sorghum cultivars Degradabilidade ruminal das silagens e produtividade de cultivares de sorgo de tipo forrageiro e granífero

    Directory of Open Access Journals (Sweden)

    José Antônio Resende

    2003-01-01

    Full Text Available Forages for feeding dairy cows should conciliate the potential for high dry matter yield per unit area, maximizing animal stocking rate, with high nutritive value, which enables decreasing the amount of concentrate feedstuffs per milk unit produced. Eighteen sorghum cultivars were cultivated 0.70 m apart and with 12 plants per linear meter. Plants harvested at the milk-to-dough stage were ensiled in the laboratory. Eleven agronomic and chemical traits were evaluated. Silage samples were incubated in situ and the effective ruminal dry matter degradation (DEF was calculated assuming a fractional passage rate of -0.04 h-1. Dry matter yield was 13.0 ± 2.3 (mean ± S.D. t ha-1. The neutral detergent fiber content was 50.3 ± 3.8 and the DEF was 48.8 ± 2.8% (dry matter basis. The average sorghum DEF was 84% of the DEF of a composite sample of 60 corn hybrids simultaneously cultivated, ensiled and ruminally incubated. The 12 forage-type cultivars were taller, more productive, had higher fiber content and lower DEF than the 6 grain and dual purpose-type cultivars. The acid detergent fiber content had the greatest correlation with DEF (r= -0.64. The linear model correlating DEF with productivity was: DEF=54.694 - 0.4449 x t of dry matter ha-1 (r²=0.14. Although there are cultivars that show high productivity and digestibility, it seems to be difficult to conciliate the maximum nutritive value with the maximum productivity of the sorghum crop.Forrageiras para alimentação de vacas leiteiras devem conciliar a alta produção de matéria seca por área, maximizando a taxa de lotação animal, com o alto valor nutritivo, capaz de reduzir a necessidade de alimentos concentrados por litro de leite produzido. Dezoito cultivares de sorgo foram cultivados com espaçamento de 0,70 m e 12 plantas por metro linear. As plantas colhidas ao atingirem o estádio leitoso-pastoso foram ensiladas em laboratório. Onze características agronômicas e químicas foram

  2. Bio-based biodegradable film to replace the standard polyethylene cover for silage conservation.

    Science.gov (United States)

    Borreani, Giorgio; Tabacco, Ernesto

    2015-01-01

    The research was aimed at studying whether the polyethylene (PE) film currently used to cover maize silage could be replaced with bio-based biodegradable films, and at determining the effects on the fermentative and microbiological quality of the resulting silages in laboratory silo conditions. Biodegradable plastic film made in 2 different formulations, MB1 and MB2, was compared with a conventional 120-μm-thick PE film. A whole maize crop was chopped; ensiled in MB1, MB2, and PE plastic bags, 12.5kg of fresh weight per bag; and opened after 170d of conservation. At silo opening, the microbial and fermentative quality of the silage was analyzed in the uppermost layer (0 to 50mm from the surface) and in the whole mass of the silo. All the silages were well fermented with little differences in fermentative quality between the treatments, although differences in the mold count and aerobic stability were observed in trial 1 for the MB1 silage. These results have shown the possibility of successfully developing a biodegradable cover for silage for up to 6mo after ensiling. The MB2 film allowed a good silage quality to be obtained even in the uppermost part of the silage close to the plastic film up to 170d of conservation, with similar results to those obtained with the PE film. The promising results of this experiment indicate that the development of new degradable materials to cover silage till 6mo after ensiling could be possible. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  3. Co-Digestion of Sugar Beet Silage Increases Biogas Yield from Fibrous Substrates

    Science.gov (United States)

    Einfalt, Daniel; Kazda, Marian

    2016-01-01

    This study tested the hypothesis that the easily degradable carbohydrates of the sugar beet silage (S) will improve the anaerobic digestion of grass silage (G) more profoundly compared to co-digestion of sugar beet silage with maize silage (M). M : S and G : S mixtures were tested in two continuous laboratory-scale AD experiments at volatile solid ratios of 1 : 0, 6 : 1, 3 : 1, and 1 : 3 at organic loading rates of 1.5 kgVS m−3 day−1. While the sugar beet effects in mixtures with maize silage were negligible, co-digestion with grass silage showed a beneficial performance. There, the specific methane production rate was 0.27 lN kg−1VS h−1at G : S ratio of 6 : 1 compared to G : S 1 : 0 with 0.14 lN kg−1VS h−1. In comparison to G : S 1 : 0, about 44% and 62% higher biogas yields were obtained at G : S 6 : 1 and 3 : 1, respectively. Also, the highest methane concentration was found in G : S at ratio of 1 : 3. Synergistic increase of methane yield was found in co-digestion in both experiments, but higher effect was realized in G : S, independently of the amount of sugar beet silage. The findings of this study emphasize the improvement of AD of grass silage by even low addition of sugar beet silage. PMID:27807538

  4. Chemical degradation of proteins in the solid state with a focus on photochemical reactions.

    Science.gov (United States)

    Mozziconacci, Olivier; Schöneich, Christian

    2015-10-01

    Protein pharmaceuticals comprise an increasing fraction of marketed products but the limited solution stability of proteins requires considerable research effort to prepare stable formulations. An alternative is solid formulation, as proteins in the solid state are thermodynamically less susceptible to degradation. Nevertheless, within the time of storage a large panel of kinetically controlled degradation reactions can occur such as, e.g., hydrolysis reactions, the formation of diketopiperazine, condensation and aggregation reactions. These mechanisms of degradation in protein solids are relatively well covered by the literature. Considerably less is known about oxidative and photochemical reactions of solid proteins. This review will provide an overview over photolytic and non-photolytic degradation reactions, and specially emphasize mechanistic details on how solid structure may affect the interaction of protein solids with light. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Prefoldin Promotes Proteasomal Degradation of Cytosolic Proteins with Missense Mutations by Maintaining Substrate Solubility.

    Directory of Open Access Journals (Sweden)

    Sophie A Comyn

    2016-07-01

    Full Text Available Misfolded proteins challenge the ability of cells to maintain protein homeostasis and can accumulate into toxic protein aggregates. As a consequence, cells have adopted a number of protein quality control pathways to prevent protein aggregation, promote protein folding, and target terminally misfolded proteins for degradation. In this study, we employed a thermosensitive allele of the yeast Guk1 guanylate kinase as a model misfolded protein to investigate degradative protein quality control pathways. We performed a flow cytometry based screen to identify factors that promote proteasomal degradation of proteins misfolded as the result of missense mutations. In addition to the E3 ubiquitin ligase Ubr1, we identified the prefoldin chaperone subunit Gim3 as an important quality control factor. Whereas the absence of GIM3 did not impair proteasomal function or the ubiquitination of the model substrate, it led to the accumulation of the poorly soluble model substrate in cellular inclusions that was accompanied by delayed degradation. We found that Gim3 interacted with the Guk1 mutant allele and propose that prefoldin promotes the degradation of the unstable model substrate by maintaining the solubility of the misfolded protein. We also demonstrated that in addition to the Guk1 mutant, prefoldin can stabilize other misfolded cytosolic proteins containing missense mutations.

  6. Microbial protein synthesis in young steers fed with grass silage and extruded corn Síntesis de proteína microbiana en novillos jóvenes alimentados con ensilaje de praderas y maíz extruido

    Directory of Open Access Journals (Sweden)

    Claudia Barchiesi-Ferrari

    2012-12-01

    Full Text Available Microbial protein synthesis is essential to meet protein requirements in ruminants. The aim of this investigation was to evaluate the effect of incorporating extruded corn (Zea mays L. into the ration of young steers fed with grass silage on microbial protein synthesis (MPS. Twenty young steers in metabolic cages were used for supplying food. The treatments consisted of combinations of extruded corn (EC and ground corn (GC EC-10 (10% EC-30% GC; EC-25 (25% EC-15% GC and EC-40 (40% EC-0% GC. These foods constituted 40% of the ration (DM-based and the remaining 60% was grass silage. The MPS was calculated through the excretion of purine derivatives (PD in the urine, using total collection of urine. The inclusion of a higher percentage of extruded corn (EC-40 increased the PD excretion (51.85 mmol d-1, P La síntesis de proteína microbiana es fundamental para satisfacer los requerimientos proteicos en rumiantes. El objetivo de esta investigation fue evaluar el efecto de incluir maíz (Zea mays L. extruido en la ración de novillos alimentados con ensilaje de pradera, sobre la sintesis de proteina microbiana (MPS. Se emplearon 20 novillos jóvenes ubicados en jaulas metabólicas para suministro de los alimentos. Los tratamientos consistieron en combinaciones de maiz extruido (EC y maiz molido (GC EC-10 (10% EC-30% GC; EC-25 (25% EC-15% GC y EC-40 (40% EC-0% GC. Estos alimentos constituyeron el 40% de la ración (base MS y el restante 60% fue ensilaje de pradera. Se estimó la MPS a través de la excreción de derivados de purinas (PD en la orina, empleando colección total de orina. La inclusión de un mayor porcentaje de EC (EC-40 incrementó la excreción de PD (51.85 mmol d-1, P < 0,05. Los valores de excreción de PD fluctuaron entre 663 y 1078 μmol BW-075 d-1. La absorción de proteína microbiana y el flujo de N duodenal también se incrementaron por una mayor inclusión de EC (EC-40, P < 0,05. Las diferencias entre los tratamientos pueden

  7. Methane emission and community composition patterns of rumen bacteria and methanogens in Holstein dairy cows as affected by silage type and dietary fat

    DEFF Research Database (Denmark)

    Poulsen, Morten; Højberg, Ole; Canibe, Nuria

    ) to investigate effects of silages with different methanogenic potential (early grass, late grass, and maize) combined with a CH4-reducing feed additive (crushed rapeseed) on bacterial and methanogenic communities in the rumen. Bacterial and methanogenic community patterns were evaluated by T-RFLP analysis of 16S...... rRNA and methyl co-enzyme M reductase (mcrA) genes, respectively. Methanogen abundances were evaluated by qPCR using two mcrA-targeting primer sets. Silage type significantly affected CH4 emissions and rumen acetate:propionate ratios, being highest for late grass and lowest for maize. Dietary fat...... significantly reduced the gross energy lost as CH4 regardless of silage type. Silage type significantly affected the bacterial community composition pattern; the grass silages favored potential hemicellulose- and cellulose-degrading bacteria, while the maize silage mainly favored potentially starch...

  8. Effects of replacing conventional corn silage with BMR corn silage

    Science.gov (United States)

    Previous research has shown that the (lignin reducing) brown mid-rib mutation in corn silage, which increases in vitro fiber digestibility, does not always improve fiber digestibility when fed as part of a TMR; however, feed intake and milk production are increased. The objectives of this experiment...

  9. The effect of silage cutting height on the nutritive value of a normal corn silage hybrid compared with brown midrib corn silage fed to lactating cows.

    Science.gov (United States)

    Kung, L; Moulder, B M; Mulrooney, C M; Teller, R S; Schmidt, R J

    2008-04-01

    A brown midrib (BMR) hybrid and a silage-specific non-BMR (7511FQ) hybrid were harvested at a normal cut height leaving 10 to 15 cm of stalk in the field. The non-BMR hybrid was also cut at a greater height leaving 45 to 50 cm of stalk. Cutting high increased the concentrations of dry matter (+4%), crude protein (+5%), net energy for lactation (+3%), and starch (+7%), but decreased the concentrations of acid detergent fiber (-9%), neutral detergent fiber (-8%), and acid detergent lignin (-13%) for 7511FQ. As expected, the BMR corn silage was 30% lower in lignin concentration than 7511FQ. After 30 h of in vitro ruminal fermentation, the digestibility of neutral detergent fiber for normal cut 7511FQ, the same hybrid cut high, and the normal cut BMR hybrid were 51.7, 51.4, and 63.5%, respectively. Twenty-seven multiparous lactating cows were fed a total mixed ration composed of the respective silages (45% of dry matter) with alfalfa haylage (5%), alfalfa hay (5%), and concentrate (45%) (to make the TMR isocaloric and isonitrogenous) in a study with a 3 x 3 Latin square design with 21-d periods. Milk production was greater for cows fed the BMR hybrid (48.8 kg/d) compared with those fed the normal cut 7511FQ (46.8 kg/d) or cut high (47.7 kg/d). Dry matter intake was not affected by treatment. Feed efficiency for cows fed the BMR silage (1.83) was greater than for those fed high-cut 7511FQ (1.75), but was not different from cows fed the normal cut 7511FQ (1.77). Cows fed the BMR silage had milk with greater concentrations of lactose but lower milk urea nitrogen than cows on other treatments. Harvesting a silage-specific, non-BMR corn hybrid at a high harvest height improved its nutritive content, but the improvement in feeding value was not equivalent to that found when cows were fed BMR corn silage.

  10. Protein synthesis and degradation during starvation-induced cardiac atrophy in rabbits

    International Nuclear Information System (INIS)

    Samarel, A.M.; Parmacek, M.S.; Magid, N.M.; Decker, R.S.; Lesch, M.

    1987-01-01

    To determine the relative importance of protein degradation in the development of starvation-induced cardiac atrophy, in vivo fractional synthetic rates of total cardiac protein, myosin heavy chain, actin, light chain 1, and light chain 2 were measured in fed and fasted rabbits by continuous infusion of [ 3 H] leucine. In addition, the rate of left ventricular protein accumulation and loss were assessed in weight-matched control and fasted rabbits. Rates of total cardiac protein degradation were then estimated as the difference between rates of synthesis and growth. Fasting produced left ventricular atrophy by decreasing the rate of left ventricular protein synthesis (34.8 +/- 1.4, 27.3 +/- 3.0, and 19.3 +/- 1.2 mg/day of left ventricular protein synthesized for 0-, 3-, and 7-day fasted rabbits, respectively). Inhibition of contractile protein synthesis was evident by significant reductions in the fractional synthetic rates of all myofibrillar protein subunits. Although fractional rates of protein degradation increased significantly within 7 days of fasting, actual amounts of left ventricular protein degraded per day were unaffected. Thus, prolonged fasting profoundly inhibits the synthesis of new cardiac protein, including the major protein constituents of the myofibril. Both this inhibition in new protein synthesis as well as a smaller but significant reduction in the average half-lives of cardiac proteins are responsible for atrophy of the heart in response to fasting

  11. In vitro degradation of the 32kDa PS II reaction centre protein

    International Nuclear Information System (INIS)

    Eckenswiller, L.C.; Greenberg, B.M.

    1989-01-01

    The 32kDa thylakoid membrane protein is an integral component of the PS II reaction centre. The protein, although stable in the dark, undergoes light dependent turnover. Light from the UV, visible and far-red spectral regions induce 32kDa protein degradation. To better understand 32kDa protein metabolism, an in vitro degradation system is being developed. It consists of isolated thylakoid membranes than contain radiolabelled protein. The 32kDa protein is actively and specifically degraded when the thylakoid preparation is exposed to UV or visible radiation. The protein is stable in the dark. The herbicides (atrazine and DCMU) inhibit degradation in the in vitro system as they do in vivo. Additionally, several methods of isolating thylakoids are being compared to optimize the 32kDa protein degradation reaction. The preparations will be evaluated based on their ability to permit light dependent degradation of the 32kDa protein without affecting the other membrane components

  12. Fermentation Characteristics and Microbial Diversity of Tropical Grass-legumes Silages

    Directory of Open Access Journals (Sweden)

    Roni Ridwan

    2015-04-01

    Full Text Available Calliandra calothyrsus preserved in silage is an alternative method for improving the crude protein content of feeds for sustainable ruminant production. The aim of this research was to evaluate the quality of silage which contained different levels of C. calothyrsus by examining the fermentation characteristics and microbial diversity. Silage was made in a completely randomized design consisting of five treatments with three replications i.e.: R0, Pennisetum purpureum 100%; R1, P. purpureum 75%+C. calothyrsus 25%;, R2, P. purpureum 50%+C. calothyrsus 50%; R3, P. purpureum 25%+C. calothyrsus 75%; and R4, C. calothyrsus 100%. All silages were prepared using plastic jar silos (600 g and incubated at room temperature for 30 days. Silages were analyzed for fermentation characteristics and microbial diversity. Increased levels of C. calothyrsus in silage had a significant effect (p<0.01 on the fermentation characteristics. The microbial diversity index decreased and activity was inhibited with increasing levels of C. calothyrsus. The microbial community indicated that there was a population of Lactobacillus plantarum, L. casei, L. brevis, Lactococcus lactis, Chryseobacterium sp., and uncultured bacteria. The result confirmed that silage with a combination of grass and C. calothyrsus had good fermentation characteristics and microbial communities were dominated by L. plantarum.

  13. Occurrence of mycotoxins and yeasts and moulds identification in corn silages in tropical climate.

    Science.gov (United States)

    Carvalho, B F; Ávila, C L S; Krempser, P M; Batista, L R; Pereira, M N; Schwan, R F

    2016-05-01

    This study was aimed to identify yeasts and moulds as well as to detect mycotoxin in corn silages in southern Minas Gerais, Brazil. Corn silages from 36 farms were sampled to analyse dry matter, crude protein, ether extract, ash, neutral detergent fibre, nonfibre carbohydrates and mycotoxins contents, yeasts and moulds population, pH and temperature values. The mycotoxins found in high frequency were aflatoxin in 77·7% of analysed samples, ochratoxin (33·3%) and zearalenone (22·2%). There was no significant correlation between the mycotoxin concentration and the presence of moulds. The pH was negatively correlated with ochratoxin concentration. Aspergillus fumigatus was identified in all silages that presented growth of moulds. Ten different yeast species were identified using the culture-dependent method: Candida diversa, Candida ethanolica, Candida rugosa, Issatchenkia orientalis, Kluyveromyces marxianus, Pichia manshurica, Pichia membranifaciens, Saccharomyces cerevisiae, Trichosporon asahii and Trichosporon japonicum. Another six different yeast species were identified using the culture-independent method. A high mycotoxin contamination rate (91·6% of the analysed silages) was observed. The results indicated that conventional culturing and PCR-DGGE should be combined to optimally describe the microbiota associated with corn silage. This study provides information about the corn silage fermentation dynamics and our findings are relevant to optimization of this silage fermentation. © 2016 The Society for Applied Microbiology.

  14. Nutritional value of grass silage of mombasa associated with additives agroindustrial

    Directory of Open Access Journals (Sweden)

    Euclides Reuter de Oliveira

    2014-06-01

    Full Text Available It was aimed to evaluate the chemical composition of grass silage-mombasa associated with different additives in four times of opening the silo. The experiment was conducted in UFGD. After harvesting the forage, biomass in natura crushed, was taken to the lab, homogenized and enriched on the basis of natural mass, with the following additives: 5% wheat bran, 5% of waste (broken grain and soy ice cream cone of soybean, 5% urea in natural matter and the witness (without additive.The silos were opened after (unprocessed material, 15, 30 and 45 days, for the analysis of chemical composition. The data obtained were analyzed through the statistical programme SISVAR and averages were compared to 5% of probability, by Skott-Knot. The grass silage- mombasa without additive presented major (P0.05 of grass silage- mombasa associated with 5 of urea in 15 days and 45 of silage. The grass silage-mombasa with 5% urea showed the highest crude protein content at time 0 and differed from other treatments. The silage of mombasa associated with 5% urea provided greater in vitro digestibility of dry matter to 15 days of silage.

  15. Wasted cabbage (Brassica oleracea silages treated with different levels of ground corn andsilage inoculant

    Directory of Open Access Journals (Sweden)

    Adauton Vilela de Rezende

    2015-08-01

    Full Text Available Our objective was to evaluate the chemical composition, fermentation profile, and aerobic stability of cabbage silages treated with ground corn and inoculant. The evaluated treatments were: addition of 200, 300, 400, 500, and 600 g of ground corn per kilogram of cabbage (fresh matter basis, with or without a bacterial inoculant composed of Lactobacillus plantarumand Pediococcus pentosaceus. As expected, ground corn additions increased the dry matter (DM content of cabbage silage, and high values were observed for the highest level of addition (540 g kg−1. Conversely, the crude protein, neutral detergent fiber, acid detergent fiber, and lignin contents decreased with ground corn additions. The in vitro dry matter digestibility coefficients increased slightly with ground corn additions, but all cabbage silages had digestibility higher than 740 g kg−1 of DM. In the fermentation process, the pH values of cabbage silages increased linearly because of the high levels of ground corn addition. Cabbage ensiled with 200 and 300 g kg−1 of ground corn had high ammonia N production and fermentative losses (effluent and gas. Cabbage silage treated with 600 g kg−1 of ground corn had lower maximum pH values during aerobic exposure, but all silages had constant temperature during aerobic exposure. The ensiling of wasted cabbage is possible and we recommend the application of 400 g kg−1ground corn to improve the silage quality, whereas the use of the inoculant is unnecessary.

  16. Parâmetros bromatológicos e fermentativos das silagens de milho e girassol Bromatological and fermentative parameters of corn and sunflower silages

    Directory of Open Access Journals (Sweden)

    Rosana Aparecida Possenti

    2005-10-01

    Full Text Available Foi desenvolvido no Instituto de Zootecnia, Agência Paulista de Tecnologia dos Agronegócios (APTA, São Paulo um experimento com o objetivo de avaliar quimicamente a qualidade das silagens e a degradação ruminal da matéria seca (MS, fibra detergente neutra (FDN e proteína bruta (PB das silagens de milho e girassol. Na avaliação químico-bromatológica das silagens, utilizou-se delineamento estatístico inteiramente ao acaso com nove repetições por tratamento (silagem de milho ou girassol e, para a degradação in situ, utilizou-se delineamento em parcelas subdivididas, com três repetições. A silagem de girassol apresentou maiores concentrações de proteína bruta (11,6 vs 9,4%, extrato etéreo (10,1 vs 3,2%, fibra detergente ácida (42,7 vs 31,9%e lignina (9,4 vs 3,7%, e menores teores de MS (22,0 vs 34,6% que a silagem de milho, respectivamente. Apresentou também maiores teores de nitrogênio amoniacal (10,7 vs 5,8 % e ácido acético (3,0 vs 0,79% e menores concentrações de ácido lático (3,7 vs 11,3% que a silagem de milho. As taxas de degradação efetiva da MS, FDN e PB da silagem de girassol foram menores do que as da silagem de milho. A silagem de milho apresentou características mais favoráveis ao processo de ensilagem.At the Instituto de Zootecnia, APTA, São Paulo State-Brazil, it was carried out an experiment to evaluated the chemical quality and ruminal degradation for dry matter (DM, neutral detergent fiber (NDF and protein (CP of sunflower and corn silages. For chemical evaluation of the silages, a total randomized design with nine replicates was used. For in situ degradability was used a split-plot design with three replicates, in which the plots were the times of incubation and the subplot the silage. Sunflower silage showed higher concentration of crude protein (11.6 vs 9.4%, ether extract (10.1 vs 3.2%, acid detergent fiber (42.7 vs 31.9% and lignin (9.4 vs 3.7%, and lower concentration of dry matter than

  17. Sequence and 3D structure based analysis of TNT degrading proteins in Arabidopsis thaliana.

    Science.gov (United States)

    Bhattacherjee, Amrita; Mandal, Rahul Shubhra; Das, Santasabuj; Kundu, Sudip

    2014-03-01

    TNT, accidentally released at several manufacturing sites, contaminates ground water and soil. It has a toxic effect to algae and invertebrate, and chronic exposure to TNT also causes harmful effects to human. On the other hand, many plants including Arabidopsis thaliana have the ability to metabolize TNT either completely or at least to a reduced less toxic form. In A. thaliana, the enzyme UDP glucosyltransferase (UDPGT) can further conjugate the reduced forms 2-HADNT and 4-HADNT (2-hydroxylamino-4, 6- dinitrotoluene and 4-hydroxylamino-2, 6- dinitrotoluene) of TNT. Based on the experimental analysis, existing literature and phylogenetic analysis, it is evident that among 107 UDPGT proteins only six are involved in the TNT degrading process. A total of 13 UDPGT proteins including five of these TNT degrading proteins fall within the same group of phylogeny. Thus, these 13 UDPGT proteins have been classified into two groups, TNT-degrading and TNT-non-degrading proteins. To understand the differences in TNT-degrading capacities; using homology modeling we first predicted two structures, taking one representative sequence from both the groups. Next, we performed molecular docking of the modeled structure and TNT reduced form 2-hydroxylamino-4, 6- dinitrotoluene (2-HADNT). We observed that while the Trp residue located within the active site region of the TNT- degrading protein showed π-Cation interaction; such type of interaction was absent in TNT-non-degrading protein, as the respective Trp residue lay outside of the pocket in this case. We observed the conservation of this π-Cation interaction during MD simulation of TNT-degrading protein. Thus, the position and the orientation of the active site residue Trp could explain the presence and absence of TNT-degrading capacity of the UDPGT proteins.

  18. A Critical Appraisal of Quantitative Studies of Protein Degradation in the Framework of Cellular Proteostasis

    Directory of Open Access Journals (Sweden)

    Beatriz Alvarez-Castelao

    2012-01-01

    Full Text Available Protein homeostasis, proteostasis, is essential to understand cell function. Protein degradation is a crucial component of the proteostatic mechanisms of the cell. Experiments on protein degradation are nowadays present in many investigations in the field of molecular and cell biology. In the present paper, we focus on the different experimental approaches to study protein degradation and present a critical appraisal of the results derived from steady-state and kinetic experiments using detection of unlabelled and labelled protein methodologies with a proteostatic perspective. This perspective allows pinpointing the limitations in interpretation of results and the need of further experiments and/or controls to establish “definitive evidence” for the role of protein degradation in the proteostasis of a given protein or the entire proteome. We also provide a spreadsheet for simple calculations of mRNA and protein decays for mimicking different experimental conditions and a checklist for the analysis of experiments dealing with protein degradation studies that may be useful for researchers interested in the area of protein turnover.

  19. Degradation of protein translation machinery by amino acid starvation-induced macroautophagy

    DEFF Research Database (Denmark)

    Gretzmeier, Christine; Eiselein, Sven; Johnson, Gregory R.

    2017-01-01

    , unbiased approaches relying on quantitative mass spectrometry-based proteomics. Macroautophagy is induced by rapamycin treatment, and by amino acid and glucose starvation in differentially, metabolically labeled cells. Protein dynamics are linked to image-based models of autophagosome turnover. Depending...... on the inducing stimulus, protein as well as organelle turnover differ. Amino acid starvation-induced macroautophagy leads to selective degradation of proteins important for protein translation. Thus, protein dynamics reflect cellular conditions in the respective treatment indicating stimulus-specific pathways...

  20. Determination of region-specific data of yield and quality of alternatives to silage maize in fodder crops – field trails with forage gras and clover grass mixtures, Sorghum as well as whole plant silage of grain

    Directory of Open Access Journals (Sweden)

    Wosnitza, Andrea

    2014-02-01

    Full Text Available This project should generate current regional results over a period of three years about the parameter yield and quality of alternative fodder crops to maize; this includes grass and clover grass mixtures, silage maize, varieties of Sorghum/millets and whole plant silages of wheat, rye and triticale. The tested silage maize showed the highest and most reliable average dry matter yield with 23 tons per hectare, with a very low variance. The Sorghum and millet varieties had dry matter yields of 3 to 5 tons per hectare below the silage maize yield but with individual values fluctuating in a broad range within years and locations. With values far below 28% the dry matter contents were not suitable for ensiling. The grass and clover grass mixtures are good, stable and established alternatives to maize for silage. They achieved high yields comparable with these of Sorghum but stable and with a highly suitable dry matter content for ensiling. The yield of the whole plant silages was up to 22% lower compared with maize. So none of the alternative crops can compete with the high level yield of silage maize in its favoured region, therefore would be a combination of two crops recommended. But some individual locally adapted mixtures or varieties of the alternative crops reached nearly 80% of the maize yield. Silage maize showed the highest level of the net energy content for lactation (NEL, followed by the values of the fodder crops and the whole plant silages. The Sorghum varieties showed the lowest NEL value of all tested cultures. The highest crude protein showed the fodder crops contents. Silage maize, Sorghum and the whole plant silages had values lying nearly around the 50% mark of the fodder crops.

  1. In situ degrability of dry matter of sheep fed with corn silage with or without Bt gene

    Directory of Open Access Journals (Sweden)

    Camila Memari Trava

    2012-12-01

    Full Text Available Silage corn is a food widely used, composing the bulk of feed for ruminants, because its present high nutritional value. Since the release by CNTBio seeds of modified genetically corn, many of it began to use transgenic silage corn, which was inserted into genetic code the Bt (Bacillus thuringiensis gene that expresses a toxic protein to caterpillar pests of corn, so occurs the reduction of production costs and pesticide use. Due to rapid expansion of transgenic maize and your wide use in animal feed by producers, the aim of this study was to evaluate the degrability in situ in animal rumen, in order to estimate the nutritional value of silage corn with and without the Bt gene. The experiment was conducted at the Institute of Animal Science Nova Odessa-SP. Were used four rumen fistulated sheep housed in individual pens for 56 days, including four periods. The animals were fed with silage with two varieties of plant corn to silage - DKB and AG, and their isogenic counterparts with the Bt gene, comprising four treatments. The degradability determination of dry matter (DM digestibility of silage corn treatments was determined by means of nylon bag in situ. After removal, the bags were washed and incubated, then placed in forced-circulation at 55°C to constant weight to determine the DM concentration. Data from in situ degradation of DM was adjusted in the mathematical model proposed by Ørskov and McDonald (1979. For the degradability of DM, the fraction “a” showed the interaction (p <0.05, where the variety DKB do not showed difference (p> 0.05 for the gene insertion. For AG, showed a slight decrease (p <0.05 when compared to its isogenic counterpart with the gene (35.68% and 37.85% respectively, means that the Bt gene reduced the solubility of DM of this fraction for AG range. The fact of the variety AG with and without the gene have suffered lower solubility of DM when compared the DKB with and without the gene is due to the fact of being

  2. Toscana virus NSs protein promotes degradation of double-stranded RNA-dependent protein kinase.

    Science.gov (United States)

    Kalveram, Birte; Ikegami, Tetsuro

    2013-04-01

    Toscana virus (TOSV), which is transmitted by Phlebotomus spp. sandflies, is a major etiologic agent of aseptic meningitis and encephalitis in the Mediterranean. Like other members of the genus Phlebovirus of the family Bunyaviridae, TOSV encodes a nonstructural protein (NSs) in its small RNA segment. Although the NSs of Rift Valley fever virus (RVFV) has been identified as an important virulence factor, which suppresses host general transcription, inhibits transcription from the beta interferon promoter, and promotes the proteasomal degradation of double-stranded RNA-dependent protein kinase (PKR), little is known about the functions of NSs proteins encoded by less-pathogenic members of this genus. In this study we report that TOSV is able to downregulate PKR with similar efficiency as RVFV, while infection with the other phleboviruses-i.e., Punta Toro virus, sandfly fever Sicilian virus, or Frijoles virus-has no effect on cellular PKR levels. In contrast to RVFV, however, cellular transcription remains unaffected during TOSV infection. TOSV NSs protein promotes the proteasome-dependent downregulation of PKR and is able to interact with kinase-inactive PKR in infected cells.

  3. The Arabidopsis CROWDED NUCLEI genes regulate seed germination by modulating degradation of ABI5 protein.

    Science.gov (United States)

    Zhao, Wenming; Guan, Chunmei; Feng, Jian; Liang, Yan; Zhan, Ni; Zuo, Jianru; Ren, Bo

    2016-07-01

    In Arabidopsis, the phytohormone abscisic acid (ABA) plays a vital role in inhibiting seed germination and in post-germination seedling establishment. In the ABA signaling pathway, ABI5, a basic Leu zipper transcription factor, has important functions in the regulation of seed germination. ABI5 protein localizes in nuclear bodies, along with AFP, COP1, and SIZ1, and was degraded through the 26S proteasome pathway. However, the mechanisms of ABI5 nuclear body formation and ABI5 protein degradation remain obscure. In this study, we found that the Arabidopsis CROWDED NUCLEI (CRWN) proteins, predicted nuclear matrix proteins essential for maintenance of nuclear morphology, also participate in ABA-controlled seed germination by regulating the degradation of ABI5 protein. During seed germination, the crwn mutants are hypersensitive to ABA and have higher levels of ABI5 protein compared to wild type. Genetic analysis suggested that CRWNs act upstream of ABI5. The observation that CRWN3 colocalizes with ABI5 in nuclear bodies indicates that CRWNs might participate in ABI5 protein degradation in nuclear bodies. Moreover, we revealed that the extreme C-terminal of CRWN3 protein is necessary for its function in the response to ABA in germination. Our results suggested important roles of CRWNs in ABI5 nuclear body organization and ABI5 protein degradation during seed germination. © 2015 Institute of Botany, Chinese Academy of Sciences.

  4. Muscle protein degradation and amino acid metabolism during prolonged knee-extensor exercise in humans

    DEFF Research Database (Denmark)

    Van Hall, Gerrit; Saltin, B; Wagenmakers, A J

    1999-01-01

    to a substantial increase in net muscle protein degradation, and that a lowering of the starting muscle glycogen content leads to a further increase. The carbon atoms of the branched-chain amino acids (BCAA), glutamate, aspartate and asparagine, liberated by protein degradation, and the BCAA and glutamate......The aim of this study was to investigate whether prolonged one-leg knee-extensor exercise enhances net protein degradation in muscle with a normal or low glycogen content. Net amino acid production, as a measure of net protein degradation, was estimated from leg exchange and from changes...... in the concentrations of amino acids that are not metabolized in skeletal muscle. Experiments were performed at rest and during one-leg knee-extensor exercise in six subjects having one leg with a normal glycogen content and the other with a low glycogen content. Exercise was performed for 90 min at a workload of 60...

  5. Degradation of extracytoplasmic catalysts for protein folding in Bacillus subtilis

    NARCIS (Netherlands)

    Krishnappa, Laxmi; Monteferrante, Carmine G; Neef, Jolanda; Dreisbach, Annette; van Dijl, Jan Maarten

    The general protein secretion pathway of Bacillus subtilis has a high capacity for protein export from the cytoplasm, which is exploited in the biotechnological production of a wide range of enzymes. These exported proteins pass the membrane in an unfolded state, and accordingly, they have to fold

  6. Effects of grain source, grain processing, and protein degradability on rumen kinetics and microbial protein synthesis in Boer kids.

    Science.gov (United States)

    Brassard, M-E; Chouinard, P Y; Berthiaume, R; Tremblay, G F; Gervais, R; Martineau, R; Cinq-Mars, D

    2015-11-01

    Microbial protein synthesis in the rumen would be optimized when dietary carbohydrates and proteins have synchronized rates and extent of degradation. The aim of this study was to evaluate the effect of varying ruminal degradation rate of energy and nitrogen sources on intake, nitrogen balance, microbial protein yield, and kinetics of nutrients in the rumen of growing kids. Eight Boer goats (38.2 ± 3.0 kg) were used. The treatments were arranged in a split-plot Latin square design with grain sources (barley or corn) forming the main plots (squares). Grain processing methods and levels of protein degradability formed the subplots in a 2 × 2 factorial arrangement for a total of 8 dietary treatments. The grain processing method was rolling for barley and cracking for corn. Levels of protein degradability were obtained by feeding untreated soybean meal (SBM) or heat-treated soybean meal (HSBM). Each experimental period lasted 21 d, consisting of a 10-d adaptation period, a 7-d digestibility determination period, and a 4-d rumen evacuation and sampling period. Kids fed with corn had higher purine derivatives (PD) excretion when coupled with SBM compared with HSBM and the opposite occurred with barley-fed kids ( ≤ 0.01). Unprocessed grain offered with SBM led to higher PD excretion than with HSBM whereas protein degradability had no effect when processed grain was fed ( ≤ 0.03). Results of the current experiment with high-concentrate diets showed that microbial N synthesis could be maximized in goat kids by combining slowly fermented grains (corn or unprocessed grains) with a highly degradable protein supplement (SBM). With barley, a more rapidly fermented grain, a greater microbial N synthesis was observed when supplementing a low-degradable protein (HSBM).

  7. ORGANIC MATTER AND CRUDE PROTEIN DEGRADATION SYNCHRONY IN DIETS SELECTED BY RANGE GOATS.

    Directory of Open Access Journals (Sweden)

    Rafael Ramírez Orduña

    2010-09-01

    Full Text Available The study was carried out with the aim to asses the synchrony of organic matter and crude protein degradation in the rumen of diets selected by range goats through two years. Five esophageal cannulated adult male goats were used to collect extrusa samples during summer (August 9–13 and autumn (November 29 –December 3 of 2006, winter (February 20 – 24, spring (April 29 –May 5, summer (September 10–15 and autumn (December 4–8 of 2007 and winter (February 20 – 25 and spring (May 9 –13 of 2008. Extrusa samples were subjected to chemical analysis to determine organic matter (OM, crude protein (CP in situ and in vitro true digestibility of dry matter. OM and CP intake were estimated by total fecal collection. Effective extent of degradation of the OM and CP was calculated hourly and total 24 hours. From the hourly quantity of OM and CP degraded, a synchrony index of CP to OM was calculated, and from the total 24 hours degradation, degraded organic matter intake and crude protein intake were also estimated. Sampling date was the main effect that determined the variation of diet OM and CP degradation parameters. Degraded crude protein intake as a proportion of degraded OM was affected by sampling date and was correlated to rainfall. During winter of the first year degraded crude protein intake was below the requirements for maintenance or to promote growth for range goats weighing 40 kg. Even though, synchrony index between OM and CP degradation was affected by sampling date goats maintained a high synchrony index throughout the years.

  8. Effects of gamma irradiation on chemical composition and ruminal protein degradation of canola meal

    Energy Technology Data Exchange (ETDEWEB)

    Shawrang, P. [Agriculture, Medical and Industrial Research School, Nuclear Science and Technology Research Institute, Atomic Energy Organization of Iran, P.O. Box 31485-498, Karaj (Iran, Islamic Republic of); Department of Animal Science, Faculty of Agriculture, Tehran University P.O. Box 4111, Karaj (Iran, Islamic Republic of)], E-mail: parvinshawrang@yahoo.co.uk; Nikkhah, A.; Zare-Shahneh, A. [Department of Animal Science, Faculty of Agriculture, Tehran University P.O. Box 4111, Karaj (Iran, Islamic Republic of); Sadeghi, A.A. [Department of Animal Science, Faculty of Agriculture, Science and Research Branch, Islamic Azad University, P.O. Box 14515-4933, Tehran (Iran, Islamic Republic of); Raisali, G. [Radiation Applications Research School, Nuclear Science and Technology Research Institute, Atomic Energy Organization of Iran, P.O. Box 11365-3486, Tehran (Iran, Islamic Republic of); Moradi-Shahrebabak, M. [Department of Animal Science, Faculty of Agriculture, Tehran University P.O. Box 4111, Karaj (Iran, Islamic Republic of)

    2008-07-15

    Gamma irradiation of canola meal (at doses of 25, 50 and 75 kGy) could alter its ruminal protein degradation characteristics by cross-linking of the polypeptide chains. This processing resulted in decrease (linear effect, P<0.001) of ruminal protein degradation and increase (linear effect, P<0.001) of intestinal protein digestibility. The results showed that gamma irradiation at doses higher than 25 kGy can be used as a cross-linking agent to improve protein properties of supplements in ruminant nutrition.

  9. Effects of gamma irradiation on chemical composition and ruminal protein degradation of canola meal

    International Nuclear Information System (INIS)

    Shawrang, P.; Nikkhah, A.; Zare-Shahneh, A.; Sadeghi, A.A.; Raisali, G.; Moradi-Shahrebabak, M.

    2008-01-01

    Gamma irradiation of canola meal (at doses of 25, 50 and 75 kGy) could alter its ruminal protein degradation characteristics by cross-linking of the polypeptide chains. This processing resulted in decrease (linear effect, P<0.001) of ruminal protein degradation and increase (linear effect, P<0.001) of intestinal protein digestibility. The results showed that gamma irradiation at doses higher than 25 kGy can be used as a cross-linking agent to improve protein properties of supplements in ruminant nutrition

  10. Colorimetric determination of in vitro feed protein degradation

    African Journals Online (AJOL)

    bag technique. Diazotized f ish meal, maize gluten meal and soya-bean were incubated with strained rumen fluid and a suitable growth medium. Only 49o/o and 18% of the colour bound to soya-bean was bound to fish meal and corn gluten meal, respectively. Degradation values expressed as a percen- tage of the total ...

  11. Silage extracts used to study the mode of action of silage inoculants in ruminants

    Science.gov (United States)

    Silage microbial inoculants can enhance animal performance, but the mechanisms involved in these effects are not clear. Our hypothesis was that an extractable factor from inoculated silage enhances rumen microbial activity. One alfalfa haylage (58% DM) and two corn silages (30% and 50% DM) were made...

  12. Critical lysine residues of Klf4 required for protein stabilization and degradation

    Energy Technology Data Exchange (ETDEWEB)

    Lim, Key-Hwan; Kim, So-Ra; Ramakrishna, Suresh; Baek, Kwang-Hyun, E-mail: baek@cha.ac.kr

    2014-01-24

    Highlights: • Klf4 undergoes the 26S proteasomal degradation by ubiquitination on its multiple lysine residues. • Essential Klf4 ubiquitination sites are accumulated between 190–263 amino acids. • A mutation of lysine at 232 on Klf4 elongates protein turnover. • Klf4 mutants dramatically suppress p53 expression both under normal and UV irradiated conditions. - Abstract: The transcription factor, Krüppel-like factor 4 (Klf4) plays a crucial role in generating induced pluripotent stem cells (iPSCs). As the ubiquitination and degradation of the Klf4 protein have been suggested to play an important role in its function, the identification of specific lysine sites that are responsible for protein degradation is of prime interest to improve protein stability and function. However, the molecular mechanism regulating proteasomal degradation of the Klf4 is poorly understood. In this study, both the analysis of Klf4 ubiquitination sites using several Klf4 deletion fragments and bioinformatics predictions showed that the lysine sites which are signaling for Klf4 protein degradation lie in its N-terminal domain (aa 1–296). The results also showed that Lys32, 52, 232, and 252 of Klf4 are responsible for the proteolysis of the Klf4 protein. These results suggest that Klf4 undergoes proteasomal degradation and that these lysine residues are critical for Klf4 ubiquitination.

  13. Nutritive value and fermentation quality of palisadegrass and stylo mixed silages.

    Science.gov (United States)

    da Silva, Juliana S; Ribeiro, Karina G; Pereira, Odilon G; Mantovani, Hilário C; Cecon, Paulo R; Pereira, Rosana C; Silva, Janaina de L

    2018-01-01

    The nutritive value and fermentation quality of palisadegrass (Brachiaria brizantha cv. Xaraes) and stylo (Stylosanthes capitata × S. macrocephala cv. Campo Grande) mixed silages were evaluated. The experiment was analyzed in a factorial scheme (5 × 2) in a completely randomized design using increasing levels of stylo (0, 25, 50, 75 and 100% on a fresh matter basis) on palisadegrass silages, with and without microbial inoculants (MI). With the increased ratio of stylo in mixed silages, dry matter (DM), crude protein (CP), acid detergent fiber (ADF), and lignin content increased in silages. The presence of MI promoted lower DM content, and higher neutral detergent fiber corrected for ash and protein, ADF and lignin content. The acid detergent insoluble nitrogen content and the lactic acid bacteria populations were not affected by treatments. The in vitroDM digestibility was affected by the interaction of levels of the stylo and MI. The pH, NH 3 -N/total nitrogen and butyric acid concentrations decreased with increasing levels of stylo. Better nutritive value and quality of fermentation was found in the silage containing higher proportions of this stylo mixed with palisadegrass. The microbial inoculant evaluated did not alter the nutritive value or quality of the fermentation of the silages in this experiment. © 2017 Japanese Society of Animal Science.

  14. Degradation of structurally characterized proteins injected into HeLa cells. Basic measurements

    International Nuclear Information System (INIS)

    Rogers, S.W.; Rechsteiner, M.

    1988-01-01

    Thirty-five proteins of known x-ray structure were labeled by chloramine-T radioiodination or by reaction with 125I-Bolton-Hunter reagent and introduced into HeLa cells using red cell-mediated microinjection. Degradation rates of the injected proteins were then determined over the next 50 h by measuring the release of soluble isotope to the culture medium. Control experiments demonstrated that the measured rates were not compromised by proteolysis within RBCs, the presence of unfused RBCs, or degradation of protein released from RBCs to the medium. Degradation of some injected proteins was faster during the first 12 h after fusion than at later times, apparently a response of HeLa cells to trypsinization. However, all proteins exhibited first-order degradation rates between 24 and 48 h post injection. Except for seven proteins, stabilities measured during this interval were unaffected by the labeling procedure. Reductive methylation was used to choose among the seven discordant values, and half-lives for the 35 proteins ranged from 16 h for lysozyme to 214 h for yeast alcohol dehydrogenase. Since half-lives for six of the injected proteins closely match values obtained by in vivo measurements, we consider our estimates of the metabolic stabilities of the injected proteins to be generally accurate. Therefore, the half-lives obtained by microinjection should prove useful in the search for relationships between protein structure and intracellular stability

  15. Polyubiquitin chain assembly and organisation determine the dynamics of protein activation and degradation

    Directory of Open Access Journals (Sweden)

    Lan K. Nguyen

    2014-01-01

    Full Text Available Protein degradation via ubiquitination is a major proteolytic mechanism in cells. Once a protein is destined for degradation, it is tagged by multiple ubiquitin molecules. The synthesised polyubiquitin chains can be recognised by the 26S proteosome where proteins are degraded. These chains form through multiple ubiquitination cycles that are similar to multi-site phosphorylation cycles. As kinases and phosphatases, two opposing enzymes (E3 ligases and deubiquitinases DUBs catalyse (deubiquitination cycles. Although multi-ubiquitination cycles are fundamental mechanisms of controlling protein concentrations within a cell, their dynamics have never been explored. Here, we fill this knowledge gap. We show that under permissive physiological conditions, the formation of polyubiquitin chain of length greater than two and subsequent degradation of the ubiquitinated protein, which is balanced by protein synthesis, can display bistable, switch-like responses. Interestingly, the occurrence of bistability becomes pronounced, as the chain grows, giving rise to all-or-none regulation at the protein levels. We give predictions of protein distributions under bistable regime awaiting experimental verification. Importantly, we show for the first time that sustained oscillations can robustly arise in the process of formation of ubiquitin chain, largely due to the degradation of the target protein. This new feature is opposite to the properties of multi-site phosphorylation cycles, which are incapable of generating oscillation if the total abundance of interconverted protein forms is conserved. We derive structural and kinetic constraints for the emergence of oscillations, indicating that a competition between different substrate forms and the E3 and DUB is critical for oscillation. Our work provides the first detailed elucidation of the dynamical features brought about by different molecular setups of the polyubiquitin chain assembly process responsible for

  16. An in vitro model to study interactions between Escherichia coli and lactic acid bacterial inoculants for silage in rumen fluid.

    Science.gov (United States)

    Weinberg, Z G; Chen, Y; Volchinski, V; Sela, S; Ogunade, I M; Adesogan, A

    2016-07-01

    Previous studies have shown that silages treated with lactic acid bacteria (LAB) inoculants enhance ruminants' performance. The objective of the current experiments was to develop an in vitro model to study interactions between LAB silage inoculants and inoculated silages and Escherichia coli (EC) in rumen fluid (RF). Our hypothesis was that some inoculants inhibit EC in RF. For that purpose buffered RF was incubated under anaerobic conditions at 39°C with commercial strains of LAB silage inoculants or with laboratory corn and wheat silages treated with these LAB, an EC strain and with various ruminant feed ingredients. The EC strain was originally isolated from cattle manure and tagged with a plasmid expressing the green fluorescence protein and kanamycin and streptomycin resistance. Results indicate that the LAB or the treated silages did not suppress EC numbers in the RF. When the pH of the RF decreased below 5·0 the EC disappeared. We conclude that both LAB inoculants for silage and EC survived in RF for several days; however, the inoculants and silages treated with such inoculants did not inhibit EC in RF in vitro. Forage crops, silage and hay are initial stages of the food chain for humans. Cattle harbours and sheds enterobacteria regularly, some strains of which are pathogens. These can contaminate forage crops through field fertilization with cattle manure. The objective of this study was to develop an in vitro model to test whether lactic acid bacteria, which are used in silage inoculants, alone or in treated silages can inhibit Escherichia coli in rumen fluid. This study presents safety aspects and it is also part of a broad research effort aimed at finding out how LAB silage inoculants and inoculated silages enhance ruminant performance or exert probiotic effects in ruminants. © 2016 The Society for Applied Microbiology.

  17. Steers performance fed diets with Alexander grass silage (Urochloa plantaginea x sorghum silage (Sorghum bicolor (L. Moench

    Directory of Open Access Journals (Sweden)

    Flânia Mônego Argenta

    2014-02-01

    Full Text Available This study aimed to evaluate the parameters related to feedlot beef steers fed with Alexander grass silage or sorghum silage as roughage source. Twelve animals with initial average of weight and age of 245 kg and 20 months, respectively, belonging to Nellore (N, Charolais (C, 21/32 N 11/32 C, 21/32 C 11/32 N; 43/64 N 21/64 C and 43/64 C 21/64 N genetic groups were used. The ration was composed by a roughage:concentrate relation of 50:50 (dry matter basis, offered at ease. The dry matter, crude protein, ether extract, neutral detergent fiber and acid detergent fiber daily intake didn’t differ between roughage sources, being the medium values of 10.18; 1.28; 0.20; 4.96 and 2.51 kg/day, respectively. However, the ether extract intake by metabolic size and by corporal weight percentage were higher for the one that consumed sorghum silage, obtaining values of 2.74 and 2.36 g/metabolic size and 0.06 and 0.05 kg/100 kg of live weight. The same behavior was observed for digestible energy intake, presenting values of 364.87 and 302.83 g/metabolic size and 8.10 and 7.02 kg/100 kg of live weight. The daily average gain, final corporal score and feed conversion weren’t influenced by roughage type, being the values of 1.31 kg; 4.05 points and 7.78 kg of DM/kg of weight gain, respectively. The Alexander grass silage is a feasible alternative as its substitution in relation to sorghum silage.

  18. Effects of Synchronicity of Carbohydrate and Protein Degradation on Rumen Fermentation Characteristics and Microbial Protein Synthesis

    Directory of Open Access Journals (Sweden)

    J. K. Seo

    2013-03-01

    Full Text Available A series of in vitro studies were carried out to determine i the effects of enzyme and formaldehyde treatment on the degradation characteristics of carbohydrate and protein sources and on the synchronicity of these processes, and ii the effects of synchronizing carbohydrate and protein supply on rumen fermentation and microbial protein synthesis (MPS in in vitro experiments. Untreated corn (C and enzyme-treated corn (EC were combined with soy bean meal with (ES and without (S enzyme treatment or formaldehyde treatment (FS. Six experimental feeds (CS, CES, CFS, ECS, ECES and ECFS with different synchrony indices were prepared. Highly synchronous diets had the greatest dry matter (DM digestibility when untreated corn was used. However, the degree of synchronicity did not influence DM digestibility when EC was mixed with various soybean meals. At time points of 12 h and 24 h of incubation, EC-containing diets showed lower ammonia-N concentrations than those of C-containing diets, irrespective of the degree of synchronicity, indicating that more efficient utilization of ammonia-N for MPS was achieved by ruminal microorganisms when EC was offered as a carbohydrate source. Within C-containing treatments, the purine base concentration increased as the diets were more synchronized. This effect was not observed when EC was offered. There were significant effects on VFA concentration of both C and S treatments and their interactions. Similar to purine concentrations, total VFA production and individual VFA concentration in the groups containing EC as an energy source was higher than those of other groups (CS, CES and CFS. The results of the present study suggested that the availability of energy or the protein source are the most limiting factors for rumen fermentation and MPS, rather than the degree of synchronicity.

  19. Supply of nutrients and productive responses in dairy cows given diets based on restrictively fermented silage

    Directory of Open Access Journals (Sweden)

    P. HUHTANEN

    2008-12-01

    Full Text Available The objective of this paper is to review research which has evaluated the feeding of dairy cows with diets containing large proportions of grass silage. In Finland, milk production systems evolved are based on the use of restrictively fermented silages. Higher potential yields, smaller production risks than with cereal grains, short grazing period and high digestibility of grasses grown in northern latitudes have facilitated this development. Factors affecting nutrient supply from these diets are discussed. Digestibility is determined mainly by the stage of maturity at harvesting and it is not markedly affected by the level of energy and protein supplementation. Intake of grass silage is influenced both by digestibility and fermentation characteristics. Efficiency of microbial synthesis is high in animals given diets based on restrictively fermented silage but rumen fermentation pattern is characterised by low molar proportions of propionate. Production responses to additional concentrate are relatively small, especially when the amount of concentrate exceeds 10 kg day-1. High substitution of silage dry matter (DM, negative associative effects on digestion and partitioning of energy towards body tissues account for small production responses. Protein supplementation has consistently increased milk protein yield but responses do not appear to be related to the level of milk production, silage crude protein content, amount of concentrate or stage of lactation. The new protein evaluation system provides an accurate prediction of protein yield with the typical Finnish dairy cow diets. The high slopes (ca. 0.5 between protein supply and milk protein yield within experiments suggest that protein supply is suboptimal and protein supplements are used with a high efficiency.;

  20. Glucose Deprivation Triggers Protein Kinase C-dependent β-Catenin Proteasomal Degradation*

    Science.gov (United States)

    Choi, Seung-Won; Song, Jun-Kyu; Yim, Ye-Seal; Yun, Ho-Geun; Chun, Kyung-Hee

    2015-01-01

    Autophagy is a conserved process that contributes to cell homeostasis. It is well known that induction mainly occurs in response to nutrient starvation, such as starvation of amino acids and insulin, and its mechanisms have been extensively characterized. However, the mechanisms behind cellular glucose deprivation-induced autophagy are as of now poorly understood. In the present study, we determined a mechanism by which glucose deprivation induced the PKC-dependent proteasomal degradation of β-catenin, leading to autophagy. Glucose deprivation was shown to cause a sub-G1 transition and enhancement of the LC3-II protein levels, whereas β-catenin protein underwent degradation in a proteasome-dependent manner. Moreover, the inhibition of GSK3β was unable to abolish the glucose deprivation-mediated β-catenin degradation or up-regulation of LC3-II protein levels, which suggested GSK3β-independent protein degradation. Intriguingly, the inhibition of PKCα using a pharmacological inhibitor and transfection of siRNA for PKCα was observed to effectively block glucose deprivation-induced β-catenin degradation as well as the increase in LC3-II levels and the accumulation of a sub-G1 population. Together, our results demonstrated a molecular mechanism by which glucose deprivation can induce the GSK3β-independent protein degradation of β-catenin, leading to autophagy. PMID:25691573

  1. Light-induced protein nitration and degradation with HONO emission

    Science.gov (United States)

    Meusel, Hannah; Elshorbany, Yasin; Kuhn, Uwe; Bartels-Rausch, Thorsten; Reinmuth-Selzle, Kathrin; Kampf, Christopher J.; Li, Guo; Wang, Xiaoxiang; Lelieveld, Jos; Pöschl, Ulrich; Hoffmann, Thorsten; Su, Hang; Ammann, Markus; Cheng, Yafang

    2017-10-01

    Proteins can be nitrated by air pollutants (NO2), enhancing their allergenic potential. This work provides insight into protein nitration and subsequent decomposition in the presence of solar radiation. We also investigated light-induced formation of nitrous acid (HONO) from protein surfaces that were nitrated either online with instantaneous gas-phase exposure to NO2 or offline by an efficient nitration agent (tetranitromethane, TNM). Bovine serum albumin (BSA) and ovalbumin (OVA) were used as model substances for proteins. Nitration degrees of about 1 % were derived applying NO2 concentrations of 100 ppb under VIS/UV illuminated conditions, while simultaneous decomposition of (nitrated) proteins was also found during long-term (20 h) irradiation exposure. Measurements of gas exchange on TNM-nitrated proteins revealed that HONO can be formed and released even without contribution of instantaneous heterogeneous NO2 conversion. NO2 exposure was found to increase HONO emissions substantially. In particular, a strong dependence of HONO emissions on light intensity, relative humidity, NO2 concentrations and the applied coating thickness was found. The 20 h long-term studies revealed sustained HONO formation, even when concentrations of the intact (nitrated) proteins were too low to be detected after the gas exchange measurements. A reaction mechanism for the NO2 conversion based on the Langmuir-Hinshelwood kinetics is proposed.

  2. Context Memory Formation Requires Activity-Dependent Protein Degradation in the Hippocampus

    Science.gov (United States)

    Cullen, Patrick K.; Ferrara, Nicole C.; Pullins, Shane E.; Helmstetter, Fred J.

    2017-01-01

    Numerous studies have indicated that the consolidation of contextual fear memories supported by an aversive outcome like footshock requires de novo protein synthesis as well as protein degradation mediated by the ubiquitin-proteasome system (UPS). Context memory formed in the absence of an aversive stimulus by simple exposure to a novel…

  3. Role of Proteasome-Dependent Protein Degradation in Long-Term Operant Memory in "Aplysia"

    Science.gov (United States)

    Lyons, Lisa C.; Gardner, Jacob S.; Gandour, Catherine E.; Krishnan, Harini C.

    2017-01-01

    We investigated the in vivo role of protein degradation during intermediate (ITM) and long-term memory (LTM) in "Aplysia" using an operant learning paradigm. The proteasome inhibitor MG-132 inhibited the induction and molecular consolidation of LTM with no effect on ITM. Remarkably, maintenance of steady-state protein levels through…

  4. Calculating the Degradation Rate of Individual Proteins Using Xenopus Extract Systems.

    Science.gov (United States)

    McDowell, Gary S; Philpott, Anna

    2018-05-16

    The Xenopus extract system has been used extensively as a simple, quick, and robust method for assessing the stability of proteins against proteasomal degradation. In this protocol, methods are provided for assessing the half-life of in vitro translated radiolabeled proteins using Xenopus egg or embryo extracts. © 2019 Cold Spring Harbor Laboratory Press.

  5. Apparent in vivo nutrient digestibility of maize silages in horses

    Directory of Open Access Journals (Sweden)

    Adriana PÍŠOVÁ

    2016-12-01

    Full Text Available The aim of the experiment was to analyzed coefficients of digestibility of the maize silage nutrients in horse feeding. Horses of Slovak warmblood breed engaged in the research were 2.5 ± 0.5 years old. Horses in boxes without bedding to avoid eating it and excrement contamination were stabled. Horses individually with ad libitum access to water were housed. The boxes with trough for corn silage were equiped. The corn silage contained 362.3 g of dry matter, 74.1 g of crude protein, 27.1 g of crude fat and 948.2 g of organic matter in kg of dry matter. The feed twice a day, in the morning and in the evening was given. The feed intake and leftovers were weighed every day. Excrements immediately after excreting to avoid urine contamination into plastic containers were collected and weighed. The content of nutrients in the feed intake and in the excrements were analysed. In analysed samples the gravimetric dry matter amount by drying at 103 ± 2 °C, crude protein content by Kjeldahl method, fat content by extraction method according to Soxhlett – Henkel, ash content by burning the sample at 530 ± 20 °C in muffle furnace and organic matter content by calculation were determined. Coefficients of apparent digestibility in percent from taken in and excreted nutrients were calculated. In maize silage for horse feeding an average coefficient of digestibility of dry matter 61.94%, of crude protein 56.03%, of crude fat 72.34% and of organic matter 65.19% was found.

  6. Optically degradable dendrons for temporary adhesion of proteins to DNA.

    Science.gov (United States)

    Kostiainen, Mauri A; Kotimaa, Juha; Laukkanen, Marja-Leena; Pavan, Giovanni M

    2010-06-18

    Experimental studies and molecular dynamics modeling demonstrate that multivalent dendrons can be used to temporarily glue proteins and DNA together with high affinity. We describe N-maleimide-cored polyamine dendrons that can be conjugated with free cysteine residues on protein surfaces through 1,4-conjugate addition to give one-to-one protein-polymer conjugates. We used a genetically engineered cysteine mutant of class II hydrophobin (HFBI) and a single-chain Fragment variable (scFv) antibody as model proteins for the conjugation reactions. The binding affinity of the protein-dendron conjugates towards DNA was experimentally assessed by using the ethidium bromide displacement assay. The binding was found to depend on the generation of the dendron, with the second generation having a stronger affinity than the first generation. Thermodynamic parameters of the binding were obtained from molecular dynamics modeling, which showed that the high binding affinity for each system is almost completely driven by a strong favorable binding enthalpy that is opposed by unfavorable binding entropy. A short exposure to UV (lambda approximately 350 nm) can cleave the photolabile o-nitrobenzyl-linked binding ligands from the surface of the dendron, which results in loss of the multivalent binding interactions and triggers the release of the DNA and protein. The timescale of the release is very rapid and the binding partners can be efficiently released after 3 min of UV exposure.

  7. Effects of whey and molasses as silage additives on potato hash ...

    African Journals Online (AJOL)

    p2492989

    20 mg copper; 20 mg iodine; 189 g calcium; 50 g phosphorus; 6000 000 I.U. ..... of corn silage by pineapple by-products on ruminal degradability in beef cattle. .... between the herbage and the course of fermentation in ensiling green fodder.

  8. Fermentative characteristics and nutritional value of elephant grass silage added with dehydrated banana peel

    Directory of Open Access Journals (Sweden)

    Lara Maria Santos Brant

    2017-04-01

    Full Text Available The aim of this study was to evaluate the fermentative losses and nutritional value of elephant grass silages with the increasing of inclusion levels of dehydrated banana peel. The experiment was conducted in a completely randomized design, with six treatments and four replications, being the silage exclusively from elephant grass, and five levels of inclusion of banana peel to the elephant grass silage, as the following - 5; 10; 15; 20 and 25%, being added based on natural matter. The addition of the banana peel in the silage reduced linearly (p < 0.05 the pH, the ammoniacal nitrogen and the losses of the fermentative process. In addition, the inclusion of banana peel increased linearly (p < 0.05 the dry matter and non-fibrous carbohydrates. On the other hand, the neutral detergent fiber and the acid detergent fiber were linearly reduced with the inclusion of the banana peel (p < 0.05, but there was no change in the dry matter digestibility in situ. The inclusion of dehydrated banana peel in elephant grass silage reduces the losses of the fermentation process with more consistent results at the 25% inclusion level, however, it reduces the silage nutritional value due to fibrous and protein quality.

  9. Corn yield for silage and grains in different integrated crop-livestock systems

    Directory of Open Access Journals (Sweden)

    Laíse da Silveira Pontes

    Full Text Available Abstract In this study, the objective was to assess the influence of two doses of N (90 and 180 kg N ha-1, added to the winter pastures, two integrated crop-livestock systems (ICLS, with and without trees and five positions between the tree rows, on the corn (Zea mays L. quality and productivity, for silage and grain. Adopting the complete randomized block design, the treatments included three replicates. In 2006, following the 14 x 3 m spacing (currently with 158 trees ha-1 the trees were planted in 6 out of the 12 paddocks. While the corn was implemented during summer of 2013/2014, cattle grazing on the annual pasture was done during the prior winter, in both ICLS. Corn for silage was reaped at the R5 phenological stage, whereas for grains it was done at 176 days post seeding. For silage, the corn plants were grinded and then stocked in the experimental mini PVC silos. The silage varied slightly in quality along the positions between the tree rows. The differences observed between N levels in the dry matter, crude protein (CP and grain productivity are expressions of the residual effects of the winter fertilization. Silage quality was improved by the shade effect which minimized the acid detergent fiber and raised the CP, although it reduced the corn production for silage and grains by 52%. Some feasible techniques to reduce these losses are discussed.

  10. Does altered protein metabolism interfere with postmortem degradation analysis for PMI estimation?

    Science.gov (United States)

    Zissler, A; Ehrenfellner, B; Foditsch, E E; Monticelli, F C; Pittner, S

    2018-03-02

    An accurate estimation of the postmortem interval (PMI) is a central aspect in forensic routine. Recently, a novel approach based on the analysis of postmortem muscle protein degradation has been proposed. However, a number of questions remain to be answered until sensible application of this method to a broad variety of forensic cases is possible. To evaluate whether altered in vivo protein metabolism interferes with postmortem degradation patterns, we conducted a comparative study. We developed a standardized animal degradation model in rats, and collected additional muscle samples from animals recovering from muscle injury and from rats with developed disuse muscle atrophy after induced spinal cord injury. All samples were analyzed by SDS-PAGE and Western blot, labeling well-characterized muscle proteins. Tropomyosin was found to be stable throughout the investigated PMI and no alterations were detected in regenerating and atrophic muscles. In contrast, significant predictable postmortem changes occurred in desmin and vinculin protein band patterns. While no significant deviations from native patterns were detected in at-death samples of disuse muscle atrophy, interestingly, samples of rats recovering from muscle injury revealed additional desmin and vinculin degradation bands that did not occur in this form in any of the examined postmortem samples regardless of PMI. It remains to be investigated whether in vivo-altered metabolism influences postmortem degradation kinetics or if such muscle samples undergo postmortem degradation in a regular fashion.

  11. The mitochondrial translocator protein, TSPO, inhibits HIV-1 envelope glycoprotein biosynthesis via the endoplasmic reticulum-associated protein degradation pathway.

    Science.gov (United States)

    Zhou, Tao; Dang, Ying; Zheng, Yong-Hui

    2014-03-01

    The HIV-1 Env glycoprotein is folded in the endoplasmic reticulum (ER), which is necessary for viral entry and replication. Currently, it is still unclear how this process is regulated. The glycoprotein folding in the ER is controlled by the ER-associated protein degradation (ERAD) pathway, which specifically targets misfolded proteins for degradation. Previously, we reported that HIV-1 replication is restricted in the human CD4(+) T cell line CEM.NKR (NKR). To understand this mechanism, we first analyzed cellular protein expression in NKR cells and discovered that levels of the mitochondrial translocator protein TSPO were upregulated by ∼64-fold. Notably, when NKR cells were treated with TSPO antagonist PK-11195, Ro5-4864, or diazepam, HIV restriction was completely disrupted, and TSPO knockdown by short hairpin RNAs (shRNAs) achieved a similar effect. We next analyzed viral protein expression, and, interestingly, we discovered that Env expression was specifically inhibited. Both TSPO knockdown and treatment with TSPO antagonist could restore Env expression in NKR cells. We further discovered that Env proteins were rapidly degraded and that kifunensine, an ERAD pathway inhibitor, could restore Env expression and viral replication, indicating that Env proteins were misfolded and degraded through the ERAD pathway in NKR cells. We also knocked out the TSPO gene in 293T cells using CRISPR/Cas9 (clustered, regularly interspaced, short palindromic repeat [CRISPR]/CRISPR-associated-9) technology and found that TSPO could similarly inhibit Env expression in these cells. Taken together, these results demonstrate that TSPO inhibits Env protein expression through the ERAD pathway and suggest that mitochondria play an important role in regulating the Env folding process. The HIV-1 Env glycoprotein is absolutely required for viral infection, and an understanding of its expression pathway in infected cells will identify new targets for antiretroviral therapies. Env proteins

  12. Inclusion of brown midrib dwarf pearl millet silage in the diet of lactating dairy cows.

    Science.gov (United States)

    Harper, M T; Melgar, A; Oh, J; Nedelkov, K; Sanchez, G; Roth, G W; Hristov, A N

    2018-06-01

    Brown midrib brachytic dwarf pearl millet (Pennisetum glaucum) forage harvested at the flag leaf visible stage and subsequently ensiled was investigated as a partial replacement of corn silage in the diet of high-producing dairy cows. Seventeen lactating Holstein cows were fed 2 diets in a crossover design experiment with 2 periods of 28 d each. Both diets had forage:concentrate ratios of 60:40. The control diet (CSD) was based on corn silage and alfalfa haylage, and in the treatment diet, 20% of the corn silage dry matter (corresponding to 10% of the dietary dry matter) was replaced with pearl millet silage (PMD). The effects of partial substitution of corn silage with pearl millet silage on dry matter intake, milk yield, milk components, fatty acid profile, apparent total-tract digestibility of nutrients, N utilization, and enteric methane emissions were analyzed. The pearl millet silage was higher in crude protein and neutral detergent fiber and lower in lignin and starch than the corn silage. Diet did not affect dry matter intake or energy-corrected milk yield, which averaged 46.7 ± 1.92 kg/d. The PMD treatment tended to increase milk fat concentration, had no effect on milk fat yield, and increased milk urea N. Concentrations and yields of milk protein and lactose were not affected by diet. Apparent total-tract digestibility of dry matter decreased from 66.5% in CSD to 64.5% in PMD. Similarly, organic matter and crude protein digestibility was decreased by PMD, whereas neutral- and acid-detergent fiber digestibility was increased. Total milk trans fatty acid concentration was decreased by PMD, with a particular decrease in trans-10 18:1. Urinary urea and fecal N excretion increased with PMD compared with CSD. Milk N efficiency decreased with PMD. Carbon dioxide emission was not different between the diets, but PMD increased enteric methane emission from 396 to 454 g/d and increased methane yield and intensity. Substituting corn silage with brown midrib dwarf

  13. Assessment of dietary ratios of red clover and corn silages on milk production and milk quality in dairy cows.

    Science.gov (United States)

    Moorby, J M; Ellis, N M; Davies, D R

    2016-10-01

    Twenty-four multiparous Holstein-Friesian dairy cows were used in a replicated 3×3 Latin square changeover design experiment to test the effects of changing from corn (Zea mays) silage to red clover (Trifolium pratense) silage in graded proportions on feed intakes, milk production, and whole-body N and P partitioning. Three dietary treatments with ad libitum access to 1 of 3 forage mixtures plus a standard allowance of 4kg/d dairy concentrates were offered. The 3 treatment forage mixtures were, on a dry matter (DM) basis: (1) R10: 90% corn silage and 10% red clover silage, (2) R50: 50% corn silage and 50% red clover silage, and (3) R90: 10% corn silage and 90% red clover silage. In each of 3 experimental periods, there were 21d for adaptation to diets, and 7d for measurements. Diet crude protein intakes increased, and starch intakes decreased, as the silage mixture changed from 90% corn to 90% red clover, although the highest forage DM intakes and milk yields were achieved on diet R50. Although milk fat yields were unaffected by diet, milk protein yields were highest with the R 0250 diet. Whole-body partitioning of N was measured in a subset of cows (n=9), and both the daily amount and proportion of N consumed that was excreted in feces and urine increased as the proportion of red clover silage in the diet increased. However, the apparent efficiency of utilization of feed N for milk protein production decreased from 0.33g/g for diet R10 to 0.25g/g for diet R90. The urinary excretion of purine derivatives (sum of allantoin and uric acid) tended to increase, suggesting greater flow of microbial protein from the rumen, as the proportion of red clover silage in the diet increased, and urinary creatinine excretion was affected by diet. Fecal shedding of E. coli was not affected by dietary treatment. In conclusion, even though microbial protein flow may have been greatest from the R 0450 diet, optimum feed intakes and milk yields were achieved on a diet that contained a

  14. Aditivos químicos ou biológicos na ensilagem de cana-de-açúcar: 2. parâmetros ruminais e degradabilidade da matéria seca e das frações fibrosas Chemical and biological additives in sugar cane silages: 2. ruminal parameters and DM and fiber degradabilities

    Directory of Open Access Journals (Sweden)

    Patrick Schmidt

    2007-10-01

    on ruminal fermentation parameters in bovines fed with diets containing these silages. Five rumen-cannulated Nelore steers were allotted to a 5 x 5 latin square design. The steers were placed in metabolic cages and fed diets with 65% forage (%DM. Five silages were evaluated (wet basis: Control sugar cane, no additives; Urea sugar cane + 0.5% of urea; Benzoate sugar cane + 0.1% of sodium benzoate; LP - sugar cane inoculated with Lactobacillus plantarum (1 x 10(6 cfu/g forage; LB sugar cane inoculated with L. buchneri (3.6 x 10(5 cfu/g forage. During 90 days, before the animal feeding, the forage was stocked in vertical silos.Ruminal parameters were slightly affected by the silages and a strong effect of sampling time. Averages of molar concentration of acetic, propionic and butyric acids were 60.9, 19.3, and 10.2 mM, respectively. Ruminal environment proportionate by the formulated diets with sugar cane sugar silages was satisfactory and similar to traditionally observed in diets containing sugar cane. The additive used in the ensiling influenced, in a no-significant form, the ruminal degradability of DM an OM, but it did not alter the ruminal degradability of the fiber fraction. The additives applied to the sugar cane resulted in slightly alterations? for most evaluated variable. Although the ruminal degradability of silages was little affected by the additive use, the observed values were similar to the observed for the sugar cane in natura.

  15. Proteomes and Ubiquitylomes Analysis Reveals the Involvement of Ubiquitination in Protein Degradation in Petunias1

    Science.gov (United States)

    Liu, Juanxu; Wei, Qian; Wang, Rongmin; Yang, Weiyuan; Ma, Yueyue; Chen, Guoju

    2017-01-01

    Petal senescence is a complex programmed process. It has been demonstrated previously that treatment with ethylene, a plant hormone involved in senescence, can extensively alter transcriptome and proteome profiles in plants. However, little is known regarding the impact of ethylene on posttranslational modification (PTM) or the association between PTM and the proteome. Protein degradation is one of the hallmarks of senescence, and ubiquitination, a major PTM in eukaryotes, plays important roles in protein degradation. In this study, we first obtained reference petunia (Petunia hybrida) transcriptome data via RNA sequencing. Next, we quantitatively investigated the petunia proteome and ubiquitylome and the association between them in petunia corollas following ethylene treatment. In total, 51,799 unigenes, 3,606 proteins, and 2,270 ubiquitination sites were quantified 16 h after ethylene treatment. Treatment with ethylene resulted in 14,448 down-regulated and 6,303 up-regulated unigenes (absolute log2 fold change > 1 and false discovery rate petunia. Several putative ubiquitin ligases were up-regulated at the protein and transcription levels. Our results showed that the global proteome and ubiquitylome were negatively correlated and that ubiquitination could be involved in the degradation of proteins during ethylene-mediated corolla senescence in petunia. Ethylene regulates hormone signaling transduction pathways at both the protein and ubiquitination levels in petunia corollas. In addition, our results revealed that ethylene increases the ubiquitination levels of proteins involved in endoplasmic reticulum-associated degradation. PMID:27810942

  16. Silage alcohols in dairy cow nutrition

    DEFF Research Database (Denmark)

    Raun, Birgitte Marie Løvendahl

    Corn silages with high propanol concentrations has been suspected to cause reduced feed intake and health problems for dairy cows in the post-pattum transition period. With the increasing use of hetero fermentative inoculants to support corn silage fermentation it is likely that silage concentrat......Corn silages with high propanol concentrations has been suspected to cause reduced feed intake and health problems for dairy cows in the post-pattum transition period. With the increasing use of hetero fermentative inoculants to support corn silage fermentation it is likely that silage...... of alcohols will lead to high alcohol concentrations in peripheral bood for a considerable period. Increased hepatic NEFA uptake in the postpartum transition period may result in even further decreased hepatic capacity for alcohol metabolism making post-partum transition cows especially vulnerable to high...... alcohol intakes. In order to evaluate the impact of alcohol fermentation in corn silages on dairy cow performance, the main purpose of this thesis was first to investigate the concentrations and composition of alcohols in typical field corn silages, and second to study how transition and lactating dairy...

  17. New inoculants on maize silage fermentation

    Directory of Open Access Journals (Sweden)

    Fábia Giovana do Val de Assis

    2014-08-01

    Full Text Available The objective of this study was to evaluate the effect of bacterial inoculants at two inoculation rates on chemical and biological characteristics of maize silage. The treatments consisted of two inoculating rates (5 and 6 log cfu g-1 of forage for each strain of lactic acid bacteria (LAB identified as Lactobacillus buchneri, L. hilgardii, or L. plantarum. The maize was ensiled in experimental PVC silos. Samples were taken for the determination of the contents of dry matter (DM, crude protein (CP, neutral detergent fiber (NDF, water-soluble carbohydrates (WSC, organic acids and alcohols, for the evaluation of the populations of lactic acid bacteria, yeasts, filamentous fungi, and for the determination of pH values during ensilage and after 30 or 90 days of fermentation. The doses of inoculants did not promote significant differences on the evaluated characteristics. There was effect of inoculants on acetic acid, 1.2-propanediol, LAB population, filamentous fungi, and pH value. No significant influence of the treatments with inoculants was observed in the variables DM, WSC, CP, lactic acid concentrations, or ethanol. The maximum temperature, i.e., the time to achieve the maximum temperature (TMT and aerobic stability (AS, was not influencied by treatments. However, a decrease in maximum temperature, an increase in TMT, and improvement in the AS were observed after 90 days of fermentation. These results proved the advantage of microbial inoculation. The treatments influenced LAB populations and filamentous fungi, but no effect was observed on the yeast population. The best inoculation dose is 6 cfu g-1 of forage because it provides higher reduction of filamentous fungi in maize silage, thereby decreasing the aerobic deterioration by these microorganisms.

  18. Use of food wastes for the production of lactic silage

    Directory of Open Access Journals (Sweden)

    Ricardo Murray Martínez

    2012-02-01

    Full Text Available The goal of the present work was to produce lactic silage from food wastes. A factorial 2³ experimental design was applied using the following factors and levels: yogurt inoculum concentration (1 and 15%, sucrose (2 and 15% and temperature (22 and 35 ºC and as response variable, the soluble nitrogen content (SNC at the end of the fermentation was considered. The best SNC output was for the treatment with 1 % of inoculum, 2 % of sucrose and temperature of 22ºC. The increase of SNC with regards to its initial content, from 0.17 % to 1.67 % for protein contents (PC 5 % represented 263 %. It was possible to produce a lactic silage and keep it stable for up to 30 days, which was enough storage time before being sent to a drying process for future use in animal feeding or compost.

  19. Long-term aversive taste memory requires insular and amygdala protein degradation.

    Science.gov (United States)

    Rodriguez-Ortiz, Carlos J; Balderas, Israela; Saucedo-Alquicira, Fernando; Cruz-Castañeda, Paulina; Bermudez-Rattoni, Federico

    2011-03-01

    Some reports have shown that the ubiquitin-proteasome system (UPS) is necessary to degrade repressor factors to produce new proteins essential to memory consolidation. Furthermore, recent evidence suggests that memory updating also relies on protein degradation through the UPS. To evaluate whether degradation of proteins is part of the cellular events needed for long-term storage of taste aversion, we injected lactacystin--an UPS inhibitor--into the amygdala and/or insular cortex 30 min before the first or second training trials. The results revealed that degradation of proteins in either the amygdala or insular cortex suffices for long-term stabilization of first-time encounter taste aversion. On the other hand, lactacystin applied in the insula, but not in the amygdala, before the second training prevented long-term storage of updated information. Our results support that degradation of proteins by means of the UPS is required every time taste aversion is to be stored in long-term memory. Copyright © 2010 Elsevier Inc. All rights reserved.

  20. Monoubiquitination of Tob/BTG family proteins competes with degradation-targeting polyubiquitination

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, Toru, E-mail: toru@ims.u-tokyo.ac.jp [Division of Oncology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Kim, Minsoo [Division of Bacterial Infection, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Kozuka-Hata, Hiroko [Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Watanabe, Masato [Department of Medical Genome Science, School of Frontier Sciences, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa 277-8562 (Japan); Oyama, Masaaki [Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Tsumoto, Kouhei [Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Department of Medical Genome Science, School of Frontier Sciences, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa 277-8562 (Japan); Yamamoto, Tadashi, E-mail: tyamamot@ims.u-tokyo.ac.jp [Division of Oncology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Cell Signal Unit, Okinawa Institute of Science and Technology, 1919-1 Onna-son, Kunigami, Okinawa 904-0412 (Japan)

    2011-05-27

    Highlights: {yields} Tob/BTG family proteins are monoubiquitinated in the absence of E3s in vitro. {yields} Monoubiquitination sites of Tob are identified by mass spectrometry. {yields} The monoubiquitination event correlates with lower levels of polyubiquitination. -- Abstract: Tob belongs to the anti-proliferative Tob/BTG protein family. The expression level of Tob family proteins is strictly regulated both transcriptionally and through post-translational modification. Ubiquitin (Ub)/proteosome-dependent degradation of Tob family proteins is critical in controlling cell cycle progression and DNA damage responses. Various Ub ligases (E3s) are responsible for degradation of Tob protein. Here, we show that Tob family proteins undergo monoubiquitination even in the absence of E3s in vitro. Determination of the ubiquitination site(s) in Tob by mass spectrometric analysis revealed that two lysine residues (Lys48 and Lys63) located in Tob/BTG homology domain are ubiquitinated. A mutant Tob, in which both Lys48 and Lys63 are substituted with alanine, is more strongly polyubiquitinated than wild-type Tob in vivo. These data suggest that monoubiquitination of Tob family proteins confers resistance against polyubiquitination, which targets proteins for degradation. The strategy for regulating the stability of Tob family proteins suggests a novel role for monoubiquitination.

  1. Monoubiquitination of Tob/BTG family proteins competes with degradation-targeting polyubiquitination

    International Nuclear Information System (INIS)

    Suzuki, Toru; Kim, Minsoo; Kozuka-Hata, Hiroko; Watanabe, Masato; Oyama, Masaaki; Tsumoto, Kouhei; Yamamoto, Tadashi

    2011-01-01

    Highlights: → Tob/BTG family proteins are monoubiquitinated in the absence of E3s in vitro. → Monoubiquitination sites of Tob are identified by mass spectrometry. → The monoubiquitination event correlates with lower levels of polyubiquitination. -- Abstract: Tob belongs to the anti-proliferative Tob/BTG protein family. The expression level of Tob family proteins is strictly regulated both transcriptionally and through post-translational modification. Ubiquitin (Ub)/proteosome-dependent degradation of Tob family proteins is critical in controlling cell cycle progression and DNA damage responses. Various Ub ligases (E3s) are responsible for degradation of Tob protein. Here, we show that Tob family proteins undergo monoubiquitination even in the absence of E3s in vitro. Determination of the ubiquitination site(s) in Tob by mass spectrometric analysis revealed that two lysine residues (Lys48 and Lys63) located in Tob/BTG homology domain are ubiquitinated. A mutant Tob, in which both Lys48 and Lys63 are substituted with alanine, is more strongly polyubiquitinated than wild-type Tob in vivo. These data suggest that monoubiquitination of Tob family proteins confers resistance against polyubiquitination, which targets proteins for degradation. The strategy for regulating the stability of Tob family proteins suggests a novel role for monoubiquitination.

  2. The F-box protein FBXO44 mediates BRCA1 ubiquitination and degradation.

    Science.gov (United States)

    Lu, Yunzhe; Li, Jiezhi; Cheng, Dongmei; Parameswaran, Balaji; Zhang, Shaohua; Jiang, Zefei; Yew, P Renee; Peng, Junmin; Ye, Qinong; Hu, Yanfen

    2012-11-30

    BRCA1 mutations account for a significant proportion of familial breast and ovarian cancers. In addition, reduced BRCA1 protein is associated with sporadic cancer cases in these tissues. At the cellular level, BRCA1 plays a critical role in multiple cellular functions such as DNA repair and cell cycle checkpoint control. Its protein level is regulated in a cell cycle-dependent manner. However, regulation of BRCA1 protein stability is not fully understood. Our earlier study showed that the amino terminus of BRCA1 harbors a degron sequence that is sufficient and necessary for conferring BRCA1 degradation. In the current study, we used mass spectrometry to identify Skp1 that regulates BRCA1 protein stability. Small interfering RNA screening that targets all human F-box proteins uncovered FBXO44 as an important protein that influences BRCA1 protein level. The Skp1-Cul1-F-box-protein44 (SCF(FBXO44)) complex ubiquitinates full-length BRCA1 in vitro. Furthermore, the N terminus of BRCA1 mediates the interaction between BRCA1 and FBXO44. Overexpression of SCF(FBXO44) reduces BRCA1 protein level. Taken together, our work strongly suggests that SCF(FBXO44) is an E3 ubiquitin ligase responsible for BRCA1 degradation. In addition, FBXO44 expression pattern in breast carcinomas suggests that SCF(FBXO44)-mediated BRCA1 degradation might contribute to sporadic breast tumor development.

  3. The F-box Protein FBXO44 Mediates BRCA1 Ubiquitination and Degradation*

    Science.gov (United States)

    Lu, Yunzhe; Li, Jiezhi; Cheng, Dongmei; Parameswaran, Balaji; Zhang, Shaohua; Jiang, Zefei; Yew, P. Renee; Peng, Junmin; Ye, Qinong; Hu, Yanfen

    2012-01-01

    BRCA1 mutations account for a significant proportion of familial breast and ovarian cancers. In addition, reduced BRCA1 protein is associated with sporadic cancer cases in these tissues. At the cellular level, BRCA1 plays a critical role in multiple cellular functions such as DNA repair and cell cycle checkpoint control. Its protein level is regulated in a cell cycle-dependent manner. However, regulation of BRCA1 protein stability is not fully understood. Our earlier study showed that the amino terminus of BRCA1 harbors a degron sequence that is sufficient and necessary for conferring BRCA1 degradation. In the current study, we used mass spectrometry to identify Skp1 that regulates BRCA1 protein stability. Small interfering RNA screening that targets all human F-box proteins uncovered FBXO44 as an important protein that influences BRCA1 protein level. The Skp1-Cul1-F-box-protein44 (SCFFBXO44) complex ubiquitinates full-length BRCA1 in vitro. Furthermore, the N terminus of BRCA1 mediates the interaction between BRCA1 and FBXO44. Overexpression of SCFFBXO44 reduces BRCA1 protein level. Taken together, our work strongly suggests that SCFFBXO44 is an E3 ubiquitin ligase responsible for BRCA1 degradation. In addition, FBXO44 expression pattern in breast carcinomas suggests that SCFFBXO44-mediated BRCA1 degradation might contribute to sporadic breast tumor development. PMID:23086937

  4. Crude protein changes on grassland along a degradation gradient ...

    African Journals Online (AJOL)

    The aim with this investigation was to quantify the impact of different veld condition classes, viz. poor, moderate and good on soil-water utilization (SWU: crude protein produced per unit of evapotranspiration), during four growing seasons (2000/01 – 2003/2004). Evapotranspiration was determined by quantifying the ...

  5. Crude protein changes on grassland along a degradation gradient ...

    African Journals Online (AJOL)

    Evapotranspiration was determined by quantifying the soil-water balance equation with the aid of runoff plots and soil-water content measurements. Crude protein ... The study shows that it is important to keep grassland in optimal condition to utilize limited soil water for sustainable plant and therefore animal production.

  6. Short communication In situ ruminal degradability of protein feeds ...

    African Journals Online (AJOL)

    2016-12-28

    Dec 28, 2016 ... ISSN 0375-1589 (print), ISSN 2221-4062 (online) ... protein at a passage rate of 2% per hour was higher in meals than in grains. .... beef cattle reduced the consumption rate, but improved the feed conversion rate, increasing.

  7. GT Biplot Analysis for Silage Potential, Nutritive Value, Gas and Methane Production of Stay-Green Grain Sorghum Shoots

    OpenAIRE

    Kaplan, Mahmut; Arslan, Mustafa; Kale, Hasan; Kara, Kanber; Kokten, Kagan

    2017-01-01

    Abstract This study was conducted to investigate the possible silage of stay-green sorghum genotypes using GT biplot analysis. Following the grain harvest, 41 sorghum genotypes were chopped to make silage. Biochemical analyses were performed after 60 d of silage. The results revealed that green herbage yields varied between 13.40-65.96 t ha−1, pH between 3.92-4.25, dry matter ratios between 24.26-35.83%, crude protein ratios between 3.44-7.03%, acid detergent fiber (ADF) ratios between 27.46-...

  8. Composição química, digestibilidade e cinética de degradação ruminal das silagens de híbridos de girassol em diferentes épocas de semeadura Effects of sowing dates on chemical composition, digestibility and ruminal degradation kinetics of silages from sunflower hybrids

    Directory of Open Access Journals (Sweden)

    Renius Mello

    2006-08-01

    Full Text Available Objetivou-se estimar o valor nutritivo das silagens de quatro híbridos de girassol - Rumbosol-91 (forrageiro, M-734 (duplo-propósito, C-11 (duplo-propósito e BRS-191 (granífero - semeados em três épocas: outubro (antecipada, novembro (normal e dezembro (tardia - por meio de análises químicas, da digestibilidade e da cinética de degradação ruminal. O delineamento experimental foi em blocos ao acaso, com arranjo fatorial 4 x 3 (quatro híbridos x três épocas de semeadura e três repetições. Os teores de MS variaram de 23,2 a 43,0%; o pH, de 4,0 a 5,1; e o N-NH3%/NT, de 5,3 a 16,8%. Os teores de CZ oscilaram de 6,8 a 8,8% e aumentaram da semeadura em outubro para dezembro. O Rumbosol-91 apresentou teores menores de EE e maiores de N na parede celular, de N indisponível e de carboidratos totais (CHOT. Os teores de PB aumentaram e os de CHOT diminuíram de outubro para dezembro. A semeadura de novembro e o C-11 apresentaram menores teores de FDN, FDNcp e FDA. O C-11 semeado em novembro apresentou menor teor de LDA. A semeadura de outubro e o Rumbosol-91 tiveram maior proporção de LDA%/FDN. O Rumbosol-91 apresentou maior DIVMS e DIVFDN. A semeadura de novembro e os híbridos C-11 e M-734 apresentaram maior teor de NDT. O desaparecimento da MS pode ser representado por um único perfil de degradação. A proporção de FDN indegradável (I aumentou da semeadura em outubro para dezembro, com maiores valores para o BRS-191 semeado em dezembro. A extensão (Vf1 e a taxa (k1 de degradação da fração solúvel de rápida digestão diminuíram da semeadura em outubro para dezembro, com maiores valores para o Rumbosol-91 semeado em outubro. A relação entre análise química, digestibilidade, NDT e cinética de degradação ruminal foi melhor para os híbridos M-734 e C-11 semeados em novembro.The objective of this trial was to estimate chemical composition, digestibility and ruminal degradation kinetics of silages from four sunflower

  9. Precision-feeding dairy heifers a high rumen-degradable protein diet with different proportions of dietary fiber and forage-to-concentrate ratios.

    Science.gov (United States)

    Lascano, G J; Koch, L E; Heinrichs, A J

    2016-09-01

    The objective of this experiment was to determine the effects of feeding a high-rumen-degradable protein (RDP) diet when dietary fiber content is manipulated within differing forage-to-concentrate ratio (F:C) on nutrient utilization of precision-fed dairy heifers. Six cannulated Holstein heifers (486.98±15.07kg of body weight) were randomly assigned to 2 F:C, low- (45% forage; LF) and high-forage (90% forage; HF) diets and to a fiber proportion sequence [33% grass hay and wheat straw (HS), 67% corn silage (CS; low fiber); 50% HS, 50% CS (medium fiber); and 67% HS, 33% CS (high fiber)] within forage proportion administered according to a split-plot, 3×3 Latin square design (16-d periods). Heifers fed LF had greater apparent total-tract organic matter digestibility coefficients (dC), neutral detergent fiber, and cellulose than those fed LC diets. Substituting CS with HS resulted in a linear reduction in dry matter, organic matter, and cellulose dC. Nitrogen dC was not different between F:C or with increasing proportions of HS in diets, but N retention tended to decrease linearly as HS was increased in the diets. Predicted microbial protein flow to the duodenum decreased linearly with HS addition and protozoa numbers HS interacted linearly, exhibiting a decrease as HS increased for LF, whereas no effects were observed for HF. Blood urea N increased linearly as HS was incorporated. The LF-fed heifers had a greater ruminal volatile fatty acids concentration. We noted a tendency for a greater dry matter, and a significantly higher liquid fraction turnover rate for HF diets. There was a linear numerical increase in the liquid and solid fraction turnover rate as fiber was added to the diets. Rumen fermentation parameters and fractional passages (solid and liquid) rates support the reduction in dC, N retention, and microbial protein synthesis observed as more dietary fiber is added to the rations of dairy heifers precision-fed a constant proportion of rumen-degradable

  10. Hydrolytic bacteria in mesophilic and thermophilic degradation of plant biomass

    Energy Technology Data Exchange (ETDEWEB)

    Zverlov, Vladimir V.; Hiegl, Wolfgang; Koeck, Daniela E.; Koellmeier, Tanja; Schwarz, Wolfgang H. [Department of Microbiology, Technische Universitaet Muenchen, Freising-Weihenstephan (Germany); Kellermann, Josef [Max Planck Institute for Biochemistry, Am Klopferspitz, Martinsried (Germany)

    2010-12-15

    Adding plant biomass to a biogas reactor, hydrolysis is the first reaction step in the chain of biological events towards methane production. Maize silage was used to enrich efficient hydrolytic bacterial consortia from natural environments under conditions imitating those in a biogas plant. At 55-60 C a more efficient hydrolyzing culture could be isolated than at 37 C. The composition of the optimal thermophilic bacterial consortium was revealed by sequencing clones from a 16S rRNA gene library. A modified PCR-RFLP pre-screening method was used to group the clones. Pure anaerobic cultures were isolated. 70% of the isolates were related to Clostridium thermocellum. A new culture-independent method for identification of cellulolytic enzymes was developed using the isolation of cellulose-binding proteins. MALDI-TOF/TOF analysis and end-sequencing of peptides from prominent protein bands revealed cellulases from the cellulosome of C. thermocellum and from a major cellulase of Clostridium stercorarium. A combined culture of C. thermocellum and C. stercorarium was shown to excellently degrade maize silage. A spore preparation method suitable for inoculation of maize silage and optimal hydrolysis was developed for the thermophilic bacterial consortium. This method allows for concentration and long-term storage of the mixed culture for instance for inoculation of biogas fermenters. (Copyright copyright 2010 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  11. Intake and digestibility of sorghum (Sorghum bicolor, L. Moench silages with different tannin contents in sheep

    Directory of Open Access Journals (Sweden)

    Alex de Matos Teixeira

    2014-01-01

    Full Text Available The purpose of this study was to evaluate the voluntary intake and digestibility of three sorghum (Sorghum bicolor, L. Moench hybrid silages in sheep. The hybrids used were H1 -BRS 655 (CMSXS 222 A × CMSXS 235 R, with tannin; H2 -(ATF54 A × CMSXS 235 R, without tannin; and H3 -BRS 610 (CMSXS 232 A × CMSXS 234 R, without tannin. The intake and digestibility of dry matter (DM, gross energy (GE, neutral detergent fiber (NDF, acid detergent fiber (ADF and crude protein (CP were measured. Eighteen crossbred sheep weighing 59.4 kg (±8.3 were used in the trial. A completely randomized design with three treatments (hybrids and six repetitions (sheep was used. There were no differences in the DM intake or apparent digestibility among the hybrids. Silage of hybrid BRS 610 displayed higher digestibility coefficients for CP, NDF, ADF, and GE compared with the other silages, which did not differ from each other. The neutral detergent fiber, ADF and digestible energy (DE intakes were similar among the hybrids silages. All of the hybrids resulted in a positive N balance in sheep. The levels of DE were superior in hybrid silage BRS 610 in comparison with the other hybrids. Sorghum hybrid BRS 610 silage exhibited superior nutritional value compared with the other hybrids, which is most likely in part due to the absence of tannins. Sorghum silage made with hybrid BRS 610 (CMSXS 232 A × CMSXS 234 R presents superior gross energy, crude protein, neutral detergent fiber and acid detergent fiber digestibility coefficients, as well as greater digestible energy levels than BRS 655 (CMSXS 222 A × CMSXS 235 R and (ATF54 A × CMSXS 235 R.

  12. Identifiability study of the proteins degradation model, based on ADM1, using simultaneous batch experiments

    DEFF Research Database (Denmark)

    Flotats, X.; Palatsi, J.; Ahring, Birgitte Kiær

    2006-01-01

    are not inhibiting the hydrolysis process. The ADM1 model adequately expressed the consecutive steps of hydrolysis and acidogenesis, with estimated kinetic values corresponding to a fast acidogenesis and slower hydrolysis. The hydrolysis was found to be the rate limiting step of anaerobic degradation. Estimation...... of yield coefficients based on the relative initial slopes of VFA profiles obtained in a simple batch experiment produced satisfactory results. From the identification study, it was concluded that it is possible to determine univocally the related kinetic parameter values for protein degradation...... if the evolution of amino acids is measured in simultaneous batch experiments, with different initial protein and amino acids concentrations....

  13. Milk production is unaffected by replacing barley or sodium hydroxide wheat with maize cob silage in rations for dairy cows

    DEFF Research Database (Denmark)

    Hymøller, Lone; Hellwing, Anne Louise Frydendahl; Lund, Peter

    2014-01-01

    . The energy-corrected milk yield was unaffected by treatment. The fat content of the milk on the MCS ration was not different from the SHW ration, whereas it was higher on the barley ration. The protein content of the milk decreased when MCS was used in the ration compared with barley and SHW. From ruminal......Starch is an important energy-providing nutrient for dairy cows that is most commonly provided from cereal grains. However, ruminal fermentation of large amounts of easily degradable starch leads to excessive production and accumulation of volatile fatty acids (VFA). VFA not only play a vital role...... in the energy metabolism of dairy cows but are also the main cause of ruminal acidosis and depressed feed intake. The aim of the present study was to compare maize cob silage (MCS) as an energy supplement in rations for dairy cows with highly rumen-digestible rolled barley and with sodium hydroxide wheat (SHW...

  14. Rumen volatile fatty acids and milk composition from cows fed hay, haylage, or urea-treated corn silage.

    Science.gov (United States)

    Schingoethe, D J; Voelker, H H; Beardsley, G L; Parsons, J G

    1976-05-01

    Alfalfa-brome hay, haylage, .5% urea-treated corn silage, or .5% urea plus 1% dried whey-treated corn silage was fed as the only forage to one of four groups of 10 lactating cows per group for a lactation trial of 10 wk. Rumen samples were collected via stomach tube 3 to 4 h after the morning feeding. The pH of the rumen samples from cows fed hay was higher than for cows fed haylage, urea-treated corn silage, and urea-whey corn silage, 6.69 versus 6.36, 6.40, and 6.50. Total volatile fatty acids and propionate were highest from cows fed urea-whey corn silage and were higher on all three fermented forages than cows fed hay. Acetate/propionate ratio was highest from cows fed hay and lowest from cows fed corn silages. Butyrate was highest from cows fed haylage or hay. Milk protein composition was not affected by ration although nonprotein nitrogen of milk was highest from cows fed the urea-treated corn silages. Oleic acid and total unsaturated fatty acids were lowest in milk fat from cows fed hay while palmitic acid was highest from cows fed hay and haylage. These results suggest that type of forage fed may cause small changes in rumen fermentation and in milk composition. The importance of these changes is unknown but may affect properties of dairy products produced from this milk.

  15. Methane production, nutrient digestion, ruminal fermentation, N balance, and milk production of cows fed timothy silage- or alfalfa silage-based diets.

    Science.gov (United States)

    Hassanat, F; Gervais, R; Massé, D I; Petit, H V; Benchaar, C

    2014-10-01

    The objective of this study was to investigate the effects of changing forage source in dairy cow diets from timothy silage (TS) to alfalfa silage (AS) on enteric CH₄ emissions, ruminal fermentation characteristics, digestion, milk production, and N balance. Nine ruminally cannulated lactating cows were used in a replicated 3 × 3 Latin square design (32-d period) and fed (ad libitum) a total mixed ration (TMR; forage:concentrate ratio of 60:40, dry matter basis), with the forage portion consisting of either TS (0% AS; 0% AS and 54.4% TS in the TMR), a 50:50 mixture of both silages (50% AS; 27.2% AS and 27.2% TS in the TMR), or AS (100% AS; 54.4% AS and 0% TS in the TMR). Compared with TS, AS contained less (36.9 vs. 52.1%) neutral detergent fiber but more (20.5 vs. 13.6%) crude protein (CP). In sacco 24-h ruminal degradability of organic matter (OM) was higher for AS than for TS (73.5 vs. 66.9%). Replacement of TS with AS in the diet entailed increasing proportions of corn grain and bypass protein supplement at the expense of soybean meal. As the dietary proportion of AS increased, CP and starch concentrations increased, whereas fiber content declined in the TMR. Dry matter intake increased linearly with increasing AS proportions in the diet. Apparent total-tract digestibility of OM and gross energy remained unaffected, whereas CP digestibility increased linearly and that of fiber decreased linearly with increasing inclusion of AS in the diet. The acetate-to-propionate ratio was not affected, whereas ruminal concentration of ammonia (NH₃) and molar proportion of branched-chain VFA increased as the proportion of AS in the diet increased. Daily CH₄ emissions tended to increase (476, 483, and 491 g/d for cows fed 0% AS, 50% AS, and 100% AS, respectively) linearly as cows were fed increasing proportions of AS. Methane production adjusted for dry matter intake (average=19.8 g/kg) or gross energy intake (average=5.83%) was not affected by increasing AS inclusion

  16. ESCRT-dependent degradation of ubiquitylated plasma membrane proteins in plants.

    Science.gov (United States)

    Isono, Erika; Kalinowska, Kamila

    2017-12-01

    To control the abundance of plasma membrane receptors and transporters is crucial for proper perception and response to extracellular signals from surrounding cells and the environment. Posttranslational modification of plasma membrane proteins, especially ubiquitin conjugation or ubiquitylation, is key for the determination of stability for many transmembrane proteins localized on the cell surface. The targeted degradation is ensured by a complex network of proteins among which the endosomal sorting complex required for transport (ESCRT) plays a central role. This review focuses on progresses made in recent years on the understanding of the function of the ESCRT machinery in the degradation of ubiquitylated plasma membrane proteins in plants. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Iron-Binding Protein Degradation by Cysteine Proteases of Naegleria fowleri

    Directory of Open Access Journals (Sweden)

    Moisés Martínez-Castillo

    2015-01-01

    Full Text Available Naegleria fowleri causes acute and fulminant primary amoebic meningoencephalitis. This microorganism invades its host by penetrating the olfactory mucosa and then traveling up the mesaxonal spaces and crossing the cribriform plate; finally, the trophozoites invade the olfactory bulbs. During its invasion, the protozoan obtains nutrients such as proteins, lipids, carbohydrates, and cationic ions (e.g., iron, calcium, and sodium from the host. However, the mechanism by which these ions are obtained, particularly iron, is poorly understood. In the present study, we evaluated the ability of N. fowleri to degrade iron-binding proteins, including hololactoferrin, transferrin, ferritin, and hemoglobin. Zymography assays were performed for each substrate under physiological conditions (pH 7 at 37°C employing conditioned medium (CM and total crude extracts (TCEs of N. fowleri. Different degradation patterns with CM were observed for hololactoferrin, transferrin, and hemoglobin; however, CM did not cause ferritin degradation. In contrast, the TCEs degraded only hololactoferrin and transferrin. Inhibition assays revealed that cysteine proteases were involved in this process. Based on these results, we suggest that CM and TCEs of N. fowleri degrade iron-binding proteins by employing cysteine proteases, which enables the parasite to obtain iron to survive while invading the central nervous system.

  18. Molecular chaperones in targeting misfolded proteins for ubiquitin-dependent degradation

    DEFF Research Database (Denmark)

    Kriegenburg, Franziska; Ellgaard, Lars; Hartmann-Petersen, Rasmus

    2012-01-01

    The accumulation of misfolded proteins presents a considerable threat to the health of individual cells and has been linked to severe diseases, including neurodegenerative disorders. Considering that, in nature, cells often are exposed to stress conditions that may lead to aberrant protein...... conformational changes, it becomes clear that they must have an efficient quality control apparatus to refold or destroy misfolded proteins. In general, cells rely on molecular chaperones to seize and refold misfolded proteins. If the native state is unattainable, misfolded proteins are targeted for degradation...... via the ubiquitin-proteasome system. The specificity of this proteolysis is generally provided by E3 ubiquitin-protein ligases, hundreds of which are encoded in the human genome. However, rather than binding the misfolded proteins directly, most E3s depend on molecular chaperones to recognize...

  19. Utilization of kura clover-reed canarygrass silage versus alfalfa silage by lactating dairy cows.

    Science.gov (United States)

    Kammes, K L; Heemink, G B H; Albrecht, K A; Combs, D K

    2008-08-01

    The mixture of kura clover (Trifolium ambiguum M. Bieb.) and reed canarygrass (Phalaris arundinacea L.) has proven to be extremely persistent in the northern United States, but information about dairy cow performance on this mixture is lacking. Twenty lactating Holstein cows were used in a crossover design to compare dry matter (DM) intake and milk production from diets containing kura clover-reed canarygrass silage (KRS) or alfalfa (Medicago sativa L.) silage (AS). Forages were cut, wilted, ensiled in horizontal plastic bags, and allowed to ferment for at least 50 d before beginning the feeding experiment. The KRS was approximately 40% kura clover and 60% reed canarygrass. Treatments were total mixed rations formulated with either 57% of total DM from 1) AS or 2) KRS. Experimental periods were 28 d, with the first 14 d for diet adaptation and the last 14 d for measurement of intake and milk production. The neutral detergent fiber (NDF) concentrations of AS and KRS were 37.3 and 47.3%, respectively. The fermentation analyses indicated that both silages underwent a restricted fermentation, producing primarily lactic acid and some acetic acid. Dry matter intake (24.2 vs. 22.8 kg) and 4% fat-corrected milk (32.8 vs. 30.9 kg) were significantly higher for cows fed AS than for cows fed KRS. Cows consumed less NDF (6.7 vs. 8.0 kg) and less digestible NDF (3.0 vs. 4.4 kg) when fed AS diets compared with KRS diets, but the pool of ruminally undegraded NDF was similar (3.7 kg) between diets. Cows produced 1.5 kg of milk/kg of DM consumed regardless of the diet, indicating that digestible NDF of KRS was utilized with similar efficiency as the cell wall constituents of AS, but the intake of cows fed KRS may have been limited by rumen fill. Milk fat concentration tended to be higher for cows fed AS, but the milk true protein concentration and yields of fat and protein did not differ by treatment. Milk urea nitrogen content was higher when cows consumed AS (16.4 mg/ d

  20. Reduced synaptic vesicle protein degradation at lysosomes curbs TBC1D24/sky-induced neurodegeneration.

    Science.gov (United States)

    Fernandes, Ana Clara; Uytterhoeven, Valerie; Kuenen, Sabine; Wang, Yu-Chun; Slabbaert, Jan R; Swerts, Jef; Kasprowicz, Jaroslaw; Aerts, Stein; Verstreken, Patrik

    2014-11-24

    Synaptic demise and accumulation of dysfunctional proteins are thought of as common features in neurodegeneration. However, the mechanisms by which synaptic proteins turn over remain elusive. In this paper, we study Drosophila melanogaster lacking active TBC1D24/Skywalker (Sky), a protein that in humans causes severe neurodegeneration, epilepsy, and DOOR (deafness, onychdystrophy, osteodystrophy, and mental retardation) syndrome, and identify endosome-to-lysosome trafficking as a mechanism for degradation of synaptic vesicle-associated proteins. In fly sky mutants, synaptic vesicles traveled excessively to endosomes. Using chimeric fluorescent timers, we show that synaptic vesicle-associated proteins were younger on average, suggesting that older proteins are more efficiently degraded. Using a genetic screen, we find that reducing endosomal-to-lysosomal trafficking, controlled by the homotypic fusion and vacuole protein sorting (HOPS) complex, rescued the neurotransmission and neurodegeneration defects in sky mutants. Consistently, synaptic vesicle proteins were older in HOPS complex mutants, and these mutants also showed reduced neurotransmission. Our findings define a mechanism in which synaptic transmission is facilitated by efficient protein turnover at lysosomes and identify a potential strategy to suppress defects arising from TBC1D24 mutations in humans. © 2014 Fernandes et al.

  1. Consumo e digestibilidade aparente da matéria seca, proteína e energia bruta, e balanço de nitrogênio das silagens de cinco genótipos de milho Consumption and apparent digestibility of dry matter, crude protein and crude energy, and balance of nitrogen of silages of five maize genotypes

    Directory of Open Access Journals (Sweden)

    G.A.R. Freitas

    2003-08-01

    Full Text Available Quantificaram-se o consumo voluntário e a digestibilidade aparente da matéria seca, proteína bruta, energia bruta e balanço de nitrogênio das silagens de cinco genótipos de milho (HT01, HT47C, HT129, AG 5011 e BR 3123. Foram utilizados 15 carneiros alojados em gaiolas metabólicas para coleta total de fezes e urina. O delineamento experimental foi o inteiramente ao acaso com cinco tratamentos e seis repetições. Não foram observadas diferenças entre os genótipos quanto ao consumo e digestibilidade da MS, da EB e da PB (P>0,05. Os consumos de MS, EB e PB digestíveis e energia metabolizável também não foram diferentes entre os híbridos (P>0,05. Quanto às relações consumo de energia digestível/consumo de MS e consumo de energia metabolizável/consumo de MS, o genótipo AG5011 foi semelhante ao HT01 (P>0,05 e superior aos demais (P0,05. Todos os genótipos produziram silagens de bom valor nutritivo, entretanto o genótipo AG5011 apresentou maior eficiência na utilização da energia (PThe voluntary intake and the apparent digestibility of dry matter, crude protein and crude energy and the nitrogen balance, of silages of five maize genotypes (HT01, HT47C, HT129, AG5011 and BR3123 were quantified. Fifteen sheep were stored in metabolic cages for total collection of feces and urine. A complete randomized design, with five treatments and six repetitions was used. The intake and digestibility of dry matter, crude protein and crude energy did not differ (P>0.05 among treatments. The intake of digestible dry matter, crude protein, crude energy and metabolic energy did not differ (P>0.05 among the genotypes. The ratios of digestible energy intake/dry matter intake and metabolic energy intake/dry matter intake of the genotype AG5011 silage was similar (P>0.05 to HT01, and higher than the other genotypes (P<0.05. All genotypes showed similar and positive nitrogen balance, and all of them produced silages of good nutritional value

  2. Desempenho de bezerros de corte alimentados com diferentes fontes protéicas associadas à silagem de sorgo colhida em duas alturas de corte Performance of beef calves fed with different protein sources associated with silage of sorghum harvested in two different heights

    Directory of Open Access Journals (Sweden)

    Fernando Miranda Vargas Júnior

    2003-06-01

    Full Text Available Foram avaliadas três fontes protéicas (uréia - U, farinha penas - Fp e farelo soja - Fs associadas a duas silagens de sorgo (silagens de sorgo corte alto - SCA e corte baixo - SCB por intermédio do desempenho de bezerros de corte, confinados por 84 dias. Foram utilizados 36 bezerros mestiços (Charolês/RedAngus/Nelore, castrados, com cinco meses de idade e pesando em média 146,44 kg. Avaliaram-se o ganho de peso médio diário (GMD; o consumo de matéria seca expressos em kg/dia (CMS, percentagem de peso vivo (CMSPV e peso metabólico (CMSPM; e a conversão alimentar (CA. O delineamento foi inteiramente casualizado por intermédio de um fatorial 3x2. Houve efeito significativo na interação silagem versus fonte protéica para a variável GMD, com o maior GMD para SCAFp (1,61 kg/animal/dia versus a SCBFs (1,32 kg/animal/dia. Os demais parâmetros não diferiram estatisticamente quando analisada a interação ou isoladamente a silagem ou fonte protéica. A silagem de sorgo corte alto promoveu numericamente um maior GMD e CMS.Three protein sources (urea - U; feather meal - Fm; soybean meal - Sm associated to two sorghum silage (sorghum at high cutting - SHC; and sorghum at low cutting - SLC were evaluated by measuring beef calf performance, confined for 84 days. Thirty-six crossbred (Charolês/RedAngus/Nelore castrated calves, with five months of age and average weight alive on the 146.44 kg, were used. Average daily weight gain (DWG; the dry matter intake expressed in kg/day (DMI, percentage live weight (DMILW and percentage of metabolic weight (DMIMW and the feed : gain ratio (FGR were evaluated. The treatments followed a 3x2 factorial in a completely randomized design. There was significant effect in the interaction silage versus protein source for variable DWG, with the higher DWG for SHCFm (1.61 kg/animal/day versus SLCSm (1.32 kg/animal/day. The other parameters did not differ when the interaction or separately the silage or protein

  3. Protein degradation in preimplantation mouse embryos and the lethality of tritiated amino acids

    International Nuclear Information System (INIS)

    Wielbold, J.L.

    1982-01-01

    The role of protein degradation in preimplantation development in the mouse was studied. Proteins of morulae and blastocysts (M and B) cultured in vitro after labeling for 1 hour (h) in 3 H-leucine exhibit a mean half-life (t 1 / 2 ) of 8.1 h. The t 1 / 2 tends to increase (9.5 h) when 10% fetal calf serum is added to the chase medium. This decrease in protein degradation in the presence of serum is associated with an increase in the percentage of B that are hatching (P 3 H-leucine in their proteins than did Day 4 embryos remaining in culture (P<0.02), while Day 4 embryos in a Day 3 uterus retained the same amount of radioactivity as did Day 4 embryos in culture. This differential effect of uterine environment was also seen when Day 4 embryos were transferred to recipients. More fetuses developed to term when the recipient was in Day 3 of PSP (50.8%) than when the recipient was in Day 4 PSP (25.9%, P<0.001), regardless of the age of the recipient. Age of the recipient does affect the percentage of transferred embryos developing to term. Thus, protein degradation may vary with the stage of embryo development and the conditions to which the embryos are exposed. However, even low levels of incorporated tritiated leucine can have lethal effects on the embryos and compromise the validity of the protein half-lives determined

  4. Cellular Cholesterol Regulates Ubiquitination and Degradation of the Cholesterol Export Proteins ABCA1 and ABCG1*

    Science.gov (United States)

    Hsieh, Victar; Kim, Mi-Jurng; Gelissen, Ingrid C.; Brown, Andrew J.; Sandoval, Cecilia; Hallab, Jeannette C.; Kockx, Maaike; Traini, Mathew; Jessup, Wendy; Kritharides, Leonard

    2014-01-01

    The objective of this study was to examine the influence of cholesterol in post-translational control of ABCA1 and ABCG1 protein expression. Using CHO cell lines stably expressing human ABCA1 or ABCG1, we observed that the abundance of these proteins is increased by cell cholesterol loading. The response to increased cholesterol is rapid, is independent of transcription, and appears to be specific for these membrane proteins. The effect is mediated through cholesterol-dependent inhibition of transporter protein degradation. Cell cholesterol loading similarly regulates degradation of endogenously expressed ABCA1 and ABCG1 in human THP-1 macrophages. Turnover of ABCA1 and ABCG1 is strongly inhibited by proteasomal inhibitors and is unresponsive to inhibitors of lysosomal proteolysis. Furthermore, cell cholesterol loading inhibits ubiquitination of ABCA1 and ABCG1. Our findings provide evidence for a rapid, cholesterol-dependent, post-translational control of ABCA1 and ABCG1 protein levels, mediated through a specific and sterol-sensitive mechanism for suppression of transporter protein ubiquitination, which in turn decreases proteasomal degradation. This provides a mechanism for acute fine-tuning of cholesterol transporter activity in response to fluctuations in cell cholesterol levels, in addition to the longer term cholesterol-dependent transcriptional regulation of these genes. PMID:24500716

  5. The Polerovirus silencing suppressor P0 targets ARGONAUTE proteins for degradation.

    Science.gov (United States)

    Baumberger, Nicolas; Tsai, Ching-Hsui; Lie, Miranda; Havecker, Ericka; Baulcombe, David C

    2007-09-18

    Plant and animal viruses encode suppressor proteins of an adaptive immunity mechanism in which viral double-stranded RNA is processed into 21-25 nt short interfering (si)RNAs. The siRNAs guide ARGONAUTE (AGO) proteins so that they target viral RNA. Most viral suppressors bind long dsRNA or siRNAs and thereby prevent production of siRNA or binding of siRNA to AGO. The one exception is the 2b suppressor of Cucumoviruses that binds to and inhibits AGO1. Here we describe a novel suppressor mechanism in which a Polerovirus-encoded F box protein (P0) targets the PAZ motif and its adjacent upstream sequence in AGO1 and mediates its degradation. F box proteins are components of E3 ubiquitin ligase complexes that add polyubiquitin tracts on selected lysine residues and thereby mark a protein for proteasome-mediated degradation. With P0, however, the targeted degradation of AGO is insensitive to inhibition of the proteasome, indicating that the proteasome is not involved. We also show that P0 does not block a mobile signal of silencing, indicating that the signal molecule does not have AGO protein components. The ability of P0 to block silencing without affecting signal movement may contribute to the phloem restriction of viruses in the Polerovirus group.

  6. Fabrication of Flexible, Fully Organic, Degradable Energy Storage Devices Using Silk Proteins.

    Science.gov (United States)

    Pal, Ramendra K; Kundu, Subhas C; Yadavalli, Vamsi K

    2018-03-21

    Flexible and thin-film devices are of great interest in epidermal and implantable bioelectronics. The integration of energy storage and delivery devices such as supercapacitors (SCs) with properties such as flexibility, miniaturization, biocompatibility, and degradability are sought for such systems. Reducing e-waste and using sustainable materials and processes are additional desirable qualities. Herein, a silk protein-based biocompatible and degradable thin-film microSC (μSC) is reported. A protein carrier with the conducting polymer poly(3,4-ethylenedioxythiophene) polystyrene sulfonate and reduced graphene oxide dopant is used as a photopatternable biocomposite ink. Active electrodes are fabricated using photolithography under benign conditions, using only water as the solvent. These electrodes are printed on flexible protein sheets to form degradable, organic devices with a benign agarose-NaCl gel electrolyte. High capacitance, power density, cycling stability over 500 cycles, and the ability to power a light-emitting diode are shown. The device is flexible, can sustain cyclic mechanical stresses over 450 cycles, and retain capacitive properties over several days in liquid. Significantly, the μSCs are cytocompatible and completely degraded over the period of ∼1 month. By precise control of the device configuration, these silk protein-based, all-polymer organic devices can be designed to be tunably transient and provide viable alternatives for powering flexible and implantable bioelectronics.

  7. In situ ruminal crude protein degradability of by-products from cereals, oilseeds and animal origin

    NARCIS (Netherlands)

    Habib, G.; Khan, N.A.; Ali, M.; Bezabih, M.

    2013-01-01

    The aim of this study was to establish a database on in situ ruminal crude protein (CP) degradability characteristics of by-products from cereal grains, oilseeds and animal origin commonly fed to ruminants in Pakistan and South Asian Countries. The oilseed by-products were soybean meal, sunflower

  8. Delaying aging and the aging-associated decline in protein homeostasis by inhibition of tryptophan degradation

    NARCIS (Netherlands)

    van der Goot, Annemieke T.; Zhu, Wentao; Vazquez-Manrique, Rafael P.; Seinstra, Renee I.; Dettmer, Katja; Michels, Helen; Farina, Francesca; Krijnen, Jasper; Melki, Ronald; Buijsman, Rogier C.; Silva, Mariana Ruiz; Thijssen, Karen L.; Kema, Ido P.; Neri, Christian; Oefner, Peter J.; Nollen, Ellen A. A.

    2012-01-01

    Toxicity of aggregation-prone proteins is thought to play an important role in aging and age-related neurological diseases like Parkinson and Alzheimer's diseases. Here, we identify tryptophan 2,3-dioxygenase (tdo-2), the first enzyme in the kynurenine pathway of tryptophan degradation, as a

  9. Degradation and de novo synthesis of D1 protein and psbA ...

    Indian Academy of Sciences (India)

    This shows that synthesis of D1 protein is not the only component involved in the recovery process. Our events, which ... transcript levels in the green alga Chlamydomonas reinhardtii in ..... and Gaba V 1996 Accelerated degradation of the D2 ...

  10. Differential expression in Phanerochaete chrysosporium of membrane- associated proteins relevant to lignin degradation

    Science.gov (United States)

    Semarjit Shary; Alexander N. Kapich; Ellen A. Panisko; Jon K. Magnuson; Daniel Cullen; Kenneth E. Hammel

    2008-01-01

    Fungal lignin-degrading systems likely include membrane-associated proteins that participate in diverse processes such as uptake and oxidation of lignin fragments, production of ligninolytic secondary metabolites, and defense of the mycelium against ligninolytic oxidants. Little is known about the nature or regulation of these membrane-associated components. We grew...

  11. Impact of adding Saccharomyces strains on fermentation, aerobic stability, nutritive value, and select lactobacilli populations in corn silage.

    Science.gov (United States)

    Duniere, L; Jin, L; Smiley, B; Qi, M; Rutherford, W; Wang, Y; McAllister, T

    2015-05-01

    Bacterial inoculants can improve the conservation and nutritional quality of silages. Inclusion of the yeast Saccharomyces in the diet of dairy cattle has also been reported to be beneficial. The present study assessed the ability of silage to be used as a means of delivering Saccharomyces strains to ruminants. Two strains of Saccharomyces cerevisiae (strain 1 and 3)and 1 strain of Saccharomyces paradoxus (strain 2) were inoculated (10(3) cfu/g) individually onto corn forage that was ensiled in mini silos for 90 d. Fermentation characteristics, aerobic stability, and nutritive value of silages were determined and real-time quantitative PCR (RT-qPCR) was used to quantify S. cerevisiae, S.paradoxus, total Saccharomyces, fungal, and bacterial populations. Fermentation characteristics of silage inoculated with S1 were similar to control silage. Although strain 3 inoculation increased ash and decreased OM contents of silage (P = 0.017), no differences were observed in nutrient composition or fermentation profiles after 90 d of ensiling. Inoculation with Saccharomyces had no detrimental effect on the aerobic stability of silage. In vitro DM disappearance, gas production, and microbial protein synthesis were not affected by yeast inoculation.Saccharomyces strain 1 was quantified throughout ensiling, whereas strain 2 was detected only immediately after inoculation. Saccharomyces cerevisiae strain 3 was quantified until d 7 and detectable 90 d after ensiling. All inoculants were detected and quantified during aerobic exposure. Inoculation with Saccharomyces did not alter lactobacilli populations. Saccharomycetales were detected by RT-qPCR throughout ensiling in all silages. Both S. cerevisiae and S. paradoxus populations increased during aerobic exposure, demonstrating that the density of these yeast strains would increase between the time that silage was removed from storage and the time it was fed.

  12. Ruminal Degradation Characteristics and Small Intestinal Digestibility of Rumen Undegraded Protein of Six Feed Ingredients%6种饲料原料瘤胃降解特性和瘤胃非降解蛋白质的小肠消化率

    Institute of Scientific and Technical Information of China (English)

    赵连生; 牛俊丽; 徐元君; 王芳; 郑琛; 李发第; 郭江鹏; 卜登攀

    2017-01-01

    This experiment was conducted to determine the ruminal degradation characteristics and small intestinal digestibility of rumen undegraded protein (RUP) of six feed ingredients for dairy cows from Xinjiang, including corn silage, cottonseed hulls, alfalfa meal, alfalfa hay, grape seed meal and tomato sauce residue.Three lactating Holstein cows fitted with permanent rumen fistulas were selected to estimate the ruminal degradation characteristics of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) and small intestinal digestibility and intestinal digestible crude protein (IDCP) content of RUP by nylon-bag technique and modified three-step in vitro method.The results showed as follows:1) DM effective degradability of alfalfa meal and corn silage was higher, which was significantly higher than that of alfalfa hay and tomato sauce residue (P alfalfa meal > corn silage > alfalfa hay > cottonseed hulls > grape seed meal with significant differences among feed ingredients (P alfalfa meal > cottonseed hulls > alfalfa hay > tomato sauce residue > grape seed meal, and the differences among feed ingredients were significant (P cottonseed hulls > alfalfa hay > tomato sauce residue > alfalfa meal > grape seed meal with significant differences among feed ingredients (P苜蓿草粉>玉米青贮>苜蓿干草>棉籽壳>葡萄籽粕,各原料间差异显著(P苜蓿草粉>棉籽壳>苜蓿干草>番茄酱渣>葡萄籽粕,各组饲料原料间差异显著(P棉籽壳>苜蓿干草>番茄酱渣>苜蓿草粉>葡萄籽粕,各组饲料原料间差异显著(P0.05),显著高于依次降低的玉米青贮、番茄酱渣、葡萄籽粕、棉籽壳(P<0.05).综上所述,不同饲料原料具有不同的瘤胃降解特性,进入小肠IDCP的含量也不同.玉米青贮的DM、NDF和ADF在瘤胃的有效降解率较高,苜蓿草粉RUP的Idg较高,苜蓿草粉和苜蓿干草的IDCP含量较高.

  13. ERManI (Endoplasmic Reticulum Class I α-Mannosidase) Is Required for HIV-1 Envelope Glycoprotein Degradation via Endoplasmic Reticulum-associated Protein Degradation Pathway.

    Science.gov (United States)

    Zhou, Tao; Frabutt, Dylan A; Moremen, Kelley W; Zheng, Yong-Hui

    2015-09-04

    Previously, we reported that the mitochondrial translocator protein (TSPO) induces HIV-1 envelope (Env) degradation via the endoplasmic reticulum (ER)-associated protein degradation (ERAD) pathway, but the mechanism was not clear. Here we investigated how the four ER-associated glycoside hydrolase family 47 (GH47) α-mannosidases, ERManI, and ER-degradation enhancing α-mannosidase-like (EDEM) proteins 1, 2, and 3, are involved in the Env degradation process. Ectopic expression of these four α-mannosidases uncovers that only ERManI inhibits HIV-1 Env expression in a dose-dependent manner. In addition, genetic knock-out of the ERManI gene MAN1B1 using CRISPR/Cas9 technology disrupts the TSPO-mediated Env degradation. Biochemical studies show that HIV-1 Env interacts with ERManI, and between the ERManI cytoplasmic, transmembrane, lumenal stem, and lumenal catalytic domains, the catalytic domain plays a critical role in the Env-ERManI interaction. In addition, functional studies show that inactivation of the catalytic sites by site-directed mutagenesis disrupts the ERManI activity. These studies identify ERManI as a critical GH47 α-mannosidase in the ER-associated protein degradation pathway that initiates the Env degradation and suggests that its catalytic domain and enzymatic activity play an important role in this process. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Estimation of Relationship Between In Situ and In Vitro Rumen Protein Degradability of Extruded Full Fat Soybean

    Directory of Open Access Journals (Sweden)

    Arzu Erol Tunç

    2017-10-01

    Full Text Available The objectives of this study were to estimate the protein degradability of extruded full fat soybean (ESB by in situ (nylon bag and in vitro enzymatic method and to develop an equation in order predict in situ degradability from in vitro values. In the study enzymatic technique; hydrolysis after 1 h (INV1 and after 24 h (INV24 by a purified protease extracted from Streptomyces griseus in a borate-phosphate buffer at pH 8 was used as in vitro method. Relationship between in situ effective protein degradability (INSE and in vitro degradability after 1 and 24 hours incubations (INV1 and INV24 were determined. In situ protein degradability was measured at 0, 2, 4, 8, 16, 24, and 48 and at 72 h incubations in the rumen of 3 Holstein cows. In the study INSE, INV1 and INV24 were determined as 58.05, 20.24 and 41.46% respectively. Despite there were differences between in situ and in vitro protein degradability values, correlation coefficients between in situ and in vitro protein degradability of ESB were high and regression equations for estimation of in situ from in vitro were found significant. As conclusion in vitro enzymatic protein degradability (INV1 and INV24 can be used for estimation of in situ effective protein degradability of extruded full fat soybean.

  15. The central domain of yeast transcription factor Rpn4 facilitates degradation of reporter protein in human cells.

    Science.gov (United States)

    Morozov, A V; Spasskaya, D S; Karpov, D S; Karpov, V L

    2014-10-16

    Despite high interest in the cellular degradation machinery and protein degradation signals (degrons), few degrons with universal activity along species have been identified. It has been shown that fusion of a target protein with a degradation signal from mammalian ornithine decarboxylase (ODC) induces fast proteasomal degradation of the chimera in both mammalian and yeast cells. However, no degrons from yeast-encoded proteins capable to function in mammalian cells were identified so far. Here, we demonstrate that the yeast transcription factor Rpn4 undergoes fast proteasomal degradation and its central domain can destabilize green fluorescent protein and Alpha-fetoprotein in human HEK 293T cells. Furthermore, we confirm the activity of this degron in yeast. Thus, the Rpn4 central domain is an effective interspecies degradation signal. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  16. Pathogenic prion protein is degraded by a manganese oxide mineral found in soils

    Science.gov (United States)

    Russo, F.; Johnson, C.J.; McKenzie, D.; Aiken, Judd M.; Pedersen, J.A.

    2009-01-01

    Prions, the aetiological agents of transmissible spongiform encephalopathies, exhibit extreme resistance to degradation. Soil can retain prion infectivity in the environment for years. Reactive soil components may, however, contribute to the inactivation of prions in soil. Members of the birnessite family of manganese oxides (MnO2) rank among the strongest natural oxidants in soils. Here, we report the abiotic degradation of pathogenic prion protein (PrPTSE) by a synthetic analogue of naturally occurring birnessite minerals. Aqueous MnO2 suspensions degraded the PrPTSE as evidenced by decreased immunoreactivity and diminished ability to seed protein misfolding cyclic amplification reactions. Birnessite-mediated PrPTSE degradation increased as a solution's pH decreased, consistent with the pH-dependence of the redox potential of MnO2. Exposure to 5.6 mg MnO2 ml-1 (PrPTSE:MnO2=1 : 110) decreased PrPTSE levels by ???4 orders of magnitude. Manganese oxides may contribute to prion degradation in soil environments rich in these minerals. ?? 2009 SGM.

  17. HIV-1 accessory proteins VPR and Vif modulate antiviral response by targeting IRF-3 for degradation

    International Nuclear Information System (INIS)

    Okumura, Atsushi; Alce, Tim; Lubyova, Barbora; Ezelle, Heather; Strebel, Klaus; Pitha, Paula M.

    2008-01-01

    The activation of IRF-3 during the early stages of viral infection is critical for the initiation of the antiviral response; however the activation of IRF-3 in HIV-1 infected cells has not yet been characterized. We demonstrate that the early steps of HIV-1 infection do not lead to the activation and nuclear translocation of IRF-3; instead, the relative levels of IRF-3 protein are decreased due to the ubiquitin-associated proteosome degradation. Addressing the molecular mechanism of this effect we show that the degradation is independent of HIV-1 replication and that virion-associated accessory proteins Vif and Vpr can independently degrade IRF-3. The null mutation of these two genes reduced the capacity of the HIV-1 virus to down modulate IRF-3 levels. The degradation was associated with Vif- and Vpr-mediated ubiquitination of IRF-3 and was independent of the activation of IRF-3. N-terminal lysine residues were shown to play a critical role in the Vif- and Vpr-mediated degradation of IRF-3. These data implicate Vif and Vpr in the disruption of the initial antiviral response and point to the need of HIV-1 to circumvent the antiviral response during the very early phase of replication

  18. A Role of Protein Degradation in Memory Consolidation after Initial Learning and Extinction Learning in the Honeybee ("Apis mellifera")

    Science.gov (United States)

    Felsenberg, Johannes; Dombrowski, Vincent; Eisenhardt, Dorothea

    2012-01-01

    Protein degradation is known to affect memory formation after extinction learning. We demonstrate here that an inhibitor of protein degradation, MG132, interferes with memory formation after extinction learning in a classical appetitive conditioning paradigm. In addition, we find an enhancement of memory formation when the same inhibitor is…

  19. Degradation of the disease-associated prion protein by a serine protease from lichens

    Science.gov (United States)

    Johnson, C.J.; Bennett, J.P.; Biro, S.M.; Duque-Velasquez, J.C.; Rodriguez, C.M.; Bessen, R.A.; Rocke, T.E.; Bartz, Jason C.

    2011-01-01

    The disease-associated prion protein (PrP(TSE)), the probable etiological agent of the transmissible spongiform encephalopathies (TSEs), is resistant to degradation and can persist in the environment. Lichens, mutualistic symbioses containing fungi, algae, bacteria and occasionally cyanobacteria, are ubiquitous in the environment and have evolved unique biological activities allowing their survival in challenging ecological niches. We investigated PrP(TSE) inactivation by lichens and found acetone extracts of three lichen species (Parmelia sulcata, Cladonia rangiferina and Lobaria pulmonaria) have the ability to degrade prion protein (PrP) from TSE-infected hamsters, mice and deer. Immunoblots measuring PrP levels and protein misfolding cyclic amplification indicated at least two logs of reductions in PrP(TSE). Degradative activity was not found in closely related lichen species or in algae or a cyanobacterium that inhabit lichens. Degradation was blocked by Pefabloc SC, a serine protease inhibitor, but not inhibitors of other proteases or enzymes. Additionally, we found that PrP levels in PrP(TSE)-enriched preps or infected brain homogenates are also reduced following exposure to freshly-collected P. sulcata or an aqueous extract of the lichen. Our findings indicate that these lichen extracts efficiently degrade PrP(TSE) and suggest that some lichens could have potential to inactivate TSE infectivity on the landscape or be a source for agents to degrade prions. Further work to clone and characterize the protease, assess its effect on TSE infectivity and determine which organism or organisms present in lichens produce or influence the protease activity is warranted.

  20. HUWE1 and TRIP12 Collaborate in Degradation of Ubiquitin-Fusion Proteins and Misframed Ubiquitin

    DEFF Research Database (Denmark)

    Poulsen, Esben G; Steinhauer, Cornelia; Lees, Michael

    2012-01-01

    In eukaryotic cells an uncleavable ubiquitin moiety conjugated to the N-terminus of a protein signals the degradation of the fusion protein via the proteasome-dependent ubiquitin fusion degradation (UFD) pathway. In yeast the molecular mechanism of the UFD pathway has been well characterized...... in degradation of the UFD substrate Ub(G76V)-YFP. The most significant hits from the screen were the E3 ubiquitin-protein ligase HUWE1, as well as PSMD7 and PSMD14 that encode proteasome subunits. Accordingly, knock down of HUWE1 led to an increase in the steady state level and a retarded degradation of the UFD...... substrate. Knock down of HUWE1 also led to a stabilization of the physiological UFD substrate UBB(+1). Precipitation experiments revealed that HUWE1 is associated with both the Ub(G76V)-YFP substrate and the 26S proteasome, indicating that it functions late in the UFD pathway. Double knock down of HUWE1...

  1. Two-stage digestion of renewable raw materials. Applying bioleaching for utilizing grass silage; Zweiphasige Vergaerung nachwachsender Rohstoffe. Einsatz des Bioleaching-Verfahrens zur Verwertung von Grassilage

    Energy Technology Data Exchange (ETDEWEB)

    Zielonka, S.; Lemmer, A.; Oechsner, H. [Hohenheim Univ., Stuttgart (Germany). Landesanstalt fuer Landwirtschaftliches Maschinen- und Bauwesen; Jungbluth, T. [Hohenheim Univ., Stuttgart (Germany). Inst. fuer Agrartechnik

    2007-07-01

    Currently renewable raw materials are being used in full scale biogas plants as co-substrates. Using grass silage frequently caused technical problems till now. Within the framework of this project, a process to digest grass silage as a single substrate is being developed. An intermittently operating two-stage process is used. As far as the degree of degradation and methane yields are concerned, good and promising results have been achieved. (orig.)

  2. Application of green fluorescent protein for monitoring phenol-degrading strains

    Directory of Open Access Journals (Sweden)

    Ana Milena Valderrama F.

    2001-07-01

    Full Text Available Several methods have been developed for detecting microorganisms in environmental samples. Some systems for incorporating reporter genes, such as lux or the green fluorescent protein (GFP gene, have been developed recently This study describes gfp gene marking of a phenol degrading strain, its evaluation and monitoring in a bioreactor containing refinery sour water. Tagged strains were obtained having the same physiological and metabolic characteristics as the parent strain. Fluorescent expression was kept stable with no selection for more than 50 consecutive generations and tagged strains were recovered from the bioreactor after forty-five days of phenol-degradation treatment.

  3. Effect of additives on the physical and chemical characteristics of sugar cane silage

    Directory of Open Access Journals (Sweden)

    Patricia do Rosario Rodrigues

    2015-08-01

    Full Text Available The objective of this study was to verify the efficiency of different additives on chemical composition, pH, ethanol production, content of volatile fatty acids (VFAs, nutritional losses during fermentation, and changes in fibrous fractions, in the levels of non-fibrous and total carbohydrates during the sugar cane silage fermentation process with different additives. The treatments consisted of control (no additive; corn meal, at 10% of natural matter; molasses, at 10% of natural matter; urea, at 2% of natural matter; and microbial inoculant for sugarcane silage (Lactobacillus plantarum, Kera-Sil® in a proportion of 2 g L-1 of water using a 2 liter solution per ton of ensilage. The experimental design was a completely randomized design with five treatments and five replications. The urea treatment provided the best preparation of silage, taking into account the pH and bromatological composition when compared to silages made with the other tested additives, and the control. The sugarcane silage showed a loss of 5.86% on average of dry matter, not differing from others additives used. There was an increase in crude protein content when urea was used. There was no difference between the treatment for fiber losses in neutral detergent and total digestible nutrients.

  4. Development of a new lactic acid bacterial inoculant for fresh rice straw silage

    Directory of Open Access Journals (Sweden)

    Jong Geun Kim

    2017-07-01

    Full Text Available Objective Effects of newly isolated Lactobacillus plantarum on the fermentation and chemical composition of fresh rice straw silage was evaluated in this study. Methods Lactic acid bacteria (LAB from good crop silage were screened by growing them in MRS broth and a minimal medium with low carbohydrate content. Selected LAB (LAB 1821 were Gram-positive, rods, catalase negative, and were identified to be Lactobacillus plantarum based on their biochemical characteristics and a 16S rRNA analysis. Fresh rice straw was ensiled with two isolated LAB (1821 and 1841, two commercial inoculants (HM/F and P1132 and no additive as a control. Results After 2 months of storage at ambient temperature, rice straw silages treated with additives were well-preserved, the pH values and butyric and acetic acid contents were lower, and the lactic acid content and lactic/acetic acid ratio were higher than those in the control (p0.05 effect on acid detergent fiber or neutral detergent fiber contents. Crude protein (CP content and in vitro DM digestibility (IVDMD increased after inoculation of LAB 1821 (p<0.05. Conclusion LAB 1821 increased the CP, IVDMD, lactic acid content and ratio of lactic acid to acetic acid in rice straw silage and decreased the pH, acetic acid, NH3-N, and butyric acid contents. Therefore, adding LAB 1821 improved the fermentation quality and feed value of rice straw silage.

  5. Unfolded protein response and activated degradative pathways regulation in GNE myopathy.

    Directory of Open Access Journals (Sweden)

    Honghao Li

    Full Text Available Although intracellular beta amyloid (Aβ accumulation is known as an early upstream event in the degenerative course of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE myopathy, the process by which Aβdeposits initiate various degradative pathways, and their relationship have not been fully clarified. We studied the possible secondary responses after amyloid beta precursor protein (AβPP deposition including unfolded protein response (UPR, ubiquitin proteasome system (UPS activation and its correlation with autophagy system. Eight GNE myopathy patients and five individuals with normal muscle morphology were included in this study. We performed immunofluorescence and immunoblotting to investigate the expression of AβPP, phosphorylated tau (p-tau and endoplasmic reticulum molecular chaperones. Proteasome activities were measured by cleavage of fluorogenic substrates. The expression of proteasome subunits and linkers between proteasomal and autophagy systems were also evaluated by immunoblotting and relative quantitative real-time RT-PCR. Four molecular chaperones, glucose-regulated protein 94 (GRP94, glucose-regulated protein 78 (GRP78, calreticulin and calnexin and valosin containing protein (VCP were highly expressed in GNE myopathy. 20S proteasome subunits, three main proteasome proteolytic activities, and the factors linking UPS and autophagy system were also increased. Our study suggests that AβPP deposition results in endoplasmic reticulum stress (ERS and highly expressed VCP deliver unfolded proteins from endoplasmic reticulum to proteosomal system which is activated in endoplasmic reticulum associated degradation (ERAD in GNE myopathy. Excessive ubiquitinated unfolded proteins are exported by proteins that connect UPS and autophagy to autophagy system, which is activated as an alternative pathway for degradation.

  6. Degradation of Human PDZ-Proteins by Human Alphapapillomaviruses Represents an Evolutionary Adaptation to a Novel Cellular Niche.

    Science.gov (United States)

    Van Doorslaer, Koenraad; DeSalle, Rob; Einstein, Mark H; Burk, Robert D

    2015-06-01

    In order to complete their life cycle, papillomaviruses have evolved to manipulate a plethora of cellular pathways. The products of the human Alphapapillomavirus E6 proteins specifically interact with and target PDZ containing proteins for degradation. This viral phenotype has been suggested to play a role in viral oncogenesis. To analyze the association of HPV E6 mediated PDZ-protein degradation with cervical oncogenesis, a high-throughput cell culture assay was developed. Degradation of an epitope tagged human MAGI1 isoform was visualized by immunoblot. The correlation between HPV E6-induced degradation of hMAGI1 and epidemiologically determined HPV oncogenicity was evaluated using a Bayesian approach within a phylogenetic context. All tested oncogenic types degraded the PDZ-containing protein hMAGI1d; however, E6 proteins isolated from several related albeit non-oncogenic viral types were equally efficient at degrading hMAGI1. The relationship between both traits (oncogenicity and PDZ degradation potential) is best explained by a model in which the potential to degrade PDZ proteins was acquired prior to the oncogenic phenotype. This analysis provides evidence that the ancestor of both oncogenic and non-oncogenic HPVs acquired the potential to degrade human PDZ-containing proteins. This suggests that HPV E6 directed degradation of PDZ-proteins represents an ancient ecological niche adaptation. Phylogenetic modeling indicates that this phenotype is not specifically correlated with oncogenic risk, but may act as an enabling phenotype. The role of PDZ protein degradation in HPV fitness and oncogenesis needs to be interpreted in the context of Alphapapillomavirus evolution.

  7. Degradation of Human PDZ-Proteins by Human Alphapapillomaviruses Represents an Evolutionary Adaptation to a Novel Cellular Niche

    Science.gov (United States)

    Van Doorslaer, Koenraad; DeSalle, Rob; Einstein, Mark H.; Burk, Robert D.

    2015-01-01

    In order to complete their life cycle, papillomaviruses have evolved to manipulate a plethora of cellular pathways. The products of the human Alphapapillomavirus E6 proteins specifically interact with and target PDZ containing proteins for degradation. This viral phenotype has been suggested to play a role in viral oncogenesis. To analyze the association of HPV E6 mediated PDZ-protein degradation with cervical oncogenesis, a high-throughput cell culture assay was developed. Degradation of an epitope tagged human MAGI1 isoform was visualized by immunoblot. The correlation between HPV E6-induced degradation of hMAGI1 and epidemiologically determined HPV oncogenicity was evaluated using a Bayesian approach within a phylogenetic context. All tested oncogenic types degraded the PDZ-containing protein hMAGI1d; however, E6 proteins isolated from several related albeit non-oncogenic viral types were equally efficient at degrading hMAGI1. The relationship between both traits (oncogenicity and PDZ degradation potential) is best explained by a model in which the potential to degrade PDZ proteins was acquired prior to the oncogenic phenotype. This analysis provides evidence that the ancestor of both oncogenic and non-oncogenic HPVs acquired the potential to degrade human PDZ-containing proteins. This suggests that HPV E6 directed degradation of PDZ-proteins represents an ancient ecological niche adaptation. Phylogenetic modeling indicates that this phenotype is not specifically correlated with oncogenic risk, but may act as an enabling phenotype. The role of PDZ protein degradation in HPV fitness and oncogenesis needs to be interpreted in the context of Alphapapillomavirus evolution. PMID:26086730

  8. Degradation of Human PDZ-Proteins by Human Alphapapillomaviruses Represents an Evolutionary Adaptation to a Novel Cellular Niche.

    Directory of Open Access Journals (Sweden)

    Koenraad Van Doorslaer

    2015-06-01

    Full Text Available In order to complete their life cycle, papillomaviruses have evolved to manipulate a plethora of cellular pathways. The products of the human Alphapapillomavirus E6 proteins specifically interact with and target PDZ containing proteins for degradation. This viral phenotype has been suggested to play a role in viral oncogenesis. To analyze the association of HPV E6 mediated PDZ-protein degradation with cervical oncogenesis, a high-throughput cell culture assay was developed. Degradation of an epitope tagged human MAGI1 isoform was visualized by immunoblot. The correlation between HPV E6-induced degradation of hMAGI1 and epidemiologically determined HPV oncogenicity was evaluated using a Bayesian approach within a phylogenetic context. All tested oncogenic types degraded the PDZ-containing protein hMAGI1d; however, E6 proteins isolated from several related albeit non-oncogenic viral types were equally efficient at degrading hMAGI1. The relationship between both traits (oncogenicity and PDZ degradation potential is best explained by a model in which the potential to degrade PDZ proteins was acquired prior to the oncogenic phenotype. This analysis provides evidence that the ancestor of both oncogenic and non-oncogenic HPVs acquired the potential to degrade human PDZ-containing proteins. This suggests that HPV E6 directed degradation of PDZ-proteins represents an ancient ecological niche adaptation. Phylogenetic modeling indicates that this phenotype is not specifically correlated with oncogenic risk, but may act as an enabling phenotype. The role of PDZ protein degradation in HPV fitness and oncogenesis needs to be interpreted in the context of Alphapapillomavirus evolution.

  9. In vitro estimation of rumen protein degradability using 35S to label the bacterial mass

    International Nuclear Information System (INIS)

    Khristov, A.; Aleksandrov, S.; Aleksiev, I.

    1994-01-01

    An experiment was carried out in order to simplify a previously developed 15 N-method for in vitro estimation of rumen protein degradability. Casein (Cas), whole soybeans (Sb) heated at 120 o C for 20 min (SbTherm) and sunflower (Sfl) were incubated at 39 o C for 4 hours in a water bathshaker with the following media: McDougall's buffer, strained and enriched with particle associated bacteria rumen fluid (2:1), rapidly (maltose, sucrose, glucose) and more slowly (pectin, soluble starch) degradable carbohydrates with final concentration of 815 mg/100 ml and 21.7 μCi/100 ml of 35 S (from Na 2 35 SO 4 ). After the incubation had been ceased, a bacterial fraction was isolated through differential centrifugation and specific activity of bacterial (Bac) and high speed total solids (TS) nitrogen was measured. The ratio was used to calculate bacterial mass in TS and through the Kjeldahl nitrogen concentration in TS - the net bacterial growth (against control vessels without protein). The level of ammonia-N in the supernate after blank correction was used to find the ammonia-N released from protein degradation. The data showed that the rate (and extend) of degradation for the Cas (as a standard protein) was lower compared to those obtained through the 15 N-method but it was higher than the rate derived through another in vitro method. The Cas equivalent of the Sb was higher than the figure we found in a previous experiment with solvent extracted soybean meal suggesting that the 35 S-method underestimated the degradability of the Cas. After being tested on a wider range of foodstuffs, the proposed 35 S-method might be considered as an alternative procedure which is less laborous than the 15 N-method. (author)

  10. Rumen microbial protein synthesis and nitrogen efficiency as affected by tanniferous and non-tanniferous forage legumes incubated individually or together in Rumen Simulation Technique.

    Science.gov (United States)

    Grosse Brinkhaus, Anja; Bee, Giuseppe; Schwarm, Angela; Kreuzer, Michael; Dohme-Meier, Frigga; Zeitz, Johanna O

    2018-03-01

    A limited availability of microbial protein can impair productivity in ruminants. Ruminal nitrogen efficiency might be optimised by combining high-quality forage legumes such as red clover (RC), which has unfavourably high ruminal protein degradability, with tanniferous legumes like sainfoin (SF) and birdsfoot trefoil (BT). Silages from SF and from BT cultivars [Bull (BB) and Polom (BP)] were incubated singly or in combination with RC using the Rumen Simulation Technique (n = 6). The tanniferous legumes, when compared to RC, changed the total short-chain fatty acid profile by increasing propionate proportions at the expense of butyrate. Silage from SF contained the most condensed tannins (CTs) (136 g CT kg -1 dry matter) and clearly differed in various traits from the BT and RC silages. The apparent nutrient degradability (small with SF), microbial protein synthesis, and calculated content of potentially utilisable crude protein (large with SF) indicated that SF had the greatest efficiency in ruminal protein synthesis. The effects of combining SF with RC were mostly linear. The potential of sainfoin to improve protein supply, demonstrated either individually or in combination with a high-performance forage legume, indicates its potential usefulness in complementing protein-deficient ruminant diets and high-quality forages rich in rumen-degradable protein. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  11. Effects of Streptococcus bovis Isolated from Bovine Rumen on the Fermentation Characteristics and Nutritive Value of Tanzania Grass Silage

    Directory of Open Access Journals (Sweden)

    Anderson de Moura Zanine

    2016-01-01

    Full Text Available This study aimed to evaluate the effects of Streptococcus bovis on the fermentation characteristics and nutritive value of Tanzania grass silage. Tanzania grass was chopped and left untreated (U or treated with Streptococcus bovis JB1 at 1 × 106 colony-forming units per gram (cfu/g of fresh forage or Streptococcus bovis HC5 at 1 × 106 cfu/g of fresh forage and packed into sixtuplicate laboratory silos. The largest number of enterobacteria, molds and yeast (M&Y occurred in untreated silages and the smallest populations of enterobacteria and M&Y and the largest numbers of lactic acid bacteria (LAB, at 9.81 and 9.87 log cfu/g, were observed in Streptococcus bovis JB1 and HC5, respectively (P<0.05. Silages treated with JB1 and HC5 had lower (P<0.05 silage pHs and concentrations of ammoniacal nitrogen (NH3-N than untreated silages. The application of Streptococcus bovis JB1 and HC5 resulted in fewer losses through gases and effluents (P<0.05, which resulted in greater dry matter recovery (DMR and crude protein recovery (CPR (P<0.05. Streptococcus bovis JB1 and HC5 improved the fermentative profile and increased the concentration of crude protein and DMR and CPR in Tanzania grass silage.

  12. Effects of fish silage on growth and biochemical characteristics of fresh water microalga Scenedesmus sp. MB 23

    Directory of Open Access Journals (Sweden)

    Jasmin Kaippilliparambil Abdulsamad

    2017-08-01

    Full Text Available Scenedesmus sp. MB 23 was cultivated in fish silage to study the effects of different concentrations on the growth and biochemical characteristics, particularly the protein, carbohydrate and lipid properties. Fish silage with 12% concentration was most effective for the growth and biomass production of Scenedesmus sp. The microalga reached maximum cell density (2433.89 × 104 cells/mL, chlorophyll-a concentration (2.766 μg/mL, specific growth rate (0.48/d and biomass (2.73 g/L on this medium. In mass culture, enhanced production of protein (123.87 mg/g dry weight of alga, carbohydrate (44.904 mg/g dry weight of alga and lipid (84.21 mg/g dry weight of alga was found using 9% fish silage. The effective reduction (up to 90% in the concentrations of nitrate, phosphorus and ammonia in the final fish silage medium proved the removal efficiency of Scenedesmus sp. The enhanced production of Scenedesmus sp. MB 23 indicated that effective bioremediation of fish waste can be conducted using algal mass production in fish silage. The study also proved that microalgae grown in fish silage have great industrial potential and can be used as a source of feed and biofuel.

  13. Phosphorylation-dependent signaling controls degradation of DNA mismatch repair protein PMS2.

    Science.gov (United States)

    Hinrichsen, Inga; Weßbecher, Isabel M; Huhn, Meik; Passmann, Sandra; Zeuzem, Stefan; Plotz, Guido; Biondi, Ricardo M; Brieger, Angela

    2017-12-01

    MutLα, a heterodimer consisting of MLH1 and PMS2, plays an important role in DNA mismatch repair and has been shown to be additionally involved in several other important cellular mechanisms. Previous work indicated that AKT could modulate PMS2 stability by phosphorylation. Still, the mechanisms of regulation of MutLα remain unclear. The stability of MutLα subunits was investigated by transiently overexpression of wild type and mutant forms of MLH1 and PMS2 using immunoblotting for measuring the protein levels after treatment. We found that treatment with the cell-permeable serine/threonine phosphatase inhibitor, Calyculin, leads to degradation of PMS2 when MLH1 or its C-terminal domain is missing or if amino acids of MLH1 essential for PMS2 interaction are mutated. In addition, we discovered that the C-terminal tail of PMS2 is relevant for this Calyculin-dependent degradation. A direct involvement of AKT, which was previously described to be responsible for PMS2 degradation, could not be detected. The multi-kinase inhibitor Sorafenib, in contrast, was able to avoid the degradation of PMS2 which postulates that cellular phosphorylation is involved in this process. Together, we show that pharmacologically induced phosphorylation by Calyculin can induce the selective proteasome-dependent degradation of PMS2 but not of MLH1 and that the PMS2 degradation could be blocked by Sorafenib treatment. Curiously, the C-terminal Lynch Syndrome-variants MLH1 L749P and MLH1 Y750X make PMS2 prone to Calyculin induced degradation. Therefore, we conclude that the specific degradation of PMS2 may represent a new mechanism to regulate MutLα. © 2017 Wiley Periodicals, Inc.

  14. REPLACEMENT OF CORN SILAGE BY CHOPPED SUGARCANE PLUS WHOLE COTTONSEED ON RUMINAL PARAMETERS, MICROBIAL PROTEIN PRODUCTION, AND NUTRIENT DIGESTIBILITY IN LACTATING COWS SUBSTITUIÇÃO DE SILAGEM DE MILHO POR CANA-DE-AÇÚCAR E CAROÇO DE ALGODÃO NOS PARÂMETROS RUMINAIS, SÍNTESE DE PROTEÍNA MICROBIANA E UTILIZAÇÃO DOS NUTRIENTES EM VACAS LACTANTES

    Directory of Open Access Journals (Sweden)

    Alexandre Vaz Pires

    2008-04-01

    Full Text Available Five multiparous Holstein cows averaging 525.91 kg BW and 85 days in milk were assigned in a 5 x 5 Latin square to evaluate the effects of corn silage replacement by sugarcane plus whole cottonseed. Cows were cannulated in the rumen and proximal duodenum and were fed a 50:50 concentrate:forage diet (DM basis. Treatments were defined by the replacement of corn silage by chopped sugarcane (whole plant plus whole cottonseed in the following proportions: 100:0 (control; 75:25; 50:50; 25:75; 0:100 in the DM basis. Dry matter intake (DMI of cows fed 100, 75, and 50% corn silage diets was higher (P<0.05 than DMI of cows fed 25 and 0% corn silage. Ruminal degradability and total tract digestibility of neutral detergent fiber and acid detergent fiber were not affected (P>0.05 by treatment diets. Total digestibilities of crude protein in diets containing up to 50% of sugarcane were lower than in diets containing 75 or 100% of sugarcane. Ruminal ammonia concentration decreased (P<0.05 with sugarcane and whole cottonseed inclusion. Concentrations of acetic, propionic and butyric acids as well as total volatile fatty acids concentration were higher (P<0.05 with sugarcane inclusion up to 50% level. Corn silage can be replaced by sugar cane plus whole cottonseed up to 50% level of forage in the diet without negative effect on nutrient utilization by dairy cows producing up to 18 kg milk/day. KEY WORDS: Digestibility, NDF, volatile fatty acid. Utilizaram-se cinco vacas holandesas pluríparas (525,91 kg PV e m

  15. Effects of different fertilizers on quantity and quality of silage corn

    Directory of Open Access Journals (Sweden)

    A. Di Francia

    2010-02-01

    Full Text Available Different fertilizers, ammonia sulfate and urea (MIN, Fertil 12.5 (ORG and Azoslow (ORG-MIN were compared in a silage corn crop. Total biomass yield was above 55.0 t ha-1 for all fertilizers. Azoslow showed the highest dry matter content (40.8%. No differences among the fertilizers were found in protein content (4.9% on average at waxy ripening, with the control showing the lowest value (3.9%. There were also no differences in silage quality among the fertilizers. The N budget was estimated in order to quantify the residual nitrogen amounts at harvest and the efficiency of fertilizers.

  16. Kinetics of ruminal degradation of alfalfa and Tifton-85 hays, and of corn SilageCinética de degradação ruminal dos fenos de alfafa e Tifton-85 e da silagem de milho

    Directory of Open Access Journals (Sweden)

    Geraldo Tadeu dos Santos

    2011-07-01

    Full Text Available The shunt portosystemic or portosystemic deviation (PSD are unique or multiples vascular communications between the systemic circulaton and the portal circulation, that permit blood flow reaches the circulatory system without first passing trhought the hepatic metabolization. May be acquired or congenital and can also be classified as intrahepatic located within the liver or extrahepatic located outside the liver parenquima. The acquired form is usually associated with intra-hepatic disorders. They usually suggest tortuous vessels that communicate with the caudal vena cava in the region of the left kidney. The congenital form is associated with genetic lineage and one of the most affected is Maltese breed. This case report describes the diagnostis and treatment of a eight year-old female Maltese dog presenting extrahepatic Portosystemic Shunt. The patient showed signs of hepatic encephalopathy, such as restlessnees, weakness, deambulation, head tremor and impaired visual. Complementary exam demonstrated: postprandial glicemia near the lower limit of reference, alkaline phosphatase (ALP and alanine aminotransferase (ALT increased and hipoalbuminemia. Ultrasonography revealed the presence of vesical calculus and bilateral kidney, liver decreases and increased echogenicity, gallbladder with anechoic content and high cellularity may suggest liver/ colangiohepatopatia and extrahepatic DPS was detected. The use of Doppler assisted in identifying the location of the bypass communication and the turbulence detecting the extrahepatic DPS. Protein-restricted diet and antibiotic therapy with amoxicillin achieved good results. The clinical treatment was decided make only the clinical and maintain the quality of life of the patient. Objetivou-se avaliar a degradabilidade ruminal e a taxa de degradação in situ da matéria seca (MS, da proteína bruta (PB e da fibra em detergente neutro (FDN, dos fenos de alfafa e Tifton-85 e da silagem de milho. Foram

  17. Generation of Oxygen Free Radicals by Proflavine: Implication in Protein Degradation

    Directory of Open Access Journals (Sweden)

    Mansour K.M. Gatasheh

    2017-07-01

    Full Text Available Proflavine, an acridine dye, is a known DNA intercalating agent. In the present study, we show that proflavine alone on photoillumination can generate reactive oxygen species (ROS. These proflavine-derived ROS cause damage to proteins, and this effect is enhanced when the divalent metal ion Cu (II is included in the reaction. Bathocuproine, a specific Cu (I sequestering agent, when present in the reaction mixture containing Cu (II, was found to inhibit the protein degradation, showing that Cu (I is an essential intermediate in the reaction. The effect of several scavengers of ROS such as superoxide dismutase, sodium azide, potassium iodide, and thiourea were examined on the protein damaging reaction. Potassium iodide was found to be the most effective in inhibiting protein damage followed by sodium azide and thiourea. Our results indicate the involvement of superoxide, singlet oxygen, triplet oxygen, and hydroxyl radicals in proflavine-induced damage to proteins.

  18. Measurement of local cerebral protein synthesis in vivo: influence of recycling of amino acids derived from protein degradation

    International Nuclear Information System (INIS)

    Smith, C.B.; Deibler, G.E.; Eng, N.; Schmidt, K.; Sokoloff, L.

    1988-01-01

    A quantitative autoradiographic method for the determination of local rates of protein synthesis in brain in vivo is being developed. The method employs L-[1- 14 C]leucine as the radiolabeled tracer. A comprehensive model has been designed that takes into account intracellular and extracellular spaces, intracellular compartmentation of leucine, and the possibility of recycling of unlabeled leucine derived from steady-state degradation of protein into the precursor pool for protein synthesis. We have evaluated the degree of recycling by measuring the ratio of the steady-state precursor pool distribution space for labeled leucine to that of unlabeled leucine. The values obtained were 0.58 in whole brain and 0.47 in liver. These results indicate that there is significant recycling of unlabeled amino acids derived from steady-state protein degradation in both tissues. Any method for the determination of rates of cerebral protein synthesis in vivo with labeled tracers that depends on estimation of precursor pool specific activity in tissue from measurements in plasma must take this recycling into account

  19. A STRUCTURAL OVERVIEW OF GH61 PROTEINS – FUNGAL CELLULOSE DEGRADING POLYSACCHARIDE MONOOXYGENASES

    Directory of Open Access Journals (Sweden)

    Leila Lo Leggio

    2012-09-01

    Full Text Available Recent years have witnessed a spurt of activities in the elucidation of the molecular function of a class of proteins with great potential in biomass degradation. GH61 proteins are of fungal origin and were originally classified in family 61 of the glycoside hydrolases. From the beginning they were strongly suspected to be involved in cellulose degradation because of their expression profiles, despite very low detectable endoglucanase activities. A major breakthrough came from structure determination of the first members, establishing the presence of a divalent metal binding site and a similarity to bacterial proteins involved in chitin degradation. A second breakthrough came from the identification of cellulase boosting activity dependent on the integrity of the metal binding site. Finally very recently GH61 proteins were demonstrated to oxidatively cleave crystalline cellulose in a Cu and reductant dependant manner. This mini-review in particular focuses on the contribution that structure elucidation has made in the understanding of GH61 molecular function and reviews the currently known structures and the challenges remaining ahead for exploiting this new class of enzymes to the full.

  20. A structural overview of GH61 proteins – fungal cellulose degrading polysaccharide monooxygenases

    Directory of Open Access Journals (Sweden)

    Leila Lo Leggio

    2012-09-01

    Full Text Available Recent years have witnessed a spurt of activities in the elucidation of the molecular function of a class of proteins with great potential in biomass degradation. GH61 proteins are of fungal origin and were originally classified in family 61 of the glycoside hydrolases. From the beginning they were strongly suspected to be involved in cellulose degradation because of their expression profiles, despite very low detectable endoglucanase activities. A major breakthrough came from structure determination of the first members, establishing the presence of a divalent metal binding site and a similarity to bacterial proteins involved in chitin degradation. A second breakthrough came from the identification of cellulase boosting activity dependent on the integrity of the metal binding site. Finally very recently GH61 proteins were demonstrated to oxidatively cleave crystalline cellulose in a Cu and reductant dependant manner. This mini-review in particular focuses on the contribution that structure elucidation has made in the understanding of GH61 molecular function and reviews the currently known structures and the challenges remaining ahead for exploiting this new class of enzymes to the full.

  1. HUWE1 and TRIP12 collaborate in degradation of ubiquitin-fusion proteins and misframed ubiquitin.

    Directory of Open Access Journals (Sweden)

    Esben G Poulsen

    Full Text Available In eukaryotic cells an uncleavable ubiquitin moiety conjugated to the N-terminus of a protein signals the degradation of the fusion protein via the proteasome-dependent ubiquitin fusion degradation (UFD pathway. In yeast the molecular mechanism of the UFD pathway has been well characterized. Recently the human E3 ubiquitin-protein ligase TRIP12 was connected with the UFD pathway, but little is otherwise known about this system in mammalian cells. In the present work, we utilized high-throughput imaging on cells transfected with a targeted siRNA library to identify components involved in degradation of the UFD substrate Ub(G76V-YFP. The most significant hits from the screen were the E3 ubiquitin-protein ligase HUWE1, as well as PSMD7 and PSMD14 that encode proteasome subunits. Accordingly, knock down of HUWE1 led to an increase in the steady state level and a retarded degradation of the UFD substrate. Knock down of HUWE1 also led to a stabilization of the physiological UFD substrate UBB(+1. Precipitation experiments revealed that HUWE1 is associated with both the Ub(G76V-YFP substrate and the 26S proteasome, indicating that it functions late in the UFD pathway. Double knock down of HUWE1 and TRIP12 resulted in an additive stabilization of the substrate, suggesting that HUWE1 and TRIP12 function in parallel during UFD. However, even when both HUWE1 and TRIP12 are downregulated, ubiquitylation of the UFD substrate was still apparent, revealing functional redundancy between HUWE1, TRIP12 and yet other ubiquitin-protein ligases.

  2. Protection of a protein against irradiation-induced degradation by additives in the solid state

    International Nuclear Information System (INIS)

    Shalaev, E.; Reddy, R.; Kimball, R.N.; Weinschenk, M.F.; Guinn, M.; Margulis, L.

    2003-01-01

    The impact of ionizing radiation on a globular protein (porcine somatotropin, pST) in the solid state was studied using rate of dissolution, high-performance liquid chromatography, and Electron spin resonance (ESR) in the presence of different additives. o-Vanillin stabilized pST against irradiation-induced degradation whereas effects of trolox and isopropyl alcohol were less significant. Stabilization effect of o-vanillin has been related to the energy transfer from pST molecules to the additive which was facilitated by formation of covalent bonds between o-vanillin and pST molecules. Anticorrelation between the level of free radicals and chemical degradation (i.e. degradation increased with decrease in a free radical level) was observed in the presence of o-vanillin

  3. Gold nanoparticles enhance the X-ray-induced degradation of human centrin 2 protein

    Energy Technology Data Exchange (ETDEWEB)

    Brun, Emilie [Laboratoire de Chimie Physique, CNRS UMR 8000, Universite Paris-Sud 11, Bat. 350, 91405 Orsay Cedex (France); Duchambon, Patricia; Blouquit, Yves [INSERM U759, Imagerie Integrative, Campus Universitaire d' Orsay, Bat. 112, Institut Curie, Centre de Recherche, Laboratoire R. Latarjet, Campus Universitaire d' Orsay, 91405 Orsay Cedex (France); Keller, Gerard [UMR CNRS 8612, Physico-Chimie-Pharmacotechnie-Biopharmacie, Universite Paris 11, Faculte de Pharmacie, 5 rue Jean-Baptiste Clement, 92296 Chatenay-Malabry (France); Sanche, Leon [Groupe en Sciences des Radiations, Departement de Medecine Nucleaire et Radiobiologie, Faculte de Medecine, Universite de Sherbrooke, Sherbrooke, Quebec, Canada J1H 5N4 (Canada); Sicard-Roselli, Cecile [Laboratoire de Chimie Physique, CNRS UMR 8000, Universite Paris-Sud 11, Bat. 350, 91405 Orsay Cedex (France)], E-mail: cecile.sicard@u-psud.fr

    2009-03-15

    In the war against cancer, radiotherapy is a prominent tool but counterbalanced by the fact that it also induces damages in healthy tissues. Nanotechnologies could open a new possibility to decrease these side effects. In particular, gold nanoparticles (GNPs) could be used as radio-sensitizers. As the role of proteins in the processes leading to cell death cannot be neglected, their radio-sensitization by GNPs is of great interest. This is particularly true in the case of the human centrin 2 protein, which has been proposed to be involved in DNA repair processes. To investigate this effect, we quantified for the first time the degradation of this protein in a gold colloidal solution when submitted to X-rays. We showed that the X-ray-induced degradation of the human centrin 2 protein is enhanced 1.5-fold in the presence of GNPs, even though no covalent bond exists between protein and GNPs. Among the conditions tested, the maximum enhancement was found with the higher GNP:protein ratio of 2x10{sup -4} and with the higher X-ray energy of 49 keV.

  4. Gold nanoparticles enhance the X-ray-induced degradation of human centrin 2 protein

    International Nuclear Information System (INIS)

    Brun, Emilie; Duchambon, Patricia; Blouquit, Yves; Keller, Gerard; Sanche, Leon; Sicard-Roselli, Cecile

    2009-01-01

    In the war against cancer, radiotherapy is a prominent tool but counterbalanced by the fact that it also induces damages in healthy tissues. Nanotechnologies could open a new possibility to decrease these side effects. In particular, gold nanoparticles (GNPs) could be used as radio-sensitizers. As the role of proteins in the processes leading to cell death cannot be neglected, their radio-sensitization by GNPs is of great interest. This is particularly true in the case of the human centrin 2 protein, which has been proposed to be involved in DNA repair processes. To investigate this effect, we quantified for the first time the degradation of this protein in a gold colloidal solution when submitted to X-rays. We showed that the X-ray-induced degradation of the human centrin 2 protein is enhanced 1.5-fold in the presence of GNPs, even though no covalent bond exists between protein and GNPs. Among the conditions tested, the maximum enhancement was found with the higher GNP:protein ratio of 2x10 -4 and with the higher X-ray energy of 49 keV

  5. Ruminal and intestinal protein degradability of various seaweed species measured in situ in dairy cows

    DEFF Research Database (Denmark)

    Tayyab, Usama; Novoa-Garrido, Margarita; Roleda, Michael Y.

    2016-01-01

    The use of seaweeds in animal diets is not new. However, little is known about the feed value of seaweed, both in terms of chemical composition and protein digestibility, and regarding variation between species and season. In this study, eight seaweed species of the genus Acrosiphonia, Alaria......, Laminaria, Mastocarpus, Palmaria, Pelvetia, Porphyra, and Ulva were sampled in spring (March) and autumn (October and November) 2014 at the coast of Bodø in Northern Norway, and were analysed for chemical composition, in situ rumen degradability and total tract crude protein (CP) digestibility. Ash content...... for Pelvetia (90 g/kg DM). Spring samples were higher in CP than autumn samples. The effective degradability estimated at 5% rumen passage rate (ED5) of CP varied between species (P Ulva (240 g...

  6. Dma1-dependent degradation of SIN proteins during meiosis in Schizosaccharomyces pombe.

    Science.gov (United States)

    Krapp, Andrea; Simanis, Viesturs

    2014-07-15

    The Schizosaccharomyces pombe septation initiation network (SIN) is required for cytokinesis during vegetative growth and for spore formation during meiosis. Regulation of the SIN during mitosis has been studied extensively, but less is known about its meiotic regulation. Here, we show that several aspects of SIN regulation differ between mitosis and meiosis. First, the presence of GTP-bound Spg1p is not the main determinant of the timing of Cdc7p and Sid1p association with the spindle pole body (SPB) during meiosis. Second, the localisation dependencies of SIN proteins differ from those in mitotic cells, suggesting a modified functional organisation of the SIN during meiosis. Third, there is stage-specific degradation of SIN components in meiosis; Byr4p is degraded after meiosis I, whereas the degradation of Cdc7p, Cdc11p and Sid4p occurs after the second meiotic division and depends upon the ubiquitin ligase Dma1p. Finally, Dma1p-dependent degradation is not restricted to the SIN, as we show that Dma1p is needed for the degradation of Mcp6p (also known as Hrs1p) during meiosis I. Taken together, these data suggest that stage-specific targeted proteolysis plays an important role in regulating meiotic progression. © 2014. Published by The Company of Biologists Ltd.

  7. Protein oxidation and degradation during proliferative senescence of human MRC-5 fibroblasts.

    Science.gov (United States)

    Sitte, N; Merker, K; von Zglinicki, T; Grune, T

    2000-03-01

    One of the highlights of age-related changes of cellular metabolism is the accumulation of oxidized proteins. The aging process on a cellular level can be treated either as the ongoing proliferation until a certain number of cell divisions is reached (the Hayflick limit) or as the aging of nondividing cells, that is, the age-related changes in cells without proliferation. The present investigation was undertaken to reveal the changes in protein turnover, proteasome activity, and protein oxidation status during proliferative senescence. We were able to demonstrate that the activity of the cytosolic proteasomal system declines dramatically during the proliferative senescence of human MRC-5 fibroblasts. Regardless of the loss in activity, it could be demonstrated that there are no changes in the transcription and translation of proteasomal subunits. This decline in proteasome activity was accompanied by an increased concentration of oxidized proteins. Cells at higher proliferation stages were no longer able to respond with increased degradation of endogenous [(35)S]-Met-radiolabeled proteins after hydrogen peroxide- or quinone-induced oxidative stress. It could be demonstrated that oxidized proteins in senescent human MRC-5 fibroblasts are not as quickly removed as they are in young cells. Therefore, our study demonstrates that the accumulation of oxidized proteins and decline in protein turnover and activity of the proteasomal system are not only a process of postmitotic aging but also occur during proliferative senescence and result in an increased half-life of oxidized proteins.

  8. New strategy for renal fibrosis: Targeting Smad3 proteins for ubiquitination and degradation.

    Science.gov (United States)

    Wang, Xin; Feng, Shaozhen; Fan, Jinjin; Li, Xiaoyan; Wen, Qiong; Luo, Ning

    2016-09-15

    Smad3 is a critical signaling protein in renal fibrosis. Proteolysis targeting chimeric molecules (PROTACs) are small molecules designed to degrade target proteins via ubiquitination. They have three components: (1) a recognition motif for E3 ligase; (2) a linker; and (3) a ligand for the target protein. We aimed to design a new PROTAC to prevent renal fibrosis by targeting Smad3 proteins and using hydroxylated pentapeptide of hypoxia-inducible factor-1α as the recognition motif for von Hippel-Lindau (VHL) ubiquitin ligase (E3). Computer-aided drug design was used to find a specific ligand targeting Smad3. Surface plasmon resonance (SPR) was used to verify and optimize screening results. Synthesized PROTAC was validated by two-stage mass spectrometry. The PROTAC's specificity for VHL (E3 ligase) was proved with two human renal carcinoma cell lines, 786-0 (VHL(-)) and ACHN (VHL(+)), and its anti-fibrosis effect was tested in renal fibrosis cell models. Thirteen small molecular compounds (SMCs) were obtained from the Enamine library using GLIDE molecular docking program. SPR results showed that #8 SMC (EN300-72284) combined best with Smad3 (KD=4.547×10(-5)M). Mass spectrometry showed that synthesized PROTAC had the correct peptide molecular weights. Western blot showed Smad3 was degraded by PROTAC with whole-cell lysate of ACHN but not 786-0. Degradation, but not ubiquitination, of Smad3 was inhibited by proteasome inhibitor MG132. The upregulation of fibronectin and Collagen I induced by TGF-β1 in both renal fibroblast and mesangial cells were inhibited by PROTAC. The new PROTAC might prevent renal fibrosis by targeting Smad3 for ubiquitination and degradation. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Nck adaptor proteins link Tks5 to invadopodia actin regulation and ECM degradation.

    Science.gov (United States)

    Stylli, Stanley S; Stacey, T T I; Verhagen, Anne M; Xu, San San; Pass, Ian; Courtneidge, Sara A; Lock, Peter

    2009-08-01

    Invadopodia are actin-based projections enriched with proteases, which invasive cancer cells use to degrade the extracellular matrix (ECM). The Phox homology (PX)-Src homology (SH)3 domain adaptor protein Tks5 (also known as SH3PXD2A) cooperates with Src tyrosine kinase to promote invadopodia formation but the underlying pathway is not clear. Here we show that Src phosphorylates Tks5 at Y557, inducing it to associate directly with the SH3-SH2 domain adaptor proteins Nck1 and Nck2 in invadopodia. Tks5 mutants unable to bind Nck show reduced matrix degradation-promoting activity and recruit actin to invadopodia inefficiently. Conversely, Src- and Tks5-driven matrix proteolysis and actin assembly in invadopodia are enhanced by Nck1 or Nck2 overexpression and inhibited by Nck1 depletion. We show that clustering at the plasma membrane of the Tks5 inter-SH3 region containing Y557 triggers phosphorylation at this site, facilitating Nck recruitment and F-actin assembly. These results identify a Src-Tks5-Nck pathway in ECM-degrading invadopodia that shows parallels with pathways linking several mammalian and pathogen-derived proteins to local actin regulation.

  10. Lysine-Less Variants of Spinal Muscular Atrophy SMN and SMNΔ7 Proteins Are Degraded by the Proteasome Pathway

    Directory of Open Access Journals (Sweden)

    Raúl Sánchez-Lanzas

    2017-12-01

    Full Text Available Spinal muscular atrophy is due to mutations affecting the SMN1 gene coding for the full-length protein (survival motor neuron; SMN and the SMN2 gene that preferentially generates an exon 7-deleted protein (SMNΔ7 by alternative splicing. To study SMN and SMNΔ7 degradation in the cell, we have used tagged versions at the N- (Flag or C-terminus (V5 of both proteins. Transfection of those constructs into HeLa cells and treatment with cycloheximide showed that those protein constructs were degraded. Proteasomal degradation usually requires prior lysine ubiquitylation. Surprisingly, lysine-less variants of both proteins tagged either at N- (Flag or C-terminus (V5 were also degraded. The degradation of the endogenous SMN protein, and the protein constructs mentioned above, was mediated by the proteasome, as it was blocked by lactacystin, a specific and irreversible proteasomal inhibitor. The results obtained allowed us to conclude that SMN and SMNΔ7 proteasomal degradation did not absolutely require internal ubiquitylation nor N-terminal ubiquitylation (prevented by N-terminal tagging. While the above conclusions are firmly supported by the experimental data presented, we discuss and justify the need of deep proteomic techniques for the study of SMN complex components (orphan and bound turn-over to understand the physiological relevant mechanisms of degradation of SMN and SMNΔ7 in the cell.

  11. Degradation of brown adipocyte purine nucleotides regulates uncoupling protein 1 activity

    Directory of Open Access Journals (Sweden)

    Tobias Fromme

    2018-02-01

    Full Text Available Objective: Non-shivering thermogenesis in mammalian brown adipose tissue depends on thermogenic uncoupling protein 1. Its activity is triggered by free fatty acids while purine nucleotides mediate inhibition. During activation, it is thought that free fatty acids overcome purine-mediated inhibition. We measured the cellular concentration and the release of purine nucleotide metabolites to uncover a possible role of purine nucleotide degradation in uncoupling protein 1 activation. Methods: With mass spectrometry, purine nucleotide metabolites were quantified in cellular homogenates and supernatants of cultured primary brown adipocytes. We also determined oxygen consumption in response to a β-adrenergic agonist. Results: Upon adrenergic activation, brown adipocytes decreased the intracellular concentration of inhibitory nucleotides (ATP, ADP, GTP and GDP and released the respective degradation products. At the same time, an increase in cellular calcium occurred. None of these phenomena occurred in white adipocytes or myotubes. The brown adipocyte expression of enzymes implicated in purine metabolic remodeling is altered upon cold exposure. Pharmacological and genetic interference of purine metabolism altered uncoupling protein 1 mediated uncoupled respiration. Conclusion: Adrenergic stimulation of brown adipocytes lowers the intracellular concentration of purine nucleotides, thereby contributing to uncoupling protein 1 activation. Keywords: Purine nucleotides, Uncoupling protein 1, Brown adipose tissue, Non-shivering thermogenesis, HILIC-MS/MS, Guanosine monophosphate reductase

  12. Methods for the visualization and analysis of extracellular matrix protein structure and degradation.

    Science.gov (United States)

    Leonard, Annemarie K; Loughran, Elizabeth A; Klymenko, Yuliya; Liu, Yueying; Kim, Oleg; Asem, Marwa; McAbee, Kevin; Ravosa, Matthew J; Stack, M Sharon

    2018-01-01

    This chapter highlights methods for visualization and analysis of extracellular matrix (ECM) proteins, with particular emphasis on collagen type I, the most abundant protein in mammals. Protocols described range from advanced imaging of complex in vivo matrices to simple biochemical analysis of individual ECM proteins. The first section of this chapter describes common methods to image ECM components and includes protocols for second harmonic generation, scanning electron microscopy, and several histological methods of ECM localization and degradation analysis, including immunohistochemistry, Trichrome staining, and in situ zymography. The second section of this chapter details both a common transwell invasion assay and a novel live imaging method to investigate cellular behavior with respect to collagen and other ECM proteins of interest. The final section consists of common electrophoresis-based biochemical methods that are used in analysis of ECM proteins. Use of the methods described herein will enable researchers to gain a greater understanding of the role of ECM structure and degradation in development and matrix-related diseases such as cancer and connective tissue disorders. © 2018 Elsevier Inc. All rights reserved.

  13. Modulation of intracellular protein degradation by SSB1-SIS1 chaperon system in yeast S. cerevisiae.

    Science.gov (United States)

    Ohba, M

    1997-06-09

    In prokaryotes, DnaK-DnaJ chaperon is involved in the protein degradation catalyzed by proteases La and ClpA/B complex as shown in E. coli. To extend this into eukaryotic cells, we examined the effects of hsp70 genes, SSA1 and SSB1, and DnaJ genes, SIS1 and YDJ1, on the growth of proteasome subunit mutants of the yeast S. cerevisiae. The results identified SSB1 and SIS1 as a pair of chaperon genes specifically involved in efficient protein turnover in the yeast, whose overexpression suppressed the growth defects caused by the proteasome mutations. Moreover, a single amino acid substitution in the putative peptide-binding site of SSB1 protein profoundly enhanced the suppression activity, indicating that the activity is mediated by the peptide-binding activity of this chaperon. Thus SSB1, with its partner DnaJ, SIS1, modulates the efficiency of protein turnover through its chaperon activity.

  14. Retinoblastoma protein co-purifies with proteasomal insulin-degrading enzyme: Implications for cell proliferation control

    Energy Technology Data Exchange (ETDEWEB)

    Radulescu, Razvan T., E-mail: ratura@gmx.net [Molecular Concepts Research (MCR), Muenster (Germany); Duckworth, William C. [Department of Medicine, Phoenix VA Health Care System, Phoenix, AZ (United States); Levy, Jennifer L. [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States); Fawcett, Janet, E-mail: janet.fawcett@va.gov [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States)

    2010-04-30

    Previous investigations on proteasomal preparations containing insulin-degrading enzyme (IDE; EC 3.4.24.56) have invariably yielded a co-purifying protein with a molecular weight of about 110 kDa. We have now found both in MCF-7 breast cancer and HepG2 hepatoma cells that this associated molecule is the retinoblastoma tumor suppressor protein (RB). Interestingly, the amount of RB in this protein complex seemed to be lower in HepG2 vs. MCF-7 cells, indicating a higher (cytoplasmic) protein turnover in the former vs. the latter cells. Moreover, immunofluorescence showed increased nuclear localization of RB in HepG2 vs. MCF-7 cells. Beyond these subtle differences between these distinct tumor cell types, our present study more generally suggests an interplay between RB and IDE within the proteasome that may have important growth-regulatory consequences.

  15. Life and death of proteins after protease cleavage: protein degradation by the N-end rule pathway.

    Science.gov (United States)

    Dissmeyer, Nico; Rivas, Susana; Graciet, Emmanuelle

    2018-05-01

    Contents Summary 929 I. conservation and diversity of N-end rule pathways 929 II. Defensive functions of the N-end rule pathway in plants 930 III. Proteases and degradation by the N-end rule pathway 930 IV. New proteomics approaches for the identification of N-end rule substrates 932 V. Concluding remarks 932 Acknowledgements 934 References 934 SUMMARY: The N-end rule relates the stability of a protein to the identity of its N-terminal residue and some of its modifications. Since its discovery in the 1980s, the repertoire of N-terminal degradation signals has expanded, leading to a diversity of N-end rule pathways. Although some of these newly discovered N-end rule pathways remain largely unexplored in plants, recent discoveries have highlighted roles of N-end rule-mediated protein degradation in plant defense against pathogens and in cell proliferation during organ growth. Despite this progress, a bottleneck remains the proteome-wide identification of N-end rule substrates due to the prerequisite for endoproteolytic cleavage and technical limitations. Here, we discuss the recent diversification of N-end rule pathways and their newly discovered functions in plant defenses, stressing the role of proteases. We expect that novel proteomics techniques (N-terminomics) will be essential for substrate identification. We review these methods, their limitations and future developments. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

  16. Determination of the Use of Lactobacillus plantarum and Propionibacterium freudenreichii Application on Fermentation Profile and Chemical Composition of Corn Silage

    Directory of Open Access Journals (Sweden)

    Norafizah Abdul Rahman

    2017-01-01

    Full Text Available Corn was inoculated with Lactobacillus plantarum and Propionibacterium freudenreichii subsp. shermanii either independently or as a mixture at ensiling, in order to determine the effect of bacterial additives on corn silage quality. Grain corn was harvested at 32–37% of dry matter and ensiled in a 4 L laboratory silo. Forage was treated as follows: bacterial types: B0 (without bacteria-control, B1 (L. plantarum, B2 (P. freudenreichii subsp. shermanii, and B3 (combination of L. plantarum and P. freudenreichii subsp. shermanii. Each 2 kg of chopped forage was treated with 10 mL of bacterial culture and allowed to ferment for 27 days. The first experiment determined the most suitable wavelength for detection of bacteria (490 nm and 419 nm for B1 and B2, resp. and the preferable inoculation size (1 × 105 cfu/g. The second experiment analysed the effect of B1 and B2 applied singly or as a mixture on the fermentation characteristics and quality of corn silage. L. plantarum alone increased crude protein (CP and reduced pH rapidly. In a mixture with P. freudenreichii, the final pH was the lowest compared to other treatments. As a mixture, inclusion of bacteria resulted in silage with lower digestibility than control. Corn silage treated with L. plantarum or P. freudenreichii either alone or mixed together produced desirable silage properties; however, this was not significantly better than untreated silage.

  17. Storage quality and marketability potential of bagged silage for ...

    African Journals Online (AJOL)

    Bagged sole maize and maize–cowpea silages in three bag sizes were assessed at 42 and 282 d post-ensiling for sensory and chemical quality. A survey of dairy farmers on silage use and preferences was conducted in the Chikwaka communal area and Marirangwe small-scale commercial farming area. Only silage ...

  18. Lactating cow response to lucerne silage inoculated with Lactobacillus plantarum

    Science.gov (United States)

    It is unclear why bacterial silage inoculants improve milk production in lactating dairy cattle. However, recent in vitro results suggest that inoculated silage effects on milk production may be tied to greater production of rumen microorganisms. Our objective was to determine if alfalfa silage trea...

  19. Physicochemical Characteristics, in Vitro Fermentation Indicators, Gas Production Kinetics, and Degradability of Solid Herbal Waste as Alternative Feed Source for Ruminants

    Directory of Open Access Journals (Sweden)

    A. N. Kisworo

    2017-08-01

    Full Text Available The aims of this research were to study the nutrient and secondary metabolite contents of solid herbal wastes (SHW that were preserved by freeze drying, sun drying and silage, as well as to analyze their effects on in vitro fermentation indicators i.e., gas production kinetics and degradability of solid herbal waste. Physical and chemical properties on three forms of SHW (sun dry, freeze dry, and silage were characterized and then an in vitro gas production experiment was performed to determine the kinetics of gas production, methane production, NH3, microbial protein, and SHW degradability. Polyethylene glycol (PEG was added to the three treatments to determine the biological activity of tannins. Results showed that all three preparations of SHW still contained high nutrient and plant secondary metabolite contents. Gas production, methane, NH3, microbial protein, in vitro degradability of dry matter (IVDMD and organic matter (IVDOM of SHW silage were lower (P<0.05 compared to sun dry and freeze dry. These results were apparently due to the high content of secondary metabolites especially tannin. It can be concluded that solid herbal wastes (SHW can be used as an alternative feed ingredients for ruminants with attention to the content of secondary metabolites that can affect the process of fermentation and digestibility in the rumen.

  20. Ubiquitination is absolutely required for the degradation of hypoxia-inducible factor - 1 alpha protein in hypoxic conditions

    International Nuclear Information System (INIS)

    Wang, Ronghai; Zhang, Ping; Li, Jinhang; Guan, Hongzai; Shi, Guangjun

    2016-01-01

    The hypoxia-inducible factor (HIF) is recognized as the master regulator of hypoxia response. HIF-α subunits expression are tightly regulated. In this study, our data show that ts20 cells still expressed detectable E1 protein even at 39.5° C for 12 h, and complete depletion of E1 protein expression at 39.5° C by siRNA enhanced HIF-1α and P53 protein expression. Further inhibition of E1 at 39.5 °C by siRNA, or E1 inhibitor Ube1-41 completely blocked HIF-1α degradation. Moreover, immunoprecipitations of co-transfection of HA-ubiquitin and FLAG–HIF–1α plasmids directly confirmed the involvement of ubiquitin in the hypoxic degradation of HIF-1α. Additionally, hypoxic HIF-1 α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization. Taken together, our data suggest that constitutive HIF-1α protein degradation in hypoxia is absolutely ubiquitination-dependent, and unidentified E3 ligase may exist for this degradation pathway. - Highlights: • HIF-1α protein is constitutively degraded in hypoxic conditions. • Requirement of ubiquitination for HIF-1α degradation in hypoxia. • Hypoxic HIF-1α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization.

  1. Ubiquitination is absolutely required for the degradation of hypoxia-inducible factor - 1 alpha protein in hypoxic conditions

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ronghai [Department of Urology, Linzi District People' s Hospital, Zibo, 255400 (China); Zhang, Ping, E-mail: zpskx001@163.com [Department of Gynecology, Qingdao Municipal Hospital, Qingdao, 266011 (China); Li, Jinhang [Department of Gynecology, Qingdao Municipal Hospital, Qingdao, 266011 (China); Guan, Hongzai [Laboratory Department, School of Medicine, Qingdao University, Qingdao, 266071 (China); Shi, Guangjun, E-mail: qdmhshigj@yahoo.com [Department of Hepatobiliary Surgery, Qingdao Municipal Hospital, Qingdao, 266071 (China)

    2016-01-29

    The hypoxia-inducible factor (HIF) is recognized as the master regulator of hypoxia response. HIF-α subunits expression are tightly regulated. In this study, our data show that ts20 cells still expressed detectable E1 protein even at 39.5° C for 12 h, and complete depletion of E1 protein expression at 39.5° C by siRNA enhanced HIF-1α and P53 protein expression. Further inhibition of E1 at 39.5 °C by siRNA, or E1 inhibitor Ube1-41 completely blocked HIF-1α degradation. Moreover, immunoprecipitations of co-transfection of HA-ubiquitin and FLAG–HIF–1α plasmids directly confirmed the involvement of ubiquitin in the hypoxic degradation of HIF-1α. Additionally, hypoxic HIF-1 α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization. Taken together, our data suggest that constitutive HIF-1α protein degradation in hypoxia is absolutely ubiquitination-dependent, and unidentified E3 ligase may exist for this degradation pathway. - Highlights: • HIF-1α protein is constitutively degraded in hypoxic conditions. • Requirement of ubiquitination for HIF-1α degradation in hypoxia. • Hypoxic HIF-1α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization.

  2. Dairy Replacement Performance Fed Various Recipes of Pangola (Digitaria eriantha) Silage Ensiled with Irradiated Silage Innoculant

    International Nuclear Information System (INIS)

    Prasanpanich, Somkiert; Boon-ek, Lerchart; Piadang, Nattayana

    2006-09-01

    Various kinds of Pangola silage ensiled with molasses and 2 types of silage innoculant (wild and mutant types). There were 6 recipes consisting of Treatment 1 : Pangola silage, Treatment 2: Pangola ensiled with molasses 4 % w/w, Treatment 3 : Pangola ensiled with 106 cfu wild type innoculant /gm Pangola, Treatment 4 : Pangola ensiled with 106 cfu mutant type innoculant/gm Pangola, Treatment 5 : Pangola ensiled with 106 cfu wild type innoculant/gm Pangola and molasses 4 % w/w, Treatment 6 : Pangola ensiled with 106 cfu mutant type innoculant/gm Pangola and molasses 4 % w/w. After 21 days of fermentation, silage sample was taken for nutritive value analysis. Preference testing on all silage recipes was studied using 18 dairy yearlings, aged 10-12 months old and weighed 200-300 kg, at 6 treatments with 3 animals each for 21 days in a Complete Randomized Design. The results did show that Treatment 1 had higher dry matter intake than those consisted of silage innoculant. However, when taking body weight changes and blood metabolites into account, Treatment 4 : Pangola ensiled with 106 cfu mutant type innoculant/gm Pangola, has an advantage on the ease of making process. Then, it is concluded that Treatment 4 is the most appropriate method to improve silage quality.

  3. A Compound Sensor for Simultaneous Measurement of Packing Density and Moisture Content of Silage.

    Science.gov (United States)

    Meng, Delun; Meng, Fanjia; Sun, Wei; Deng, Shuang

    2017-12-28

    Packing density and moisture content are important factors in investigating the ensiling quality. Low packing density is a major cause of loss of sugar content. The moisture content also plays a determinant role in biomass degradation. To comprehensively evaluate the ensiling quality, this study focused on developing a compound sensor. In it, moisture electrodes and strain gauges were embedded into an ASABE Standard small cone for the simultaneous measurements of the penetration resistance (PR) and moisture content (MC) of silage. In order to evaluate the performance of the designed sensor and the theoretical analysis being used, relevant calibration and validation tests were conducted. The determination coefficients are 0.996 and 0.992 for PR calibration and 0.934 for MC calibration. The validation indicated that this measurement technique could determine the packing density and moisture content of the silage simultaneously and eliminate the influence of the friction between the penetration shaft and silage. In this study, we not only design a compound sensor but also provide an alternative way to investigate the ensiling quality which would be useful for further silage research.

  4. Fermentation characteristics and nutritional quality of elephant grass silage added the buriti fruit peel

    Directory of Open Access Journals (Sweden)

    Raimundo Ribeiro Ferreira

    2017-05-01

    Full Text Available The objective of this research was to evaluated the inclusion of buriti fruit peel as additive on the fermentation profile, losses, chemical composition and degradability of elephant grass silage. We used a completely randomized design with five levels of buriti fruit peel (0, 50, 100, 200 and 400 g kg-1. The silos were opened after 28 days of storage. In situ degradability monitoring was conducted using a split-plot design in which four animals represented the blocks and silage supplemented with five levels of buriti fruit peel represented the treatments. The use of the additive in elephant silage increased dry matter (DM (P < 0.001, ether extract (EE (P < 0.001, ash (P < 0.001, neutral detergent fiber (NDF (P < 0.001 and acid detergent fiber (ADF (P= 0.0000. The pH (P= 0.0000, N-NH3 (P = 0.024 and there was a decrease in gas losses (P < 0.001, effluent losses and dry matter recovery (P = 0.218 not were influenced by the addition of buriti fruit peel. The inclusion of buriti fruit peel linearly reduced the ruminal degradability DM of soluble fraction (a (P < 0.001 and potentially degradable insoluble fraction (b (P < 0.001. The DM content increase with the addition of the buriti fruit peel to the elephant grass silage promotes improvements in the fermentation process, reduces losses of nutrients and ruminal disappearance of dry matter and does not significantly change the chemical composition with the inclusion of 166.7 g kg-1 of the buriti fruit peel.

  5. Studies to Prevent Degradation of Recombinant Fc-Fusion Protein Expressed in Mammalian Cell Line and Protein Characterization

    Directory of Open Access Journals (Sweden)

    Sanjukta Chakrabarti

    2016-06-01

    Full Text Available Clipping of recombinant proteins is a major issue in animal cell cultures. A recombinant Fc-fusion protein, VEGFR1(D1–D3-Fc expressed in CHOK1SV GS-KO cells was observed to be undergoing clippings in lab scale cultures. Partial cleaving of expressed protein initiated early on in cell culture and was observed to increase over time in culture and also on storage. In this study, a few parameters were explored in a bid to inhibit clipping in the fusion protein The effects of culture temperature, duration of culture, the addition of an anti-clumping agent, ferric citrate and use of protease inhibitor cocktail on inhibition of proteolysis of the Fc fusion were studied. Lowering of culture temperature from 37 to 30 °C alone appears to be the best solution for reducing protein degradation from the quality, cost and regulatory points of view. The obtained Fc protein was characterized and found to be in its stable folded state, exhibiting a high affinity for its ligand and also biological and functional activities.

  6. Copper-mediated oxidative degradation of catecholamines and oxidative damage of protein

    Energy Technology Data Exchange (ETDEWEB)

    Goncalves, P.R.; Harria, M.I.N.; Felix, J.M.; Hoffmann, M.E. [Universidade Estadual de Campinas, SP (Brazil). Inst. de Biologia

    1997-12-31

    Full text. Degradative oxidation of catecholamines has been a matter of large interest in recent years due to the evidences associating their autoxidation with the etiology of neurotoxic and cardiotoxic processes. In this work we present data on the degradative oxidation of catecholamines of physiological importance: isoproterenol (IP), epinephrine (EP), norepinephrine (NEP), deoxyepinephrine (DEP) and dopamine (DA). The degradative oxidation of the catecholamines was followed by measurement of spectral changes and oxygen consumption by neutral aqueous solutions. The data show that Cu{sup 2+} strongly accelerated the rate of catecholamine oxidation, following the decreasing order; EP>DEP>IP>NEP>DA. The production of superoxide anion radical during catecholamine oxidation was very slow, even in the presence of Cu{sup 2+}. The ability of IP to induce damages on bovine serum albumin (BSA) was determined by measuring the formation of carbonyl-groups in the protein, detected by reduction with tritiated Na BH{sub 4}. The incubation of BSA with IP (50-500{mu}M), in the presence of 100{mu}M Cu{sup 2+} leaded to an increased and dose dependent {sup 3} H-incorporation by the oxidized protein. The production of oxidative damage by IP/Cu{sup 2+} was accompanied by marked BSA fragmentation, detected by SDS-polyacrylamide gel dependent (25-400{mu}M IP) des appearance of the original BSA band and appearance of smaller fragments spread in the gel, when incubation has been done in the presence of 100{mu}M Cu{sup 2+}. These results suggest that copper-catalysed oxidative degradation of proteins induced by catecholamines might be critically involved in the toxic action of these molecules

  7. MEAT QUALITY FROM CHAROLAIS BULLS FED DIETS WITH DIFFERENT LEVELS OF CORN SILAGE INCLUSION

    Directory of Open Access Journals (Sweden)

    G. Cozzi

    2007-06-01

    Full Text Available A sample of 6 intensive beef farms was selected according to the feeding plan adopted during the fattening period of Charolais bulls. Two farms did not include any corn silage in the diet (CS0, while corn silage represented 22% of the dietary DM in the second group of 2 farms (CS22, and it raised up to 44% of the dietary DM in the last 2 farms (CS44. Five bulls were randomly selected from each farm to be slaughtered in the same abattoir. Bulls age was similar across treatments but the CS44 bulls had a lower carcass weight (396 kg than the other two treatments (436 and 446 kg for CS0 and CS22, respectively. Carcass fleshiness (SEUROP and fatness scores were not affected by the level of corn silage in the diet. Meat quality was evaluated on a joint sample of the m. Longissimus thoracis, excised from the 5th to the 9th rib of each right half carcass 24 h post-mortem, after an ageing period of 10 d vacuum packaged at 4°C. Meat chemical analysis showed no variations in pH, DM, intramuscular fat and protein content due to the different silage inclusion in the diet. Only the cholesterol content was progressively reduced in the meat of bulls fed increasing quantities of corn silage according to a significant negative linear trend. Meat colour, cooking losses and shear force values were not affected by the diet. Therefore, based on these findings there are no substantial arguments against the use of a large amount of corn silage in the fattening diets of Charolais bulls.

  8. The effect of supplementation of grass silage with rapeseed meal or Gasol-treated barley on the performance of growing cattle

    Directory of Open Access Journals (Sweden)

    Pekka Huhtanen

    1985-05-01

    Full Text Available Grass silage was offered ad libitum to 16 Ayrshire cattle in a 2 x 2 factorial experiment lasting 196 days. The silage was supplemented with barley preserved with 0.85 % of propionic acid (PAB or 1.28 % Gasol solution (GB 45 g DM/kg W0.75. Four of the animals in PAB and GB groups received no protein supplement(NPS and four of the animals in both groups received extracted rapeseed meal as protein supplement(RSM. RSM was fed according to UDP (undegradable feed protein requirements. The average age of the animals at the beginning of the experiment was 86 days and the average liveweight 106 kg. The average daily gains were 1066 and 1070 g/d for PAB and GB groups and 1012 and 1124 g/d (P < 0.01 for NPS and RSM groups. In feed intake and feed conversion there were no significant differences between the groups. Supplementation of silage barley diet with RSM increased the digestibility of organic matter from 71.1 to 73.5 % (P < 0.05, of crude protein from 62.8 to 67.4 % (P < 0.05 and of crude fibre from 57.2 to 60.2 % (P < 0.05. Both barleys were well preserved and there was no deterioration during the storage. The degradation rates of DM, crude protein and starch determined by nylon bag method were lower in GB than in PAB. Crude protein disappearances in 9 hours were 46.6 % for GB and 76.4 % for PAB. The utilization of absorbed protein calculated by factorial method averaged 0.566 ± 0.01 in the four different groups. Plasma urea N level was higher (P < 0.05 in the RSM than in the NPS group. The proportion of acetic acid in the rumen VFA was lower (P < 0.05 and that of propionic acid higher (P < 0.01 in the RSM than in the NPS group. PAB resulted in a higher (P < 0.05 proportion of propionic acid in rumen VFA than did GB.

  9. Quantifying protein synthesis and degradation in Arabidopsis by dynamic 13CO2 labeling and analysis of enrichment in individual amino acids in their free pools and in protein.

    Science.gov (United States)

    Ishihara, Hirofumi; Obata, Toshihiro; Sulpice, Ronan; Fernie, Alisdair R; Stitt, Mark

    2015-05-01

    Protein synthesis and degradation represent substantial costs during plant growth. To obtain a quantitative measure of the rate of protein synthesis and degradation, we supplied (13)CO2 to intact Arabidopsis (Arabidopsis thaliana) Columbia-0 plants and analyzed enrichment in free amino acids and in amino acid residues in protein during a 24-h pulse and 4-d chase. While many free amino acids labeled slowly and incompletely, alanine showed a rapid rise in enrichment in the pulse and a decrease in the chase. Enrichment in free alanine was used to correct enrichment in alanine residues in protein and calculate the rate of protein synthesis. The latter was compared with the relative growth rate to estimate the rate of protein degradation. The relative growth rate was estimated from sequential determination of fresh weight, sequential images of rosette area, and labeling of glucose in the cell wall. In an 8-h photoperiod, protein synthesis and cell wall synthesis were 3-fold faster in the day than at night, protein degradation was slow (3%-4% d(-1)), and flux to growth and degradation resulted in a protein half-life of 3.5 d. In the starchless phosphoglucomutase mutant at night, protein synthesis was further decreased and protein degradation increased, while cell wall synthesis was totally inhibited, quantitatively accounting for the inhibition of growth in this mutant. We also investigated the rates of protein synthesis and degradation during leaf development, during growth at high temperature, and compared synthesis rates of Rubisco large and small subunits of in the light and dark. © 2015 American Society of Plant Biologists. All Rights Reserved.

  10. Delaying aging and the aging-associated decline in protein homeostasis by inhibition of tryptophan degradation

    Science.gov (United States)

    van der Goot, Annemieke T.; Zhu, Wentao; Vázquez-Manrique, Rafael P.; Seinstra, Renée I.; Dettmer, Katja; Michels, Helen; Farina, Francesca; Krijnen, Jasper; Melki, Ronald; Buijsman, Rogier C.; Ruiz Silva, Mariana; Thijssen, Karen L.; Kema, Ido P.; Neri, Christian; Oefner, Peter J.; Nollen, Ellen A. A.

    2012-01-01

    Toxicity of aggregation-prone proteins is thought to play an important role in aging and age-related neurological diseases like Parkinson and Alzheimer’s diseases. Here, we identify tryptophan 2,3-dioxygenase (tdo-2), the first enzyme in the kynurenine pathway of tryptophan degradation, as a metabolic regulator of age-related α-synuclein toxicity in a Caenorhabditis elegans model. Depletion of tdo-2 also suppresses toxicity of other heterologous aggregation-prone proteins, including amyloid-β and polyglutamine proteins, and endogenous metastable proteins that are sensors of normal protein homeostasis. This finding suggests that tdo-2 functions as a general regulator of protein homeostasis. Analysis of metabolite levels in C. elegans strains with mutations in enzymes that act downstream of tdo-2 indicates that this suppression of toxicity is independent of downstream metabolites in the kynurenine pathway. Depletion of tdo-2 increases tryptophan levels, and feeding worms with extra l-tryptophan also suppresses toxicity, suggesting that tdo-2 regulates proteotoxicity through tryptophan. Depletion of tdo-2 extends lifespan in these worms. Together, these results implicate tdo-2 as a metabolic switch of age-related protein homeostasis and lifespan. With TDO and Indoleamine 2,3-dioxygenase as evolutionarily conserved human orthologs of TDO-2, intervening with tryptophan metabolism may offer avenues to reducing proteotoxicity in aging and age-related diseases. PMID:22927396

  11. HECTD3 Mediates an HSP90-Dependent Degradation Pathway for Protein Kinase Clients

    Directory of Open Access Journals (Sweden)

    Zhaobo Li

    2017-06-01

    Full Text Available Inhibition of the ATPase cycle of the HSP90 chaperone promotes ubiquitylation and proteasomal degradation of its client proteins, which include many oncogenic protein kinases. This provides the rationale for HSP90 inhibitors as cancer therapeutics. However, the mechanism by which HSP90 ATPase inhibition triggers ubiquitylation is not understood, and the E3 ubiquitin ligases involved are largely unknown. Using a siRNA screen, we have identified components of two independent degradation pathways for the HSP90 client kinase CRAF. The first requires CUL5, Elongin B, and Elongin C, while the second requires the E3 ligase HECTD3, which is also involved in the degradation of MASTL and LKB1. HECTD3 associates with HSP90 and CRAF in cells via its N-terminal DOC domain, which is mutationally disrupted in tumor cells with activated MAP kinase signaling. Our data implicate HECTD3 as a tumor suppressor modulating the activity of this important oncogenic signaling pathway.

  12. Particulate matter air pollution disrupts endothelial cell barrier via calpain-mediated tight junction protein degradation

    Directory of Open Access Journals (Sweden)

    Wang Ting

    2012-08-01

    Full Text Available Abstract Background Exposure to particulate matter (PM is a significant risk factor for increased cardiopulmonary morbidity and mortality. The mechanism of PM-mediated pathophysiology remains unknown. However, PM is proinflammatory to the endothelium and increases vascular permeability in vitro and in vivo via ROS generation. Objectives We explored the role of tight junction proteins as targets for PM-induced loss of lung endothelial cell (EC barrier integrity and enhanced cardiopulmonary dysfunction. Methods Changes in human lung EC monolayer permeability were assessed by Transendothelial Electrical Resistance (TER in response to PM challenge (collected from Ft. McHenry Tunnel, Baltimore, MD, particle size >0.1 μm. Biochemical assessment of ROS generation and Ca2+ mobilization were also measured. Results PM exposure induced tight junction protein Zona occludens-1 (ZO-1 relocation from the cell periphery, which was accompanied by significant reductions in ZO-1 protein levels but not in adherens junction proteins (VE-cadherin and β-catenin. N-acetyl-cysteine (NAC, 5 mM reduced PM-induced ROS generation in ECs, which further prevented TER decreases and atteneuated ZO-1 degradation. PM also mediated intracellular calcium mobilization via the transient receptor potential cation channel M2 (TRPM2, in a ROS-dependent manner with subsequent activation of the Ca2+-dependent protease calpain. PM-activated calpain is responsible for ZO-1 degradation and EC barrier disruption. Overexpression of ZO-1 attenuated PM-induced endothelial barrier disruption and vascular hyperpermeability in vivo and in vitro. Conclusions These results demonstrate that PM induces marked increases in vascular permeability via ROS-mediated calcium leakage via activated TRPM2, and via ZO-1 degradation by activated calpain. These findings support a novel mechanism for PM-induced lung damage and adverse cardiovascular outcomes.

  13. In Vitro Ruminal Degradability of Soybean Meal Protein Protected with Natural Tannin

    Science.gov (United States)

    Prasetiyono, B. W. H. E.; Subrata, A.; Tampoebolon, B. I. M.; Surono; Widiyanto

    2018-02-01

    The influence of tannin from tea waste and gambier as natural tannin sources on ruminal protein degradability was studied in this investigation. The soybean meal was used as protein source in this investigation. There were three treatments in this investigation mainly without protection (NT); protection with tea waste (Tt); and protection with gambier (Tg). The measured parameters consisted of in vitro dry matter digestibility (IVDMD), in vitro organic matter digestibility (IVOMD), and the ruminal fermentation characteristics. Results of this experiment showed that protection with tannin from tea waste as well as gambier increased (pRUP) in Tt and Tg group was higher than that in NT group (66.29 and 69.20 vs 51.10%). The ruminal protozoa population decreased (pRUP. The ruminal protozoa population and ammonia concentration, on the other hand, were decreased by tannin protection from those tannin natural sources. The natural tannin from gambier was the most effective protection agent for soybean meal protein.

  14. Ruminal and intestinal protein degradability of various seaweed species measured in situ in dairy cows

    DEFF Research Database (Denmark)

    Tayyab, Usama; Novoa-Garrido, Margarita; Roleda, Michael Y.

    2016-01-01

    , Laminaria, Mastocarpus, Palmaria, Pelvetia, Porphyra, and Ulva were sampled in spring (March) and autumn (October and November) 2014 at the coast of Bodø in Northern Norway, and were analysed for chemical composition, in situ rumen degradability and total tract crude protein (CP) digestibility. Ash content......The use of seaweeds in animal diets is not new. However, little is known about the feed value of seaweed, both in terms of chemical composition and protein digestibility, and regarding variation between species and season. In this study, eight seaweed species of the genus Acrosiphonia, Alaria....../kg CP). Digestible rumen escape protein (DEP) varied significantly between species (P Laminaria, Mastocarpus and Palmaria can supply...

  15. Differential Regulation of Telomerase Reverse Transcriptase Promoter Activation and Protein Degradation by Histone Deacetylase Inhibition.

    Science.gov (United States)

    Qing, Hua; Aono, Jun; Findeisen, Hannes M; Jones, Karrie L; Heywood, Elizabeth B; Bruemmer, Dennis

    2016-06-01

    Telomerase reverse transcriptase (TERT) maintains telomeres and is rate limiting for replicative life span. While most somatic tissues silence TERT transcription resulting in telomere shortening, cells derived from cancer or cardiovascular diseases express TERT and activate telomerase. In the present study, we demonstrate that histone deacetylase (HDAC) inhibition induces TERT transcription and promoter activation. At the protein level in contrast, HDAC inhibition decreases TERT protein abundance through enhanced degradation, which decreases telomerase activity and induces senescence. Finally, we demonstrate that HDAC inhibition decreases TERT expression during vascular remodeling in vivo. These data illustrate a differential regulation of TERT transcription and protein stability by HDAC inhibition and suggest that TERT may constitute an important target for the anti-proliferative efficacy of HDAC inhibitors. © 2015 Wiley Periodicals, Inc.

  16. The influence of grass silage-to-maize silage ratio and concentrate composition on methane emissions, performance and milk composition of dairy cows.

    Science.gov (United States)

    Hart, K J; Huntington, J A; Wilkinson, R G; Bartram, C G; Sinclair, L A

    2015-06-01

    It is well-established that altering the proportion of starch and fibre in ruminant diets can alter ruminal and post-ruminal digestion, although quantitative evidence that this reduces enteric methane (CH4) production in dairy cattle is lacking. The objective of this study was to examine the effect of varying grass-to-maize silage ratio (70 : 30 and 30 : 70 DM basis), offered ad libitum, with either a concentrate that was high in starch or fibre, on CH4 production, intake, performance and milk composition of dairy cows. A total of 20 cows were allocated to one of the four experimental diets in a two-by-two factorial design run as a Latin square with each period lasting 28 days. Measurements were conducted during the final 7 days of each period. Cows offered the high maize silage ration had a higher dry matter intake (DMI), milk yield, milk energy output and lower CH4 emissions when expressed per kg DMI and per unit of ingested gross energy, but there was no difference in total CH4 production. Several of the milk long-chain fatty acids (FA) were affected by forage treatment with the most notable being an increase in 18:0, 18:1 c9, 18:2 c9 c12 and total mono unsaturated FA, observed in cows offered the higher inclusion of maize silage, and an increase in 18:3 c9 c12 c15 when offered the higher grass silage ration. Varying the composition of the concentrate had no effect on DMI or milk production; however, when the high-starch concentrate was fed, milk protein concentration and milk FAs, 10:0, 14:1, 15:0, 16:1, increased and 18:0 decreased. Interactions were observed for milk fat concentration, being lower in cows offered high-grass silage and high-fibre concentrates compared with the high-starch concentrate, and FA 17:0, which was the highest in milk from cows fed the high-grass silage diet supplemented with the high-starch concentrate. In conclusion, increasing the proportion of maize silage in the diets of dairy cows increased intake and performance, and reduced CH

  17. A critical role for protein degradation in the nucleus accumbens core in cocaine reward memory.

    Science.gov (United States)

    Ren, Zhen-Yu; Liu, Meng-Meng; Xue, Yan-Xue; Ding, Zeng-Bo; Xue, Li-Fen; Zhai, Suo-Di; Lu, Lin

    2013-04-01

    The intense associative memories that develop between cocaine-paired contexts and rewarding stimuli contribute to cocaine seeking and relapse. Previous studies have shown impairment in cocaine reward memories by manipulating a labile state induced by memory retrieval, but the mechanisms that underlie the destabilization of cocaine reward memory are unknown. In this study, using a Pavlovian cocaine-induced conditioned place preference (CPP) procedure in rats, we tested the contribution of ubiquitin-proteasome system-dependent protein degradation in destabilization of cocaine reward memory. First, we found that polyubiquitinated protein expression levels and polyubiquitinated N-ethylmaleimide-sensitive fusion (NSF) markedly increased 15 min after retrieval while NSF protein levels decreased 1 h after retrieval in the synaptosomal membrane fraction in the nucleus accumbens (NAc) core. We then found that infusion of the proteasome inhibitor lactacystin into the NAc core prevented the impairment of memory reconsolidation induced by the protein synthesis inhibitor anisomycin and reversed the effects of anisomycin on NSF and glutamate receptor 2 (GluR2) protein levels in the synaptosomal membrane fraction in the NAc core. We also found that lactacystin infusion into the NAc core but not into the shell immediately after extinction training sessions inhibited CPP extinction and reversed the extinction training-induced decrease in NSF and GluR2 in the synaptosomal membrane fraction in the NAc core. Finally, infusions of lactacystin by itself into the NAc core immediately after each training session or before the CPP retrieval test had no effect on the consolidation and retrieval of cocaine reward memory. These findings suggest that ubiquitin-proteasome system-dependent protein degradation is critical for retrieval-induced memory destabilization.

  18. L-Alanylglutamine inhibits signaling proteins that activate protein degradation, but does not affect proteins that activate protein synthesis after an acute resistance exercise.

    Science.gov (United States)

    Wang, Wanyi; Choi, Ran Hee; Solares, Geoffrey J; Tseng, Hung-Min; Ding, Zhenping; Kim, Kyoungrae; Ivy, John L

    2015-07-01

    Sustamine™ (SUS) is a dipeptide composed of alanine and glutamine (AlaGln). Glutamine has been suggested to increase muscle protein accretion; however, the underlying molecular mechanisms of glutamine on muscle protein metabolism following resistance exercise have not been fully addressed. In the present study, 2-month-old rats climbed a ladder 10 times with a weight equal to 75 % of their body mass attached at the tail. Rats were then orally administered one of four solutions: placebo (PLA-glycine = 0.52 g/kg), whey protein (WP = 0.4 g/kg), low dose of SUS (LSUS = 0.1 g/kg), or high dose of SUS (HSUS = 0.5 g/kg). An additional group of sedentary (SED) rats was intubated with glycine (0.52 g/kg) at the same time as the ladder-climbing rats. Blood samples were collected immediately after exercise and at either 20 or 40 min after recovery. The flexor hallucis longus (FHL), a muscle used for climbing, was excised at 20 or 40 min post exercise and analyzed for proteins regulating protein synthesis and degradation. All supplements elevated the phosphorylation of FOXO3A above SED at 20 min post exercise, but only the SUS supplements significantly reduced the phosphorylation of AMPK and NF-kB p65. SUS supplements had no effect on mTOR signaling, but WP supplementation yielded a greater phosphorylation of mTOR, p70S6k, and rpS6 compared with PLA at 20 min post exercise. However, by 40 min post exercise, phosphorylation of mTOR and rpS6 in PLA had risen to levels not different than WP. These results suggest that SUS blocks the activation of intracellular signals for MPB, whereas WP accelerates mRNA translation.

  19. Influence of species and preservations on the quality and safety of grass silages

    Directory of Open Access Journals (Sweden)

    Jiří Skládanka

    2010-01-01

    Full Text Available The paper evaluates the quality of model silages made of wilted grass biomass and treated with silage additives. Grass species used for the production of silages were Lolium perenne, Festulolium pabulare and Festulolium braunii harvested in the first cut at the stage of earing. The assessed grass species were wilted after the cut for an identical time 36 hours (2008, resp. 24 hours (2009. The treatment was made either with a chemical preparation (formic acid, propionic acid, ammonium formate and/or with a bio­lo­gi­cal inoculant (Enterococcus faecium, Lactobacillus plantarum, Pediococcus acidilactici, Lactobacillus salivarius, cellulase, hemicellulase and amylase. The amount of the chemical ingredient was 4 l . t−1 and the amount of the biological additive was 10 g . t−1.The biomass was after wilting ensilaged in con­tai­ners whose diameter and height were 0.15 m and 0.64 m, respectively. After 60 days of ensilaging, the silages were assessed for pH, organic acids content, ethanol content and acidity of water extract (AWE; organic nutrients assessed in the silages were crude fibre (CF, neutral detergent fibre (NDF, acid detergent fibre (ADF, crude protein (CP and digestibility of organic matter (DOM. Hygienic safety was assessed from the contents of zearalenon, fumonisin and aflatoxin mycotoxins. The high (P < 0.05 dry matter (DM content in Festulolium pabulare silages indicates that the species tends to rapid wilting. The higher DM content reflected in lower biomass losses (P < 0.05. The lowest pH values (P < 0.05 were detected in silages made of Festulolium braunii. The fact relates to the higher content of lactic acid in the prepared microsilages. The use of ensiling additives affected the quality of extracts. Namely the application of the biological additive led to the increased content (P < 0.05 of not only lactic acid but acetic acid too. Titrable acidity was not affected by the ensiling additives. As to the

  20. Ruminal degradation and intestinal digestibility of protein and amino acids in high-protein feedstuffs commonly used in dairy diets.

    Science.gov (United States)

    Paz, H A; Klopfenstein, T J; Hostetler, D; Fernando, S C; Castillo-Lopez, E; Kononoff, P J

    2014-10-01

    A study was conducted to determine the rumen degradation and intestinal digestibility of crude protein (CP) and AA, and AA composition of the rumen-undegradable protein (RUP) from 3 sources of blood meal (BM1, BM2, and BM3), canola meal (CM), low-fat distillers dried grains with solubles (LFDG), soybean meal (SBM), and expeller soybean meal (ESBM). Two Holstein cows fitted with ruminal and proximal duodenal cannulas were used for in situ incubation of 16h and for the mobile bag technique. To correct for bacterial contamination of the RUP, 2 methods were used: purines and DNA as bacterial markers. Ruminal degradations of CP were 85.3, 29.8, 40.7, 75.7, 76.9, 68.8, and 37.0 ± 3.93% for BM1, BM2, BM3, CM, LFDG, SBM, and ESBM, respectively. Ruminal degradation of both total essential AA and nonessential AA followed a similar pattern to that of CP across feedstuffs. Based on the ratio of AA concentration in the RUP to AA concentration in the original feedstuff, ruminal incubation decreased (ratio 1) the concentrations of Ile and Met across feedstuffs. Compared with purines, the use of DNA as bacterial marker resulted in a higher estimate of bacterial CP contamination for CM and lower estimates for LFDG and ESBM. Intestinal digestibility of RUP could not be estimated for BM1, BM3, and SBM due to insufficient recovery of residue. For the remaining feedstuffs, intestinal digestibility of RUP was highest for ESBM, followed by BM2 and LFDG, and lowest for CM: 98.8, 87.9, 89.7, and 72.4 ± 1.40%, respectively. Intestinal absorbable dietary protein was higher for BM2 compared with CM and LFDG, at 61.7, 17.9, and 20.7 ± 2.73% CP, respectively. As prices fluctuate, intestinal absorbable protein or AA may be used as a tool to aid in the selection among feedstuffs with different protein quality. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  1. CNPY2 inhibits MYLIP-mediated AR protein degradation in prostate cancer cells.

    Science.gov (United States)

    Ito, Saya; Ueno, Akihisa; Ueda, Takashi; Nakagawa, Hideo; Taniguchi, Hidefumi; Kayukawa, Naruhiro; Fujihara-Iwata, Atsuko; Hongo, Fumiya; Okihara, Koji; Ukimura, Osamu

    2018-04-03

    The androgen receptor (AR) is a ligand-dependent transcription factor that promotes prostate cancer (PC) cell growth through control of target gene expression. This report suggests that Canopy FGF signaling regulator 2 (CNPY2) controls AR protein levels in PC cells. We found that AR was ubiquitinated by an E3 ubiquitin ligase, myosin regulatory light chain interacting protein (MYLIP) and then degraded through the ubiquitin-proteasome pathway. CNPY2 decreased the ubiquitination activity of MYLIP by inhibition of interaction between MYLIP and UBE2D1, an E2 ubiquitin ligase. CNPY2 up-regulated gene expression of AR target genes such as KLK3 gene which encodes the prostate specific antigen (PSA) and promoted cell growth of PC cells. The cell growth inhibition by CNPY2 knockdown was rescued by AR overexpression. Furthermore, positive correlation of expression levels between CNPY2 and AR/AR target genes was observed in tissue samples from human prostate cancer patients. Together, these results suggested that CNPY2 promoted cell growth of PC cells by inhibition of AR protein degradation through MYLIP-mediated AR ubiquitination.

  2. Saturated fatty acid palmitate negatively regulates autophagy by promoting ATG5 protein degradation in meniscus cells.

    Science.gov (United States)

    Mallik, Aritra; Yammani, Raghunatha R

    2018-07-20

    Obesity and associated metabolic factors are major risk factors for the development of osteoarthritis. Previously, we have shown that the free fatty acid palmitate induces endoplasmic reticulum (ER) stress and induces apoptosis in meniscus cells. However, the molecular mechanisms involved in these effects are not clearly understood. In our current study, we found that palmitate inhibits autophagy by modulating the protein levels of autophagy-related genes-5 (ATG5) that is associated with decreased lipidation of LC3 and increased activation of cleaved caspase 3. Pretreatment of meniscus cells with 4-phenyl butyric acid, a small molecule chemical chaperone that alleviates ER stress, or with MG-132, a proteasome inhibitor, restored normal levels of ATG5 and autophagosome formation, and decreased expression of cleaved caspase 3. Thus, our data suggest that palmitate downregulates autophagy in meniscus cells by degrading ATG5 protein via ER-associated protein degradation, and thus promotes apoptosis. This is the first study to demonstrate that palmitate-induced endoplasmic reticulum stress negatively regulates autophagy. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  3. NAD(P)H quinone oxidoreductase 1 inhibits the proteasomal degradation of homocysteine-induced endoplasmic reticulum protein

    Energy Technology Data Exchange (ETDEWEB)

    Maeda, Tomoji, E-mail: t-maeda@nichiyaku.ac.jp [Department of Neuroscience, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuta, Yahaba-Cho, Shiwagun, Iwate, 028-3603 (Japan); Tanabe-Fujimura, Chiaki; Fujita, Yu; Abe, Chihiro; Nanakida, Yoshino; Zou, Kun; Liu, Junjun; Liu, Shuyu [Department of Neuroscience, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuta, Yahaba-Cho, Shiwagun, Iwate, 028-3603 (Japan); Nakajima, Toshihiro [Institute of Medical Science, Tokyo Medical University, 6-1-1 Shinjyuku, Shinjyuku, Tokyo, Tokyo, 160-8402 (Japan); Komano, Hiroto, E-mail: hkomano@iwate-med.ac.jp [Department of Neuroscience, School of Pharmacy, Iwate Medical University, 2-1-1 Nishitokuta, Yahaba-Cho, Shiwagun, Iwate, 028-3603 (Japan)

    2016-05-13

    Homocysteine-induced endoplasmic reticulum (ER) protein (Herp) is an ER stress-inducible key regulatory component of ER-associated degradation (ERAD) that has been implicated in insulin hypersecretion in diabetic mouse models. Herp expression is tightly regulated. Additionally, Herp is a highly labile protein and interacts with various proteins, which are characteristic features of ubiquitinated protein. Previously, we reported that ubiquitination is not required for Herp degradation. In addition, we found that the lysine residues of Herp (which are ubiquitinated by E3 ubiquitin ligase) are not sufficient for regulation of Herp degradation. In this study, we found that NAD(P)H quinone oxidoreductase 1 (NQO1)-mediated targeting of Herp to the proteasome was involved in Herp degradation. In addition, we found that Herp protein levels were markedly elevated in synoviolin-null cells. The E3 ubiquitin ligase synoviolin is a central component of ERAD and is involved in the degradation of nuclear factor E2-related factor-2 (Nrf2), which regulates cellular reactive oxygen species. Additionally, NQO1 is a target of Nrf2. Thus, our findings indicated that NQO1 could stabilize Herp protein expression via indirect regulation of synoviolin. -- Highlights: •Herp interacts with NQO1. •NQO1 regulates Herp degradation.

  4. NAD(P)H quinone oxidoreductase 1 inhibits the proteasomal degradation of homocysteine-induced endoplasmic reticulum protein

    International Nuclear Information System (INIS)

    Maeda, Tomoji; Tanabe-Fujimura, Chiaki; Fujita, Yu; Abe, Chihiro; Nanakida, Yoshino; Zou, Kun; Liu, Junjun; Liu, Shuyu; Nakajima, Toshihiro; Komano, Hiroto

    2016-01-01

    Homocysteine-induced endoplasmic reticulum (ER) protein (Herp) is an ER stress-inducible key regulatory component of ER-associated degradation (ERAD) that has been implicated in insulin hypersecretion in diabetic mouse models. Herp expression is tightly regulated. Additionally, Herp is a highly labile protein and interacts with various proteins, which are characteristic features of ubiquitinated protein. Previously, we reported that ubiquitination is not required for Herp degradation. In addition, we found that the lysine residues of Herp (which are ubiquitinated by E3 ubiquitin ligase) are not sufficient for regulation of Herp degradation. In this study, we found that NAD(P)H quinone oxidoreductase 1 (NQO1)-mediated targeting of Herp to the proteasome was involved in Herp degradation. In addition, we found that Herp protein levels were markedly elevated in synoviolin-null cells. The E3 ubiquitin ligase synoviolin is a central component of ERAD and is involved in the degradation of nuclear factor E2-related factor-2 (Nrf2), which regulates cellular reactive oxygen species. Additionally, NQO1 is a target of Nrf2. Thus, our findings indicated that NQO1 could stabilize Herp protein expression via indirect regulation of synoviolin. -- Highlights: •Herp interacts with NQO1. •NQO1 regulates Herp degradation.

  5. Novel function of the endoplasmic reticulum degradation-enhancing α-mannosidase-like proteins in the human hepatitis B virus life cycle, mediated by the middle envelope protein.

    Science.gov (United States)

    Lazar, Catalin; Uta, Mihaela; Petrescu, Stefana Maria; Branza-Nichita, Norica

    2017-02-01

    Cells replicating the human hepatitis B virus (HBV) express high levels of degradation-enhancing α-mannosidase-like proteins (EDEMs), a family of proteins involved in the endoplasmic reticulum associated degradation, one of the pathways activated during the unfolded protein response. Owing to their α-1,2 mannosidase activity, the EDEM1-3 proteins are able to process the N-linked glycans of misfolded or incompletely folded proteins, providing the recognition signal for their subsequent degradation. The HBV small (S), medium (M), and large (L) surface proteins bear an N-linked glycosylation site in the common S domain that is partially occupied in all proteins. The M protein contains an additional site in its preS2 domain, which is always functional. Here, we report that these oligosaccharides are processed by EDEMs, more efficiently by EDEM3, which induces degradation of L and S proteins, accompanied by a reduction of subviral particles production. In striking contrast, M not only is spared from degradation but its trafficking is also accelerated leading to an improved secretion. This unusual behavior of the M protein requires strictly the mannose trimming of the preS2 N-linked glycan. Furthermore, we show that HBV secretion is significantly inhibited under strong endoplasmic reticulum stress conditions when M expression is prevented by mutagenesis of the viral genome. These observations unfold unique properties of the M protein in the HBV life cycle during unfolded protein response and point to alternative mechanisms employed by EDEMs to alleviate this stress in case of necessity by promoting glycoprotein trafficking rather than degradation. © 2016 John Wiley & Sons Ltd.

  6. Effective rumen degradation of dry matter, crude protein and neutral detergent fibre in forage determined by near infrared reflectance spectroscopy

    DEFF Research Database (Denmark)

    Ohlsson, C; Houmøller, L P; Weisbjerg, Martin Riis

    2007-01-01

    The objective of the present study was to examine if near infrared reflectance spectroscopy (NIRS) could be used to predict degradation parameters and effective degradation from scans of original forage samples. Degradability of dry matter (DM), crude protein (CP) and neutral detergent fibre (NDF......) of 61 samples of perennial ryegrass (Lolium perenne L.) and orchardgrass (Dactylis glomerata L.) was tested by using the in situ technique. The grass samples were harvested at three different stages, early vegetative growth, early reproductive growth and late reproductive growth. Degradability...

  7. GCK-MODY diabetes associated with protein misfolding, cellular self-association and degradation.

    Science.gov (United States)

    Negahdar, Maria; Aukrust, Ingvild; Johansson, Bente B; Molnes, Janne; Molven, Anders; Matschinsky, Franz M; Søvik, Oddmund; Kulkarni, Rohit N; Flatmark, Torgeir; Njølstad, Pål Rasmus; Bjørkhaug, Lise

    2012-11-01

    GCK-MODY, dominantly inherited mild fasting hyperglycemia, has been associated with >600 different mutations in the glucokinase (GK)-encoding gene (GCK). When expressed as recombinant pancreatic proteins, some mutations result in enzymes with normal/near-normal catalytic properties. The molecular mechanism(s) of GCK-MODY due to these mutations has remained elusive. Here, we aimed to explore the molecular mechanisms for two such catalytically 'normal' GCK mutations (S263P and G264S) in the F260-L270 loop of GK. When stably overexpressed in HEK293 cells and MIN6 β-cells, the S263P- and G264S-encoded mutations generated misfolded proteins with an increased rate of degradation (S263P>G264S) by the protein quality control machinery, and a propensity to self-associate (G264S>S263P) and form dimers (SDS resistant) and aggregates (partly Triton X-100 insoluble), as determined by pulse-chase experiments and subcellular fractionation. Thus, the GCK-MODY mutations S263P and G264S lead to protein misfolding causing destabilization, cellular dimerization/aggregation and enhanced rate of degradation. In silico predicted conformational changes of the F260-L270 loop structure are considered to mediate the dimerization of both mutant proteins by a domain swapping mechanism. Thus, similar properties may represent the molecular mechanisms for additional unexplained GCK-MODY mutations, and may also contribute to the disease mechanism in other previously characterized GCK-MODY inactivating mutations. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. Ubiquitin Ligase gp78 Targets Unglycosylated Prion Protein PrP for Ubiquitylation and Degradation

    OpenAIRE

    Shao, Jia; Choe, Vitnary; Cheng, Haili; Tsai, Yien Che; Weissman, Allan M.; Luo, Shiwen; Rao, Hai

    2014-01-01

    Prion protein PrP is a central player in several devastating neurodegenerative disorders, including mad cow disease and Creutzfeltd-Jacob disease. Conformational alteration of PrP into an aggregation-prone infectious form PrPSc can trigger pathogenic events. How levels of PrP are regulated is poorly understood. Human PrP is known to be degraded by the proteasome, but the specific proteolytic pathway responsible for PrP destruction remains elusive. Here, we demonstrate that the ubiquitin ligas...

  9. In-Vitro gas production technique as for feed evaluation: volume of gas production and feed degradability

    International Nuclear Information System (INIS)

    Asih Kurniawati

    2007-01-01

    In-vitro gas production technique can be used to predict feed quality. The effect of molasses supplementation as a source of degradable carbohydrate to protein source red clover silage has been done using this technique. Data showed there were positive correlation between total volume gas produced and feed degradability (r = 0.96), between total volume gas produced and microbial biomass (r = 0,96). Dry matter degradability, dry matter degraded, microbial biomass production and efficiency of nitrogen utilization, highly significant (P<0,01) increased due to increasing of degradable carbohydrate. The addition of 0.3 g molasses gave the best result whereas the addition of 0.15 g and 0.225 g have better effect than 0.0625 g molasses addition and red clover only. This result suggested that In-vitro production technique can be used as tool for feed evaluation. (author)

  10. Two waves of proteasome-dependent protein degradation in the hippocampus are required for recognition memory consolidation.

    Science.gov (United States)

    Figueiredo, Luciana S; Dornelles, Arethuza S; Petry, Fernanda S; Falavigna, Lucio; Dargél, Vinicius A; Köbe, Luiza M; Aguzzoli, Cristiano; Roesler, Rafael; Schröder, Nadja

    2015-04-01

    Healthy neuronal function and synaptic modification require a concert of synthesis and degradation of proteins. Increasing evidence indicates that protein turnover mediated by proteasome activity is involved in long-term synaptic plasticity and memory. However, its role in different phases of memory remains debated, and previous studies have not examined the possible requirement of protein degradation in recognition memory. Here, we show that the proteasome inhibitor, lactacystin (LAC), infused into the CA1 area of the hippocampus at two specific time points during consolidation, impairs 24-retention of memory for object recognition in rats. Administration of LAC after retrieval did not affect retention. These findings provide the first evidence for a requirement of proteasome activity in recognition memory, indicate that protein degradation in the hippocampus is necessary during selective time windows of memory consolidation, and further our understanding of the role of protein turnover in memory formation. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Clofibrate-induced increases in peroxisomal proteins: effect on synthesis, degradation, and mRNA activity

    International Nuclear Information System (INIS)

    Mortensen, R.M.

    1983-01-01

    The effect of clofibrate on the polypeptide composition of peroxisomes was determined. A simple method was developed for the isolation of peroxisomes with a purity of 90-95% using sedimentation in a metrizamide gradient. The specific activities of HD did not change with clofibrate treatment so that the increases in enzyme activities are solely due to increases in protein amounts. The hepatic concentration of HD increased 63 times. The HD synthesis rate, as measured by the incorporation of [ 3 H]leucine, increased 74 times, so that the increase in the synthesis was sufficient to account for the increase in protein. Clofibrate caused no discernible change in the degradation rate of HD labeled with [ 14 C]bicarbonate. The half-life of HD was approximately 2 days. The translatable mRBA coding for HD increased 55 times. This value is not significantly different from the increase in HD protein or in HD synthesis. This observation was also true for several other peroxisomal proteins. Therefore, clofibrate causes an increase in the mRNA activity, which increases the synthesis of HD leading to an accumulation of protein and enzyme activity. The kinetics of the clofibrate-induced changes in HD synthesis rate, protein level, and enzymatic activity was analyzed using a simple model which included the half-lives of the drug, mRNA, and protein. The best fit of the model to the data gave an mRNA half-life of 10 hours and a protein half-life of 1.8 days, with no significant change by clofibrate

  12. Effects of delayed wrapping of baled silage

    Science.gov (United States)

    Use of baled silage allows greater flexibility for harvest management when weather does not allow drying and harvesting forage as dry hay. However, timely wrapping on the day of baling can be difficult if significant numbers of bales need to be wrapped, or if a mechanical breakdown occurs. Researc...

  13. Hydrothermal degradation of cellulosic matter to sugars and their fermentative conversion to protein

    International Nuclear Information System (INIS)

    Bobleter, O.; Niesner, R.; Roehr, M.

    1976-01-01

    For the hydrothermal degradation of cellulosic matter, an apparatus was developed in which water is used as extraction medium. Samples, 0.15 g each, of pure cellulose (filter paper), natural straw, and 14 C-labeled straw were treated at temperatures of between 200 and 275 0 C. Of the inserted cellulose, 65.7 percent was recovered at the optimum temperature as sugars and hydroxymethylfurfural. It was possible to degrade the straw selectively: at lower temperatures, the hemicellulose part of the plant matter was converted to xylose and arabinose; and then at higher temperatures, the cellulose was converted to glucose and cellobiose. At the same time, a certain amount of the sugars was transformed to furfural compounds. The growth behavior of the yeast Candida utilis (strain Weissenbach) was analyzed, using cellobiose, xylose, and glucose (standard) as carbon sources. The growth curves applying cellobiose were nearly identical to those of glucose. Xylose showed lower productivity than the hexoses. The main products of the hydrothermal degradation can, therefore, be used favorably as nutritive substances for this protein-producing yeast

  14. Postmortem muscle protein degradation in humans as a tool for PMI delimitation.

    Science.gov (United States)

    Pittner, Stefan; Ehrenfellner, Bianca; Monticelli, Fabio C; Zissler, Angela; Sänger, Alexandra M; Stoiber, Walter; Steinbacher, Peter

    2016-11-01

    Forensic estimation of time since death relies on diverse approaches, including measurement and comparison of environmental and body core temperature and analysis of insect colonization on a dead body. However, most of the applied methods have practical limitations or provide insufficient results under certain circumstances. Thus, new methods that can easily be implemented into forensic routine work are required to deliver more and discrete information about the postmortem interval (PMI). Following a previous work on skeletal muscle degradation in the porcine model, we analyzed human postmortem skeletal muscle samples of 40 forensic cases by Western blotting and casein zymography. Our results demonstrate predictable protein degradation processes in human muscle that are distinctly associated with temperature and the PMI. We provide information on promising degradation markers for certain periods of time postmortem, which can be useful tools for time since death delimitation. In addition, we discuss external influencing factors such as age, body mass index, sex, and cause of death that need to be considered in future routine application of the method in humans.

  15. Indian hedgehog signaling triggers Nkx3.2 protein degradation during chondrocyte maturation

    Science.gov (United States)

    Choi, Seung-Won; Jeong, Da-Un; Kim, Jeong-Ah; Lee, Boyoung; Joeng, Kyu Sang; Long, Fanxin; Kim, Dae-Won

    2015-01-01

    The Indian hedgehog (Ihh) pathway plays an essential role in facilitating chondrocyte hypertrophy and bone formation during skeletal development. Nkx3.2 is initially induced in chondrocyte precursor cells, maintained in early-stage chondrocytes, and down-regulated in terminal-stage chondrocytes. Consistent with these expression patterns, Nkx3.2 has been shown to enhance chondrocyte differentiation and cell survival, while inhibiting chondrocyte hypertrophy and apoptosis. Thus, in this work, we investigate whether Nkx3.2, an early stage chondrogenic factor, can be regulated by Ihh, a key regulator for chondrocyte hypertrophy. Here, we show that Ihh signaling can induce proteasomal degradation of Nkx3.2. In addition, we found that Ihh can suppress levels of Lrp (Wnt co-receptor) and Sfrp (Wnt antagonist) expression, which, in turn, may selectively enhance Lrp-independent non-canonical Wnt pathways in chondrocyte. In agreement with these findings, Ihh-induced Nkx3.2 degradation requires Wnt5a, which is capable of triggering Nkx3.2 degradation. Finally, we found that Nkx3.2 protein levels in chondrocytes are remarkably elevated in mice defective in Ihh signaling by deletion of either Ihh or Smoothened. Thus, these results suggest that Ihh/Wnt5a signaling may play a role in negative regulation of Nkx3.2 for appropriate progression of chondrocyte hypertrophy during chondrogenesis. PMID:22507129

  16. Demonstrating the Effect of Forage Source on the Carbon Footprint of a Canadian Dairy Farm Using Whole-Systems Analysis and the Holos Model: Alfalfa Silage vs. Corn Silage

    Directory of Open Access Journals (Sweden)

    Shannan M. Little

    2017-11-01

    Full Text Available Before recommending a feeding strategy for greenhouse gas (GHG mitigation, it is important to conduct a holistic assessment of all related emissions, including from those arising from feed production, digestion of these feeds, managing the resulting manure, and other on-farm production processes and inputs. Using a whole-systems approach, the Holos model, and experimentally measured data, this study compares the effects of alfalfa silage- versus corn silage-based diets on GHG estimates in a simulated Canadian dairy production system. When all emissions and sources are accounted for, the differences between the two forage systems in terms of overall net GHG emissions were minimal. Utilizing the functional units of milk, meat, and total energy in food products generated by the system, the comparison demonstrates very little difference between the two silage production systems. However, the corn silage system generated 8% fewer emissions per kg of protein in food products as compared to the alfalfa silage system. Exploratory analysis of the impact of the two silage systems on soil carbon showed alfalfa silage has greater potential to store carbon in the soil. This study reinforces the need to utilize a whole-systems approach to investigate the interrelated effects of management choices. Reported GHG reduction factors cannot be simply combined additively because the interwoven effects of management choices cascade through the entire system, sometimes with counter-intuitive outcomes. It is necessary to apply this whole-systems approach before implementing changes in management intended to reduce GHG emissions and improve sustainability.

  17. Intercropping of corn, brachiaria grass and leguminous plants: productivity, quality and composition of silages

    Directory of Open Access Journals (Sweden)

    Patrícia Monteiro Costa

    2012-10-01

    Full Text Available The present study was carried out with the objective to evaluate the productive and qualitative characteristics of forages produced in systems of intercropping of corn, brachiaria grass and different leguminous plants. Productivity, bromatological composition and the fermentative profile of the silages from the following treatments were evaluated: corn in exclusive cultivation (CEC; intercropping of corn with brachiaria grass (CB; intercropping of corn, brachiaria grass and Calopogonium mucunoides (CBCal; intercropping of corn, brachiaria grass and Macrotyloma axillare (CBMac; and intercropping of corn, brachiaria grass and Stylozanthes capitata (CBSty. The experimental design utilized was completely randomized. For each type of cultivation, five plots or replications of three linear meters were harvested, and the material was separated. The variables assessed were: dry matter productivity per area; dry matter productivity of corn per area; crude protein production per area and productivity of total digestible nutrients per area. The material originated from the cultures was ensiled, with dry matter between 28 and 32%. After, the material was placed and compacted appropriately in bucket silos. A sample was collected from each replication for determination of the contents of DM, crude protein (CP, ether extract (EE, lignin, neutral and acid detergent fibers (NDF and ADF and TDN. A fraction of the sample of silages from each treatment was compressed for extraction of the juice and determination of the silage quality. There was difference between the forms of cultivation for the dry matter production per hectare. The CEC with production of 11920.1 kg DM/ha did not differ from CB (8997.41 kg DM/ha or CBCal (10452.10 kg DM/ha; however, it was superior to CBMac (8429.75 kg DM/ha and to CBSty (8164.83 kg DM/ha. The contents of DM, CP, NDF, ADF, lignin and TDN did not differ between the silages from the different treatments. All the silages presented

  18. SUPPLY OF Tithonia diversifolia SILAGE ALONE OR MIXED WITH CASSAVA BRAN IN BROILERS DIET

    Directory of Open Access Journals (Sweden)

    Jaime Andrés Betancourt

    2017-08-01

    Full Text Available Tithonia diversifolia and cassava bran are two food resources that noted for their protein and energy levels, respectively. The aim of this study was to evaluate the response performance of broilers fed diets containing T. diversifolia silage alone or mixed with cassava bran. We used 150 females chickens, which were housed in 15 pens and each pen was assigned to the treatments according to a randomized experimental design. The treatments consisted of three diets that were supplied to birds between 35 and 56 d of age: a control where the birds were fed only concentrate and two other treatments in which the birds were supplied concentrate of control treatment and additionally offered T. diversifolia silage either alone or mixed with cassava bran. Silage supply did not affect live weight at 56 d of age, but resulted in a reduction in concentrate consumption (P<0.05 and the carcass weight (P<0.05. It was concluded that T. diversifolia silage can be used to decrease the consumption of concentrate, but is necessary to carry out a proper habituation to avoid decrease in dry matter intake.

  19. Effect of inclusion of citrus pulp inxaraés grass silage

    Directory of Open Access Journals (Sweden)

    Júnior Issamu Yasuoka

    2016-01-01

    Full Text Available The objective of this study was to evaluate the effect of inclusion of different levels of pelleted citrus pulp (PCP on the quality of xaraés grass silage. Xaraés grass was ensiled at 54 days of growth with 0, 10, 20 and 30% PCP and divided into 20 experimental silos (five repetitions/treatment. A completely randomized design was adopted. The silos were opened after 67 days for the determination of dry matter (DM, crude protein (CP, neutral detergent fiber (NDF, acid detergent fiber (ADF, pH, and titratable acidity of the silages. The inclusion of PCP did not influence DM content, but reduced CP content. NDF and ADF content decreased with the addition of citrus pulp. Regarding pH, a significant difference was only observed for the inclusion of 30% PCP. Titratable acidity in the silage was lower in the absence of PCP (0% compared to the treatment with 30% (P<0.05. The inclusion of 30% PCP in xaraés grass silage is indicated since it improves the quality of the fibrous fraction, while maintaining acceptable pH values.

  20. Transient degradation of NF-κB proteins in macrophages after interaction with mast cell granules

    Directory of Open Access Journals (Sweden)

    Noriko Ito

    1998-01-01

    Full Text Available The exposure of the macrophage cell line, J774 to mast cell granules (MCG led to the form ation of altered nuclear transcription factor proteins (NFκBx, which had faster electrophoretic mobility than the p50 homodimer of NF-κB, but retained comparable DNA binding capacity. Antibodies to N-terminal peptides of p50, p52, p65 or c-Rel supershifted only a fraction of NF-κBx. Western blot analyses revealed that nuclear p65 and c-Rel were progressively degraded after exposure to MCG, whereas nuclear p50 appeared to be unaffected. In contrast, cytoplasmic p50, p65, c-Rel as well as IkBα remained intact after MCG treatment, although p52 was clearly degraded. In comparison to J774 cells, incubation of m ouse peritoneal macrophages with MCG resulted in more extensive alterations to NF-κB proteins. The alterations in NF-κB proteins did not affect the expression of inducible nitric oxide synthase (iNOS or TNF-α mRNA in J774 cells. These data indicate that exposure of J774 cells to MCG leads to generation of altered nuclear p52, p65 and c-Rel, which retain intact N-terminal peptides, specific oligonucleotide binding and transactivating activity. On the other hand, in peritoneal macrophages, MCG induce more extensive modifications to NF-κB proteins with associated inhibition of iNOS or TNF-α mRNA expression.

  1. Hepatitis C virus induces E6AP-dependent degradation of the retinoblastoma protein.

    Directory of Open Access Journals (Sweden)

    Tsubasa Munakata

    2007-09-01

    Full Text Available Hepatitis C virus (HCV is a positive-strand RNA virus that frequently causes persistent infections and is uniquely associated with the development of hepatocellular carcinoma. While the mechanism(s by which the virus promotes cancer are poorly defined, previous studies indicate that the HCV RNA-dependent RNA polymerase, nonstructural protein 5B (NS5B, forms a complex with the retinoblastoma tumor suppressor protein (pRb, targeting it for degradation, activating E2F-responsive promoters, and stimulating cellular proliferation. Here, we describe the mechanism underlying pRb regulation by HCV and its relevance to HCV infection. We show that the abundance of pRb is strongly downregulated, and its normal nuclear localization altered to include a major cytoplasmic component, following infection of cultured hepatoma cells with either genotype 1a or 2a HCV. We further demonstrate that this is due to NS5B-dependent ubiquitination of pRb and its subsequent degradation via the proteasome. The NS5B-dependent ubiquitination of pRb requires the ubiquitin ligase activity of E6-associated protein (E6AP, as pRb abundance was restored by siRNA knockdown of E6AP or overexpression of a dominant-negative E6AP mutant in cells containing HCV RNA replicons. E6AP also forms a complex with pRb in an NS5B-dependent manner. These findings suggest a novel mechanism for the regulation of pRb in which the HCV NS5B protein traps pRb in the cytoplasm, and subsequently recruits E6AP to this complex in a process that leads to the ubiquitination of pRb. The disruption of pRb/E2F regulatory pathways in cells infected with HCV is likely to promote hepatocellular proliferation and chromosomal instability, factors important for the development of liver cancer.

  2. Chitin Degradation Proteins Produced by the Marine Bacterium Vibrio harveyi Growing on Different Forms of Chitin.

    Science.gov (United States)

    Svitil, A L; Chadhain, S; Moore, J A; Kirchman, D L

    1997-02-01

    Relatively little is known about the number, diversity, and function of chitinases produced by bacteria, even though chitin is one of the most abundant polymers in nature. Because of the importance of chitin, especially in marine environments, we examined chitin-degrading proteins in the marine bacterium Vibrio harveyi. This bacterium had a higher growth rate and more chitinase activity when grown on (beta)-chitin (isolated from squid pen) than on (alpha)-chitin (isolated from snow crab), probably because of the more open structure of (beta)-chitin. When exposed to different types of chitin, V. harveyi excreted several chitin-degrading proteins into the culture media. Some chitinases were present with all of the tested chitins, while others were unique to a particular chitin. We cloned and identified six separate chitinase genes from V. harveyi. These chitinases appear to be unique based on DNA restriction patterns, immunological data, and enzyme activity. This marine bacterium and probably others appear to synthesize separate chitinases for efficient utilization of different forms of chitin and chitin by-products.

  3. LINGO-1 promotes lysosomal degradation of amyloid-β protein precursor

    Directory of Open Access Journals (Sweden)

    Rian de Laat

    2015-03-01

    Full Text Available Sequential proteolytic cleavages of amyloid-β protein precursor (AβPP by β-secretase and γ-secretase generate amyloid β (Aβ peptides, which are thought to contribute to Alzheimer's disease (AD. Much of this processing occurs in endosomes following endocytosis of AβPP from the plasma membrane. However, this pathogenic mode of processing AβPP may occur in competition with lysosomal degradation of AβPP, a common fate of membrane proteins trafficking through the endosomal system. Following up on published reports that LINGO-1 binds and promotes the amyloidogenic processing of AβPP we have examined the consequences of LINGO-1/AβPP interactions. We report that LINGO-1 and its paralogs, LINGO-2 and LINGO-3, decrease processing of AβPP in the amyloidogenic pathway by promoting lysosomal degradation of AβPP. We also report that LINGO-1 levels are reduced in AD brain, representing a possible pathogenic mechanism stimulating the generation of Aβ peptides in AD.

  4. Autoantibodies to myelin basic protein catalyze site-specific degradation of their antigen.

    Science.gov (United States)

    Ponomarenko, Natalia A; Durova, Oxana M; Vorobiev, Ivan I; Belogurov, Alexey A; Kurkova, Inna N; Petrenko, Alexander G; Telegin, Georgy B; Suchkov, Sergey V; Kiselev, Sergey L; Lagarkova, Maria A; Govorun, Vadim M; Serebryakova, Marina V; Avalle, Bérangère; Tornatore, Pete; Karavanov, Alexander; Morse, Herbert C; Thomas, Daniel; Friboulet, Alain; Gabibov, Alexander G

    2006-01-10

    Autoantibody-mediated tissue destruction is among the main features of organ-specific autoimmunity. This report describes "an antibody enzyme" (abzyme) contribution to the site-specific degradation of a neural antigen. We detected proteolytic activity toward myelin basic protein (MBP) in the fraction of antibodies purified from the sera of humans with multiple sclerosis (MS) and mice with induced experimental allergic encephalomyelitis. Chromatography and zymography data demonstrated that the proteolytic activity of this preparation was exclusively associated with the antibodies. No activity was found in the IgG fraction of healthy donors. The human and murine abzymes efficiently cleaved MBP but not other protein substrates tested. The sites of MBP cleavage determined by mass spectrometry were localized within immunodominant regions of MBP. The abzymes could also cleave recombinant substrates containing encephalytogenic MBP(85-101) peptide. An established MS therapeutic Copaxone appeared to be a specific abzyme inhibitor. Thus, the discovered epitope-specific antibody-mediated degradation of MBP suggests a mechanistic explanation of the slow development of neurodegeneration associated with MS.

  5. The N-end rule pathway catalyzes a major fraction of the protein degradation in skeletal muscle

    Science.gov (United States)

    Solomon, V.; Lecker, S. H.; Goldberg, A. L.

    1998-01-01

    In skeletal muscle, overall protein degradation involves the ubiquitin-proteasome system. One property of a protein that leads to rapid ubiquitin-dependent degradation is the presence of a basic, acidic, or bulky hydrophobic residue at its N terminus. However, in normal cells, substrates for this N-end rule pathway, which involves ubiquitin carrier protein (E2) E214k and ubiquitin-protein ligase (E3) E3alpha, have remained unclear. Surprisingly, in soluble extracts of rabbit muscle, we found that competitive inhibitors of E3alpha markedly inhibited the 125I-ubiquitin conjugation and ATP-dependent degradation of endogenous proteins. These inhibitors appear to selectively inhibit E3alpha, since they blocked degradation of 125I-lysozyme, a model N-end rule substrate, but did not affect the degradation of proteins whose ubiquitination involved other E3s. The addition of several E2s or E3alpha to the muscle extracts stimulated overall proteolysis and ubiquitination, but only the stimulation by E3alpha or E214k was sensitive to these inhibitors. A similar general inhibition of ubiquitin conjugation to endogenous proteins was observed with a dominant negative inhibitor of E214k. Certain substrates of the N-end rule pathway are degraded after their tRNA-dependent arginylation. We found that adding RNase A to muscle extracts reduced the ATP-dependent proteolysis of endogenous proteins, and supplying tRNA partially restored this process. Finally, although in muscle extracts the N-end rule pathway catalyzes most ubiquitin conjugation, it makes only a minor contribution to overall protein ubiquitination in HeLa cell extracts.

  6. THE INFLUENCE OF ADDITIVES FOR QUALITATIVE PARAMETERS OF SILAGE FEEDS

    Directory of Open Access Journals (Sweden)

    F. LÁD

    2007-05-01

    Full Text Available We observed the influence of silage additives for choice qualitative parameters at clover-grass silages in working conditions. We evaluated total classification and categorization to quality classes according to fermentative process. It has been found out positive effect of the silage additives for fermentative class and for total silage quality of silages. This positive effect has been more considerable at the classification to the fermentative classes at clover-grass silages. The high content of crude fibre decreased fermentative results and total silage quality at test clovergrass silages. The greatest (deterioration influence for clasification to total quality class has crude fibre content. It is see from correlation coefficient at clover-grass silages r = 0.75 (P < 0.05. The weak dependence r = 0.37 (P < 0.05 was detected between fermentative class and acetic acid content. It was detected large dependence between fermentative class and butyric acid content r = 0.73 (P < 0.05. The silages with additives had better degree of proteolysis and higher value of lactic acid. The additive improved fermentative quality. The result of better fermentative process was reduction of crude fibre values with maintainance of energy content.

  7. Activity dependent protein degradation is critical for the formation and stability of fear memory in the amygdala.

    Directory of Open Access Journals (Sweden)

    Timothy J Jarome

    Full Text Available Protein degradation through the ubiquitin-proteasome system [UPS] plays a critical role in some forms of synaptic plasticity. However, its role in memory formation in the amygdala, a site critical for the formation of fear memories, currently remains unknown. Here we provide the first evidence that protein degradation through the UPS is critically engaged at amygdala synapses during memory formation and retrieval. Fear conditioning results in NMDA-dependent increases in degradation-specific polyubiquitination in the amygdala, targeting proteins involved in translational control and synaptic structure and blocking the degradation of these proteins significantly impairs long-term memory. Furthermore, retrieval of fear memory results in a second wave of NMDA-dependent polyubiquitination that targets proteins involved in translational silencing and synaptic structure and is critical for memory updating following recall. These results indicate that UPS-mediated protein degradation is a major regulator of synaptic plasticity necessary for the formation and stability of long-term memories at amygdala synapses.

  8. A novel mosquito ubiquitin targets viral envelope protein for degradation and reduces virion production during dengue virus infection.

    Science.gov (United States)

    Troupin, Andrea; Londono-Renteria, Berlin; Conway, Michael J; Cloherty, Erin; Jameson, Samuel; Higgs, Stephen; Vanlandingham, Dana L; Fikrig, Erol; Colpitts, Tonya M

    2016-09-01

    Dengue virus (DENV) is a mosquito-borne flavivirus that causes significant human disease and mortality in the tropics and subtropics. By examining the effects of virus infection on gene expression, and interactions between virus and vector, new targets for prevention of infection and novel treatments may be identified in mosquitoes. We previously performed a microarray analysis of the Aedes aegypti transcriptome during infection with DENV and found that mosquito ubiquitin protein Ub3881 (AAEL003881) was specifically and highly down-regulated. Ubiquitin proteins have multiple functions in insects, including marking proteins for proteasomal degradation, regulating apoptosis and mediating innate immune signaling. We used qRT-PCR to quantify gene expression and infection, and RNAi to reduce Ub3881 expression. Mosquitoes were infected with DENV through blood feeding. We transfected DENV protein expression constructs to examine the effect of Ub3881 on protein degradation. We used site-directed mutagenesis and transfection to determine what amino acids are involved in Ub3881-mediated protein degradation. Immunofluorescence, Co-immunoprecipitation and Western blotting were used to examine protein interactions and co-localization. The overexpression of Ub3881, but not related ubiquitin proteins, decreased DENV infection in mosquito cells and live Ae. aegypti. The Ub3881 protein was demonstrated to be involved in DENV envelope protein degradation and reduce the number of infectious virions released. We conclude that Ub3881 has several antiviral functions in the mosquito, including specific viral protein degradation. Our data highlights Ub3881 as a target for future DENV prevention strategies in the mosquito transmission vector. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  9. Leucine supplementation stimulates protein synthesis and reduces degradation signal activation in muscle of newborn pigs during acute endotoxemia

    Science.gov (United States)

    Sepsis disrupts skeletal muscle proteostasis and mitigates the anabolic response to leucine (Leu) in muscle of mature animals. We have shown that Leu stimulates muscle protein synthesis (PS) in healthy neonatal piglets. To determine if supplemental Leu can stimulate PS and reduce protein degradation...

  10. Interplay between Molecular Chaperones and the Ubiquitin-Proteasome System in Targeting of Misfolded Proteins for Degradation

    DEFF Research Database (Denmark)

    Poulsen, Esben Guldahl

    interacting with purified 26S proteasomes, and the subsequent characterization of two novel proteasome interacting proteins. The third study was aimed at analyzing the chaperone-assisted pathway leading to degradation of misfolded kinetochore proteins in S. pombe. In this study chaperones, E2s, E3s and DUBs...

  11. Alteration of cardiac glycoside positive inotropic action by modulators of protein synthesis and degradation

    International Nuclear Information System (INIS)

    Nosek, T.M.; Adams, R.J.

    1986-01-01

    Numerous membrane bound and cytoplasmic proteins participate in the cardiac expression of the positive inotropic action (PIA) of digitalis glycosides including the Na,K-ATPase (NKA). Exposure of the myocardium to an inhibitor of protein synthesis (cycloheximide, CYC) or of protein degradation (leupeptin, LEU) alters the PIA of ouabain in isolated, paced guinea pig papillary muscles (PM) in opposite ways. In vivo exposure to CYC for 3 hr resulted in a 30% depression of the in vitro PIA of ouabain at 1.7μM compared to control. In vivo exposure to LEU for 1 hr resulted in a 47% enhancement of the in vitro PIA of 1.7μM ouabain. Neither drug had an apparent effect on the ouabain PIA ED50. Neither CYC nor LEU exposure to PM in vitro affect resting or developed tension or the response of skinned PM to calcium. The mechanisms of the PIA alterations by CYC or LEU do not involve a direct effect on the digitalis receptor. Exposure of isolated cardiac sarcolemma enriched in NKA to 10-100μM CYC or LEU did not affect NKA activity or 3 H-ouabain binding. Although direct physicochemical effects of CYC or LEU may be involved in the alterations of the ouabain PIA, it is possible that modulation of the cellular levels or turnover rate of short-lived proteins may affect cardiac regulation of the digitalis PIA

  12. Ciliopathy proteins regulate paracrine signaling by modulating proteasomal degradation of mediators

    Science.gov (United States)

    Liu, Yangfan P.; Tsai, I-Chun; Morleo, Manuela; Oh, Edwin C.; Leitch, Carmen C.; Massa, Filomena; Lee, Byung-Hoon; Parker, David S.; Finley, Daniel; Zaghloul, Norann A.; Franco, Brunella; Katsanis, Nicholas

    2014-01-01

    Cilia are critical mediators of paracrine signaling; however, it is unknown whether proteins that contribute to ciliopathies converge on multiple paracrine pathways through a common mechanism. Here, we show that loss of cilopathy-associated proteins Bardet-Biedl syndrome 4 (BBS4) or oral-facial-digital syndrome 1 (OFD1) results in the accumulation of signaling mediators normally targeted for proteasomal degradation. In WT cells, several BBS proteins and OFD1 interacted with proteasomal subunits, and loss of either BBS4 or OFD1 led to depletion of multiple subunits from the centrosomal proteasome. Furthermore, overexpression of proteasomal regulatory components or treatment with proteasomal activators sulforaphane (SFN) and mevalonolactone (MVA) ameliorated signaling defects in cells lacking BBS1, BBS4, and OFD1, in morphant zebrafish embryos, and in induced neurons from Ofd1-deficient mice. Finally, we tested the hypothesis that other proteasome-dependent pathways not known to be associated with ciliopathies are defective in the absence of ciliopathy proteins. We found that loss of BBS1, BBS4, or OFD1 led to decreased NF-κB activity and concomitant IκBβ accumulation and that these defects were ameliorated with SFN treatment. Taken together, our data indicate that basal body proteasomal regulation governs paracrine signaling pathways and suggest that augmenting proteasomal function might benefit ciliopathy patients. PMID:24691443

  13. Human Adenovirus Infection Causes Cellular E3 Ubiquitin Ligase MKRN1 Degradation Involving the Viral Core Protein pVII.

    Science.gov (United States)

    Inturi, Raviteja; Mun, Kwangchol; Singethan, Katrin; Schreiner, Sabrina; Punga, Tanel

    2018-02-01

    Human adenoviruses (HAdVs) are common human pathogens encoding a highly abundant histone-like core protein, VII, which is involved in nuclear delivery and protection of viral DNA as well as in sequestering immune danger signals in infected cells. The molecular details of how protein VII acts as a multifunctional protein have remained to a large extent enigmatic. Here we report the identification of several cellular proteins interacting with the precursor pVII protein. We show that the cellular E3 ubiquitin ligase MKRN1 is a novel precursor pVII-interacting protein in HAdV-C5-infected cells. Surprisingly, the endogenous MKRN1 protein underwent proteasomal degradation during the late phase of HAdV-C5 infection in various human cell lines. MKRN1 protein degradation occurred independently of the HAdV E1B55K and E4orf6 proteins. We provide experimental evidence that the precursor pVII protein binding enhances MKRN1 self-ubiquitination, whereas the processed mature VII protein is deficient in this function. Based on these data, we propose that the pVII protein binding promotes MKRN1 self-ubiquitination, followed by proteasomal degradation of the MKRN1 protein, in HAdV-C5-infected cells. In addition, we show that measles virus and vesicular stomatitis virus infections reduce the MKRN1 protein accumulation in the recipient cells. Taken together, our results expand the functional repertoire of the HAdV-C5 precursor pVII protein in lytic virus infection and highlight MKRN1 as a potential common target during different virus infections. IMPORTANCE Human adenoviruses (HAdVs) are common pathogens causing a wide range of diseases. To achieve pathogenicity, HAdVs have to counteract a variety of host cell antiviral defense systems, which would otherwise hamper virus replication. In this study, we show that the HAdV-C5 histone-like core protein pVII binds to and promotes self-ubiquitination of a cellular E3 ubiquitin ligase named MKRN1. This mutual interaction between the pVII and

  14. C1q protein binds to the apoptotic nucleolus and causes C1 protease degradation of nucleolar proteins.

    Science.gov (United States)

    Cai, Yitian; Teo, Boon Heng Dennis; Yeo, Joo Guan; Lu, Jinhua

    2015-09-11

    In infection, complement C1q recognizes pathogen-congregated antibodies and elicits complement activation. Among endogenous ligands, C1q binds to DNA and apoptotic cells, but whether C1q binds to nuclear DNA in apoptotic cells remains to be investigated. With UV irradiation-induced apoptosis, C1q initially bound to peripheral cellular regions in early apoptotic cells. By 6 h, binding concentrated in the nuclei to the nucleolus but not the chromatins. When nucleoli were isolated from non-apoptotic cells, C1q also bound to these structures. In vivo, C1q exists as the C1 complex (C1qC1r2C1s2), and C1q binding to ligands activates the C1r/C1s proteases. Incubation of nucleoli with C1 caused degradation of the nucleolar proteins nucleolin and nucleophosmin 1. This was inhibited by the C1 inhibitor. The nucleoli are abundant with autoantigens. C1q binding and C1r/C1s degradation of nucleolar antigens during cell apoptosis potentially reduces autoimmunity. These findings help us to understand why genetic C1q and C1r/C1s deficiencies cause systemic lupus erythematosus. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. Pu-Erh Tea Extract Induces the Degradation of FET Family Proteins Involved in the Pathogenesis of Amyotrophic Lateral Sclerosis

    Directory of Open Access Journals (Sweden)

    Yang Yu

    2014-01-01

    Full Text Available FET family proteins consist of fused in sarcoma/translocated in liposarcoma (FUS/TLS, Ewing's sarcoma (EWS, and TATA-binding protein-associated factor 15 (TAF15. Mutations in the copper/zinc superoxide dismutase (SOD1, TAR DNA-binding protein 43 (TDP-43, and FET family proteins are associated with the development of amyotrophic lateral sclerosis (ALS, a fatal neurodegenerative disease. There is currently no cure for this disease and few effective treatments are available. Epidemiological studies indicate that the consumption of tea is associated with a reduced risk of developing neurodegenerative diseases. The results of this study revealed that components of a pu-erh tea extract (PTE interacted with FET family proteins but not with TDP-43 or SOD1. PTE induced the degradation of FET family proteins but had no effects on TDP-43 or SOD1. The most frequently occurring ALS-linked FUS/TLS mutant protein, R521C FUS/TLS, was also degraded in the presence of PTE. Furthermore, ammonium chloride, a lysosome inhibitor, but not lactacystin, a proteasome inhibitor, reduced the degradation of FUS/TLS protein by PTE. PTE significantly reduced the incorporation of R521C FUS/TLS into stress granules under stress conditions. These findings suggest that PTE may have beneficial health effects, including preventing the onset of FET family protein-associated neurodegenerative diseases and delaying the progression of ALS by inhibiting the cytoplasmic aggregation of FET family proteins.

  16. Calpain activation by ROS mediates human ether-a-go-go-related gene protein degradation by intermittent hypoxia.

    Science.gov (United States)

    Wang, N; Kang, H S; Ahmmed, G; Khan, S A; Makarenko, V V; Prabhakar, N R; Nanduri, J

    2016-03-01

    Human ether-a-go-go-related gene (hERG) channels conduct delayed rectifier K(+) current. However, little information is available on physiological situations affecting hERG channel protein and function. In the present study we examined the effects of intermittent hypoxia (IH), which is a hallmark manifestation of sleep apnea, on hERG channel protein and function. Experiments were performed on SH-SY5Y neuroblastoma cells, which express hERG protein. Cells were exposed to IH consisting of alternating cycles of 30 s of hypoxia (1.5% O2) and 5 min of 20% O2. IH decreased hERG protein expression in a stimulus-dependent manner. A similar reduction in hERG protein was also seen in adrenal medullary chromaffin cells from IH-exposed neonatal rats. The decreased hERG protein was associated with attenuated hERG K(+) current. IH-evoked hERG protein degradation was not due to reduced transcription or increased proteosome/lysomal degradation. Rather it was mediated by calcium-activated calpain proteases. Both COOH- and NH2-terminal sequences of the hERG protein were the targets of calpain-dependent degradation. IH increased reactive oxygen species (ROS) levels, intracellular Ca(2+) concentration ([Ca(2+)]i), calpain enzyme activity, and hERG protein degradation, and all these effects were prevented by manganese-(111)-tetrakis-(1-methyl-4-pyridyl)-porphyrin pentachloride, a membrane-permeable ROS scavenger. These results demonstrate that activation of calpains by ROS-dependent elevation of [Ca(2+)]i mediates hERG protein degradation by IH. Copyright © 2016 the American Physiological Society.

  17. Masseter muscle myofibrillar protein synthesis and degradation in an experimental critical illness myopathy model.

    Directory of Open Access Journals (Sweden)

    Hazem Akkad

    Full Text Available Critical illness myopathy (CIM is a debilitating common consequence of modern intensive care, characterized by severe muscle wasting, weakness and a decreased myosin/actin (M/A ratio. Limb/trunk muscles are primarily affected by this myopathy while cranial nerve innervated muscles are spared or less affected, but the mechanisms underlying these muscle-specific differences remain unknown. In this time-resolved study, the cranial nerve innervated masseter muscle was studied in a unique experimental rat intensive care unit (ICU model, where animals were exposed to sedation, neuromuscular blockade (NMB, mechanical ventilation, and immobilization for durations varying between 6 h and 14d. Gel electrophoresis, immunoblotting, RT-PCR and morphological staining techniques were used to analyze M/A ratios, myofiber size, synthesis and degradation of myofibrillar proteins, and levels of heat shock proteins (HSPs. Results obtained in the masseter muscle were compared with previous observations in experimental and clinical studies of limb muscles. Significant muscle-specific differences were observed, i.e., in the masseter, the decline in M/A ratio and muscle fiber size was small and delayed. Furthermore, transcriptional regulation of myosin and actin synthesis was maintained, and Akt phosphorylation was only briefly reduced. In studied degradation pathways, only mRNA, but not protein levels of MuRF1, atrogin-1 and the autophagy marker LC3b were activated by the ICU condition. The matrix metalloproteinase MMP-2 was inhibited and protective HSPs were up-regulated early. These results confirm that the cranial nerve innervated masticatory muscles is less affected by the ICU-stress response than limb muscles, in accordance with clinical observation in ICU patients with CIM, supporting the model' credibility as a valid CIM model.

  18. Effects of cutting height and maturity on the nutritive value of corn silage for lactating cows.

    Science.gov (United States)

    Neylon, J M; Kung, L

    2003-06-01

    We studied the effect of increasing the cutting height of whole-plant corn at the time of harvest from 12.7 (NC) to 45.7 (HC) cm on yield and nutritive value of silage for dairy cows. Three leafy corn silage hybrids were harvested at NC and HC at about 34% dry matter (E) and 41% DM (L) and ensiled in laboratory silos. Increasing the height of cutting lowered yields of harvested DM/ha. In addition, the concentrations of DM and starch were higher but the concentrations of lactic acid, crude protein, neutral detergent fiber (NDF), and acid detergent fiber were lower in HC than in NC. The concentration of acid detergent lignin was also lower in HC, but only in corn harvested at E. In vitro digestion (30 h) of NDF was greater in HC (50.7%) than NC (48.3%). Calculated yield of milk per tonne of forage DM was greater for HC than for NC at E but not at L. In a lactation experiment, increasing the height of cutting of another leafy corn silage hybrid, TMF29400, in general also resulted in similar changes in nutrient composition as just described. When fed to lactating dairy cows, HC corn silage resulted in tendencies for greater NDF digestion in the total tract, higher milk production and improved feed efficiency, but there were no differences in 3.5% fat corrected milk between treatments. Results of this study suggest that increasing the cutting height of whole plant corn at harvest can improve the nutritive value of corn silage for lactating dairy cows.

  19. Feed intake, growth performance and digestibility in goats fed whole corn plant silage and Napier grass

    Directory of Open Access Journals (Sweden)

    Khaing, K.T.

    2015-06-01

    Full Text Available Shortage and inconsistent quality of forage in developing countries are the major constraints to the development of ruminant sector. To overcome these problems, feeding of ruminants with conserved forages is an important feeding strategy to ensure the success of ruminant production in the third world countries. The use of whole corn plant as silage has drawn many attention due to high protein efficiency, relatively high digestible energy and total digestible nutrients. Thus, the objective of this study was to determine feed intake, growth performance and nutrients digestibility in goats fed different inclusion level of whole corn plant silage to Napier grass based diets. Fifteen male Boer cross goats around six months old and approximately 18.54 ? 1.83 kg of body weight were used as experimental animals. The goats were assigned into five treatment groups consisted of different proportions of Napier grass (G and whole plant corn silage (CS ?T1:100/0 G/CS; T2:75/25 G/CS; T3:50/50 G/CS; T4:25/75 G/CS and T5:0/100 G/CS. The increase of corn silage to Napier grass proportion demonstrates increase in dry matter intake and growth performance in the goats. The highest nutrient digestibility was observed in T5:0/100 G/CS and T3:50/50 G/CS. It can be concluded that high proportion of corn silage to grass diets had resulted in increases in feed intake and growth performance of goats. Feeding the animals with T5 and T3 resulted in high nutrient utilization compared to other treatments. However, the highest growth performance was observed in animals that were fed with T5 diets.

  20. Effect of feeding a by-product feed-based silage on nutrients intake, apparent digestibility, and nitrogen balance in sheep.

    Science.gov (United States)

    Seok, J S; Kim, Y I; Lee, Y H; Choi, D Y; Kwak, W S

    2016-01-01

    Literature is lacking on the effects of feeding by-product feed (BF)-based silage on rumen fermentation parameters, nutrient digestion and nitrogen (N) retention in sheep. Therefore, this study was conducted to determine the effect of replacing rye straw with BF-based silage as a roughage source on ruminal parameters, total-tract apparent nutrient digestibility, and N balance in sheep. The by-product feed silage was composed of spent mushroom substrate (SMS) (45 %), recycled poultry bedding (RPB) (21 %), rye straw (11 %), rice bran (10.8 %), corn taffy residue (10 %), protected fat (1.0 %), bentonite (0.6 %), and mixed microbial additive (0.6 %). Six sheep were assigned randomly to either the control (concentrate mix + rye straw) or a treatment diet (concentrate mix + BF-based silage). Compared with the control diet, feeding a BF-based silage diet resulted in similar ruminal characteristics (pH, acetate, propionate, and butyrate concentrations, and acetate: propionate ratio), higher (p < 0.05) ruminal NH3-N, higher (p < 0.05) ether extract digestibility, similar crude protein digestibility, lower (p < 0.05) dry matter, fiber, and crude ash digestibilities, and higher (p < 0.05) N retention (g/d). The BF-based silage showed similar energy value, higher protein metabolism and utilization, and lower fiber digestion in sheep compared to the control diet containing rye straw.

  1. Chemical biology based on target-selective degradation of proteins and carbohydrates using light-activatable organic molecules.

    Science.gov (United States)

    Toshima, Kazunobu

    2013-05-01

    Proteins and carbohydrates play crucial roles in a wide range of biological processes, including serious diseases. The development of novel and innovative methods for selective control of specific proteins and carbohydrates functions has attracted much attention in the field of chemical biology. In this account article, the development of novel chemical tools, which can degrade target proteins and carbohydrates by irradiation with a specific wavelength of light under mild conditions without any additives, is introduced. This novel class of photochemical agents promise bright prospects for finding not only molecular-targeted bioprobes for understanding of the structure-activity relationships of proteins and carbohydrates but also novel therapeutic drugs targeting proteins and carbohydrates.

  2. Tau protein degradation is catalyzed by the ATP/ubiquitin-independent 20S proteasome under normal cell conditions

    OpenAIRE

    Grune, Tilman; Botzen, Diana; Engels, Martina; Voss, Peter; Kaiser, Barbara; Jung, Tobias; Grimm, Stefanie; Ermak, Gennady; Davies, Kelvin J. A.

    2010-01-01

    Tau is the major protein exhibiting intracellular accumulation in Alzheimer disease. The mechanisms leading to its accumulation are not fully understood. It has been proposed that the proteasome is responsible for degrading tau but, since proteasomal inhibitors block both the ubiquitin-dependent 26S proteasome and the ubiqutin-independent 20S proteasome pathways, it is not clear which of these pathways is involved in tau degradation. Some involvement of the ubiquitin ligase, CHIP in tau degra...

  3. Storage stability of cauliflower soup powder: The effect of lipid oxidation and protein degradation reactions.

    Science.gov (United States)

    Raitio, Riikka; Orlien, Vibeke; Skibsted, Leif H

    2011-09-15

    Soups based on cauliflower soup powders, prepared by dry mixing of ingredients and rapeseed oil, showed a decrease in quality, as evaluated by a sensory panel, during the storage of the soup powder in the dark for up to 12weeks under mildly accelerated conditions of 40°C and 75% relative humidity. Antioxidant, shown to be effective in protecting the rapeseed bulk oil, used for the powder preparation, had no effect on storage stability of the soup powder. The freshly prepared soup powder had a relatively high concentration of free radicals, as measured by electron spin resonance spectroscopy, which decreased during storage, and most remarkably during the first two weeks of storage, with only marginal increase in lipid hydroperoxides as primary lipid oxidation products, and without any increase in secondary lipid oxidation products. Analyses of volatiles by SPME-GC-MS revealed a significant increase in concentrations of 2-methyl- and 3-methyl butanals, related to Maillard reactions, together with an increase in 2-acetylpyrrole concentration. The soup powders became more brown during storage, as indicated by a decreasing Hunter L-value, in accord with non-enzymatic browning reactions. A significant increase in the concentrations of dimethyl disulfide in soup powder headspace indicated free radical-initiated protein oxidation. Protein degradation, including Maillard reactions and protein oxidation, is concluded to be more important than lipid oxidation in determining the shelf-life of dry cauliflower soup powder. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. A maize spermine synthase 1 PEST sequence fused to the GUS reporter protein facilitates proteolytic degradation.

    Science.gov (United States)

    Maruri-López, Israel; Rodríguez-Kessler, Margarita; Rodríguez-Hernández, Aída Araceli; Becerra-Flora, Alicia; Olivares-Grajales, Juan Elías; Jiménez-Bremont, Juan Francisco

    2014-05-01

    Polyamines are low molecular weight aliphatic compounds involved in various biochemical, cellular and physiological processes in all organisms. In plants, genes involved in polyamine biosynthesis and catabolism are regulated at transcriptional, translational, and posttranslational level. In this research, we focused on the characterization of a PEST sequence (rich in proline, glutamic acid, serine, and threonine) of the maize spermine synthase 1 (ZmSPMS1). To this aim, 123 bp encoding 40 amino acids of the C-terminal region of the ZmSPMS1 enzyme containing the PEST sequence were fused to the GUS reporter gene. This fusion was evaluated in Arabidopsis thaliana transgenic lines and onion monolayers transient expression system. The ZmSPMS1 PEST sequence leads to specific degradation of the GUS reporter protein. It is suggested that the 26S proteasome may be involved in GUS::PEST fusion degradation in both onion and Arabidopsis. The PEST sequences appear to be present in plant spermine synthases, mainly in monocots. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  5. Bidirectional gene sequences with similar homology to functional proteins of alkane degrading bacterium pseudomonas fredriksbergensis DNA

    International Nuclear Information System (INIS)

    Megeed, A.A.

    2011-01-01

    The potential for two overlapping fragments of DNA from a clone of newly isolated alkanes degrading bacterium Pseudomonas frederiksbergensis encoding sequences with similar homology to two parts of functional proteins is described. One strand contains a sequence with high homology to alkanes monooxygenase (alkB), a member of the alkanes hydroxylase family, and the other strand contains a sequence with some homology to alcohol dehydrogenase gene (alkJ). Overlapping of the genes on opposite strands has been reported in eukaryotic species, and is now reported in a bacterial species. The sequence comparisons and ORFS results revealed that the regulation and the genes organization involved in alkane oxidation represented in Pseudomonas frederiksberghensis varies among the different known alkane degrading bacteria. The alk gene cluster containing homologues to the known alkane monooxygenase (alkB), and rubredoxin (alkG) are oriented in the same direction, whereas alcohol dehydrogenase (alkJ) is oriented in the opposite direction. Such genomes encode messages on both strands of the DNA, or in an overlapping but different reading frames, of the same strand of DNA. The possibility of creating novel genes from pre-existing sequences, known as overprinting, which is a widespread phenomenon in small viruses. Here, the origin and evolution of the gene overlap to bacteriophages belonging to the family Microviridae have been investigated. Such a phenomenon is most widely described in extremely small genomes such as those of viruses or small plasmids, yet here is a unique phenomenon. (author)

  6. Recombinant protein production facility for fungal biomass-degrading enzymes using the yeast Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Mireille eHaon

    2015-09-01

    Full Text Available Filamentous fungi are the predominant source of lignocellulolytic enzymes used in industry for the transformation of plant biomass into high-value molecules and biofuels. The rapidity with which new fungal genomic and post-genomic data are being produced is vastly outpacing functional studies. This underscores the critical need for developing platforms dedicated to the recombinant expression of enzymes lacking confident functional annotation, a prerequisite to their functional and structural study. In the last decade, the yeast Pichia pastoris has become increasingly popular as a host for the production of fungal biomass-degrading enzymes, and particularly carbohydrate-active enzymes (CAZymes. This study aimed at setting-up a platform to easily and quickly screen the extracellular expression of biomass-degrading enzymes in Pichia pastoris. We first used three fungal glycoside hydrolases that we previously expressed using the protocol devised by Invitrogen to try different modifications of the original protocol. Considering the gain in time and convenience provided by the new protocol, we used it as basis to set-up the facility and produce a suite of fungal CAZymes (glycoside hydrolases, carbohydrate esterases and auxiliary activity enzyme families out of which more than 70% were successfully expressed. The platform tasks range from gene cloning to automated protein purifications and activity tests, and is open to the CAZyme users’ community.

  7. RING finger protein 121 facilitates the degradation and membrane localization of voltage-gated sodium channels

    Science.gov (United States)

    Ogino, Kazutoyo; Low, Sean E.; Yamada, Kenta; Saint-Amant, Louis; Zhou, Weibin; Muto, Akira; Asakawa, Kazuhide; Nakai, Junichi; Kawakami, Koichi; Kuwada, John Y.; Hirata, Hiromi

    2015-01-01

    Following their synthesis in the endoplasmic reticulum (ER), voltage-gated sodium channels (NaV) are transported to the membranes of excitable cells, where they often cluster, such as at the axon initial segment of neurons. Although the mechanisms by which NaV channels form and maintain clusters have been extensively examined, the processes that govern their transport and degradation have received less attention. Our entry into the study of these processes began with the isolation of a new allele of the zebrafish mutant alligator, which we found to be caused by mutations in the gene encoding really interesting new gene (RING) finger protein 121 (RNF121), an E3-ubiquitin ligase present in the ER and cis-Golgi compartments. Here we demonstrate that RNF121 facilitates two opposing fates of NaV channels: (i) ubiquitin-mediated proteasome degradation and (ii) membrane localization when coexpressed with auxiliary NaVβ subunits. Collectively, these results indicate that RNF121 participates in the quality control of NaV channels during their synthesis and subsequent transport to the membrane. PMID:25691753

  8. Propanol in maize silage at Danish dairy farms

    DEFF Research Database (Denmark)

    Raun, Birgitte Marie Løvendahl; Kristensen, Niels Bastian

    2010-01-01

    The objective of the present study was to investigate the prevalence maize silage containing propanol, the seasonal variation in propanol content of maize silage, and correlations between propanol and other fermentation products in maize silage collected from 20 randomly selected Danish dairy farms...... farms, the maize silage had ≥5 g propanol/kg DM. The present study indicates that dairy cows in Denmark are commonly exposed to propanol and that approximately 20% of the dairy cows will have an intake in the range of 75-100 g propanol/d under common feeding conditions....

  9. The balance of protein expression and degradation: an ESCRTs point of view.

    Science.gov (United States)

    Babst, Markus; Odorizzi, Greg

    2013-08-01

    Endosomal sorting complexes required for transport (ESCRTs) execute the biogenesis of late endosomal multivesicular bodies (MVBs). The ESCRT pathway has traditionally been viewed as a means by which transmembrane proteins are degraded in vacuoles/lysosomes. More recent studies aimed at understanding the broader functions of ESCRTs have uncovered unexpected links with pathways that control cellular metabolism. Central to this communication is TORC1, the kinase complex that controls many of the catabolic and anabolic systems. The connection between TORC1 activity and ESCRTs allows cells to quickly adapt to the stress of nutrient limitations until the longer-term autophagic pathway is activated. Increasing evidence also points to ESCRTs regulating RNA interference (RNAi) pathways that control translation. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Stage of lactation and corresponding diets affect in situ protein degradation by dairy cows.

    Science.gov (United States)

    Schadt, I; Mertens, D R; Van Soest, P J; Azzaro, G; Licitra, G

    2014-12-01

    The influence of stage of lactation and corresponding diets on rates of protein degradation (kd) is largely unstudied. Study objectives were to measure and compare in situ ruminal kd of crude protein (CP) and estimate rumen CP escape (rumen-undegradable protein; RUP) of selected feeds by cows at 3 stages of lactation fed corresponding diets, and to determine the incubation times needed in an enzymatic in vitro procedure, using 0.2 units of Streptomyces griseus protease per percent of true CP, that predicted in situ RUP. Residue CP was measured after in situ fermentation for 4, 8, 12, 24, 36, 48, and 72 h of 5 protein sources and 3 total mixed rations, which were fed to the in situ cows. Two nonlactating (dry) cows and 2 cows each at 190 (mid) and 90 (peak) days of lactation were used. Each pair of cows was offered free-choice diets that differed in composition to meet their corresponding nutrient requirements. Diets had decreasing proportions of forages and contained (dry matter basis) 11.9, 15.1 and 16.4% CP and 54.3, 40.3 and 35.3% neutral detergent fiber, for dry, mid, and peak TMR (TMR1, TMR2, and TMR3), respectively. Intakes were 10.3, 21.4, and 23.8kg of dry matter/d, respectively. Kinetic CP fractions (extractable, potentially degradable, undegradable, or slowly degradable) were unaffected by treatment. Lag time and kd varied among feeds. The kd was faster for all feeds (0.136/h) when incubated in dry-TMR1 cows compared with mid-TMR2 (0.097/h) or peak-TMR3 (0.098/h) cows, and no differences in lag time were detected. Calculated RUP, using estimated passage rates for each cow based on intake, differed between dry-TMR1 (0.382) and mid-TMR2 (0.559) or peak-TMR3 (0.626) cows, with a tendency for mid-TMR2 to be different from peak-TMR3. Using the average kd and lag time obtained from dry-TMR1 to calculate RUP for mid-TMR2 and peak-TMR3 cows using their passage rates reduced RUP values by 6.3 and 9.5 percentage units, respectively. Except for that of herring meal

  11. Effect of conventional and extrusion pelleting on in situ ruminal degradability of starch, protein, and fibre in cattle

    DEFF Research Database (Denmark)

    Razzaghi, Ali; Larsen, Mogens; Lund, Peter

    2016-01-01

    +50% sugar beet pulp (SBP), or 50% maize+50% SBP. Meals were pelleted by either conventional pelleting, or by cooking extrusion using two distinct settings giving pellets with either high density (HD) or low density (LD). Ruminal degradation of starch, crude protein (CP) and NDF, and intestinal...... affected ruminal degradability of starch, protein, and NDF differently depending on both type of cereal and composition of the concentrate mixture.......>Pelleting>Meal). In contradiction, ESD for pure wheat and wheat mixtures was reduced, though differences were minor. Conventional pelleting reduced the effective protein degradability (EPD) for pure wheat, but extrusion did not further affect the EPD. In contrast, the most intense processing with extrusion LD increased EPD...

  12. Chemical composition and fermentation characteristics of elephant grass silage with biodiesel industry co-products

    OpenAIRE

    Cleef,Eric Haydt Castello Branco van; Silva Filho,José Cleto da; Neiva Júnior,Arnaldo Prata; Patiño Pardo,René Maurício; Rêgo,Aníbal Coutinho do; Gonçalves,Josemir de Souza

    2012-01-01

    The objective of this research was to evaluate the effect of three concentrations (3, 6, and 9%) of forage turnip (Raphanus sativus) and physic nut (Jatropha curcas) cakes on dry matter, crude protein, ether extract, neutral detergent fiber, acid detergent fiber, lignin, acid detergent insoluble nitrogen neutral detergent insoluble nitrogen contents, in vitro dry matter digestibility, pH values and concentrations of N-NH3 in elephant grass silages. It was used an entirely randomized design in...

  13. Effects of Dietary Supplementation with Hainanmycin on Protein Degradation and Populations of Ammonia-producing Bacteria

    Directory of Open Access Journals (Sweden)

    Z. B. Wang

    2013-05-01

    Full Text Available An in vitro fermentation was conducted to determine the effects of hainanmycin on protein degradation and populations of ammonia-producing bacteria. The substrates (DM basis for in vitro fermentation consisted of alfalfa hay (31.7%, Chinese wild rye grass hay (28.3%, ground corn grain (24.5%, soybean meal (15.5% with a forage: concentrate of 60:40. Treatments were the control (no additive and hainanmycin supplemented at 0.1 (H0.1, 1 (H1, 10 (H10, and 100 mg/kg (H100 of the substrates. After 24 h of fermentation, the highest addition level of hainanmycin decreased total VFA concentration and increased the final pH. The high addition level of hainanmycin (H1, H10, and H100 reduced (p0.05. After 24 h of fermentation, H10 and H100 increased (p<0.05 concentrations of peptide nitrogen and AA nitrogen and proteinase activity, and decreased (p<0.05 NH3-N concentration and deaminase activity compared with control. Peptidase activitives were not affected by hainanmycin. Hainanmycin supplementation only inhibited the growth of Butyrivibrio fibrisolvens, which is one of the species of low deaminative activity. Hainanmycin supplementation also decreased (p<0.05 relative population sizes of hyper-ammonia-producing species, except for H0.1 on Clostridium aminophilum. It was concluded that dietary supplementation with hainanmycin could improve ruminal fermentation and modify protein degradation by changing population size of ammonia-producing bacteria in vitro; and the addition level of 10 mg/kg appeared to achieve the best results.

  14. Nutritional value of sorghum silage of different purposes

    Directory of Open Access Journals (Sweden)

    Arthur Behling Neto

    Full Text Available ABSTRACT Sorghum is a crop that stands out as an alternative to corn due to lower soil fertility demand and increased tolerance to drought. Lack of information about the qualitative behaviour of sorghum hinders the recommendation of different purpose sorghum cultivars. The goal was to evaluate the chemical composition and in vitro digestibility of different purpose sorghum cultivar silages, at two cropping seasons. The trial was conducted at the Plant Production Department, Federal Institute of Education, Science and Technology of Rondônia, Colorado do Oeste campus, and chemical analyses and in vitro incubation were performed at the Laboratory of Animal Nutrition, Federal University of Mato Grosso, Cuiabá campus. The experimental design was a randomized block with a split-plot arrangement and four replications. Plot treatments consisted of six different purpose sorghum cultivars (BRS 308 and BRS 310, grain sorghum; BR 655 and BRS 610, forage sorghum; and BRS 506 and CMSXS 647, sweet sorghum. Split-plot treatments consisted of two cropping periods (first crop and second crop. Forage sorghum cultivar BRS 655 demonstrated higher non-fiber carbohydrate content and lower potentially digestible fibre content than the other cultivars did. Sweet sorghum cultivars had higher levels of water soluble carbohydrates and non-protein nitrogen based on protein, lower indigestible neutral detergent fibre content at second crop, and higher in vitro dry matter digestibility than the other cultivars. The silages of sweet sorghum cultivars BRS 506 and CMSXS 647, and forage sorghum cultivar BRS 655 presented higher nutritional values.

  15. Ubiquitination and degradation of the hominoid-specific oncoprotein TBC1D3 is regulated by protein palmitoylation

    Energy Technology Data Exchange (ETDEWEB)

    Kong, Chen; Lange, Jeffrey J.; Samovski, Dmitri [Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110 (United States); Su, Xiong [Department of Internal Medicine, Center for Human Nutrition Washington University School of Medicine, St. Louis, MO 63110 (United States); Liu, Jialiu [Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110 (United States); Sundaresan, Sinju [Department of Internal Medicine, Center for Human Nutrition Washington University School of Medicine, St. Louis, MO 63110 (United States); Stahl, Philip D., E-mail: pstahl@wustl.edu [Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110 (United States)

    2013-05-03

    Highlights: •Hominoid-specific oncogene TBC1D3 is targeted to plasma membrane by palmitoylation. •TBC1D3 is palmitoylated on two cysteine residues: 318 and 325. •TBC1D3 palmitoylation governs growth factors-induced TBC1D3 degradation. •Post-translational modifications may regulate oncogenic properties of TBC1D3. -- Abstract: Expression of the hominoid-specific oncoprotein TBC1D3 promotes enhanced cell growth and proliferation by increased activation of signal transduction through several growth factors. Recently we documented the role of CUL7 E3 ligase in growth factors-induced ubiquitination and degradation of TBC1D3. Here we expanded our study to discover additional molecular mechanisms that control TBC1D3 protein turnover. We report that TBC1D3 is palmitoylated on two cysteine residues: 318 and 325. The expression of double palmitoylation mutant TBC1D3:C318/325S resulted in protein mislocalization and enhanced growth factors-induced TBC1D3 degradation. Moreover, ubiquitination of TBC1D3 via CUL7 E3 ligase complex was increased by mutating the palmitoylation sites, suggesting that depalmitoylation of TBC1D3 makes the protein more available for ubiquitination and degradation. The results reported here provide novel insights into the molecular mechanisms that govern TBC1D3 protein degradation. Dysregulation of these mechanisms in vivo could potentially result in aberrant TBC1D3 expression and promote oncogenesis.

  16. Proteolytic degradation of regulator of G protein signaling 2 facilitates temporal regulation of Gq/11 signaling and vascular contraction.

    Science.gov (United States)

    Kanai, Stanley M; Edwards, Alethia J; Rurik, Joel G; Osei-Owusu, Patrick; Blumer, Kendall J

    2017-11-24

    Regulator of G protein signaling 2 (RGS2) controls signaling by receptors coupled to the G q/11 class heterotrimeric G proteins. RGS2 deficiency causes several phenotypes in mice and occurs in several diseases, including hypertension in which a proteolytically unstable RGS2 mutant has been reported. However, the mechanisms and functions of RGS2 proteolysis remain poorly understood. Here we addressed these questions by identifying degradation signals in RGS2, and studying dynamic regulation of G q/11 -evoked Ca 2+ signaling and vascular contraction. We identified a novel bipartite degradation signal in the N-terminal domain of RGS2. Mutations disrupting this signal blunted proteolytic degradation downstream of E3 ubiquitin ligase binding to RGS2. Analysis of RGS2 mutants proteolyzed at various rates and the effects of proteasome inhibition indicated that proteolytic degradation controls agonist efficacy by setting RGS2 protein expression levels, and affecting the rate at which cells regain agonist responsiveness as synthesis of RGS2 stops. Analyzing contraction of mesenteric resistance arteries supported the biological relevance of this mechanism. Because RGS2 mRNA expression often is strikingly and transiently up-regulated and then down-regulated upon cell stimulation, our findings indicate that proteolytic degradation tightly couples RGS2 transcription, protein levels, and function. Together these mechanisms provide tight temporal control of G q/11 -coupled receptor signaling in the cardiovascular, immune, and nervous systems. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Ubiquitination is absolutely required for the degradation of hypoxia-inducible factor--1 alpha protein in hypoxic conditions.

    Science.gov (United States)

    Wang, Ronghai; Zhang, Ping; Li, Jinhang; Guan, Hongzai; Shi, Guangjun

    2016-01-29

    The hypoxia-inducible factor (HIF) is recognized as the master regulator of hypoxia response. HIF-α subunits expression are tightly regulated. In this study, our data show that ts20 cells still expressed detectable E1 protein even at 39.5° C for 12 h, and complete depletion of E1 protein expression at 39.5° C by siRNA enhanced HIF-1α and P53 protein expression. Further inhibition of E1 at 39.5 °C by siRNA, or E1 inhibitor Ube1-41 completely blocked HIF-1α degradation. Moreover, immunoprecipitations of co-transfection of HA-ubiquitin and FLAG-HIF-1α plasmids directly confirmed the involvement of ubiquitin in the hypoxic degradation of HIF-1α. Additionally, hypoxic HIF-1 α degradation is independent of HAF, RACK1, sumoylation or nuclear/cytoplasmic localization. Taken together, our data suggest that constitutive HIF-1α protein degradation in hypoxia is absolutely ubiquitination-dependent, and unidentified E3 ligase may exist for this degradation pathway. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Effect of Electron Beam Irradiation on Degradability Coefficients and Ruminalpostruminal Digestibility of Dry Matter and Crude Protein of some Plant Protein Sources

    Directory of Open Access Journals (Sweden)

    gasem tahan

    2016-06-01

    Full Text Available Effect of electron beam irradiation on degradability coefficients and ruminal- postruminal digestibility of dry matter and crude protein of soybean meal, canola meal and Lathyrus sativus seed, irradiated at doses of 50, 100 and 150 kGy was investigated. Ruminal degradability of dry matter and crude protein was determined by in situ method using two cannulated Holstein heifers. Ruminal- postruminal digestibility of dry matter and crude protein was determined by in situ (nylon bag-in vitro (daisy digestor techniques. Data analyzed using SAS software as randomized completely design and the treatment means were compared using Tukey test. The results indicated that irradiation had no effect on dry matter, ether extract and ash content of feeds. In soybean meal, washout fraction and potentially degradable fraction of dry matter and crude protein was higher and lower at dose of 150 kGy irradiation than other treatments, respectively, and degradation rate constant and ruminal effective degradability of dry matter and crude protein was lower at all doses of irradiation than untreated soybean meal. In canola meal, irradiation at doses of 50 and 100 kGy decreased washout fraction and increased potentially degradable fraction of crude protein compared with untreated canola meal. In Lathyrus sativus seed, only potentially degradable fraction of dry matter and crude protein was lower at dose of 150 kGy irradiation than untreated Lathyrus sativus seed. Ruminal digestibility of crude protein decreased in soybean meal at doses of 100 and 150 kGy irradiation and for canola meal at all doses of irradiation than untreated samples. Total tract digestibility of crude protein decreased in soybean meal at dose of 150 kGy irradiation and for canola meal at all doses of irradiation than untreated samples. In Lathyrus sativus seed, ruminal-postruminal digestibility and total tract digestibility of dry matter increased at doses of 100 and 150 kGy irradiation than untreated

  19. Digestion and ruminal fermentation of cocoa pod silage based ration enriched by gliricidia and calliandra leaves on goats

    Directory of Open Access Journals (Sweden)

    Puastuti W

    2015-03-01

    Full Text Available In term of availability, cacao pod is potential for ruminant feed. According to its nutrients content, cacao pod can be used as feed fiber source. Protein sources materials must be added when cacao pod was ensilaged due to low protein content of this material. The aim of this study was to investigate digestibility value and end products of rumen fermentation of goat fed grass or cacao pod based ration. Randomized block design and 20 heads of lambs (16.95±2.36 kg to evaluated 5 type of rations: R (50% grass + 50% concentrate; S (50% cacao pod silage + 50% concentrate; SG (50% cacao pod-gliricidia silage + 50% concentrate; SK (50% cacao pod-calliandra silage + 50% concentrate dan SC (50% cacao pod-mixture of gliricidia-calliandra silage + 50% concentrate. Feeding trial was conducted for over 15 weeks. Measurements were taken on feed digestibility and rumen-fermentation end-products after 3 weeks of treatments. Results shows that nutrients digestibility was different significantly among the groups of treatments (P<0.05. Digestibillity of organic matter, NDF and energy of R ration was those of higher significantly (P<0.05 than those of other groups. N-ammonia of rumen from goat feed R ration was higher (P<0.05 than other groups. Total VFA and each component were different among the groups (P<0.05, however the value was similar among the groups of cacao pod silage rations. It is concluded that cacao pod silaged based rations enriched by Gliricidia and Calliandra leaves did not produce similar digestibility value and end products of rumen fermentation with grass based ration.

  20. 6种肉牛常用粗饲料瘤胃降解特性和瘤胃非降解蛋白质的小肠消化率%Ruminal Degradation Characteristics and Small Intestinal Digestibility of Rumen Undegraded Protein of Six Kinds of Commonly Used Roughages for Steers

    Institute of Scientific and Technical Information of China (English)

    陈艳; 张晓明; 王之盛; 彭全辉; 邹华围

    2014-01-01

    本试验旨在研究黑麦草、牛鞭草、甘薯蔓、玉米秸青贮、玉米秸秆和稻草共6种肉牛常用粗饲料的瘤胃降解特性和瘤胃非降解蛋白质( RUP)的小肠消化率。选用3头装有永久性瘤胃瘘管的宣汉阉公牛为试验动物,采用尼龙袋技术评定6种粗饲料的干物质( DM)、粗蛋白质( CP)、中性洗涤纤维( NDF)和酸性洗涤纤维( ADF)瘤胃降解率,并用改进三步法评价RUP的小肠消化率。结果表明:1)黑麦草和甘薯蔓DM有效降解率较高,并与依次降低的牛鞭草、玉米秸青贮、玉米秸秆和稻草差异显著( P<0.05)。CP有效降解率和 ADF有效降解率以黑麦草最高,且与依次降低的甘薯蔓、牛鞭草、玉米秸青贮、玉米秸秆和稻草差异显著( P<0.05)。NDF有效降解率为黑麦草>甘薯蔓>玉米秸青贮>牛鞭草>稻草>玉米秸秆,各粗饲料间差异显著(P<0.05)。2)牛鞭草、玉米秸秆和稻草RUP的小肠消化率差异不显著( P>0.05),并显著高于依次降低的玉米秸青贮、黑麦草和甘薯蔓( P<0.05)。小肠可消化粗蛋白质含量为黑麦草>甘薯蔓>牛鞭草>玉米秸青贮>玉米秸秆>稻草,各粗饲料间差异显著( P<0.05)。由此可见,不同粗饲料瘤胃降解特性不同,为小肠提供可消化粗蛋白质的潜力也不同。黑麦草的 DM、CP、NDF 和ADF在瘤胃的有效降解率最高,牛鞭草、玉米秸秆和稻草RUP的小肠消化率较高,黑麦草和甘薯蔓小肠可消化粗蛋白质含量较高。%This experiment was conducted to determine the ruminal degradation characteristics and small intesti-nal digestibility of rumen undegraded protein( RUP)of 6 kinds of commonly used roughages:ryegrass,hem-arthria compressa,sweet potato stem,corn stover silage,corn stover and rice straw. Three Xuanhan steers fit-ted with permanent ruminal cannulas were used. Nylon

  1. Effects of wheat dried distillers' grains with solubles and cinnamaldehyde on in vitro fermentation and protein degradation using the Rusitec technique.

    Science.gov (United States)

    Lia, Yangling; He, Maolong; Li, Chun; Forster, Robert; Beauchemin, Karen Anne; Yang, Wenzhu

    2012-04-01

    This study was conducted to evaluate the effect of wheat dried distillers' grains with solubles (DDGS) and cinnamaldehyde (CIN) on in vitro fermentation and microbial profiles using the rumen simulation technique. The control substrate (10% barley silage, 85% barley grain and 5% supplement, on dry matter basis) and the wheat DDGS substrate (30% wheat DDGS replaced an equal portion of barley grain) were combined with 0 and 300 mg CIN/l of culture fluid. The inclusion of DDGS increased (p fermentation pattern changed to greater acetate and less propionate proportions (p fermentability and potentially increase protein flows to the intestine. Supplementation of high-grain substrates with CIN reduced methane production and potentially increased the true protein reaching the small intestine; however, overall reduction of feed fermentation may lower the feeding value of a high-grain diet.

  2. Rift Valley fever virus NSs inhibits host transcription independently of the degradation of dsRNA-dependent Protein Kinase PKR

    OpenAIRE

    Kalveram, Birte; Lihoradova, Olga; Indran, Sabarish V.; Lokugamage, Nandadeva; Head, Jennifer A.; Ikegami, Tetsuro

    2012-01-01

    Rift Valley fever virus (RVFV) encodes one major virulence factor, the NSs protein. NSs suppresses host general transcription, including interferon (IFN)-β mRNA synthesis, and promotes degradation of the dsRNA-dependent protein kinase (PKR). We generated a novel RVFV mutant (rMP12-NSsR173A) specifically lacking the function to promote PKR degradation. rMP12-NSsR173A infection induces early phosphorylation of eIF2α through PKR activation, while retaining the function to inhibit host general tr...

  3. Memory formation for trace fear conditioning requires ubiquitin-proteasome mediated protein degradation in the prefrontal cortex.

    Directory of Open Access Journals (Sweden)

    David S Reis

    2013-10-01

    Full Text Available The cellular mechanisms supporting plasticity during memory consolidation have been a subject of considerable interest. De novo protein and mRNA synthesis in several brain areas are critical, and more recently protein degradation, mediated by the ubiquitin-proteasome system (UPS, has been shown to be important. Previous work clearly establishes a relationship between protein synthesis and protein degradation in the amygdala, but it is unclear whether cortical mechanisms of memory consolidation are similar to those in the amygdala. Recent work demonstrating a critical role for prefrontal cortex (PFC in the acquisition and consolidation of fear memory allows us to address this question. Here we use a PFC-dependent fear conditioning protocol to determine whether UPS mediated protein degradation is necessary for memory consolidation in PFC. Groups of rats were trained with auditory delay or trace fear conditioning and sacrificed 60 min after training. PFC tissue was then analyzed to quantify the amount of polyubiquinated protein. Other animals were trained with similar procedures but were infused with either a proteasome inhibitor (clasto-lactacystin β-lactone or a translation inhibitor (anisomycin in the PFC immediately after training. Our results show increased UPS-mediated protein degradation in the PFC following trace but not delay fear conditioning. Additionally, post-training proteasome or translation inhibition significantly impaired trace but not delay fear memory when tested the next day. Our results further support the idea that the PFC is critical for trace but not delay fear conditioning highlight the role of UPS-mediated degradation as critical for synaptic plasticity.

  4. Neuroprotective mechanism of Kai Xin San: upregulation of hippocampal insulin-degrading enzyme protein expression and acceleration of amyloid-beta degradation

    Directory of Open Access Journals (Sweden)

    Na Wang

    2017-01-01

    Full Text Available Kai Xin San is a Chinese herbal formula composed of Radix Ginseng , Poria , Radix Polygalae and Acorus Tatarinowii Rhizome . It has been used in China for many years for treating amnesia. Kai Xin San ameliorates amyloid-β (Aβ-induced cognitive dysfunction and is neuroprotective in vivo , but its precise mechanism remains unclear. Expression of insulin-degrading enzyme (IDE, which degrades Aβ, is strongly correlated with cognitive function. Here, we injected rats with exogenous Aβ42 (200 μM, 5 μL into the hippocampus and subsequently administered Kai Xin San (0.54 or 1.08 g/kg/d intragastrically for 21 consecutive days. Hematoxylin-eosin and Nissl staining revealed that Kai Xin San protected neurons against Aβ-induced damage. Furthermore, enzyme-linked immunosorbent assay, western blot and polymerase chain reaction results showed that Kai Xin San decreased Aβ42 protein levels and increased expression of IDE protein, but not mRNA, in the hippocampus. Our findings reveal that Kai Xin San facilitates hippocampal Aβ degradation and increases IDE expression, which leads, at least in part, to the alleviation of hippocampal neuron injury in rats.

  5. Decomposition of insoluble and hard-to-degrade animal proteins by enzyme E77 and its potential applications.

    Science.gov (United States)

    Zhao, Hui; Mitsuiki, Shinji; Takasugi, Mikako; Sakai, Masashi; Goto, Masatoshi; Kanouchi, Hiroaki; Oka, Tatsuzo

    2012-04-01

    Insoluble and hard-to-degrade animal proteins are group of troublesome proteins, such as collagen, elastin, keratin, and prion proteins that are largely generated by the meat industry and ultimately converted to industrial wastes. We analyzed the ability of the abnormal prion protein-degrading enzyme E77 to degrade insoluble and hard-to-degrade animal proteins including keratin, collagen, and elastin. The results indicate that E77 has a much higher keratinolytic activity than proteinase K and subtilisin. Maximal E77 keratinolytic activity was observed at pH 12.0 and 65 °C. E77 was also adsorbed by keratin in a pH-independent manner. E77 showed lower collagenolytic and elastinolytic specificities than proteinase K and subtilisin. Moreover, E77 treatment did not damage collagens in ovine small intestines but did almost completely remove the muscles. We consider that E77 has the potential ability for application in the processing of animal feedstuffs and sausages.

  6. Investigating ER-Associated Degradation with RNAi Screening - and Searching for Model Proteins to Do It with

    DEFF Research Database (Denmark)

    Jensen, Njal Winther

    Abstract In eukaryotes, secretory proteins are translocated into the endoplasmic reticulum (ER) for folding assistance, acquisition of posttranslational modifications and sorting. Proteins that do not obtain their native conformation are eliminated by ER-associated degradation (ERAD). ERAD...... is a sophisticated pathway that recognizes misfolded proteins and targets them for degradation by the 26S proteasome residing in the cytosol. More than 60 diseases including Alzheimer’s disease, Huntington’s disease and Parkinson’s disease have been linked to the ERAD pathway underscoring its crucial role...... for cellular homeostasis. The aim of this thesis has been to gain insight into ERAD. The experimental approach was RNAi screening, which is a fast and efficient method for initial evaluation of a large pool of genes. Since relatively few proteins routinely are used as ERAD substrates, the first goal...

  7. Comparison of protein degradation, protein oxidation, and μ-calpain activation between pale, soft, and exudative and red, firm, and nonexudative pork during postmortem aging.

    Science.gov (United States)

    Yin, Y; Zhang, W G; Zhou, G H; Guo, B

    2014-08-01

    The objective of this study was to investigate the differences in protein modifications between pale, soft, and exudative (PSE) and red, firm, and nonexudative (RFN) pork during postmortem (PM) aging. Longissimus dorsi (LD) including 8 PSE and 8 RFN muscles were individually removed from 16 carcasses. These 16 LD muscles were vacuum packaged at 24 h after slaughter and stored at 4°C for 1, 3, and 5 d. The centrifugation loss, drip loss, color, protein solubility, protein oxidation, protein degradation including desmin, troponin T, and integrin, and μ-calpain activation were determined. The pH of PSE samples was significantly lower than that of RFN samples at both 1 and 24 h PM (P 0.05). In addition, PSE pork presented a lower solubility of sarcoplasmic protein, myofibrillar protein, and total protein than RFN pork except the solubility of myofibrillar protein at d 1 (P firm, and nonexudative pork presented lower intensity of intact 80 kDa calpain and greater intensity of autolyzed 76 kDa product compared to PSE pork (P < 0.01). The results indicate that the degree of μ-calpain activation, the extent of protein degradation including desmin and integrin, and the level of protein solubility in PSE pork could contribute to its low water holding capacity during PM storage.

  8. Effect of Rapid Chilling on Beef Quality and Cytoskeletal Protein Degradation in of Chinese Yellow Crossbred Bulls

    Directory of Open Access Journals (Sweden)

    Yanwei Mao

    2012-08-01

    Full Text Available The objective of this study was to investigate the effect of rapid chilling (RC on beef quality and the degradation of cytoskeletal proteins. Twenty Chinese Yellow crossbred bulls were selected and randomly divided into two groups. RC and conventional chilling (CC were applied to left and right sides of the carcasses respectively after slaughtering. To determine whether electrical stimulation (ES treatment can alleviate the potential hazard of RC on meat quality, ES was applied to one group. The effects of RC and ES were determined by meat color, shear force and cytoskeletal protein degradation postmortem (PM. The results showed that RC decreased beef tenderness at 1 d and 3 d postmortem, but had no detrimental effect on meat color. Western blotting showed that RC decreased the degradation rate of desmin and troponin-T, but the effects weakened gradually as postmortem aging extended. Degradation rates of both desmin and troponin-T were accelerated by ES. The combination of RC and ES could improve beef color, accelerate degradation rate of cytoskeletal protein and improve beef tenderness.

  9. Plasma urea nitrogen and progesterone concentrations and follicular dynamics in ewes fed proteins of different degradability

    Directory of Open Access Journals (Sweden)

    Gustavo Bianchi Lazarin

    2012-07-01

    Full Text Available The effects of overfeeding with protein of different degradability on body condition, plasma urea nitrogen and progesterone concentrations, ovulation number and follicular dynamics were assessed in Santa Ines ewes. Twelve ewes were assigned to a randomized block design according to body weight and received overfeeding with soybean meal or with corn gluten meal or maintenance diet for 28 days before ovulation and during the next estrous cycle. Blood samples were taken on days 7, 14, 21, and 28 after the beginning of treatments for analysis of plasma urea nitrogen and on days 3, 6, 9, 12, and 15 into the estrous cycle for analysis of plasma urea nitrogen and progesterone. Follicular dynamics was monitored daily by ultrasound during one estrous cycle. Dry matter and crude protein intake, weight gain, plasma urea nitrogen concentration before ovulation, number of ovulations, diameter of the largest follicle of the 1st and of the 2nd waves and the growth rate of the largest follicle of the 1st wave were higher in the ewes that received overfeeding. The growth rate of the largest follicle of the 3rd wave was higher in the ewes fed maintenance diet. The back fat thickness, plasma urea nitrogen before ovulation and progesterone concentrations, diameter of the largest follicle of the 2nd wave and growth rate of the largest follicle of the 3rd wave were higher in ewes that received overfeeding with soybean meal. The growth rate of the largest follicle of the 1st wave was higher in ewes that received overfeeding with corn gluten meal. Overfeeding with protein-rich feeds may increase the ovulation number and with soybean meal, it may be effective in increasing plasma progesterone concentration in ewes.

  10. Trafficking and degradation pathways in pathogenic conversion of prions and prion-like proteins in neurodegenerative diseases.

    Science.gov (United States)

    Victoria, Guiliana Soraya; Zurzolo, Chiara

    2015-09-02

    Several neurodegenerative diseases such as transmissible spongiform encephalopathies, Alzheimer's and Parkinson's diseases are caused by the conversion of cellular proteins to a pathogenic conformer. Despite differences in the primary structure and subcellular localization of these proteins, which include the prion protein, α-synuclein and amyloid precursor protein (APP), striking similarity has been observed in their ability to seed and convert naïve protein molecules as well as transfer between cells. This review aims to cover what is known about the intracellular trafficking of these proteins as well as their degradation mechanisms and highlight similarities in their movement through the endocytic pathway that could contribute to the pathogenic conversion and seeding of these proteins which underlies the basis of these diseases. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Characteristics of on-demand biogas production by using sugar beet silage.

    Science.gov (United States)

    Ahmed, Sharif; Kazda, Marian

    2017-08-01

    On-demand electricity generation can be achieved by just-in-time biogas production instantly utilized in co-generation units. For this goal, easily degradable substrates like sugar beet silage have a high potential. Potential for on-demand biogas production from co-digestion of sugar beet silage (SS) with grass silage (GS) was evaluated in two experiments at organic loading rates (OLRs) of 1.5 kgVS m -3 day -1 and 2.5 kgVS m -3 day -1 , respectively. Each experiment was fed with intermittent feeding system at 8 hrs interval at the same feedstock ratios (volatile solids based) of GS:SS-1:0, 3:1 and 1:3, respectively. Modelling by Gaussian equation was performed in order to understand the effects of SS on biogas production. Addition of sugar beet silage led to maximum biogas production within a short time, but it differed significantly depending on feedstock ratios and OLRs, respectively. At OLR 1.5 kgVS m -3 day -1 , during mono fermentation of grass silage maximum biogas production rate of 0.27 l N hr -1 was reached at 2.74 hrs. Production rate did not change at feedstock ratio of GS:SS-3:1 but increased to 0.64 l N hr -1 at GS:SS-1:3 within a shorter time span (1.58 hrs). On the contrary, at OLR of 2.5 kgVS m -3 day -1 time span between feedstock input and maximum biogas production did not differ significantly (p > 0.05) among the reactors. Biogas production rates were 0.60 l N hr -1 within 2.27 hrs and 0.82 l N hr -1 within 2.30 hrs at GS:SS-3:1 and GS:SS-1:3, respectively. Surprisingly, there was no time lag between maximum biogas and methane production rates, irrespectively of OLR. This implies that once the whole microbial community is adapted to intermittent substrate input, the metabolic products are instantly utilized through the all steps of anaerobic substrate degradation. Applying this finding opens new perspectives for on-demand biogas energy production. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Effect of cumin essential oil usage on fermentation quality, aerobic stability and in vitro digetibility of alfalfa silage.

    Science.gov (United States)

    Turan, Aslı; Önenç, Sibel Soycan

    2018-03-02

    This study was carried out to determine the effects of cumin essential oil on the silage fermentation, aerobic stability and in vitro digestibility of alfalfa silages. Alfalfa was harvested at early bloom (5 th cutting) stage in October and wilted for about 3 hours. The research was carried out at three groups which were the control group where no additive control was done (CON), cumin essential oil (CMN3) with 300 mg/kg and CMN5 with 500 mg/kg cumin essential oil addition. Alfalfa was ensiled in plastic bags. The packages were stored at 8±2 °C under laboratory conditions. All groups were sampled for physical, chemical and microbiological analysis 120th day after ensiling. At the end of the ensiling period, all silages were subjected to an aerobic stability test for 7 days. In addition, enzimatic solubility of organic matter (ESOM), metabolizable energy (ME) and relative feed value (RFV) of these silages were determined. pH level decreased in the cumin groups compared to CON (P<0.05), thus inhibiting proteolytic enzymes from breaking down proteins into ammonia. In addition, it increased ESOM amount, and concordantly provided an increase of ME contents. Similarly, dry matter intake (DMI) and RFV ratio increased. After opening the silage, it kept its aerobic stability for three days. Cumin essential oil improved fermentation, and affected chemical and microbiological characteristics of silages. Especially the addition of 300 mg/kg cumin provided cell wall fractionation through stimulating the activities of enzymes responsible. It also increased the number and activity of lactic acid bacteri (LAB) through providing a development of LAB.

  13. Protein degradation and post-deboning tenderization in broiler breast meat with different degrees of muscle shortening

    Science.gov (United States)

    Deboning broiler breast fillets prior to rigor mortis negatively influences tenderness due to sarcomere shortening. The effects of sarcomere shortening on muscle protein degradation and breast meat tenderization during post-deboning aging are not well understood. The objective of this study was to m...

  14. Protein degradation rate is the dominant mechanism accounting for the differences in protein abundance of basal p53 in a human breast and colorectal cancer cell line.

    Directory of Open Access Journals (Sweden)

    Eszter Lakatos

    Full Text Available We determine p53 protein abundances and cell to cell variation in two human cancer cell lines with single cell resolution, and show that the fractional width of the distributions is the same in both cases despite a large difference in average protein copy number. We developed a computational framework to identify dominant mechanisms controlling the variation of protein abundance in a simple model of gene expression from the summary statistics of single cell steady state protein expression distributions. Our results, based on single cell data analysed in a Bayesian framework, lends strong support to a model in which variation in the basal p53 protein abundance may be best explained by variations in the rate of p53 protein degradation. This is supported by measurements of the relative average levels of mRNA which are very similar despite large variation in the level of protein.

  15. Protein degradation rate is the dominant mechanism accounting for the differences in protein abundance of basal p53 in a human breast and colorectal cancer cell line.

    Science.gov (United States)

    Lakatos, Eszter; Salehi-Reyhani, Ali; Barclay, Michael; Stumpf, Michael P H; Klug, David R

    2017-01-01

    We determine p53 protein abundances and cell to cell variation in two human cancer cell lines with single cell resolution, and show that the fractional width of the distributions is the same in both cases despite a large difference in average protein copy number. We developed a computational framework to identify dominant mechanisms controlling the variation of protein abundance in a simple model of gene expression from the summary statistics of single cell steady state protein expression distributions. Our results, based on single cell data analysed in a Bayesian framework, lends strong support to a model in which variation in the basal p53 protein abundance may be best explained by variations in the rate of p53 protein degradation. This is supported by measurements of the relative average levels of mRNA which are very similar despite large variation in the level of protein.

  16. Silage review: Factors affecting dry matter and quality losses in silages.

    Science.gov (United States)

    Borreani, G; Tabacco, E; Schmidt, R J; Holmes, B J; Muck, R E

    2018-05-01

    An overview was made of dry matter (DM) and quality losses that occur during the ensiling process from the field through the feeding phase. The aim was to review the relevant published literature of the last 15 yr focusing on developments achieved after the publication of the book Silage Science and Technology. This review discusses the factors affecting DM and quality losses in terms of field and pre-ensiling conditions, respiration and temperature at ensiling, fermentation patterns, methods of covering and weighting the silage cover, and management of aerobic deterioration. The possibility of reducing DM and quality losses during the ensiling process requires knowledge of how to measure losses on farm and establish the status of the silage during the feed-out phase, implementing the most effective management practices to avoid air exposure during conservation and reduce silage aerobic deterioration during feeding. The paper concludes with future perspectives and recommended management practices to reduce losses and increase efficiency over the whole ensiling process in view of increasing sustainability of the livestock production chain. The Authors. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

  17. Wogonin inhibits tumor angiogenesis via degradation of HIF-1α protein

    International Nuclear Information System (INIS)

    Song, Xiuming; Yao, Jing; Wang, Fei; Zhou, Mi; Zhou, Yuxin; Wang, Hu; Wei, Libin; Zhao, Li; Li, Zhiyu; Lu, Na; Guo, Qinglong

    2013-01-01

    Wogonin, a plant-derived flavone, has been shown recently to have antitumor effects. However, the mechanisms that wogonin inhibits tumor angiogenesis are not well known. In this study, we investigated the effects of wogonin on expression of hypoxia-inducible factor-1α (HIF-1α) and secretion of vascular endothelial growth factor (VEGF) in tumor cells. We found that wogonin decreased the expression of HIF-1α by affecting its stability and reduced the secretion of VEGF, which suppressed angiogenesis in cancer. Wogonin promoted the degradation of HIF-1α by increasing its prolyl hydroxylation, which depended on prolyl hydroxylase (PHD) and the von Hippel–Lindau tumor suppressor (VHL). Intriguingly, wogonin impeded the binding between heat-shock protein 90 (Hsp90) and HIF-1α. In addition, wogonin down-regulated the Hsp90 client proteins EGFR, Cdk4 and survivin, but did not affect the level of Hsp90. Wogonin also increased ubiquitination of HIF-1α and promoted its degradation in proteasome. We also found that wogonin could inhibit nuclear translocation of HIF-1α. Electrophoresis mobility shift assay (EMSA) showed that wogonin decreased the binding activity of exogenous consensus DNA oligonucleotide with HIF-1α in nuclear extracts from MCF-7 cells. Chromatin immunoprecipitation (ChIP) assay also revealed that HIF-1α directly binded to endogenous hypoxia-responsive element (HRE) and this binding was significantly decreased in MCF-7 cells treated with wogonin. Preliminary results indicated in vivo activity of wogonin against xenograft-induced angiogenesis in nude mice. Taken together, the results suggested that wogonin was a potent inhibitor of HIF-1α and provided a new insight into the mechanisms of wogonin against cancers. - Highlights: • Wogonin is an all around inhibitor of VEGF signaling. • We firstly demonstrate that wogonin inhibits secretion of VEGF by decreasing HIF-1α. • Wogonin enhances PDH and VHL expression and inhibits Hsp90 function.

  18. Wogonin inhibits tumor angiogenesis via degradation of HIF-1α protein

    Energy Technology Data Exchange (ETDEWEB)

    Song, Xiuming; Yao, Jing; Wang, Fei; Zhou, Mi; Zhou, Yuxin; Wang, Hu; Wei, Libin; Zhao, Li; Li, Zhiyu; Lu, Na, E-mail: luna555@163.com; Guo, Qinglong, E-mail: anticancer_drug@yahoo.com.cn

    2013-09-01

    Wogonin, a plant-derived flavone, has been shown recently to have antitumor effects. However, the mechanisms that wogonin inhibits tumor angiogenesis are not well known. In this study, we investigated the effects of wogonin on expression of hypoxia-inducible factor-1α (HIF-1α) and secretion of vascular endothelial growth factor (VEGF) in tumor cells. We found that wogonin decreased the expression of HIF-1α by affecting its stability and reduced the secretion of VEGF, which suppressed angiogenesis in cancer. Wogonin promoted the degradation of HIF-1α by increasing its prolyl hydroxylation, which depended on prolyl hydroxylase (PHD) and the von Hippel–Lindau tumor suppressor (VHL). Intriguingly, wogonin impeded the binding between heat-shock protein 90 (Hsp90) and HIF-1α. In addition, wogonin down-regulated the Hsp90 client proteins EGFR, Cdk4 and survivin, but did not affect the level of Hsp90. Wogonin also increased ubiquitination of HIF-1α and promoted its degradation in proteasome. We also found that wogonin could inhibit nuclear translocation of HIF-1α. Electrophoresis mobility shift assay (EMSA) showed that wogonin decreased the binding activity of exogenous consensus DNA oligonucleotide with HIF-1α in nuclear extracts from MCF-7 cells. Chromatin immunoprecipitation (ChIP) assay also revealed that HIF-1α directly binded to endogenous hypoxia-responsive element (HRE) and this binding was significantly decreased in MCF-7 cells treated with wogonin. Preliminary results indicated in vivo activity of wogonin against xenograft-induced angiogenesis in nude mice. Taken together, the results suggested that wogonin was a potent inhibitor of HIF-1α and provided a new insight into the mechanisms of wogonin against cancers. - Highlights: • Wogonin is an all around inhibitor of VEGF signaling. • We firstly demonstrate that wogonin inhibits secretion of VEGF by decreasing HIF-1α. • Wogonin enhances PDH and VHL expression and inhibits Hsp90 function.

  19. Parâmetros da degradação protéica ruminal de diferentes alimentos e rações estimados por técnica in vitro Parameters of ruminal protein degradation of different feeds and diets estimated by an in vitro method

    Directory of Open Access Journals (Sweden)

    R.P. Lana

    2007-04-01

    -elefante apresentou o menor nível de amônia e a de cepilho, o maior. As camas influenciaram o conteúdo de proteína solúvel e a DPB, que variaram de 9,0 a 14,5mg/dl e 39 a 63%, respectivamente, sem efeitos da monensina.Three experiments were carried out in order to study the parameters of ruminal protein degradation. In the first, isoproteic diets, constituted of elephant grass, ground corn and soybean meal, at five concentrate levels (0:100, 25:75, 50:50, 75:25 e 100:0, with or without monensin (5µM, were incubated in ruminal fluid of bovines. There was a decreasing linear effect of the concentrate level on ammonia concentration and degradability of crude protein, and cubic effect in soluble protein concentration, with the largest value in the diet with 25% concentrate. Monensin decreased degradability of crude protein and soluble protein concentration with no effect on ammonia production. In the second, five different roughages (corn and elephant grass silage - Pennisetum purpureum, Brachiaria haylage - Brachiaria decumbens, ammoniated Tifton 85 hay - Cynodon sp. e Tifton 85 hay. The were incubated Brachiaria haylage and the ammoniated Tifton 85 hay showed the greatest concentrations of ammonia (8.7 and 5.3mg/dl and soluble protein (5.4 and 7.0mg/dl, due to their higher crude protein content, followed by elephant grass silage and Tifton 85 hay. The degradability of crude protein ranged from 29.6 to 80.6% for Brachiaria haylage and Tifton 85 hay, and the degradability of dry matter ranged from 40.1 to 64.3% for elephant grass silage and Brachiaria haylage, respectively. The effective degradability of crude protein showed low values due to low degradation rate of the insoluble fraction. In the third, four different poultry litter (hulls coffee, shopped dry elephant grass, corn cobs and wood shavings were incubated, with or without monensin (5µM. No difference in ammonia concentration among the poultry litter samples, was observed in the absence of monensin. However, when

  20. Degradation kinetics of fisetin and quercetin in solutions affected by medium pH, temperature and co-existed proteins

    Directory of Open Access Journals (Sweden)

    Wang Jing

    2016-01-01

    Full Text Available Impacts of medium pH, temperature and coexisted proteins on the degradation of two flavonoids fisetin and quercetin were assessed by spectroscopic method in the present study. Based on the measured degradation rate constants (k, fisetin was more stable than quercetin in all cases. Increasing medium pH from 6.0 to 7.5 at 37°C enhanced respective k values of fisetin and quercetin from 8.30x10−3 and 2.81x10−2 to 0.202 and 0.375 h-1 (P<0.05. In comparison with their degradation at 37°C, fisetin and quercetin showed larger k values at higher temperature (0.124 and 0.245 h−1 at 50°C, or 0.490 and 1.42 h−1 at 65°C. Four protein products in medium could stabilize the two flavonoids (P<0.05, as these proteins at 0.10 g L-1 decreased respective k values of fisetin and quercetin to 2.28x10−2-2.98x10−2 and 4.37´10−2-5.97x10−2 h−1. Hydrophobic interaction between the proteins and the two flavonoids was evidenced responsible for the stabilization, as sodium dodecyl sulfate could destroy the stabilization significantly (P<0.05. Casein and soybean protein provided greater stabilization than whey protein isolate. It is thus concluded that higher temperature and alkaline pH can enhance flavonoid loss, whereas coexisted proteins as flavonoid stabilizers can inhibit flavonoid degradation.

  1. Silage review: Using molecular approaches to define the microbial ecology of silage.

    Science.gov (United States)

    McAllister, T A; Dunière, L; Drouin, P; Xu, S; Wang, Y; Munns, K; Zaheer, R

    2018-05-01

    Ensiling of forages was recognized as a microbial-driven process as early as the late 1800s, when it was associated with the production of "sweet" or "sour" silage. Classical microbiological plating techniques defined the epiphytic microbial populations associated with fresh forage, the pivotal role of lactic acid-producing bacteria in the ensiling process, and the contribution of clostridia, bacilli, yeast, and molds to the spoilage of silage. Many of these classical studies focused on the enumeration and characterization of a limited number of microbial species that could be readily isolated on selective media. Evidence suggested that many of the members of these microbial populations were viable but unculturable, resulting in classical studies underestimating the true microbial diversity associated with ensiling. Polymerase chain reaction-based techniques, including length heterogeneity PCR, terminal RFLP, denaturing gradient gel electrophoresis, and automated ribosomal intergenic spacer analysis, were the first molecular methods used to study silage microbial communities. Further advancements in whole comparative genomic, metagenomic, and metatranscriptomic sequencing have or are in the process of superseding these methods, enabling microbial communities during ensiling to be defined with a degree of detail that is impossible using classical microbiology. These methods have identified new microbial species in silage, as well as characterized shifts in microbial communities with forage type and composition, ensiling method, and in response to aerobic exposure. Strain- and species-specific primers have been used to track the persistence and contribution of silage inoculants to the ensiling process and the role of specific species of yeast and fungi in silage spoilage. Sampling and the methods used to isolate genetic materials for further molecular analysis can have a profound effect on results. Primer selection for PCR amplification and the presence of inhibitors

  2. Anti-Fusarium moniliforme activity and fumonisin biodegradation by corn and silage microflora

    Directory of Open Access Journals (Sweden)

    Simone B. Camilo

    2000-01-01

    Full Text Available Studies were carried out to isolate microorganisms from corn and silage screened for their ability to inhibit F. moniliforme growth (strain 113F in association with fumonisin detoxification. Among 150 isolates four Gram-positive bacilli and one yeast with inhibitory activity were selected. The inhibition zone ranged from 50 to 72.5 mm using cultures, and from 25 to 52.5mm for crude alcoholic extract. The isolates S9, S10, S69 (sporulated bacilli and SE3071 (yeast degraded 43, 48, 83 and 57% of the initial FB1 concentration, respectively. The pH increased gradually in the medium during incubation for biodegradation process.

  3. Improvement of Nutritive Value and Ruminal Fermentation of Silage by Molasses and Urea Supplementation

    Directory of Open Access Journals (Sweden)

    K. Phesatcha

    2016-08-01

    Full Text Available Leucaena silage was supplemented with different levels of molasses and urea to study its nutritive value and in vitro rumen fermentation efficiency. The ensiling study was randomly assigned according to a 3×3 factorial arrangement in which the first factor was molasses (M supplement at 0%, 1%, and 2% of crop dry matter (DM and the second was urea (U supplement as 0%, 0.5%, and 1% of the crop DM, respectively. After 28 days of ensiling, the silage samples were collected and analyzed for chemical composition. All the nine Leucaena silages were kept for study of rumen fermentation efficiency using in vitro gas production techniques. The present result shows that supplementation of U or M did not affect DM, organic matter, neutral detergent fiber, and acid detergent fiber content in the silage. However, increasing level of U supplementation increased crude protein content while M level did not show any effect. Moreover, the combination of U and M supplement decreased the content of mimosine concentration especially with M2U1 (molasses 2% and urea 1% silage. The result of the in vitro study shows that gas production kinetics, cumulation gas at 96 h and in vitro true digestibility increased with the increasing level of U and M supplementation especially in the combination treatments. Supplementation of M and U resulted in increasing propionic acid and total volatile fatty acid whereas, acetic acid, butyric acid concentrations and methane production were not changed. In addition, increasing U level supplementation increased NH3-N concentration. Result from real-time polymerase chain reaction revealed a significant effect on total bacteria, whereas F. succinogenes and R. flavefaciens population while R. albus was not affected by the M and U supplementation. Based on this study, it could be concluded that M and urea U supplementation could improve the nutritive value of Leucaena silage and enhance in vitro rumen fermentation efficiency. This study also

  4. Misfolding, degradation, and aggregation of variant proteins. The molecular pathogenesis of short chain acyl-CoA dehydrogenase (SCAD) deficiency

    DEFF Research Database (Denmark)

    Pedersen, Christina Bak; Bross, P.; Winter, V.S.

    2003-01-01

    and aggregation of variant SCAD proteins. In this study we investigated the processing of a set of disease-causing variant SCAD proteins (R22W, G68C, W153R, R359C, and Q341H) and two common variant proteins (R147W and G185S) that lead to reduced SCAD activity. All SCAD proteins, including the wild type, associate...... proteolytic degradation by mitochondrial proteases or, especially at elevated temperature, aggregation of non-native conformers. The latter finding may indicate that accumulation of aggregated SCAD proteins may play a role in the pathogenesis of SCAD deficiency.......Short chain acyl-CoA dehydrogenase (SCAD) deficiency is an inborn error of the mitochondrial fatty acid metabolism caused by rare variations as well as common susceptibility variations in the SCAD gene. Earlier studies have shown that a common variant SCAD protein (R147W) was impaired in folding...

  5. Thermolysin damages animal life through degradation of plasma proteins enhanced by rapid cleavage of serpins and activation of proteases.

    Science.gov (United States)

    Kong, Lulu; Lu, Anrui; Guan, Jingmin; Yang, Bing; Li, Muwang; Hillyer, Julián F; Ramarao, Nalini; Söderhäll, Kenneth; Liu, Chaoliang; Ling, Erjun

    2015-01-01

    Thermolysin, a metallopeptidase secreted by pathogenic microbes, is concluded as an important virulence factor due to cleaving purified host proteins in vitro. Using the silkworm Bombyx mori as a model system, we found that thermolysin injection into larvae induces the destruction of the coagulation response and the activation of hemolymph melanization, which results in larval death. Thermolysin triggers the rapid degradation of insect and mammalian plasma proteins at a level that is considerably greater than expected in vitro and/or in vivo. To more specifically explore the mechanism, thermolysin-induced changes to key proteins belonging to the insect melanization pathway were assessed as a window for observing plasma protein cleavage. The application of thermolysin induced the rapid cleavage of the melanization negative regulator serpin-3, but did not directly activate the melanization rate-limiting enzyme prophenoloxidase (PPO) or the terminal serine proteases responsible for PPO activation. Terminal serine proteases of melanization are activated indirectly after thermolysin exposure. We hypothesize that thermolysin induces the rapid degradation of serpins and the activation of proteases directly or indirectly, boosting uncontrolled plasma protein degradation in insects and mammalians. © 2014 Wiley Periodicals, Inc.

  6. Different redox sensitivity of endoplasmic reticulum associated degradation clients suggests a novel role for disulphide bonds in secretory proteins.

    Science.gov (United States)

    Medraño-Fernandez, Iria; Fagioli, Claudio; Mezghrani, Alexandre; Otsu, Mieko; Sitia, Roberto

    2014-04-01

    To maintain proteostasis in the endoplasmic reticulum (ER), terminally misfolded secretory proteins must be recognized, partially unfolded, and dislocated to the cytosol for proteasomal destruction, in a complex process called ER-associated degradation (ERAD). Dislocation implies reduction of inter-chain disulphide bonds. When in its reduced form, protein disulphide isomerase (PDI) can act not only as a reductase but also as an unfoldase, preparing substrates for dislocation. PDI oxidation by Ero1 favours substrate release and transport across the ER membrane. Here we addressed the redox dependency of ERAD and found that DTT stimulates the dislocation of proteins with DTT-resistant disulphide bonds (i.e., orphan Ig-μ chains) but stabilizes a ribophorin mutant (Ri332) devoid of them. DTT promotes the association of Ri332, but not of Ig-µ, with PDI. This discrepancy may suggest that disulphide bonds in cargo proteins can be utilized to oxidize PDI, hence facilitating substrate detachment and degradation also in the absence of Ero1. Accordingly, Ero1 silencing retards Ri332 degradation, but has little if any effect on Ig-µ. Thus, some disulphides can increase the stability and simultaneously favour quality control of secretory proteins.

  7. Polysorbates 20 and 80 used in the formulation of protein biotherapeutics: structure and degradation pathways.

    Science.gov (United States)

    Kerwin, Bruce A

    2008-08-01

    Polysorbates 20 and 80 (Tween 20 and Tween 80) are used in the formulation of biotherapeutic products for both preventing surface adsorption and as stabilizers against protein aggregation. The polysorbates are amphipathic, nonionic surfactants composed of fatty acid esters of polyoxyethylene sorbitan being polyoxyethylene sorbitan monolaurate for polysorbate 20 and polyoxyethylene sorbitan monooleate for polysorbate 80. The polysorbates used in the formulation of biopharmaceuticals are mixtures of different fatty acid esters with the monolaurate fraction of polysorbate 20 making up only 40-60% of the mixture and the monooleate fraction of polysorbate 80 making up >58% of the mixture. The polysorbates undergo autooxidation, cleavage at the ethylene oxide subunits and hydrolysis of the fatty acid ester bond. Autooxidation results in hydroperoxide formation, side-chain cleavage and eventually formation of short chain acids such as formic acid all of which could influence the stability of a biopharmaceutical product. Oxidation of the fatty acid moiety while well described in the literature has not been specifically investigated for polysorbate. This review focuses on the chemical structure of the polysorbates, factors influencing micelle formation and factors and excipients influencing stability and degradation of the polyoxyethylene and fatty acid ester linkages.

  8. SCF(SAP) controls organ size by targeting PPD proteins for degradation in Arabidopsis thaliana.

    Science.gov (United States)

    Wang, Zhibiao; Li, Na; Jiang, Shan; Gonzalez, Nathalie; Huang, Xiahe; Wang, Yingchun; Inzé, Dirk; Li, Yunhai

    2016-04-06

    Control of organ size by cell proliferation and growth is a fundamental process, but the mechanisms that determine the final size of organs are largely elusive in plants. We have previously revealed that the ubiquitin receptor DA1 regulates organ size by repressing cell proliferation in Arabidopsis. Here we report that a mutant allele of STERILE APETALA (SAP) suppresses the da1-1 mutant phenotype. We show that SAP is an F-box protein that forms part of a SKP1/Cullin/F-box E3 ubiquitin ligase complex and controls organ size by promoting the proliferation of meristemoid cells. Genetic analyses suggest that SAP may act in the same pathway with PEAPOD1 and PEAPOD2, which are negative regulators of meristemoid proliferation, to control organ size, but does so independently of DA1. Further results reveal that SAP physically associates with PEAPOD1 and PEAPOD2, and targets them for degradation. These findings define a molecular mechanism by which SAP and PEAPOD control organ size.

  9. In vivo imaging of brain ischemia using an oxygen-dependent degradative fusion protein probe.

    Directory of Open Access Journals (Sweden)

    Youshi Fujita

    Full Text Available Within the ischemic penumbra, blood flow is sufficiently reduced that it results in hypoxia severe enough to arrest physiological function. Nevertheless, it has been shown that cells present within this region can be rescued and resuscitated by restoring perfusion and through other protective therapies. Thus, the early detection of the ischemic penumbra can be exploited to improve outcomes after focal ischemia. Hypoxia-inducible factor (HIF-1 is a transcription factor induced by a reduction in molecular oxygen levels. Although the role of HIF-1 in the ischemic penumbra remains unknown, there is a strong correlation between areas with HIF-1 activity and the ischemic penumbra. We recently developed a near-infrared fluorescently labeled-fusion protein, POH-N, with an oxygen-dependent degradation property identical to the alpha subunit of HIF-1. Here, we conduct in vivo imaging of HIF-active regions using POH-N in ischemic brains after transient focal cerebral ischemia induced using the intraluminal middle cerebral artery occlusion technique in mice. The results demonstrate that POH-N enables the in vivo monitoring and ex vivo detection of HIF-1-active regions after ischemic brain injury and suggest its potential in imaging and drug delivery to HIF-1-active areas in ischemic brains.

  10. Analyzing pepsin degradation assay conditions used for allergenicity assessments to ensure that pepsin susceptible and pepsin resistant dietary proteins are distinguishable.

    Directory of Open Access Journals (Sweden)

    Rong Wang

    Full Text Available The susceptibility of a dietary protein to proteolytic degradation by digestive enzymes, such as gastric pepsin, provides information on the likelihood of systemic exposure to a structurally intact and biologically active macromolecule, thus informing on the safety of proteins for human and animal consumption. Therefore, the purpose of standardized in vitro degradation studies that are performed during protein safety assessments is to distinguish whether proteins of interest are susceptible or resistant to pepsin degradation via a study design that enables study-to-study comparison. Attempting to assess pepsin degradation under a wide-range of possible physiological conditions poses a problem because of the lack of robust and consistent data collected under a large-range of sub-optimal conditions, which undermines the needs to harmonize in vitro degradation conditions. This report systematically compares the effects of pH, incubation time, and pepsin-to-substrate protein ratio on the relative degradation of five dietary proteins: three pepsin susceptible proteins [ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco, horseradish peroxidase (HRP, hemoglobin (Hb], and two pepsin resistant proteins [lipid transfer protein (LTP and soybean trypsin inhibitor (STI]. The results indicate that proteins susceptible to pepsin degradation are readily distinguishable from pepsin-resistant proteins when the reaction conditions are within the well-characterized optima for pepsin. The current standardized in vitro pepsin resistant assay with low pH and high pepsin-to-substrate ratio fits this purpose. Using non-optimal pH and/or pepsin-to-substrate protein ratios resulted in susceptible proteins no longer being reliably degraded by this stomach enzyme, which compromises the ability of this in vitro assay to distinguish between resistant and susceptible proteins and, therefore, no longer providing useful data to an overall weight-of-evidence approach to

  11. Rumen Degradability and Post-ruminal Digestion of Dry Matter, Nitrogen and Amino Acids of Three Protein Supplements

    Directory of Open Access Journals (Sweden)

    Wei Gao

    2015-04-01

    Full Text Available This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID of dry matter, crude protein (CP, and amino acids (AA of cottonseed meal (CSM, sunflower seed meal (SFSM and distillers dried grains with solubles (DDGS by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen micro-organisms, especially for Trp, Arg, His, and Lys (p<0.01. The SID of individual RUP-AA was different within specific RUP origin (p<0.01. Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance.

  12. Rumen Degradability and Post-ruminal Digestion of Dry Matter, Nitrogen and Amino Acids of Three Protein Supplements.

    Science.gov (United States)

    Gao, Wei; Chen, Aodong; Zhang, Bowen; Kong, Ping; Liu, Chenli; Zhao, Jie

    2015-04-01

    This study evaluated the in situ ruminal degradability, and subsequent small intestinal digestibility (SID) of dry matter, crude protein (CP), and amino acids (AA) of cottonseed meal (CSM), sunflower seed meal (SFSM) and distillers dried grains with solubles (DDGS) by using the modified three-step in vitro procedure. The ruminal degradability and subsequent SID of AA in rumen-undegradable protein (RUP-AA) varied among three protein supplements. The result show that the effective degradability of DM for SFSM, CSM, and DDGS was 60.8%, 56.4%, and 41.0% and their ruminal fermentable organic matter was 60.0%, 55.9%, and 39.9%, respectively. The ruminal degradable protein (RDP) content in CP for SFSM, CSM, and DDGS was 68.3%, 39.0%, and 32.9%, respectively, at the ruminal solid passage rate of 1.84%/h. The SFSM is a good source of RDP for rumen micro-organisms; however, the SID of RUP of SFSM was lower. The DDGS and CSM are good sources of RUP for lambs to digest in the small intestine to complement ruminal microbial AA of growing lambs. Individual RUP-AA from each protein source was selectively removed by the rumen micro-organisms, especially for Trp, Arg, His, and Lys (p<0.01). The SID of individual RUP-AA was different within specific RUP origin (p<0.01). Limiting amino acid was Leu for RUP of CSM and Lys for both RUP of SFSM and DDGS, respectively. Therefore, different protein supplements with specific limitations should be selected and combined carefully in growing lambs ration to optimize AA balance.

  13. F-Box Protein FBXO22 Mediates Polyubiquitination and Degradation of CD147 to Reverse Cisplatin Resistance of Tumor Cells.

    Science.gov (United States)

    Wu, Bo; Liu, Zhen-Yu; Cui, Jian; Yang, Xiang-Min; Jing, Lin; Zhou, Yang; Chen, Zhi-Nan; Jiang, Jian-Li

    2017-01-20

    Drug resistance remains a major clinical obstacle to successful treatment of cancer. As posttranslational modification is becoming widely recognized to affect the function of oncoproteins, targeting specific posttranslational protein modification provides an attractive strategy for anticancer drug development. CD147 is a transmembrane glycoprotein contributing to chemo-resistance of cancer cells in a variety of human malignancies. Ubiquitination is an important posttranslational modification mediating protein degradation. Degradation of oncoproteins, CD147 included, emerges as an attractive alternative for tumor inhibition. However, the ubiquitination of CD147 remains elusive. Here in this study, we found that deletion of the CD147 intracellular domain (CD147-ICD) prolonged the half-life of CD147 in HEK293T cells, and we identified that CD147-ICD interacts with FBXO22 using mass spectrometry and Western blot. Then, we demonstrated that FBXO22 mediates the polyubiquitination and degradation of CD147 by recognizing CD147-ICD. While knocking down of FBXO22 prolonged the half-life of CD147 in HEK293T cells, we found that FBXO22 regulates CD147 protein turnover in SMMC-7721, Huh-7 and A549 cells. Moreover, we found that the low level of FBXO22 contributes to the accumulation of CD147 and thereafter the cisplatin resistance of A549/DDP cells. To conclude, our study demonstrated that FBXO22 mediated the polyubiquitination and degradation of CD147 by interacting with CD147-ICD, and CD147 polyubiquitination by FBXO22 reversed cisplatin resistance of tumor cells.

  14. Rift Valley fever virus NSs inhibits host transcription independently of the degradation of dsRNA-dependent protein kinase PKR.

    Science.gov (United States)

    Kalveram, Birte; Lihoradova, Olga; Indran, Sabarish V; Lokugamage, Nandadeva; Head, Jennifer A; Ikegami, Tetsuro

    2013-01-20

    Rift Valley fever virus (RVFV) encodes one major virulence factor, the NSs protein. NSs suppresses host general transcription, including interferon (IFN)-β mRNA synthesis, and promotes degradation of the dsRNA-dependent protein kinase (PKR). We generated a novel RVFV mutant (rMP12-NSsR173A) specifically lacking the function to promote PKR degradation. rMP12-NSsR173A infection induces early phosphorylation of eIF2α through PKR activation, while retaining the function to inhibit host general transcription including IFN-β gene inhibition. MP-12 NSs but not R173A NSs binds to wt PKR. R173A NSs formed filamentous structure in nucleus in a mosaic pattern, which was distinct from MP-12 NSs filament pattern. Due to early phosphorylation of eIF2α, rMP12-NSsR173A could not efficiently accumulate viral proteins. Our results suggest that NSs-mediated host general transcription suppression occurs independently of PKR degradation, while the PKR degradation is important to inhibit the phosphorylation of eIF2α in infected cells undergoing host general transcription suppression. Copyright © 2012 Elsevier Inc. All rights reserved.

  15. Novel Phosphorylation and Ubiquitination Sites Regulate Reactive Oxygen Species-dependent Degradation of Anti-apoptotic c-FLIP Protein*

    Science.gov (United States)

    Wilkie-Grantham, Rachel P.; Matsuzawa, Shu-Ichi; Reed, John C.

    2013-01-01

    The cytosolic protein c-FLIP (cellular Fas-associated death domain-like interleukin 1β-converting enzyme inhibitory protein) is an inhibitor of death receptor-mediated apoptosis that is up-regulated in a variety of cancers, contributing to apoptosis resistance. Several compounds found to restore sensitivity of cancer cells to TRAIL, a TNF family death ligand with promising therapeutic potential, act by targeting c-FLIP ubiquitination and degradation by the proteasome. The generation of reactive oxygen species (ROS) has been implicated in c-FLIP protein degradation. However, the mechanism by which ROS post-transcriptionally regulate c-FLIP protein levels is not well understood. We show here that treatment of prostate cancer PPC-1 cells with the superoxide generators menadione, paraquat, or buthionine sulfoximine down-regulates c-FLIP long (c-FLIPL) protein levels, which is prevented by the proteasome inhibitor MG132. Furthermore, pretreatment of PPC-1 cells with a ROS scavenger prevented ubiquitination and loss of c-FLIPL protein induced by menadione or paraquat. We identified lysine 167 as a novel ubiquitination site of c-FLIPL important for ROS-dependent degradation. We also identified threonine 166 as a novel phosphorylation site and demonstrate that Thr-166 phosphorylation is required for ROS-induced Lys-167 ubiquitination. The mutation of either Thr-166 or Lys-167 was sufficient to stabilize c-FLIP protein levels in PPC-1, HEK293T, and HeLa cancer cells treated with menadione or paraquat. Accordingly, expression of c-FLIP T166A or K167R mutants protected cells from ROS-mediated sensitization to TRAIL-induced cell death. Our findings reveal novel ROS-dependent post-translational modifications of the c-FLIP protein that regulate its stability, thus impacting sensitivity of cancer cells to TRAIL. PMID:23519470

  16. Genome comparison implies the role of Wsm2 in membrane trafficking and protein degradation

    Directory of Open Access Journals (Sweden)

    Guorong Zhang

    2018-04-01

    Wsm2 may combat WSMV disease through a molecular mechanism involving protein degradation and/or membrane trafficking. The 93 putative Wsm2 ancestor loci discovered in this study could serve as good candidates for future genetic isolation of the true Wsm2 locus.

  17. Prolyl hydroxylation regulates protein degradation, synthesis, and splicing in human induced pluripotent stem cell-derived cardiomyocytes.

    Science.gov (United States)

    Stoehr, Andrea; Yang, Yanqin; Patel, Sajni; Evangelista, Alicia M; Aponte, Angel; Wang, Guanghui; Liu, Poching; Boylston, Jennifer; Kloner, Philip H; Lin, Yongshun; Gucek, Marjan; Zhu, Jun; Murphy, Elizabeth

    2016-06-01

    Protein hydroxylases are oxygen- and α-ketoglutarate-dependent enzymes that catalyse hydroxylation of amino acids such as proline, thus linking oxygen and metabolism to enzymatic activity. Prolyl hydroxylation is a dynamic post-translational modification that regulates protein stability and protein-protein interactions; however, the extent of this modification is largely uncharacterized. The goals of this study are to investigate the biological consequences of prolyl hydroxylation and to identify new targets that undergo prolyl hydroxylation in human cardiomyocytes. We used human induced pluripotent stem cell-derived cardiomyocytes in combination with pulse-chase amino acid labelling and proteomics to analyse the effects of prolyl hydroxylation on protein degradation and synthesis. We identified 167 proteins that exhibit differences in degradation with inhibition of prolyl hydroxylation by dimethyloxalylglycine (DMOG); 164 were stabilized. Proteins involved in RNA splicing such as serine/arginine-rich splicing factor 2 (SRSF2) and splicing factor and proline- and glutamine-rich (SFPQ) were stabilized with DMOG. DMOG also decreased protein translation of cytoskeletal and sarcomeric proteins such as α-cardiac actin. We searched the mass spectrometry data for proline hydroxylation and identified 134 high confidence peptides mapping to 78 unique proteins. We identified SRSF2, SFPQ, α-cardiac actin, and cardiac titin as prolyl hydroxylated. We identified 29 prolyl hydroxylated proteins that showed a significant difference in either protein degradation or synthesis. Additionally, we performed next-generation RNA sequencing and showed that the observed decrease in protein synthesis was not due to changes in mRNA levels. Because RNA splicing factors were prolyl hydroxylated, we investigated splicing ± inhibition of prolyl hydroxylation and detected 369 alternative splicing events, with a preponderance of exon skipping. This study provides the first extensive

  18. Degradation of p53 by human Alphapapillomavirus E6 proteins shows a stronger correlation with phylogeny than oncogenicity.

    Directory of Open Access Journals (Sweden)

    Leiping Fu

    2010-09-01

    Full Text Available Human Papillomavirus (HPV E6 induced p53 degradation is thought to be an essential activity by which high-risk human Alphapapillomaviruses (alpha-HPVs contribute to cervical cancer development. However, most of our understanding is derived from the comparison of HPV16 and HPV11. These two viruses are relatively distinct viruses, making the extrapolation of these results difficult. In the present study, we expand the tested strains (types to include members of all known HPV species groups within the Alphapapillomavirus genus.We report the biochemical activity of E6 proteins from 27 HPV types representing all alpha-HPV species groups to degrade p53 in human cells. Expression of E6 from all HPV types epidemiologically classified as group 1 carcinogens significantly reduced p53 levels. However, several types not associated with cancer (e.g., HPV53, HPV70 and HPV71 were equally active in degrading p53. HPV types within species groups alpha 5, 6, 7, 9 and 11 share a most recent common ancestor (MRCA and all contain E6 ORFs that degrade p53. A unique exception, HPV71 E6 ORF that degraded p53 was outside this clade and is one of the most prevalent HPV types infecting the cervix in a population-based study of 10,000 women. Alignment of E6 ORFs identified an amino acid site that was highly correlated with the biochemical ability to degrade p53. Alteration of this amino acid in HPV71 E6 abrogated its ability to degrade p53, while alteration of this site in HPV71-related HPV90 and HPV106 E6s enhanced their capacity to degrade p53.These data suggest that the alpha-HPV E6 proteins' ability to degrade p53 is an evolved phenotype inherited from a most recent common ancestor of the high-risk species that does not always segregate with carcinogenicity. In addition, we identified an amino-acid residue strongly correlated with viral p53 degrading potential.

  19. Activation of chromatin degradation by a protein factor of thymocyte cytoplasm of irradiated mice

    International Nuclear Information System (INIS)

    Soldatenkov, V.A.; Filippovich, I.V.

    1986-01-01

    A cytoplasmic thymocyte fraction isolated 1 h after irradiation of mice accelerates chromatin degradation in isolated nuclei. Treatment of the cytoplasmic fraction by heat and injection of cycloheximide to mice prevent the acceleration of DNA degradation. The analysis of the chromatin degradation products and the kinetics of this process at acid and alkaline pH shows that activation of DNA degradation in thymocytes by a factor obtained from the irradiated cell cytoplasm is specific for a Ca 2+ , Mg 2+ -dependent enzyme. The time- and dose-dependent parameters of the appearance in the thymocyte cytoplasm of the factor influencing degradation of chromatin are in a good agreement with both the time of the onset of its postirradiation degradation and the dose dependence of this process

  20. Degradation of vitamin C in a product made from mango (Mangifera indica L. and whey protein

    Directory of Open Access Journals (Sweden)

    Fernando Alonso Mendoza-Corvis

    2017-01-01

    Full Text Available This study aimed to determine the kinetics of vitamin C degradation in a product made from mango pulp (Mangifera indica L. and whey protein powder, in order to determine the effect of temperature on its conservation and further evaluate the behavior of the L*, a*, b* and the total color difference (ΔE in the powder product. Vitamin C was determined by the aoac 967.21/90 method using 2,6-dichlorophenol indophenol, and the color was quantified with a HunterLab Color Flex EZ colorimeter. Vitamin C showed greater stability in the powder product ResumoEsta pesquisa procurou determinar a cinética de degradação da vitamina C em um produto em pó elaborado a base de polpa de manga (Mangifera indica L. var. Hilacha e lactisoro, com o fim de conhecer o efeito da temperatura na sua conservação e ademais avaliar o comportamento dos parâmetros L*, a*, b* e a diferença total da cor (ΔE no produto. A vitamina C determinou-se mediante o método aoac 967.21/90, com 2,6-diclorofenol indofe-nol; enquanto que a cor se quantificou com um colorímetro cor Flex EZ marca HunterLab. A vitamina C exibiu maior estabilidade no produto em pó armazenado a uma temperatura de 4 °C, stored at 4 °C with a concentration at the end of the eighth sampling week of 13.94 ± 1.2 mg/10 0 g-1 sample and showing a first order degradation kinetics with k1 values of 0.014 and 0.041 mg/100 g/week at temperatures of 4 °C and 28 °C, respec-tively. The greatest variations in color occurred in samples stored at 28 °C, indicating the influence of the temperature change on the product compo-nents. In addition, L*, a* and b* parameters were less affected under storage temperature of 4 °C, and their values correspond to a second degree polynomial.

  1. Methane production, ruminal fermentation characteristics, nutrient digestibility, nitrogen excretion, and milk production of dairy cows fed conventional or brown midrib corn silage.

    Science.gov (United States)

    Hassanat, F; Gervais, R; Benchaar, C

    2017-04-01

    The objective of this study was to examine the effect of replacing conventional corn silage (CCS) with brown midrib corn silage (BMCS) in dairy cow diets on enteric CH 4 emission, nutrient intake, digestibility, ruminal fermentation characteristics, milk production, and N excretion. Sixteen rumen-cannulated lactating cows used in a crossover design (35-d periods) were fed (ad libitum) a total mixed ration (forage:concentrate ratio = 65:35, dry matter basis) based (59% dry matter) on either CCS or BMCS. Dry matter intake and milk yield increased when cows were fed BMCS instead of CCS. Of the milk components, only milk fat content slightly decreased when cows were fed the BMCS-based diet compared with when fed the CCS-based diet (3.81 vs. 3.92%). Compared with CCS, feeding BMCS to cows increased yields of milk protein and milk fat. Ruminal pH, protozoa numbers, total VFA concentration, and molar proportions of acetate and propionate were similar between cows fed BMCS and those fed CCS. Daily enteric CH 4 emission (g/d) was unaffected by dietary treatments, but CH 4 production expressed as a proportion of gross energy intake or on milk yield basis was lower for cows fed the BMCS-based diet than for cows fed the CCS-based diet. A decline in manure N excretion and a shift in N excretion from urine to feces were observed when BMCS replaced CCS in the diet, suggesting reduced potential of manure N volatilization. Results from this study show that improving fiber quality of corn silage in dairy cow diets through using brown midrib trait cultivar can reduce enteric CH 4 emissions as well as potential emissions of NH 3 and N 2 O from manure. However, CH 4 emissions during manure storage may increase due to excretion of degradable OM when BMCS diet is fed, which merits further investigation. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  2. Monitoring protein turnover during phosphate starvation-dependent autophagic degradation using a photoconvertible fluorescent protein aggregate in tobacco BY-2 cells.

    Science.gov (United States)

    Tasaki, Maiko; Asatsuma, Satoru; Matsuoka, Ken

    2014-01-01

    We have developed a system for quantitative monitoring of autophagic degradation in transformed tobacco BY-2 cells using an aggregate-prone protein comprised of cytochrome b5 (Cyt b5) and a tetrameric red fluorescent protein (RFP). Unfortunately, this system is of limited use for monitoring the kinetics of autophagic degradation because the proteins synthesized before and after induction of autophagy cannot be distinguished. To overcome this problem, we developed a system using kikume green-red (KikGR), a photoconvertible and tetrameric fluorescent protein that changes its fluorescence from green to red upon irradiation with purple light. Using the fusion protein of Cyt b5 and KikGR together with a method for the bulk conversion of KikGR, which we had previously used to convert the Golgi-localized monomeric KikGR fusion protein, we were able to monitor both the growth and de novo formation of aggregates. Using this system, we found that tobacco cells do not cease protein synthesis under conditions of phosphate (Pi)-starvation. Induction of autophagy under Pi-starvation, but not under sugar- or nitrogen-starvation, was specifically inhibited by phosphite, which is an analog of Pi with a different oxidation number. Therefore, the mechanism by which BY-2 cells can sense Pi-starvation and induce autophagy does not involve sensing a general decrease in energy supply and a specific Pi sensor might be involved in the induction of autophagy under Pi-starvation.

  3. Nutritive value of brewers’ grain and maize silage for fattening rabbits

    Directory of Open Access Journals (Sweden)

    H. Guermah

    2016-09-01

    Full Text Available Specific knowledge of the nutritive value of raw materials is fundamental to formulate balanced diets for rabbits and allows greater use of by-products and non-conventional feedstuffs. This paper examines the feeding value of sun-dried brewers’ grain and maize silage (whole plant for fattening rabbits. Twenty-four individually caged 8-wk-old rabbits were used to determine the digestibility. Both wet products were sun-dried and ground before being incorporated into a basal diet. The inclusion level at the expense of all basal ingredients amounted to 30%. Basal diet and both experimental diets were fed ad libitum to 8 rabbits during the 4-d balance trial. The determined digestibility of protein, fat, crude fibre and neutral detergent fibre digestibility amounted to 76.2 and 77.2%; 86.5 and 99.1%; 8.1 and 8.3% and 28.0 and 13.5%, respectively, for brewers’ grain and maize silage. The digestible energy content amounted to 11.66 MJ/kg dry matter (DM (brewers’ grain and 11.10 MJ/kg DM (maize silage. Both by-products have potential as alternative feedstuff in rabbit diets. However, further experiments are necessary to determine the effect of ensilaging the whole maize plant, as a significantly lower (P<0.001 feed intake was observed.

  4. Isolation and characterization of a tomato non-specific lipid transfer protein involved in polygalacturonase-mediated pectin degradation.

    Science.gov (United States)

    Tomassen, Monic M M; Barrett, Diane M; van der Valk, Henry C P M; Woltering, Ernst J

    2007-01-01

    An important aspect of the ripening process of tomato fruit is softening. Softening is accompanied by hydrolysis of the pectin in the cell wall by pectinases, causing loss of cell adhesion in the middle lamella. One of the most significant pectin-degrading enzymes is polygalacturonase (PG). Previous reports have shown that PG in tomato may exist in different forms (PG1, PG2a, PG2b, and PGx) commonly referred to as PG isoenzymes. The gene product PG2 is differentially glycosylated and is thought to associate with other proteins to form PG1 and PGx. This association is thought to modulate its pectin-degrading activity in planta. An 8 kDa protein that is part of the tomato PG1 multiprotein complex has been isolated, purified, and functionally characterized. This protein, designated 'activator' (ACT), belongs to the class of non-specific lipid transfer proteins (nsLTPs). ACT is capable of 'converting' the gene product PG2 into a more active and heat-stable form, which increases PG-mediated pectin degradation in vitro and stimulates PG-mediated tissue breakdown in planta. This finding suggests a new, not previously identified, function for nsLTPs in the modification of hydrolytic enzyme activity. It is proposed that ACT plays a role in the modulation of PG activity during tomato fruit softening.

  5. GSK3 is required for rapalogs to induce degradation of some oncogenic proteins and to suppress cancer cell growth.

    Science.gov (United States)

    Koo, Junghui; Wang, Xuerong; Owonikoko, Taofeek K; Ramalingam, Suresh S; Khuri, Fadlo R; Sun, Shi-Yong

    2015-04-20

    The single-agent activity of rapalogs (rapamycin and its analogues) in most tumor types has been modest at best. The underlying mechanisms are largely unclear. In this report, we have uncovered a critical role of GSK3 in regulating degradation of some oncogenic proteins induced by rapalogs and cell sensitivity to rapalogs. The basal level of GSK3 activity was positively correlated with cell sensitivity of lung cancer cell lines to rapalogs. GSK3 inhibition antagonized rapamycin's growth inhibitory effects both in vitro and in vivo, while enforced activation of GSK3β sensitized cells to rapamycin. GSK3 inhibition rescued rapamcyin-induced reduction of several oncogenic proteins such as cyclin D1, Mcl-1 and c-Myc, without interfering with the ability of rapamycin to suppress mTORC1 signaling and cap binding. Interestingly, rapamycin induces proteasomal degradation of these oncogenic proteins, as evidenced by their decreased stabilities induced by rapamcyin and rescue of their reduction by proteasomal inhibition. Moreover, acute or short-time rapamycin treatment dissociated not only raptor, but also rictor from mTOR in several tested cell lines, suggesting inhibition of both mTORC1 and mTORC2. Thus, induction of GSK3-dependent degradation of these oncogenic proteins is likely secondary to mTORC2 inhibition; this effect should be critical for rapamycin to exert its anticancer activity.

  6. Failure of cellulolysis in the rumen of reindeer fed timothy silage

    Directory of Open Access Journals (Sweden)

    Monica A. Olsen

    1995-01-01

    Full Text Available Three male reindeer (Rangifer tarandus tarandus calves were brought from mountain pastures in April and fed regrowth timothy (Phleum pratense silage with 76% leaves and 24.0% dry matter (DM ad libitum. The silage contained (on DM basis 25.4% cellulose, 12.0% crude protein and 19-6% water soluble carbohydrates. After an initial period of 11 days the daily silage intake rose to almost similar values for all animals, but independently of food intake, body mass (BM increased by as much as 13.3 kg for animal R3 during the first 21 days, compared to 4.4 kg and 2.8 kg for Rl and R2, respectively. At slaughter the wet weight of the rumen contents of animal R3 constituted 30.2% of the total BM, compared to 18.5% and 19.1% in animals Rl and R2, respectively. A reduced ability of the rumen micro-biota to ferment pure cellulose in vitro was observed in R3. The ruminal pH was 7.07 and the concentration of volatile fatty acids was only 50.0 mM in R3, indicating a low rate of fermentation. The initial rates of in vitro dry matter digestibility of timothy silage and standard hay were also affected by the rumen fermentation failure in animal R3. Depressed rumen cellulolysis, which may be related to natural periods of starvation prior to the feeding experiment, could have caused the low rate of fermentation and the large rumen size observed in this animal.

  7. Chemical composition of elephant grass silages supplemented with different levels of dehydrated cashew bagasse

    Directory of Open Access Journals (Sweden)

    Danillo Glaydson Farias Guerra

    2016-04-01

    Full Text Available The objective of the present study was to evaluate the chemical composition of elephant grass silages supplemented with different levels dried cashew bagasse (DCB. Our experiment used a randomized design replicated four times, each replicate consisting of the following five treatments: 100% elephant grass; 95% elephant grass + 5% DCB; 90% elephant grass + 10% DCB; 85% elephant grass + 15% DCB; and 80% elephant grass + 20% DCB. The elephant grass was cut manually to a residual height of 5 cm at 80 days of age, and cashew bagasse was obtained from the processing of cashew stalks used in fruit pulp manufacturing in Mossoró/RN. Plastic buckets were used as experimental silos, and 90 days after ensiling the experimental silos were opened and the contents analyzed. The addition of dried cashew bagasse to silage linearly increased the levels of dried matter and crude protein by 0.59% and 0.13%, respectively, for each 1% addition (P < 0.05. The neutral detergent fiber and acid detergent content of the silages was reduced by 0.22% and 0.09%, respectively, for each 1% addition of the bagasse. The total carbohydrate content was not influenced by the bagasse addition (P > 0.05, and averaged 82.29%. The levels of non-fiber carbohydrate showed linear growth (P < 0.05 as the dehydrated cashew bagasse was added, and pH and ammoniacal nitrogen levels were reduced. The addition of the dehydrated bagasse to elephant grass silage improves its chemical composition, and it can be effectively added up to the level of 20%.

  8. Cyclin A degradation by primate cytomegalovirus protein pUL21a counters its innate restriction of virus replication.

    Directory of Open Access Journals (Sweden)

    Nicolas Caffarelli

    Full Text Available Cyclin A is critical for cellular DNA synthesis and S phase progression of the cell cycle. Human cytomegalovirus (HCMV can reduce cyclin A levels and block cellular DNA synthesis, and cyclin A overexpression can repress HCMV replication. This interaction has only been previously observed in HCMV as murine CMV does not downregulate cyclin A, and the responsible viral factor has not been identified. We previously reported that the HCMV protein pUL21a disrupted the anaphase-promoting complex (APC, but a point mutant abrogating this activity did not phenocopy a UL21a-deficient virus, suggesting that pUL21a has an additional function. Here we identified a conserved arginine-x-leucine (RxL cyclin-binding domain within pUL21a, which allowed pUL21a to interact with cyclin A and target it for proteasome degradation. Homologous pUL21a proteins from both chimpanzee and rhesus CMVs also contained the RxL domain and similarly degraded cyclin A, indicating that this function is conserved in primate CMVs. The RxL point mutation disabled the virus' ability to block cellular DNA synthesis and resulted in a growth defect similar to pUL21a-deficient virus. Importantly, knockdown of cyclin A rescued growth of UL21a-deficient virus. Together, these data show that during evolution, the pUL21a family proteins of primate CMVs have acquired a cyclin-binding domain that targets cyclin A for degradation, thus neutralizing its restriction on virus replication. Finally, the combined proteasome-dependent degradation of pUL21a and its cellular targets suggests that pUL21a may act as a novel suicide protein, targeting its protein cargos for destruction.

  9. Elephant grass clones for silage production

    Directory of Open Access Journals (Sweden)

    Rerisson José Cipriano dos Santos

    2013-02-01

    Full Text Available Ensiling warm-season grasses often requires wilting due to their high moisture content, and the presence of low-soluble sugars in these grasses usually demands the use of additives during the ensiling process. This study evaluated the bromatological composition of the fodder and silage from five Pennisetum sp. clones (IPA HV 241, IPA/UFRPE Taiwan A-146 2.114, IPA/UFRPE Taiwan A-146 2.37, Elephant B, and Mott. The contents of 20 Polyvinyl chloride (PVC silos, which were opened after 90 days of storage, were used for the bromatological analysis and the evaluation of the pH, nitrogen, ammonia, buffer capacity, soluble carbohydrates, and fermentation coefficients. The effluent losses, gases and dry matter recovery were also calculated. Although differences were observed among the clones (p < 0.05 for the concentrations of dry matter, insoluble nitrogen in acid detergents, insoluble nitrogen in neutral detergents, soluble carbohydrates, fermentation coefficients, and in vitro digestibility in the forage before ensiling, no differences were observed for most of these variables after ensiling. All of the clones were efficient in the fermentation process. The IPA/UFRPE TAIWAN A-146 2.37 clone, however, presented a higher dry matter concentration and the best fermentation coefficient, resulting in a better silage quality, compared to the other clones.

  10. FERMENTATION PROCESS CHARACTERISTICS OF DIFFERENT MAIZE SILAGE HYBRIDS

    Directory of Open Access Journals (Sweden)

    Daniel Bíro

    2009-03-01

    Full Text Available The aim of this study was to detect the fermentation process differences in different hybrid maize silage. We conserved in laboratory conditions hybrids of whole maize plants with different length of the vegetative period (FAO number. Maize hybrids for silage were harvested in the vegetation stage of the milk-wax maturity of corn and the content of dry matter was from 377.7 to 422.8 g.kg-1. The highest content of dry matter was typical for silages made from the hybrids with FAO number 310 (400.0 g.kg-1 and FAO 300a (400.4 g.kg-1. The content of desirable lactic acid ranged from 23.7 g.kg-1 of dry matter (FAO 350 to 58.9 g.kg-1 of dry matter (FAO 420. We detected the occurrence of undesirable butyric acid in silages from hybrids FAO 250, 300b, 310 and 380. The highest content of total alcohols we found in silages made from hybrid with FAO number 240 (25.2 g.kg-1 of dry matter. Ammonia contents were in tested silages from 0.153 (FAO 270 to 0.223 g.kg-1 of dry matter (FAO 240. The lowest value of silage titration acidity we analyzed in silage made from hybrid FAO 420 (3.66. We observed in maize silages with different length of plant maturity tested in the experiment differences in content of lactic acid, total alcohols, titration acidity, pH and content of fermentation products.

  11. Characterization of Enzymatic Activity of MlrB and MlrC Proteins Involved in Bacterial Degradation of Cyanotoxins Microcystins.

    Science.gov (United States)

    Dziga, Dariusz; Zielinska, Gabriela; Wladyka, Benedykt; Bochenska, Oliwia; Maksylewicz, Anna; Strzalka, Wojciech; Meriluoto, Jussi

    2016-03-16

    Bacterial degradation of toxic microcystins produced by cyanobacteria is a common phenomenon. However, our understanding of the mechanisms of these processes is rudimentary. In this paper several novel discoveries regarding the action of the enzymes of the mlr cluster responsible for microcystin biodegradation are presented using recombinant proteins. In particular, the predicted active sites of the recombinant MlrB and MlrC were analyzed using functional enzymes and their inactive muteins. A new degradation intermediate, a hexapeptide derived from linearized microcystins by MlrC, was discovered. Furthermore, the involvement of MlrA and MlrB in further degradation of the hexapeptides was confirmed and a corrected biochemical pathway of microcystin biodegradation has been proposed.

  12. Characterization of Enzymatic Activity of MlrB and MlrC Proteins Involved in Bacterial Degradation of Cyanotoxins Microcystins

    Directory of Open Access Journals (Sweden)

    Dariusz Dziga

    2016-03-01

    Full Text Available Bacterial degradation of toxic microcystins produced by cyanobacteria is a common phenomenon. However, our understanding of the mechanisms of these processes is rudimentary. In this paper several novel discoveries regarding the action of the enzymes of the mlr cluster responsible for microcystin biodegradation are presented using recombinant proteins. In particular, the predicted active sites of the recombinant MlrB and MlrC were analyzed using functional enzymes and their inactive muteins. A new degradation intermediate, a hexapeptide derived from linearized microcystins by MlrC, was discovered. Furthermore, the involvement of MlrA and MlrB in further degradation of the hexapeptides was confirmed and a corrected biochemical pathway of microcystin biodegradation has been proposed.

  13. Effects of toasting blue lupins, soybeans or barley as supplement for high-yielding organic dairy cows fed grass-clover silage ad libitum

    DEFF Research Database (Denmark)

    Mogensen, Lisbeth; Lund, Peter; Kristensen, Troels

    2008-01-01

    . The supplement under investigation was: lupins in experiment 1, barley in experiment 2 and soybeans in experiment 3. The same forage mixture of grass-clover silage (84% of DM), grass pellets (11% of DM) and straw (5% of DM) was fed ad libitum in all the experiments. Toasting decreased effective rumen protein...... degradability determined in situ for all three supplements. Compared to untreated lupins toasting of lupins tended (P = 0.10) to increase milk yield, whereas toasting of soybeans did not affect milk yield. Toasting of lupins decreased (P = 0.03) milk protein content (32.2 versus 32.7 g/kg), whereas toasting...... of soybeans did not affect milk protein content. ECM yield was significantly higher (P = 0.002) for cows fed toasted soybeans than for cows fed untreated soybeans (28.1 versus 26.4 kg ECM) whereas there was no significant effect on ECM yield from toasting lupins or barley. It can be concluded...

  14. Feasibility of Hydrothermal Pretreatment on Maize Silage for Bioethanol Production

    DEFF Research Database (Denmark)

    Xu, Jian; Thomsen, Mette Hedegaard; Thomsen, Anne Belinda

    2010-01-01

    The potential of maize silage as a feedstock to produce bioethanol was evaluated in the present study. The hydrothermal pretreatment with five different pretreatment severity factors (PSF) was employed to pretreat the maize silage and compared in terms of sugar recovery, toxic test, and ethanol...... the liquors from the five conditions were not toxic to the Baker’s yeast. Pretreatment under 195°C for 7 min had the similar PSF with that of 185°C for 15 min, and both gave the higher ethanol concentration of 19.92 and 19.98 g/L, respectively. The ethanol concentration from untreated maize silage was only 7...

  15. Fiber digestion kinetics and protein degradability characteristics of stockpiled Tifton 85 bermudagrass.

    Science.gov (United States)

    Sechler, S R; Mullenix, M K; Holland, C M; Muntifering, R B

    2017-09-01

    A 2-yr study was conducted to determine effects of N fertilization level on fiber digestion kinetics and protein degradability characteristics of stockpiled Tifton 85 bermudagrass (T85). Six 0.76-ha pastures of stockpiled T85 were cut to a 10-cm stubble height on August 1 of each yr and fertilized with 56 (56N), 112 (112N), or 168 (168N) kg N/ha (2 pastures/treatment). Fiber digestion kinetics included the 72-hr potential extent of NDF digestion (PED), rate of NDF digestion, and lag time. In yr 1 and 2, PED decreased over the stockpile season. Rates of NDF digestion did not differ ( > 0.05) among N fertilization treatments in either yr. In yr 1, rate of NDF digestion was greatest ( digestion decreased ( digestion rates were similar for November and January 21 sampling dates. Lag time was greater ( digestion ( = -0.60 and -0.25 in yr 1 and 2, respectively) was observed. There was a trend ( = 0.06) for lignin concentration to be positively correlated with lag time ( = 0.39) in yr 1, and a strong relationship was observed in yr 2 ( = 0.91; digestion in stockpiled T85 were influenced more by temporal changes over the stockpile season than by N fertilization level. Supplement formulations based on kinetic parameters of fiber digestion may require periodic adjustment to insure that energy-yielding components of NDF are sufficient to meet animal requirements throughout the stockpile season. The CP fraction in stockpiled T85 contains sufficient RDP to support fibrolytic activity and growth of ruminal microorganisms throughout the stockpile season. Toward the latter end of the season, supplementation with sources of digestible fiber and RDP could be expected to increase MP supply to the host animal.

  16. Milk Production of Lactating Cows Fed Pangola Silage Ensiled with Irradiated Silage Innoculant and Fresh Cut Pangola Grass

    International Nuclear Information System (INIS)

    Prasanpanich, Somkiert; Buaphan, Sirirat; Boon-ek, Lertchatand; Piadang, Nattayana

    2006-09-01

    Ten crossbred Holstein lactating cows of mid-lactation and similar body score condition were balanced across the 2 treatments containing Group 1: 5 cows given fresh cut Pangola grass and Group 2: 5 cows fed Pangola silage ensiled with silage innoculant, under Group Comparison Design. They were also provided with an exact amount of 1 kg meal concentrate to 1 kg of milk produced. Mineral supplement and water were supplied free choice. The results did show that the cows in Group 1 had significantly higher (P<0.05) roughage and total dry matter intake than did the cows in Group 2. However, milk production in Group 2 appeared to have a non-significant higher amount, the concentrations of Blood urea nitrogen, Blood glucose and Triiodothyronine were not significant different between groups. The body weight change increased in both groups. It can concluded that cows fed Pangola silage ensiled with silage innoculant seemed to have higher Digestibility Co-efficiency due to the high efficiency of silage innoculant fermentation. Then, silage quality would be improved by mean of silage innoculant application.

  17. Nycterohemeral eating and ruminating patterns in heifers fed grass or corn silage: analysis by finite Fourier transform.

    Science.gov (United States)

    Deswysen, A G; Dutilleul, P; Godfrin, J P; Ellis, W C

    1993-10-01

    Average daily and within-day nycterohemeral patterns of eating and ruminating behavior were determined in six Holstein-Friesian heifers (average BW = 427 kg) given ad libitum access to either corn or grass silage in a two-period crossover design. Rhythm components (number of cycles/24 h) were characterized by finite Fourier transform of the 24-h mastication activities as measured during 4 d by continuous jaw movement recordings. Average daily voluntary intake of corn silage was 8.2% greater (P = .05) than that for grass silage and was associated (P finite Fourier transform was reparameterized to express the amplitude (as periodograms) and phase of each rhythm component. Rhythm Components 1, 3, and 4 contributed primarily to explaining the total dispersion of the 24-h series of time spent eating and ruminating, for both silage types and individual heifers. Relative importance of Rhythm Component 1 of time spent eating, indicative of a main circadian pattern, was related positively to pedigree value for milk production (P = .01) and negatively to milk protein concentration (P = .09).(ABSTRACT TRUNCATED AT 250 WORDS)

  18. HIV-1 tat protein recruits CIS to the cytoplasmic tail of CD127 to induce receptor ubiquitination and proteasomal degradation

    Energy Technology Data Exchange (ETDEWEB)

    Sugden, Scott, E-mail: scott.sugden@ircm.qc.ca [The Ottawa Hospital Research Institute, 501 Smyth Road, Ottawa, Ontario, Canada K1H 8L6 (Canada); Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, 451 Smyth Road, Ottawa, Ontario, Canada K1H 8M5 (Canada); Ghazawi, Feras [The Ottawa Hospital Research Institute, 501 Smyth Road, Ottawa, Ontario, Canada K1H 8L6 (Canada); Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, 451 Smyth Road, Ottawa, Ontario, Canada K1H 8M5 (Canada); MacPherson, Paul, E-mail: pmacpherson@toh.on.ca [The Ottawa Hospital Research Institute, 501 Smyth Road, Ottawa, Ontario, Canada K1H 8L6 (Canada); Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, 451 Smyth Road, Ottawa, Ontario, Canada K1H 8M5 (Canada); Division of Infectious Diseases, The Ottawa Hospital General Campus, 501 Smyth Road, Ottawa, Ontario, Canada K1H 8L6 (Canada)

    2016-11-15

    HIV-1 Tat protein down regulates expression of the IL-7 receptor alpha-chain (CD127) from the surface of CD8 T cells resulting in impaired T cell proliferation and cytolytic capacity. We have previously shown that soluble Tat protein is taken up by CD8 T cells and interacts with the cytoplasmic tail of CD127 to induce receptor degradation. The N-terminal domain of Tat interacts with CD127 while the basic domain directs CD127 to the proteasome. We have also shown that upon IL-7 binding to its receptor, CD127 is phosphorylated resulting in CIS-mediated proteasomal degradation. Here, we show that Tat mimics this process by recruiting CIS to CD127 in the absence of IL-7 and receptor phosphorylation, leading to CD127 ubiquitination and degradation. Tat therefore acts as an adapter to induce cellular responses under conditions where they may not otherwise occur. Thusly, Tat reduces IL-7 signaling and impairs CD8 T cell survival and function. -- Highlights: •Soluble HIV-1 Tat decreases CD127 expression on CD8 T cells, causing dysfunction. •Tat induces CD127 ubiquitination without activating IL-7 signaling. •Tat binds CD127 and recruits the E3 ubiquitin ligase CIS via its basic domain. •Tat hijacks a normal cellular mechanism to degrade CD127 without IL-7 signaling.

  19. HIV-1 tat protein recruits CIS to the cytoplasmic tail of CD127 to induce receptor ubiquitination and proteasomal degradation

    International Nuclear Information System (INIS)

    Sugden, Scott; Ghazawi, Feras; MacPherson, Paul

    2016-01-01

    HIV-1 Tat protein down regulates expression of the IL-7 receptor alpha-chain (CD127) from the surface of CD8 T cells resulting in impaired T cell proliferation and cytolytic capacity. We have previously shown that soluble Tat protein is taken up by CD8 T cells and interacts with the cytoplasmic tail of CD127 to induce receptor degradation. The N-terminal domain of Tat interacts with CD127 while the basic domain directs CD127 to the proteasome. We have also shown that upon IL-7 binding to its receptor, CD127 is phosphorylated resulting in CIS-mediated proteasomal degradation. Here, we show that Tat mimics this process by recruiting CIS to CD127 in the absence of IL-7 and receptor phosphorylation, leading to CD127 ubiquitination and degradation. Tat therefore acts as an adapter to induce cellular responses under conditions where they may not otherwise occur. Thusly, Tat reduces IL-7 signaling and impairs CD8 T cell survival and function. -- Highlights: •Soluble HIV-1 Tat decreases CD127 expression on CD8 T cells, causing dysfunction. •Tat induces CD127 ubiquitination without activating IL-7 signaling. •Tat binds CD127 and recruits the E3 ubiquitin ligase CIS via its basic domain. •Tat hijacks a normal cellular mechanism to degrade CD127 without IL-7 signaling.

  20. Unique nonstructural proteins of Pneumonia Virus of Mice (PVM) promote degradation of interferon (IFN) pathway components and IFN-stimulated gene proteins.

    Science.gov (United States)

    Dhar, Jayeeta; Barik, Sailen

    2016-12-01

    Pneumonia Virus of Mice (PVM) is the only virus that shares the Pneumovirus genus of the Paramyxoviridae family with Respiratory Syncytial Virus (RSV). A deadly mouse pathogen, PVM has the potential to serve as a robust animal model of RSV infection, since human RSV does not fully replicate the human pathology in mice. Like RSV, PVM also encodes two nonstructural proteins that have been implicated to suppress the IFN pathway, but surprisingly, they exhibit no sequence similarity with their RSV equivalents. The molecular mechanism of PVM NS function, therefore, remains unknown. Here, we show that recombinant PVM NS proteins degrade the mouse counterparts of the IFN pathway components. Proteasomal degradation appears to be mediated by ubiquitination promoted by PVM NS proteins. Interestingly, NS proteins of PVM lowered the levels of several ISG (IFN-stimulated gene) proteins as well. These results provide a molecular foundation for the mechanisms by which PVM efficiently subverts the IFN response of the murine cell. They also reveal that in spite of their high sequence dissimilarity, the two pneumoviral NS proteins are functionally and mechanistically similar.

  1. NSs Virulence Factor of Rift Valley Fever Virus Engages the F-Box Proteins FBXW11 and β-TRCP1 To Degrade the Antiviral Protein Kinase PKR.

    Science.gov (United States)

    Kainulainen, Markus; Lau, Simone; Samuel, Charles E; Hornung, Veit; Weber, Friedemann

    2016-07-01

    Rift Valley fever virus (RVFV, family Bunyaviridae, genus Phlebovirus) is a relevant pathogen of both humans and livestock in Africa. The nonstructural protein NSs is a major virulence factor known to suppress the type I interferon (IFN) response by inhibiting host cell transcription and by proteasomal degradation of a major antiviral IFN effector, the translation-inhibiting protein kinase PKR. Here, we identified components of the modular SCF (Skp1, Cul1, F-box protein)-type E3 ubiquitin ligases as mediators of PKR destruction by NSs. Small interfering RNAs (siRNAs) against the conserved SCF subunit Skp1 protected PKR from NSs-mediated degradation. Consequently, RVFV replication was severely reduced in Skp1-depleted cells when PKR was present. SCF complexes have a variable F-box protein subunit that determines substrate specificity for ubiquitination. We performed an siRNA screen for all (about 70) human F-box proteins and found FBXW11 to be involved in PKR degradation. The partial stabilization of PKR by FBXW11 depletion upregulated PKR autophosphorylation and phosphorylation of the PKR substrate eIF2α and caused a shutoff of host cell protein synthesis in RVFV-infected cells. To maximally protect PKR from the action of NSs, knockdown of structurally and functionally related FBXW1 (also known as β-TRCP1), in addition to FBXW11 deletion, was necessary. Consequently, NSs was found to interact with both FBXW11 and β-TRCP1. Thus, NSs eliminates the antiviral kinase PKR by recruitment of SCF-type E3 ubiquitin ligases containing FBXW11 and β-TRCP1 as substrate recognition subunits. This antagonism of PKR by NSs is essential for efficient RVFV replication in mammalian cells. Rift Valley fever virus is a pathogen of humans and animals that has the potential to spread from Africa and the Arabian Peninsula to other regions. A major virulence mechanism is the proteasomal degradation of the antiviral kinase PKR by the viral protein NSs. Here, we demonstrate that NSs

  2. Betulinic acid selectively increases protein degradation and enhances prostate cancer-specific apoptosis: possible role for inhibition of deubiquitinase activity.

    Directory of Open Access Journals (Sweden)

    Teresita Reiner

    Full Text Available Inhibition of the ubiquitin-proteasome system (UPS of protein degradation is a valid anti-cancer strategy and has led to the approval of bortezomib for the treatment of multiple myeloma. However, the alternative approach of enhancing the degradation of oncoproteins that are frequently overexpressed in cancers is less developed. Betulinic acid (BA is a plant-derived small molecule that can increase apoptosis specifically in cancer but not in normal cells, making it an attractive anti-cancer agent. Our results in prostate cancer suggested that BA inhibited multiple deubiquitinases (DUBs, which resulted in the accumulation of poly-ubiquitinated proteins, decreased levels of oncoproteins, and increased apoptotic cell death. In normal fibroblasts, however, BA did not inhibit DUB activity nor increased total poly-ubiquitinated proteins, which was associated with a lack of effect on cell death. In the TRAMP transgenic mouse model of prostate cancer, treatment with BA (10 mg/kg inhibited primary tumors, increased apoptosis, decreased angiogenesis and proliferation, and lowered androgen receptor and cyclin D1 protein. BA treatment also inhibited DUB activity and increased ubiquitinated proteins in TRAMP prostate cancer but had no effect on apoptosis or ubiquitination in normal mouse tissues. Overall, our data suggests that BA-mediated inhibition of DUBs and induction of apoptotic cell death specifically in prostate cancer but not in normal cells and tissues may provide an effective non-toxic and clinically selective agent for chemotherapy.

  3. Effects of Supplementation of Branched-Chain Amino Acids to Reduced-Protein Diet on Skeletal Muscle Protein Synthesis and Degradation in the Fed and Fasted States in a Piglet Model

    Directory of Open Access Journals (Sweden)

    Liufeng Zheng

    2016-12-01

    Full Text Available Supplementation of branched-chain amino acids (BCAA has been demonstrated to promote skeletal muscle mass gain, but the mechanisms underlying this observation are still unknown. Since the regulation of muscle mass depends on a dynamic equilibrium (fasted losses–fed gains in protein turnover, the aim of this study was to investigate the effects of BCAA supplementation on muscle protein synthesis and degradation in fed/fasted states and the related mechanisms. Fourteen 26- (Experiment 1 and 28-day-old (Experiment 2 piglets were fed reduced-protein diets without or with supplemental BCAA. After a four-week acclimation period, skeletal muscle mass and components of anabolic and catabolic signaling in muscle samples after overnight fasting were determined in Experiment 1. Pigs in Experiment 2 were implanted with carotid arterial, jugular venous, femoral arterial and venous catheters, and fed once hourly along with the intravenous infusion of NaH13CO3 for 2 h, followed by a 6-h infusion of [1-13C]leucine. Muscle leucine kinetics were measured using arteriovenous difference technique. The mass of most muscles was increased by BCAA supplementation. During feeding, BCAA supplementation increased leucine uptake, protein synthesis, protein degradation and net transamination. The greater increase in protein synthesis than in protein degradation resulted in elevated protein deposition. Protein synthesis was strongly and positively correlated with the intramuscular net production of α-ketoisocaproate (KIC and protein degradation. Moreover, BCAA supplementation enhanced the fasted-state phosphorylation of protein translation initiation factors and inhibited the protein-degradation signaling of ubiquitin-proteasome and autophagy-lysosome systems. In conclusion, supplementation of BCAA to reduced-protein diet increases fed-state protein synthesis and inhibits fasted-state protein degradation, both of which could contribute to the elevation of skeletal muscle

  4. In vitro cytotoxicity of fungi spoiling maize silage

    DEFF Research Database (Denmark)

    Rasmussen, Rie Romme; Rasmussen, Peter Have; Larsen, Thomas Ostenfeld

    2011-01-01

    Penicillium roqueforti, Penicillium paneum, Monascus ruber, Alternaria tenuissima, Fusarium graminearum, Fusarium avenaceum, Byssochlamys nivea and Aspergillus fumigatus have previously been identified as major fungal contaminants of Danish maize silage. In the present study their metabolite....... roqueforti metabolites roquefortine C (48μg/mL), andrastin A (>50μg/mL), mycophenolic acid (>100μg/mL) and 1-hydroxyeremophil-7(11),9(10)-dien-8-one (>280μg/mL) were high. Fractionating of agar extracts identified PR-toxin as an important cytotoxic P. roqueforti metabolite, also detectable in maize silage....... The strongly cytotoxic B. nivea and P. paneum agar extracts contained patulin above the IC50 of 0.6μg/mL, however inoculated onto maize silage B. nivea and P. paneum did not produce patulin (>371μg/kg). Still B. nivea infected maize silage containing mycophenolic acid (∼50mg/kg), byssochlamic acid and other...

  5. Ensiling characteristics and feeding value of silage made from cattle ...

    African Journals Online (AJOL)

    In this investigation, silage made from cattle waste being ensiled with maize residues (ECW), was evaluated ... high concentrate ration tended to be of a better quality and scored 8,9 points out of 13in comparison ..... reclaimed fecal residue.

  6. The effect of the source of microorganisms on adaptation of hydrolytic consortia dedicated to anaerobic digestion of maize silage.

    Science.gov (United States)

    Poszytek, Krzysztof; Pyzik, Adam; Sobczak, Adam; Lipinski, Leszek; Sklodowska, Aleksandra; Drewniak, Lukasz

    2017-08-01

    The main aim of this study was to evaluate the effect of the source of microorganisms on the selection of hydrolytic consortia dedicated to anaerobic digestion of maize silage. The selection process was investigated based on the analysis of changes in the hydrolytic activity and the diversity of microbial communities derived from (i) a hydrolyzer of a commercial agricultural biogas plant, (ii) cattle slurry and (iii) raw sewage sludge, during a series of 10 passages. Following the selection process, the adapted consortia were thoroughly analyzed for their ability to utilize maize silage and augmentation of anaerobic digestion communities. The results of selection of the consortia showed that every subsequent passage of each consortium leads to their adaptation to degradation of maize silage, which was manifested by the increased hydrolytic activity of the adapted consortia. Biodiversity analysis (based on the 16S rDNA amplicon sequencing) confirmed the changes microbial community of each consortium, and showed that after the last (10th) passage all microbial communities were dominated by the representatives of Lactobacillaceae, Prevotellaceae, Veillonellaceae. The results of the functional analyses showed that the adapted consortia improved the efficiency of maize silage degradation, as indicated by the increase in the concentration of glucose and volatile fatty acids (VFAs), as well as the soluble chemical oxygen demand (sCOD). Moreover, bioaugmentation of anaerobic digestion communities by the adapted hydrolytic consortia increased biogas yield by 10-29%, depending on the origin of the community. The obtained results also indicate that substrate input (not community origin) was the driving force responsible for the changes in the community structure of hydrolytic consortia dedicated to anaerobic digestion. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. The structural determinants responsible for c-Fos protein proteasomal degradation differ according to the conditions of expression.

    Science.gov (United States)

    Ferrara, Patrizia; Andermarcher, Elisabetta; Bossis, Guillaume; Acquaviva, Claire; Brockly, Frédérique; Jariel-Encontre, Isabelle; Piechaczyk, Marc

    2003-03-13

    c-fos gene is expressed constitutively in a number of tissues as well as in certain tumor cells and is inducible, in general rapidly and transiently, in virtually all other cell types by a variety of stimuli. Its protein product, c-Fos, is a short-lived transcription factor that heterodimerizes with various protein partners within the AP-1 transcription complex via leucine zipper/leucine zipper interactions for binding to specific DNA sequences. It is mostly, if not exclusively, degraded by the proteasome. To localize the determinant(s) responsible for its instability, we have conducted a genetic analysis in which the half-lives of c-Fos mutants and chimeras made with the stable EGFP reporter protein were compared under two experimental conditions taken as example of continous and inducible expression. Those were constitutive expression in asynchronously growing Balb/C 3T3 mouse embryo fibroblasts and transient induction in the same cells undergoing the G0/G1 phase transition upon stimulation by serum. Our work shows that c-Fos is degraded faster in synchronous- than in asynchronous cells. This difference in turnover is primarily accounted for by several mechanisms. First, in asynchronous cells, a unique C-terminal destabilizer is active whereas, in serum-stimulated cells two destabilizers located at both extremities of the protein are functional. Second, heterodimerization and/or binding to DNA accelerates protein degradation only during the G0/G1 phase transition. Adding another level of complexity to turnover control, phosphorylation at serines 362 and 374, which are c-Fos phosphorylation sites largely modified during the G0/G1 phase transition, stabilizes c-Fos much more efficiently in asynchronous than in serum-stimulated cells. In both cases, the reduced degradation rate is due to inhibition of the activity of the C-terminal destabilizer. However, in serum-stimulated cells, this effect is partially masked by the activation of the N-terminal destabilizer and

  8. F-Box Protein FBXO22 Mediates Polyubiquitination and Degradation of CD147 to Reverse Cisplatin Resistance of Tumor Cells

    Directory of Open Access Journals (Sweden)

    Bo Wu

    2017-01-01

    Full Text Available Drug resistance remains a major clinical obstacle to successful treatment of cancer. As posttranslational modification is becoming widely recognized to affect the function of oncoproteins, targeting specific posttranslational protein modification provides an attractive strategy for anticancer drug development. CD147 is a transmembrane glycoprotein contributing to chemo-resistance of cancer cells in a variety of human malignancies. Ubiquitination is an important posttranslational modification mediating protein degradation. Degradation of oncoproteins, CD147 included, emerges as an attractive alternative for tumor inhibition. However, the ubiquitination of CD147 remains elusive. Here in this study, we found that deletion of the CD147 intracellular domain (CD147-ICD prolonged the half-life of CD147 in HEK293T cells, and we identified that CD147-ICD interacts with FBXO22 using mass spectrometry and Western blot. Then, we demonstrated that FBXO22 mediates the polyubiquitination and degradation of CD147 by recognizing CD147-ICD. While knocking down of FBXO22 prolonged the half-life of CD147 in HEK293T cells, we found that FBXO22 regulates CD147 protein turnover in SMMC-7721, Huh-7 and A549 cells. Moreover, we found that the low level of FBXO22 contributes to the accumulation of CD147 and thereafter the cisplatin resistance of A549/DDP cells. To conclude, our study demonstrated that FBXO22 mediated the polyubiquitination and degradation of CD147 by interacting with CD147-ICD, and CD147 polyubiquitination by FBXO22 reversed cisplatin resistance of tumor cells.

  9. Fermentation characteristics of different purpose sorghum silage

    Directory of Open Access Journals (Sweden)

    Arthur Behling Neto

    2017-08-01

    Full Text Available Sorghum stands out among other plants recommended for ensiling due to its forage composition, its resistance to drought, and its planting range. New cultivars of grain and sweet sorghum that can be used for silage production are available, but there is little information regarding their ensiling characteristics. The aim of this study was to evaluate the fermentation characteristics at the ensiling of different purpose sorghum cultivars, at two crop periods. The trial was carried out at the Plant Production Department of the Federal Institute of Education, Science and Technology of Rondônia, Colorado do Oeste campus, Rondônia, Brazil, and chemical analyses were performed at the Laboratory of Animal Nutrition, at the Federal University of Mato Grosso, Cuiabá campus, Mato Grosso, Brazil. The experimental design used was a randomized block, in split-plot design, with four replicates. The plot treatments consisted of six sorghum cultivars grown for different purposes (grain sorghum: BRS 308 and BRS 310; forage sorghum: BR 655 and BRS 610; sweet sorghum: BRS 506 and CMSXS 647. Split-plot treatments consisted of two cropping seasons (first crop and second crop. The grain sorghum cultivar BRS 310 was the only one that had suitable dry matter content for ensiling; however, it was also the only one that did not show ideal water soluble carbohydrate content for ensiling. Nevertheless, all treatments presented pH below than 4.2 and ammonia nitrogen lower than 12% of total N, which indicates that the fermentation inside the silo had proceeded well. For sweet sorghum cultivars, higher ethanol and butyric acid content were observed for the first crop than for the second crop. All evaluated sorghum cultivars can be used for silage production, but the use of sweet sorghum is recommended at the second crop.

  10. Losses and nutritional value of elephant grass silage with inclusion levels of cottonseed meal - doi: 10.4025/actascianimsci.v35i2.13736

    Directory of Open Access Journals (Sweden)

    Mauro Pereira de Figueiredo

    2013-03-01

    Full Text Available The experiment was conducted to evaluate losses and nutritive value of elephant grass silage containing increasing levels of cottonseed meal. The experimental design was completely randomized, with five treatments: 0, 7, 14, 21 and 28% inclusion of cottonseed meal, and four replications. The material was chopped and packed in PVC silos and stored for 80 days. The pH and dry matter (DM, crude protein, lignin and ether extract of the silages increased linearly with the addition of cottonseed meal, while the levels of acid detergent insoluble nitrogen, neutral detergent fiber, acid detergent fiber, cellulose and hemicellulose decreased linearly. The addition of 28% cottonseed meal reduced the in situ disappearance of DM (48h to values lower than those of silage without additives. The concentration of ammonia nitrogen and the losses from gases and effluent were reduced. The inclusion of 28% cottonseed meal in elephant grass silage containing 18.4% DM improved the fermentation characteristics of silage more efficiently by reducing the moisture content and effluent losses, starting at the 7% level of inclusion.

  11. Retraction: Myostatin Induces Degradation of Sarcomeric Proteins through a Smad3 Signaling Mechanism During Skeletal Muscle Wasting

    Science.gov (United States)

    Lokireddy, Sudarsanareddy; McFarlane, Craig; Ge, Xiaojia; Zhang, Huoming; Sze, Siu Kwan; Sharma, Mridula

    2011-01-01

    Ubiquitination-mediated proteolysis is a hallmark of skeletal muscle wasting manifested in response to negative growth factors, including myostatin. Thus, the characterization of signaling mechanisms that induce the ubiquitination of intracellular and sarcomeric proteins during skeletal muscle wasting is of great importance. We have recently characterized myostatin as a potent negative regulator of myogenesis and further demonstrated that elevated levels of myostatin in circulation results in the up-regulation of the muscle-specific E3 ligases, Atrogin-1 and muscle ring finger protein 1 (MuRF1). However, the exact signaling mechanisms by which myostatin regulates the expression of Atrogin-1 and MuRF1, as well as the proteins targeted for degradation in response to excess myostatin, remain to be elucidated. In this report, we have demonstrated that myostatin signals through Smad3 (mothers against decapentaplegic homolog 3) to activate forkhead box O1 and Atrogin-1 expression, which further promotes the ubiquitination and subsequent proteasome-mediated degradation of critical sarcomeric proteins. Smad3 signaling was dispensable for myostatin-dependent overexpression of MuRF1. Although down-regulation of Atrogin-1 expression rescued approximately 80% of sarcomeric protein loss induced by myostatin, only about 20% rescue was seen when MuRF1 was silenced, implicating that Atrogin-1 is the predominant E3 ligase through which myostatin manifests skeletal muscle wasting. Furthermore, we have highlighted that Atrogin-1 not only associates with myosin heavy and light chain, but it also ubiquitinates these sarcomeric proteins. Based on presented data we propose a model whereby myostatin induces skeletal muscle wasting through targeting sarcomeric proteins via Smad3-mediated up-regulation of Atrogin-1 and forkhead box O1. PMID:21964591

  12. Protein degradation in skeletal muscle during experimental hyperthyroidism in rats and the effect of beta-blocking agents.

    Science.gov (United States)

    Angerås, U; Hasselgren, P O

    1987-04-01

    beta-Blocking agents are increasingly used in the management of hyperthyroid patients. The effect of this treatment on increased muscle protein breakdown in the hyperthyroid state is not known. In the present study, experimental hyperthyroidism was induced in rats by daily ip injections of T3 (100 micrograms/100 g BW) during a 10-day period. Control animals received corresponding volumes of solvent. In groups of rats the selective beta-1-blocking agent metoprolol or the nonselective beta-blocker propranolol was infused by miniosmotic pumps implanted sc on the backs of the animals. Protein degradation was measured in incubated intact soleus and extensor digitorum longus muscles by determining tyrosine release into the incubation medium. The protein degradation rate in incubated extensor digitorum longus and soleus muscles was increased by 50-60% during T3 treatment. Metoprolol or propranolol did not influence muscle protein breakdown in either T3-treated or control animals. The results suggest that T3-induced increased muscle proteolysis is not mediated by beta-receptors, and muscle weakness and wasting in hyperthyroidism might not be affected by beta-blockers.

  13. BAG3 down-modulation reduces anaplastic thyroid tumor growth by enhancing proteasome-mediated degradation of BRAF protein.

    Science.gov (United States)

    Chiappetta, Gennaro; Basile, Anna; Arra, Claudio; Califano, Daniela; Pasquinelli, Rosa; Barbieri, Antonio; De Simone, Veronica; Rea, Domenica; Giudice, Aldo; Pezzullo, Luciano; De Laurenzi, Vincenzo; Botti, Gerardo; Losito, Simona; Conforti, Daniela; Turco, Maria Caterina

    2012-01-01

    Anaplastic thyroid tumors (ATC) express high levels of BAG3, a member of the BAG family of cochaperone proteins that is involved in regulating cell apoptosis through multiple mechanisms. The objective of the study was the investigation of the influence of B-cell lymphoma-2-associated athanogene 3 (BAG3) on ATC growth. We investigated the effects of BAG3 down-modulation, obtained by using a specific small interfering RNA, on in vitro and in vivo growth of the human ATC cell line 8505C. Because BRAF protein plays an important role in ATC cell growth, we analyzed the effects of BAG3 down-modulation on BRAF protein levels. Furthermore, by using a proteasome inhibitor, we verified whether BAG3-mediated regulation of BRAF levels involved a proteasome-dependent mechanism. BAG3 down-modulation significantly inhibits ATC growth in vitro and in vivo. BAG3 coimmunoprecipitates with BRAF protein, and its down-modulation results in a significant reduction of BRAF protein levels, which can be reverted by incubation with the proteasome inhibitor MG132. BAG3 protein sustains ATC growth in vitro and in vivo. The underlying molecular mechanism appears to rely on BAG3 binding to BRAF, thus protecting it from proteasome-dependent degradation. These results are in line with the reported ability of BAG3 to interfere with the proteasomal delivery of a number of other client proteins.

  14. Physical degradation of proteins in well-defined fluid flows studied within a four-roll apparatus.

    Science.gov (United States)

    Simon, S; Krause, H J; Weber, C; Peukert, W

    2011-12-01

    In most applications of biotechnology and downstream processing proteins are exposed to fluid stresses in various flow configurations which often lead to the formation of unwanted protein aggregates. In this paper we present physical degradation experiments for proteins under well-defined flow conditions in a four-roll apparatus. The flow field was characterized numerically by computational fluid dynamics (CFD) and experimentally by particle image velocimetry (PIV). The local shear strain rate as well as the local shear and elongation rate was used to characterize the hydrodynamic stress environment acting on the proteins. Lysozyme was used as a model protein and subjected to well-defined fluid stresses in high and low stress environment. By using in situ turbidity measurements during stressing the aggregate formation was monitored directly in the fluid flow. An increase in absorbance at 350 nm was attributed to a higher content of visible particles (>1 µm). In addition to lysozyme, the formation of aggregates was confirmed for two larger proteins (bovine serum albumin and alcohol dehydrogenase). Thus, the presented experimental setup is a helpful tool to monitor flow-induced protein aggregation with high reproducibility. For instance, screening experiments for formulation development of biopharmaceuticals for fill and finish operations can be performed in the lab-scale in a short time-period if the stress distributions in the application are transferred and applied in the four-roll mill. Copyright © 2011 Wiley Periodicals, Inc.

  15. Casca de algodão em substituição parcial à silagem de capim-elefante para novilhos. 2. Parâmetros ruminais e séricos, produção microbiana e excreção urinária de compostos nitrogenados Partial replacement of elephantgrass silage with cottonseed hulls. 2. Ruminal and serum metabolites, microbial protein synthesis, and urinary excretion of nitrogenous compounds in steers

    Directory of Open Access Journals (Sweden)

    Mario Luiz Chizzotti

    2005-12-01

    Full Text Available Quatro novilhos holandeses fistulados no rúmen, com peso médio de 259 kg, foram distribuídos em um quadrado latino 4 x 4 para se avaliar o efeito dos níveis de casca de algodão na dieta de novilhos sobre a concentração de nitrogênio uréico no soro (NUS e de amônia no rúmen, o pH ruminal, a excreção urinária de uréia e derivados de purinas e a produção de proteína microbiana estimada pelo método das bases purinas omasais e da excreção de derivados de purinas na urina. As dietas experimentais continham na base da matéria seca: 0, 10, 20 e 30% de casca de algodão peletizada, em substituição à silagem de capim-elefante, sendo a dieta total constituída de 60% de volumoso. Não houve efeito dos diferentes tratamentos sobre o pH e as concentrações de amônia no rúmen. A concentração de NUS e a excreção de uréia (em mg/kgPV diminuíram, enquanto a excreção de derivados de purinas na urina e a síntese de proteína microbiana no rúmen aumentaram linearmente com a inclusão da casca de algodão nas dietas. A estimativa da síntese de proteína microbiana não diferiu entre as metodologias das bases purínicas omasais e dos derivados de purina na urina. A casca de algodão mostrou-se um bom volumoso alternativo, podendo ser fornecida até o nível de 30% na MS total na dieta de novilhos de origem leiteira.Four ruminally cannulated Holstein steers averaging 259 kg of body weight were assigned to a 4x4 Latin square to study the effects of replacing elephantgrass silage with cottonseed hulls on serum urea nitrogen (SUN, ruminal metabolism, urinary excretion of nitrogenous compounds, and microbial protein synthesis measured by omasal purine bases or by urinary excretion of purine derivatives. Treatments (60% of forage contained on DM basis: 0, 10, 20 or 30% of cottonseed hulls that partially replaced elephantgrass silage in the diet. No significant differences in ruminal pH and concentration of ruminal ammonia were

  16. The establishment of a database of Italian feeds for the Cornell Net Carbohydrate and Protein System

    Directory of Open Access Journals (Sweden)

    Enzo Tartari

    2010-01-01

    Full Text Available A field application of the Cornell Net Carbohydrate and Protein System (CNCPS in Italy has been limited because thefeed bank is based on North American feedstuffs and still few laboratories are able to analyze feeds as requested by theCNCPS. Moreover, the standardization of analytical procedures is still not homogeneous among laboratories. This workwas carried out to establish a first database for feeds commonly used in Italy, providing nutritionists and producers anaccurate and current feed composition, also indicating methods and apparatus for analytical procedures potentially availablefor routine analysis. A total of 909 samples of hays, silages and raw materials (protein feeds, cereals and by-productswere analyzed through 1999 and 2002; analysis included protein solubility and degradability, protein fractions,structural carbohydrate fractions and the calculation of neutral detergent structural carbohydrates. When possible, averagedata were compared with those included in the feed bank of CNCPS ver. 3 and with those obtained by another Italianlaboratory. The main differences were observed in chemical composition of forages and silages, whose composition largelydepends on environmental conditions and physiological stage; protein feeds, cereals and by-products showed somedifferences in crude protein, soluble protein and protein fractions even in feeds of national origin.The intent to modify the feed bank values of CNCPS for establishing an Italian data base of feeds will require a collaborativestudy of many laboratories not only for forages, hays and silages samples - whose composition is greatly dependenton environmental factors and agronomic techniques - but also for protein fractions, whose values are largely influencedby even small changes in analytical techniques.

  17. Theobromine suppresses adipogenesis through enhancement of CCAAT-enhancer-binding protein β degradation by adenosine receptor A1.

    Science.gov (United States)

    Mitani, Takakazu; Watanabe, Shun; Yoshioka, Yasukiyo; Katayama, Shigeru; Nakamura, Soichiro; Ashida, Hitoshi

    2017-12-01

    Theobromine, a methylxanthine derived from cacao beans, reportedly has various health-promoting properties but molecular mechanism by which effects of theobromine on adipocyte differentiation and adipogenesis remains unclear. In this study, we aimed to clarify the molecular mechanisms of the anti-adipogenic effect of theobromine in vitro and in vivo. ICR mice (4week-old) were administered with theobromine (0.1g/kg) for 7days. Theobromine administration attenuated gains in body and epididymal adipose tissue weights in mice and suppressed expression of adipogenic-associated genes in mouse adipose tissue. In 3T3-L1 preadipocytes, theobromine caused degradation of C/EBPβ protein by the ubiquitin-proteasome pathway. Pull down assay showed that theobromine selectively interacts with adenosine receptor A1 (AR1