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Tamotsu Yoshimori - One of the best experts on this subject based on the ideXlab platform.

  • Dissection of the Autophagosome Maturation Process by a Novel Reporter Protein, Tandem Fluorescent-Tagged LC3
    Autophagy, 2007
    Co-Authors: Shunsuke Kimura, Takeshi Noda, Tamotsu Yoshimori
    Abstract:

    During the process of autophagy, autophagosomes undergo a maturation process consisting of multiple fusions with endosomes and lysosomes, which provide an acidic environment and Digestive Function to the interior of the autophagosome. Here we found that a fusion protein of monomeric red-fluorescence protein and LC3, the most widely used marker for autophagosomes, exhibits a quite different localization pattern from that of GFP-LC3. GFP-LC3 loses fluorescence due to lysosomal acidic and degradative conditions but mRFP-LC3 does not, indicating that the latter can label the autophagic compartments both before and after fusion with lysosomes. Taking advantage of this property, we devised a novel method for dissecting the maturation process of autophagosomes. mRFP-GFP tandem fluorescent-tagged LC3 (tfLC3) showed a GFP and mRFP signal before the fusion with lysosomes, and exhibited only the mRFP signal subsequently. Using this method, we provided evidence that overexpression of a dominant negative form of Rab7 prevented the fusion of autophagosomes with lysosomes, suggesting that Rab7 is involved in this step. This method will be of general utility for analysis of the autophagosome maturation process.

  • dissection of the autophagosome maturation process by a novel reporter protein tandem fluorescent tagged lc3
    Autophagy, 2007
    Co-Authors: Shunsuke Kimura, Takeshi Noda, Tamotsu Yoshimori
    Abstract:

    During the process of autophagy, autophagosomes undergo a maturation process consisting of multiple fusions with endosomes and lysosomes, which provide an acidic environment and Digestive Function to the interior of the autophagosome. Here we found that a fusion protein of monomeric Red-fluorescence protein and LC3, the most widely used marker for autophagosomes, exhibits a quite different localization pattern from that of GFP-LC3. GFP-LC3 loses fluorescence due to lysosomal acidic and degradative conditions but mRFP-LC3 does not, indicating that the latter can label the autophagic compartments both before and after fusion with lysosomes. Taking advantage of this property, we devised a novel method for dissecting the maturation process of autophagosomes. mRFP-GFP tandem fluorescent-tagged LC3 (tfLC3) showed a GFP and mRFP signal before the fusion with lysosomes, and exhibited only the mRFP signal subsequently. Using this method, we provided evidence that overexpression of a dominant negative form of Rab7 ...

R A Zinn - One of the best experts on this subject based on the ideXlab platform.

  • influences of urea fermentation potential on growth performance dietary energetics Digestive Function and blood urea n in holstein steers fed a finishing diet based on steam flaked corn where metabolizable protein supply was otherwise in excess of theoretical requirement
    Applied Animal Science, 2019
    Co-Authors: J Salinaschavira, M. F. Montano, Gilberto Lopezvalencia, A Plascencia, R A Zinn
    Abstract:

    ABSTRACT Objectives This study was conducted to examine the effect of urea fermentation potential (UFP) on Digestive Function, BUN, growth performance, and dietary energetics, in steers fed a finishing diet based on steam-flaked corn containing 15% dried distillers grains with solubles. Materials and Methods Four UFP levels were evaluated (−0.30, 0.10, 0.50, 0.90). The UFP levels were manipulated by dietary urea inclusion level of 1.20, 0.80, 0.40 and 0%, respectively (DM basis). Results and Discussion Dietary UFP did not affect (P > 0.10) ruminal digestion of feed N. Ruminal starch digestion decreased with increasing UFP (linear effect, P = 0.02). Ruminal digestion of starch and OM tended to be maximal (quadratic effect, P ≤ 0.07) at dietary UFP of 0.1%. Increasing UFP decreased flow to duodenum of nonammonia-N (linear effect, P = 0.04), and microbial N (linear effect, P = 0.05). Duodenal flow of feed N was not affected (P = 0.18) by dietary UFP. Increasing dietary UFP decreased total-tract digestion of OM (linear effect, P = 0.02), starch (linear effect, P = 0.03), and digestible energy (linear effect, P = 0.03). Increasing dietary UFP decreased ruminal pH (linear effect, P = 0.01) 1 h after feeding. Thereafter, the influence of dietary UFP on ruminal pH was not appreciable. Increasing dietary UFP decreased ruminal ammonia-N 1 h after feeding (linear effect, P Implications and Applications Even when finishing diet formulations otherwise exceed MP requirements, dietary UFP level remains a relevant factor to optimize dietary energy use. Dietary UFP levels greater than 0.50% could negatively affect digestion, gain efficiency, and dietary NE use.

  • influence of protein nutrition and virginiamycin supplementation on feedlot growth performance and Digestive Function of calf fed holstein steers
    Journal of Animal Science, 2016
    Co-Authors: J Salinaschavira, Juan Diego Navarrete, N. Torrentera, A Plascencia, A Barreras, M F Montano, R A Zinn
    Abstract:

    Two experiments were conducted to examine the influence of protein and virginiamycin (VM) supplementation on feedlot growth performance, digestion, and metabolizable AA (MAA) supply of calf-fed Holstein steers. Growth performance and dietary energetics were evaluated in 120 Holstein steers (127 ± 9 kg). During the initial 112-d feeding period, a steam-flaked corn-based diet was balanced to meet either 100% (MAB) or 87% (UREA) of MAA requirements. Diets were supplemented with or without 22.5 mg/kg VM in a 2 × 2 factorial arrangement. Subsequently (d 112 to 308), all steers received the UREA diet with or without VM. During the initial 112-d, MAB increased ADG, G:F, and dietary NE ( 0.10) across initial supplementation treatments. Overall (d 1 to 308), MAB did not affect ADG ( > 0.10) but enhanced G:F efficiency ( = 0.03) and dietary NE ( = 0.05). During the initial 112-d period and through the remainder of the experiment, VM increased G:F ( 0.10) on ruminal digestion of OM, NDF, starch, microbial efficiency, or total tract digestion of OM and NDF. The MAB increased indispensable AA flow to the small intestine ( 0.10) total tract N digestion. Extrapolating from AA supplies in the metabolism study, MAB satisfied indispensable AA requirements during the initial 112-d period, whereas the UREA diet met 73.5% and 79.2% of methionine and lysine requirements, respectively. During the subsequent periods (d 112 to 308) indispensable AA supplies exceeded theoretical requirements. We conclude that enhancements in energy utilization when diets are balanced to meet MAA requirements of calf-fed Holstein steers during the initial 112-d feedlot period remain appreciable throughout time on feed. Virginiamycin enhanced efficiency of energy utilization throughout the feedlot growing-finishing period.

  • effects of monensin and virginiamycin supplementation in finishing diets with distiller dried grains plus solubles on growth performance and Digestive Function of steers
    Journal of Applied Animal Research, 2015
    Co-Authors: M. F. Montano, N. Torrentera, J Salinaschavira, O M Manriquez, R A Zinn
    Abstract:

    Two trials were conducted to evaluate the influence of virginiamycin (26 mg/kg) and monensin (34 mg/kg) supplementation of a steam-flaked corn-based finishing diet containing 15% distiller dried grains plus solubles (DDGS) on growth performance and Digestive Function. In Trial 1, 135 steers (257 ± 21 kg) were used to evaluate treatment effects on 145-d growth performance. There were no treatment effects (P > 0.20) on dry matter intake (DMI). Antibiotic supplementation tended to increase average daily gain (ADG; 7%, P = 0.07); increased gain efficiency (11%, P < 0.01) and estimated dietary net energy (NE; 10%, P < 0.01). Carcass marbling score was greater (12%, P = 0.04) for virginiamycin vs. monensin. In Trial 2, three Holstein steers (300 ± 7 kg) were used in a 3 × 3 Latin square to evaluate treatment effects on digestion. Both monensin and virginiamycin depressed ruminal digestion of organic matter (OM; 6%, P < 0.01) and feed nitrogen (N, 15%, P = 0.03) and microbial protein synthesis (15%, P = 0.03). T...

  • effects of monensin and virginiamycin supplementation in finishing diets with distiller dried grains plus solubles on growth performance and Digestive Function of steers
    Journal of Applied Animal Research, 2015
    Co-Authors: M. F. Montano, N. Torrentera, J Salinaschavira, O M Manriquez, R A Zinn
    Abstract:

    Two trials were conducted to evaluate the influence of virginiamycin (26 mg/kg) and monensin (34 mg/kg) supplementation of a steam-flaked corn-based finishing diet containing 15% distiller dried grains plus solubles (DDGS) on growth performance and Digestive Function. In Trial 1, 135 steers (257 ± 21 kg) were used to evaluate treatment effects on 145-d growth performance. There were no treatment effects (P > 0.20) on dry matter intake (DMI). Antibiotic supplementation tended to increase average daily gain (ADG; 7%, P = 0.07); increased gain efficiency (11%, P < 0.01) and estimated dietary net energy (NE; 10%, P < 0.01). Carcass marbling score was greater (12%, P = 0.04) for virginiamycin vs. monensin. In Trial 2, three Holstein steers (300 ± 7 kg) were used in a 3 × 3 Latin square to evaluate treatment effects on digestion. Both monensin and virginiamycin depressed ruminal digestion of organic matter (OM; 6%, P < 0.01) and feed nitrogen (N, 15%, P = 0.03) and microbial protein synthesis (15%, P = 0.03). T...

  • influence of feeding enzymatically hydrolyzed yeast cell wall on growth performance and Digestive Function of feedlot cattle during periods of elevated ambient temperature
    Asian-australasian Journal of Animal Sciences, 2015
    Co-Authors: J Salinaschavira, Constantino Raymundo, C Arzola, Victor M Gonzalezvizcarra, Olga M Manriqueznunez, Martin F Montanogomez, J D Navarretereyes, R A Zinn
    Abstract:

    In experiment 1, eighty crossbred steers (239±15 kg) were used in a 229-d experiment to evaluate the effects of increasing levels of enzymatically hydrolyzed yeast (EHY) cell wall in diets on growth performance feedlot cattle during periods of elevated ambient temperature. Treatments consisted of steam-flaked corn-based diets supplemented to provide 0, 1, 2, or 3 g EHY/hd/d. There were no effects on growth performance during the initial 139-d period. However, from d 139 to harvest, when 24-h temperature humidity index averaged 80, EHY increased dry matter intake (DMI) (linear effect, p 0.10) on carcass characteristics. In experiment 2, four Holstein steers (292±5 kg) with cannulas in the rumen and proximal duodenum were used in a 4×4 Latin Square design experiment to evaluate treatments effects on characteristics of ruminal and total tract digestion in steers. There were no treatment effects (p>0.10) on ruminal pH, total volatile fatty acid, molar proportions of acetate, butyrate, or estimated methane production. Supplemental EHY decreased ruminal molar proportion of acetate (p = 0.08), increased molar proportion of propionate (p = 0.09), and decreased acetate:propionate molar ratio (p = 0.07) and estimated ruminal methane production (p = 0.09). It is concluded that supplemental EHY may enhance DMI and ADG of feedlot steers during periods of high ambient temperature. Supplemental EHY may also enhance ruminal fiber digestion and decrease ruminal acetate:propionate molar ratios in feedlot steers fed steam-flaked corn-based finishing diets.

Shunsuke Kimura - One of the best experts on this subject based on the ideXlab platform.

  • Dissection of the Autophagosome Maturation Process by a Novel Reporter Protein, Tandem Fluorescent-Tagged LC3
    Autophagy, 2007
    Co-Authors: Shunsuke Kimura, Takeshi Noda, Tamotsu Yoshimori
    Abstract:

    During the process of autophagy, autophagosomes undergo a maturation process consisting of multiple fusions with endosomes and lysosomes, which provide an acidic environment and Digestive Function to the interior of the autophagosome. Here we found that a fusion protein of monomeric red-fluorescence protein and LC3, the most widely used marker for autophagosomes, exhibits a quite different localization pattern from that of GFP-LC3. GFP-LC3 loses fluorescence due to lysosomal acidic and degradative conditions but mRFP-LC3 does not, indicating that the latter can label the autophagic compartments both before and after fusion with lysosomes. Taking advantage of this property, we devised a novel method for dissecting the maturation process of autophagosomes. mRFP-GFP tandem fluorescent-tagged LC3 (tfLC3) showed a GFP and mRFP signal before the fusion with lysosomes, and exhibited only the mRFP signal subsequently. Using this method, we provided evidence that overexpression of a dominant negative form of Rab7 prevented the fusion of autophagosomes with lysosomes, suggesting that Rab7 is involved in this step. This method will be of general utility for analysis of the autophagosome maturation process.

  • dissection of the autophagosome maturation process by a novel reporter protein tandem fluorescent tagged lc3
    Autophagy, 2007
    Co-Authors: Shunsuke Kimura, Takeshi Noda, Tamotsu Yoshimori
    Abstract:

    During the process of autophagy, autophagosomes undergo a maturation process consisting of multiple fusions with endosomes and lysosomes, which provide an acidic environment and Digestive Function to the interior of the autophagosome. Here we found that a fusion protein of monomeric Red-fluorescence protein and LC3, the most widely used marker for autophagosomes, exhibits a quite different localization pattern from that of GFP-LC3. GFP-LC3 loses fluorescence due to lysosomal acidic and degradative conditions but mRFP-LC3 does not, indicating that the latter can label the autophagic compartments both before and after fusion with lysosomes. Taking advantage of this property, we devised a novel method for dissecting the maturation process of autophagosomes. mRFP-GFP tandem fluorescent-tagged LC3 (tfLC3) showed a GFP and mRFP signal before the fusion with lysosomes, and exhibited only the mRFP signal subsequently. Using this method, we provided evidence that overexpression of a dominant negative form of Rab7 ...

Ashild Krogdahl - One of the best experts on this subject based on the ideXlab platform.

  • bt maize mon810 and non gm soybean meal in diets for atlantic salmon salmo salar l juveniles impact on survival growth performance development Digestive Function and transcriptional expression of intestinal immune and stress responses
    PLOS ONE, 2014
    Co-Authors: Jinni Gu, Anne Marie Bakke, Elin C Valen, Ingrid Lein, Ashild Krogdahl
    Abstract:

    Responses in Atlantic salmon (Salmo salar L.) juveniles (fry) fed diets containing genetically modified maize (Bt-maize, MON810) expressing Cry1Ab protein from first-feeding were investigated during a 99-day feeding trial. Four experimental diets were made; each diet contained ∼20% maize, either Bt-maize or its near-isogenic maternal line (non-GM maize). One pair was fishmeal-based while the other pair included standard (extracted) soybean meal (SBM; 16.7% inclusion level), with the intention of investigating responses to the maize varieties in healthy fish as well as in immunologically challenged fish with SBM-induced distal intestinal inflammation, respectively. Three replicate tanks of fry (0.17±0.01 g; initial mean weight ± SEM) were fed one of the four diets and samples were taken on days 15, 36, 48 and 99. Survival, growth performance, whole body composition, Digestive Function, morphology of intestine, liver and skeleton, and mRNA expression of some immune and stress response parameters in the distal intestine were evaluated. After 99 days of feeding, survival was enhanced and the intended SBM-induced inflammatory response in the distal intestine of the two groups of SBM-fed fish was absent, indicating that the juvenile salmon were tolerant to SBM. Mortality, growth performance and body composition were similar in fish fed the two maize varieties. The Bt-maize fed fish, however, displayed minor but significantly decreased Digestive enzyme activities of leucine aminopeptidase and maltase, as well as decreased concentration of gut bile salts, but significantly increased amylase activity at some sampling points. Histomorphological, radiographic and mRNA expression evaluations did not reveal any biologically relevant effects of Bt-maize in the gastrointestinal tract, liver or skeleton. The results suggest that the Cry1Ab protein or other compositional differences in GM Bt-maize may cause minor alterations in intestinal responses in juvenile salmon, but without affecting overall survival, growth performance, development or health.

  • Digestive Function and intestinal integrity in atlantic salmon salmo salar fed kernel meals and protein concentrates made from yellow or narrow leafed lupins
    Aquaculture, 2006
    Co-Authors: Stale Refstie, Brett Glencross, Thor Landsverk, Mette Sorensen, Einar Lilleeng, Wayne Hawkins, Ashild Krogdahl
    Abstract:

    Abstract This study assessed the effects of yellow lupin ( Lupinus luteus ) and narrow-leafed lupin ( L . angustifolius ) kernel meals and protein concentrates on the gastrointestinal integrity, capacity for Digestive hydrolysis, and digestibility of nutrients in Atlantic salmon. A basal diet (FM) was made from fish meal, wheat, and fish oil. Six additional diets were formulated by replacing 30% of the FM diet with lupin kernel meal made from L . l . cv. Wodjil (LKM), L . a . cv. Belara (BKM), and L . a . cv. Myallie (MKM), lupin protein concentrates made from the same L . l . (LPC) and L . a . cv. M (MPC), or extracted soybean meal (SBM). All diets were extruded. Each diet was fed to three groups of 176 g salmon kept in 1 m 2 tanks with 5.6 °C saltwater for 3 weeks prior to sampling of blood, intestinal organs, digesta, and faeces. Inclusion of lupin meals in the diets resulted in harder and more condensed feed particles. Ulcer-like lesions were observed in the stomach of fish from all feeding groups, and this was worsened by lupin in the diet, but did not appear to be pellet hardness related. No consistent altered morphology was observed in the distal intestine (DI) of fish fed the FM and lupin diets, while the DI of fish fed SBM showed consistent and typical soybean meal-induced pathomorphological changes. Plasma cholesterol was higher when feeding MKM and LKM than when feeding FM, MPC, and LPC, with intermediate levels when feeding BKM and SBM. Feeding LKM and LPC resulted in a higher weight of the GIT when related to body weight. Trypsin activity and bile acid concentration were generally higher in digesta from the pyloric (PI) and mid (MI) intestine when feeding FM and lupin diets than when feeding SBM, while the opposite was seen for trypsin activity in digesta from DI. There were no effects of diet on leucine aminopeptidase (LAP) and maltase activity in PI and MI, but in DI the activity of these brush border enzymes were significantly lowered when feeding SBM. SBM in the diet resulted in watery faeces and lowered apparent digestibility of lipid, but this was not observed when feeding the lupin diets. To conclude, the tested lupin kernel meals and protein concentrates did not alter the intestinal Function in Atlantic salmon when included at 30% of the diet. Dietary lupin was, however, involved in the worsening of ulcer-like gastric lesions.

Takeshi Noda - One of the best experts on this subject based on the ideXlab platform.

  • Dissection of the Autophagosome Maturation Process by a Novel Reporter Protein, Tandem Fluorescent-Tagged LC3
    Autophagy, 2007
    Co-Authors: Shunsuke Kimura, Takeshi Noda, Tamotsu Yoshimori
    Abstract:

    During the process of autophagy, autophagosomes undergo a maturation process consisting of multiple fusions with endosomes and lysosomes, which provide an acidic environment and Digestive Function to the interior of the autophagosome. Here we found that a fusion protein of monomeric red-fluorescence protein and LC3, the most widely used marker for autophagosomes, exhibits a quite different localization pattern from that of GFP-LC3. GFP-LC3 loses fluorescence due to lysosomal acidic and degradative conditions but mRFP-LC3 does not, indicating that the latter can label the autophagic compartments both before and after fusion with lysosomes. Taking advantage of this property, we devised a novel method for dissecting the maturation process of autophagosomes. mRFP-GFP tandem fluorescent-tagged LC3 (tfLC3) showed a GFP and mRFP signal before the fusion with lysosomes, and exhibited only the mRFP signal subsequently. Using this method, we provided evidence that overexpression of a dominant negative form of Rab7 prevented the fusion of autophagosomes with lysosomes, suggesting that Rab7 is involved in this step. This method will be of general utility for analysis of the autophagosome maturation process.

  • dissection of the autophagosome maturation process by a novel reporter protein tandem fluorescent tagged lc3
    Autophagy, 2007
    Co-Authors: Shunsuke Kimura, Takeshi Noda, Tamotsu Yoshimori
    Abstract:

    During the process of autophagy, autophagosomes undergo a maturation process consisting of multiple fusions with endosomes and lysosomes, which provide an acidic environment and Digestive Function to the interior of the autophagosome. Here we found that a fusion protein of monomeric Red-fluorescence protein and LC3, the most widely used marker for autophagosomes, exhibits a quite different localization pattern from that of GFP-LC3. GFP-LC3 loses fluorescence due to lysosomal acidic and degradative conditions but mRFP-LC3 does not, indicating that the latter can label the autophagic compartments both before and after fusion with lysosomes. Taking advantage of this property, we devised a novel method for dissecting the maturation process of autophagosomes. mRFP-GFP tandem fluorescent-tagged LC3 (tfLC3) showed a GFP and mRFP signal before the fusion with lysosomes, and exhibited only the mRFP signal subsequently. Using this method, we provided evidence that overexpression of a dominant negative form of Rab7 ...