• Title/Summary/Keyword: 43 kD protein

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Proteomics Analysis of Immunoprecipitated Proteins Associated with the Oncogenic Kinase Cot

  • Wu, Binhui;Wilmouth, Rupert C.
    • Molecules and Cells
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    • v.25 no.1
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    • pp.43-49
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    • 2008
  • Cancer Osaka thyroid, also known as Tpl-2 (Cot) is a member of the MAP3K kinase family and plays a key role in the regulation of the immune response to pro-inflammatory stimuli such as lipopolysaccharide (LPS) and tumour necrosis $factor-{\alpha}$ ($TNF-{\alpha}$). A series of Cot constructs with an N-terminal 6xHis tag were transiently expressed in HEK293 cells: $Cot_{130-399}$ (kinase domain), $Cot_{1-388}$ (N-terminal and kinase do-mains), $Cot_{1-413}$, $Cot_{1-438}$ (containing a putative PEST sequence), $Cot_{1-457}$ (containing both PEST and degron sequences) and $Cot_{1-467}$ (full-length protein). These Cot proteins were pulled down using an anti-6xHis antibody and separated by 2D electrophoresis. The gels were silver-stained and 21 proteins were detected that did not appear, or had substantially reduced intensity, in the control sample. Three of these were identified by MS and MS/MS analysis as Hsp90, Hsp70 and Grp78. Hsp90 appeared to bind to the kinase domain of Cot and this interaction was further investigated using co-immuno-precipitation with both overexpressed Cot in HEK293 cells and endogenous Cot in Hela cells.

EFFECTS OF PHOSPHORUS AND CALCIUM ON FEED INTAKE AND YIELD AND COMPOSITION OF MILK OF HOLSTEIN COWS

  • Morse, D.;Head, H.H.;Wilcox, C.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.2
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    • pp.231-237
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    • 1994
  • Three concentrations of P (0.33, 0.43 and 0.54%) and two concentrations of Ca (0.60 and 0.97%) in ration dry matter were evaluated for effects on dry matter intake and on milk yield and composition using 24 Holstein cows. Cows were arranged in a $3{\times}2$ factorial experiment as an incomplete randomized block design with three 28-day periods. Each cow consumed at least one ration with each concentration of Ca. Dry matter intake, yield of 3.5% Fat Corrected Milk, and milk composition were not affected by concentration of P, but milk yield was greater when lowest concentration of P was fed (22.8 vs. 22.1 kg/day; p<0.07). Cows fed rations containing 0.60% Ca had greater milk (22.7 vs. 21.9 kg/day; p<0.02) and 3.5% Fat Corrected Milk yields (p<0.03) and slightly greater protein content than when fed 0.97% Ca. Dietary Ca:P ratios between 1.1:1 and 2.9:1 had no effect on dry matter intake, milk yield, or composition. Concentrations of P in plasma were within the normal range for all rations. Because cows had high dry matter intake, mean daily intakes of both P and Ca were greater than required for their level of milk yield.

Utilization of Putrescine by Streptococcus pneumoniae During Growth in Choline-limited Medium

  • Ware D.;Watt J.;Swiatlo E.
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.398-405
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    • 2005
  • Polyamines such as putrescine are small, ubiquitous polycationic molecules that are required for optimal growth of eukaryotic and prokaryotic cells. These molecules have diverse effects on cell physiology and their intracellular content is regulated by de novo synthesis and uptake from the environment. The studies presented here examined the structure of a putative polyamine transporter (Pot) operon in Streptococcus pneumoniae (pneumococcus) and growth of pneumococci in medium containing putrescine substituted for choline. RT-PCR experiments demonstrated that the four genes encoding the Pot system are co-transcribed with murB, a gene involved in an intermediary step of peptidoglycan synthesis. Pneumococci grown in chemically-defined media (CDM) containing putrescine without choline enter logarithmic phase growth after 36-48 hs. However, culture density at stationary phase eventually reaches that of choline-containing medium. Cells grown in CDM-putrescine formed abnormally elongated chains in which the daughter cells failed to separate and the choline-binding protein PspA was no longer cell-associated. Experiments with CDM containing radiolabeled putrescine demonstrated that pneumococci concentrate this polyamine in cell walls. These data suggest that pneumococci can replicate without choline if putrescine is available and this polyamine may substitute for aminoalcohols in the cell wall teichoic acids.

Isolation and Identification of Microorganisms Producing the Soy Protein-Hydrolyzing Enzyme from Traditional Mejus (전통메주로부터 대두단백질 가수분해효소 생산성 미생물의 분리 및 동정)

  • Kang, Min-Jung;Kim, Seong-Ho;Joo, Hyun-Kyu;Lee, Gap-Sang;Yim, Moo-Hyun
    • Applied Biological Chemistry
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    • v.43 no.2
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    • pp.86-94
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    • 2000
  • In order to develop the enzymatic hydrolysis system concerned with taste and flavor, strains having the high hydrolyzing activity on the soy protein were selected from some traditional Mejus. Two molds and one bacterium producing enzymes which were different in character of hydrolysis were isolated and identified. Leucine and azodye enzyme activities of both M4 and M5 were relatively high among in the isolated molds. And, leucine enzyme activity of B16 was the lowest in the isolated bacteria. These strains were isolated as microorganisms having a dissimilar hydrolysis pattern on the soy protein by enzymatic reactions. Mold M4 on the culture solid media was mycelium colors of white and its sclerotia colors were changed from white to black. According to the result of slide culture, radial conidial head, subclavate vesicle, conidia of subglobose, stipes of uncolored with smooth walls and metula and phialides were existed. Because M4 was taxonomically similar to the characteristics of Aspergillus oryzae (ahlburg) species, M4 was identified and named as Aspergillus oryzae M4.Mold M5 showed white and black mycelium on the MEA medium. Mold M5 colony exhibited grayish-green color and have long(7 mm) sporangiophores at slide culture. Sporangia became brownish-gray and the wall of larger sporangia was broken to form small collars, and smaller sporangia were fomed continually from large basal membrane. Columella is globose and hyaline, and sporangiospores are ellipsoidal of small diameter$(80\;{\mu}m)$. Because M5 was taxonomically similar to the Mucor circinelloides of zygomycetes, M5 was was identified and named as Mucor circinelloides M5. Bacteria B16 colony was opaque white, circular and lobate, and had rod shaped endospore. B16 was found positive in stain, catalase, ${\beta}-glucosidse$ and V-P tests. B16 was found to utilize D-fructose, ${\alpha}-D-glucose$, maltose, D-mannose, D-raffinose, stachyose and sucrose. By the morphological and physiological results, the characteristics of B16 was thought to correspond to that of Bacillus megaterium. However, fatty acid composition was similar to Paenibacillus marcerans, requiring further study for the definite identification. Accordingly, Bacteria B16 was provisionally classified and named as Bacillus megaterium B16.

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Effects of Replacing Corn with Rice or Brown Rice on Laying Performance, Egg Quality, and Apparent Fecal Digestibility of Nutrient in Hy-Line Brown Laying Hens

  • Kim, Chan Ho;Park, Seong Bok;Kang, Hwan Ku
    • Korean Journal of Poultry Science
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    • v.43 no.2
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    • pp.97-103
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    • 2016
  • This study aimed of effects of replacing maize with rice or brown rice on laying performance, egg quality, and apparent fecal digestibility of nutrient in Hy-Line Brown laying hens. A total of 200, 25-week old Hy-Line Brown laying hens were randomly assigned to one of 5 treatments, each of which was replicated 4 times. Four experimental diets that were formulated two different grains (brown rice or rice) with two inclusion levels of (15 and 30%) as a replacement of maize. The experimental period was 8 weeks. During the experiment, hens were provided with feed and water ad libitum and were exposed to a 16:8=L:D lighting schedule. There were no differences in feed intake, egg weight, and egg mass during the 8 weeks of the feeding trial among groups. However, hen-day egg production was greater (P<0.05) for Brown rice treatment groups than that for basal and rice treatment groups. In addition, the supplementation of brown rice or rice did not have an effect on eggshell strength, eggshell thickness, and HU during of the feeding trial (Table 4). However, egg yolk color was less (P<0.05) for basal treatment groups than that for among treatment group. ATTD of dry matter (DM) and crude fat were greater (P<0.05) for brown rice 15 treatment than for rice 30 treatment. Crude protein (CP) was greater (P<0.05) for brown rice 15 treatment than for basal treatment. In conclusion, brown rice or rice grains is a good alternative energy feedstuff and can be used in laying hens, totally replacing maize, without any negative effect on the laying performance, egg quality, and nutrient digestibility.

Effects of Mutation at Two Conserved Aspartate Residues and a Serine Residue on Functions of Yeast TSA 1 (Saccharomyces cerevisiae TSA1의 보존된 아스파트산 잔기 및 세린 잔기의 변이가 과산화효소 활성 및 샤페론 활성에 미치는 영향)

  • Lee, Songmi;Cho, Eun Yi;Kim, Kanghwa
    • Microbiology and Biotechnology Letters
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    • v.45 no.1
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    • pp.81-86
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    • 2017
  • Alignment of 967 reference sequences of the typical 2-Cys peroxiredoxin family of proteins revealed that 10 amino acids were conserved, with over 99% identity. To investigate whether the conserved aspartic acid residues and serine residue affect the peroxidase and chaperone activity of the protein, we prepared yeast TSA1 mutant proteins in which aspartic acids at positions 75 and 103 were replaced by valine or asparagine, and serine at position 73 was replaced by alanine. By non-reducing SDS-PAGE, TSA1 and the S73A, D75V and D75N mutants were detected in dimeric form, whereas the D103V and D103N mutants were detected in various forms, ranging from high molecular-weight to monomeric. Compared with wild type TSA1, the D75N mutant exhibited 50% thioredoxin peroxidase activity, and the S73A and D75V mutants showed 25% activity. However, the D103V and D103N mutants showed no peroxidase activity. All proteins, except for the D103V and D103N mutants, exhibited chaperone activity at $43^{\circ}C$. Our results suggest that the two conserved aspartic acid residues and serine residue of TSA1 play important roles in its thioredoxin peroxidase activity, and D103 plays a critical role in its chaperone activity.

Effect of Replacement of Groundnut Cake with Urea-treated Neem (Azadirachta indica A. juss) Seed Kernel Cake on Nutrient Utilisation in Lambs

  • Musalia, L.M.;Anandan, S.;Sastry, V.R.B.;Katiyar, R.C.;Agrawal, D.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.9
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    • pp.1273-1277
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    • 2002
  • The effect of urea treatment on chemical composition of neem seed kernel cake (NSKC) was investigated by soaking the cake in 2.1% urea solution (1.2 l $kg^{-1}$ NSKC) for five days. The effect on utilisation of nutrients by replacing groundnut cake (GNC) (30%) with urea-treated neem seed kernel cake (UTNSKC) (33%) in a concentrate mixture fed to meet 70% of the protein requirements of lambs (8 males and 8 females), was monitored in a digestibility study. Following urea treatment of NSKC only 9.5% of urea was hydrolysed and the crude protein content of the cake was increased by 6.65%. The tannin content in depulped neem seeds was 37% catechin equivalent. Whereas feeding UTNSKC had no effect on intake of dry matter (72.5 vs 66.3 g/kg $BW^{0.75}day^{-1}$) and digestibility of crude fibre (41.3 vs 43.4%), the cake depressed (p<0.01) the percent digestibility of dry matter (63.7 vs 70.2), crude protein (63.2 vs 70.2), nitrogen free extract (73.8 vs 80.5) and gross energy (64.3 vs 69.1). Digestibility of ether extract (75.8 vs 70.9%) was higher (p<0.05) in animals offered UTNSKC. The nutritive value of the composite ration consumed by lambs offered UTNSKC was lower (p<0.01) in terms of total digestible nutrients (64.7 vs 70.2%) and digestible energy (2.8 vs 3.0 Kcal/g DM). Intake of digestible energy (199.8 vs 194.1 Kcal/kg $BW^{0.75}day^{-1}$) and retention of nitrogen (7.53 vs 8.23 g $day^{-1}$) and calcium (2.12 vs 1.84 g $day^{-1}$) were comparable on the 2 rations. Animals fed UTNSKC retained less (p<0.01) phosphorus (0.37 vs 1.05 g $day^{-1}$). The results indicate that urea treatment increased the protein level of NSKC whereas feeding the treated cake as a replacement of GNC, lowered the digestibility of nutrients and retention of phosphorus.

Dendropanax morbifera Extract Inhibits Intimal Hyperplasia in Balloon-Injured Rat Carotid Arteries by Modulating Phenotypic Changes in Vascular Smooth Muscle Cells

  • Lim, Leejin;Jo, Juyeong;Yoon, Sang Pil;Jang, Inyoub;Ki, Young-Jae;Choi, Dong-Hyun;Song, Heesang
    • Natural Product Sciences
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    • v.26 no.1
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    • pp.71-78
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    • 2020
  • The plant Dendropanax morbifera Léveille is effective folk medicines for the treatment of several conditions, such as infectious diseases, skin diseases, and other illnesses. Although the inhibitory effects of D. morbifera on the proliferation and migration of vascular smooth muscle cells (VSMCs) have been shown in our previous study, its effects in vivo remain to be elucidated. In this study, we aimed to investigate the protective effects of the extracts from D. morbifera (EDM) on neointimal hyperplasia of rat carotid artery and explore the underlying mechanisms. We observed that the ratio of intima to media thickness (I/M) was significantly decreased in the EDM-treated groups by ~80% compared to that of the control. The expression of Ki-67 and proliferating cell nuclear antigen was decreased by ~70% in the EDM-treated groups compared to that of the control. In addition, matrix metalloproteinase (MMP)2 and MMP9 significantly reduced in the neointimal layer of the EDM-treated groups. Moreover, the decreased levels of contractile phenotypic markers of VSMCs, such as α-smooth muscle actin, myocardin, and smooth muscle-myosin heavy chain, were successfully restored by EDM treatment. Furthermore, the levels of synthetic phenotypic markers, cellular retinal binding protein 1 and connexin 43 were also restored to normal levels. These results suggest that EDM inhibits vascular neointimal hyperplasia induced by balloon injury in rats via phenotypic modulation of VSMCs. Therefore, EDM may be a potential drug candidate for the prevention of restenosis.

Effect of Interaction between Nutritional Level and Breed on Performance of Broiler Production (BROILER 사료의 영양수준이 육용계종의 산육능력에 미치는 영향)

  • 오봉국;오세정
    • Korean Journal of Poultry Science
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    • v.6 no.1
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    • pp.12-23
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    • 1979
  • This experiment was carrid out to investigate the interaction between boilelr strains and nutrition levels, and the performances of four broiler strains such as Han Hyup 603, Hubbard, Anak and Filch when they were fed by four different nutrition levels (High Protein and energy; HP. HE., Medium Protein and energy; MP. ME., Low Protein ana energy; LP. LE., and low protein and energy; LLP. LLE.). The data used in this study were obtained from a total of 1200 broiler type chicks in Poultry Testing Station, Korean Poultry Association from June 16, to August 11, 1978. Differences of all characters among four nutrition levels were significant except viability and carcass rate. HP. HE and MP. ME treatments showed nearly the same performances in body weight, feed efficiency and point, spread but they were significantly superior to those of LP. LE and LLP. LLE. There were not significant differences among four strains in feed efficiency and viability but other characters, body weight, point spread and carcass rate were observed that the performance of the best strain B was significantly superior to strain D but it was not recognized significance compared with strain A, C in tile result of statisticel analysis. In the interaction between strains and nutrition levels, body weight at high and levels showed significantly differences but at low and low nutrition levels were nearly same among four strains. Therefore this study demonstrated that comparision of body weights between strains should be performed at medium nutrition level or above. Also point spread calculated as index of body weight and feed efficiency was observed that strain B at low nutrition level is excellently higher than other strains and there were little differences at low nutrition level among all strains. It was found that ]it tie differences between performances of high arid medium levels seemed to be as the reason of high fat addition for energy source to high mutrition feed, and in general superior strain showed good performance at all the nutrition levels in$.$all characters but in body weight and point spread there were significantly different responses with different nutrition level, The most superior strain B among four strains earned the most profit per bird, Although performances of high and medium nutrition levels were nearly the same, medium nutrition level also showed the most profit because the feed cost of high nutrition level was higher than that of medium nutrition level.

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Nutritional and performance viability of cactus Opuntia-based diets with different concentrate levels for Girolando lactating dairy cows

  • Inacio, Jonas Gomes;da Conceicao, Maria Gabriela;dos Santos, Djalma Cordeiro;de Oliveira, Julio Cesar Vieira;Chagas, Juana Catarina Cariri;de Oliveira Moraes, Glaucia Sabrine;dos Santos Silva, Evannielly Thuanny;de Andrade Ferreira, Marcelo
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.1
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    • pp.35-43
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    • 2020
  • Objective: The aim of this research was to evaluate the effect of different concentrate levels in diets based on cactus Opuntia Stricta (Haw.) Haw cladodes on the performance of lactating Girolando cows. Methods: The experiment involved 10 Girolando multiparous dairy cows at 512.6 kg of body weight (BW) and producing 13.2 kg milk/d, allocated into two 5×5 Latin squares. The experimental treatments consisted of control diet composed by cactus Nopalea cochenillifera. Salm-Dyck. cladodes (Nopalea), forage sorghum silage and concentrate at 20% on dry matter (DM) basis, and four concentrate levels diets (20%, 24%, 28%, and 32%) plus cactus Opuntia stricta (Haw.) Haw. cladodes (Opuntia) and forage sorghum silage. Results: Regarding cows fed control diet, the nutrients intake were greater than for cows fed with cactus Opuntia and concentrate. Regarding concentrate levels, intakes of DM, organic matter (OM), crude protein (CP), non-fiber carbohydrates (NFC), and total digestible nutrients of cows increased linearly. Organic matter, CP, and NDF digestibilities were similar in between to control diet and cactus Opuntia-based diets. The digestibility of NFC increased linearly when the concentrate was inserted. The N balance was the same for control diet and cactus Opuntia-based diets, irrespective the concentrate levels. Conclusion: For cows producing 14 kg/d with 3.5% of fat, it is recommended 32% of concentrate to be included in cactus Opuntia-based diets, and the increase in concentrate level promotes a linear increase in milk yield.