• Title/Summary/Keyword: Protein Profile

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Proteome Analysis of Recombinant CHO Cells Under Hyperosmotic Stress

  • Lee, Mun-Su;Kim, Gyeong-Uk;Kim, Yeong-Hwan;Lee, Gyun-Min
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.311-314
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    • 2003
  • Under hyperosmotic stress, rCHO cells display decreased specific growth rate $({\mu})$ and increased specific antibody productivity $(q_{Ab})$. The effects of hyperosmotic stress on batch culture cellular dynamics are not well understood. To this end, we conducted a proteome profile of rCHO cells, using 2D-gel, MALDI-TOF-MS and MS/MS. As a result, the proteome profile of rCHO cells could be established using 41 identified proteins. Based on this proteome profile of rCHO cells, we have found at least 8 differently expressed spots at hyperosmotic osmolality (450 mOsm/kg). Among these spots, two metabolic enzymes were found to be up-regulated (pyruvate kinase and GAPDH), while down-regulated protein was identified as tubulin. It shows that hyperosmotic stress can alter metabolic state, by up-regulated activities of two glycolysis enzymes, which could lead to activate the generation of metabolic energy. Tubulin expression was down-regulated, suggesting a reduction of cell division. Finally, the increased conversion energy could leads to improve overall productivity.

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Gene Expression Analysis of Acetaminophen-induced Liver Toxicity in Rat (아세트아미노펜에 의해 간손상이 유발된 랫드의 유전자 발현 분석)

  • Chung, Hee-Kyoung
    • Toxicological Research
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    • v.22 no.4
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    • pp.323-328
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    • 2006
  • Global gene expression profile was analyzed by microarray analysis of rat liver RNA after acute acetaminophen (APAP) administration. A single dose of 1g/kg body weight of APAP was given orally, and the liver samples were obtained after 24, 48 h, and 2 weeks. Histopathologic and biochemical studies enabled the classification of the APAP effect into injury (24 and 48 h) and regeneration (2 weeks) stages. The expression levels of 4900 clones on a custom rat gene microarray were analyzed and 484 clones were differentially expressed with more than a 1.625-fold difference(which equals 0.7 in log2 scale) at one or more time points. Two hundred ninety seven clones were classified as injury-specific clones, while 149 clones as regeneration-specific ones. Characteristic gene expression profiles could be associated with APAP-induced gene expression changes in lipid metabolism, stress response, and protein metabolism. We established a global gene expression profile utilizing microarray analysis in rat liver upon acute APAP administration with a full chronological profile that not only covers injury stage but also later point of regeneration stage.

Effect of Niacin Supplementation on Growth, Nutrient Utilization and Blood Biochemical Profile in Male Buffalo Calves

  • Kumar, Ravindra;Dass, R.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.10
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    • pp.1422-1428
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    • 2006
  • In order to investigate the effect of different levels of niacin supplementation on growth, nutrient utilization, their balance and blood biochemical profile, 15 male buffalo calves (9-10 months of age, $88.4{\pm}4.37kg$ average body weight) were divided into 3 equal groups each of 5 calves, following a completely randomized design, and fed individually for 120 days with wheat straw and concentrate mixture to meet their nutrient requirements. In addition calves were supplemented with 0 ppm (control, group I), 100 ppm (group II) and 200 ppm (group III) niacin. After 90 days of experimental feeding a metabolism trial was conducted to estimate the digestibility of nutrients and their balance. Fortnightly body weights were recorded to assess their growth rate and blood was collected from the jugular vein at day 0 and subsequently at 30-day intervals from all the experimental buffalo calves to study blood biochemical parameters. Results showed that intake and digestibility of dry matter, organic matter, crude protein, ether extract, total carbohydrates, neutral detergent fibre, acid detergent fibre, cellulose and hemicelluloses were statistically similar in the 3 groups. Buffalo calves in all three groups were in positive nitrogen, calcium and phosphorus balance, without showing any significant effect of the treatments. Dry matter, crude protein, digestible crude protein and total digestible nutrient intake ($g/d/kgW^{0.75}$) were similar in the control and niacin supplemented groups. Digestible crude protein (%) and total digestible nutrients (%) in the ration of the 3 groups were 8.07, 7.99, 7.92 and 56.70, 56.63, 56.74, respectively, and were comparable among the groups. The average daily gain (g) in-group II (567.50) was not significantly (p>0.05) higher than group I (500.0) and group III (510.0). Blood biochemical constituents (glucose, total protein, albumin, globulin, urea-N, insulin) showed no significant effect of niacin supplementation. However, serum cholesterol (mg/100 ml) was significantly (p<0.01) lower in the 200 ppm niacin-supplemented group than in the control and 100 ppm niacin-supplemented groups. It can be concluded that supplementation of niacin at 100 and 200 ppm in the diet of buffalo calves had no significant beneficial effect on their growth and nutrient utilization.

Protein and Amino Acid Composition of Domestic Potato Cultivars (국내산 감자 주요 품종의 아미노산 및 단백질 조성)

  • Kwon, Oh-Yun;Kim, Mi-Yeon;Son, Chan-Wok;Liu, Xi-Wen;Kim, Hyoung-Chin;Yoon, Won-Kee;Kim, Hwan-Mook;Kim, Mee-Ree
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.1
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    • pp.117-123
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    • 2008
  • The protein profiles of domestic potato cultivars were evaluated for total protein determination, amino acid composition, SDS-PAGE analysis and scanning densitometry. There were statistically significant differences in the levels of amino acids among potato cultivars. Total nitrogen amount was also significantly different among cultivars, ranging from 1.27 to 1.64%. SDS-PAGE analysis showed that there were significant differences in the content of major potato proteins such as papatin (40 kDa), trypsin inhibitor (20 kDa) and protease inhibitor (15 kDa) among cultivars (p<0.05). The amount of papatin among cultivars with a range of 22.16 to 25.81 mg/g d.w. was higher in Jopung, Shepody and Superior, whereas the amount of protease inhibitors including 15 kDa and 20 kDa was the highest in Jopung (37.0%). The Shepody contains the highest amount of papatin (25.8%) and the lowest of trypsin inhibitor (5.22%). Thus, it is suggested that Shepody is the most desirable cultivar for better nutrition based on the protein profile.

Control of Encapsulation Efficiency and Initial Burst in Polymeric Microparticle Systems

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

Genomewide Expression Profile of Forsythia Suspensa on Lipopolysaccaride-induced Activation in Microglial Cells

  • Sohn, Sung-Hwa;Ko, Eun-Jung;Kim, Yang-Seok;Shin, Min-Kyu;Hong, Moo-Chang;Bae, Hyun-Su
    • Molecular & Cellular Toxicology
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    • v.4 no.2
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    • pp.113-123
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    • 2008
  • Microglia, which is the primary immune effector cells in the central nervous system, constitutes the first line of defense against infection and injury in the brain. The goal of this study was to determine the protective (anti-inflammation) mechanisms of forsythia suspense (FS) on LPS-induced activation of BV-2 microglial cells. The effects of FS on gene expression profiles in activated BV-2 microglial cells were evaluated using microarray analysis. BV-2 microglial cells were cultured in a 100mm dish $(1{\times}10^7/dish)$ for 24hr and then pretreated with $1{\mu}g/mL$ FS or left untreated for 30 min. Next, $1{\mu}g/mL$ LPS was added to the samples and the cells were reincubated at $37^{\circ}C$ for 30 min, 1hr, and 3hr. The gene expression profiles of the BV-2 microglial cells varied depending on the FS. The oligonucleotide microarray analysis revealed that MAPK pathway-related genes such as Mitogen activated protein kinase 1 (Mapk1), RAS protein activator like 2 (Rasal2), and G-protein coupled receptor 12 (Gpr12) and nitric oxide biosynthesis-related genes such as nitric oxide synthase 1 (neuronal) adaptor protein (Nos1ap), and dimethylarginine dimethylaminohydrolase 1 (Ddah1) were down regulated in FS-treated BV-2 microglial cells. FS can affect the MAPK pathway and nitric oxide biosynthesis in BV-2 microglial cells.

Consumption and Digestion of Forages by Male Rusa (Cervus timorensis russa) Deer-the Effects of Castration and Season

  • Sookhareea, R.;Dryden, G. McL
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.8
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    • pp.1098-1106
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    • 2004
  • In a split-unit design, 12 Javan rusa (Cervus timorensis russa) stags (6 castrated and 6 entire) were used to investigate seasonal (winter, spring and summer) effects on intake, digestibility of food constituents, volatile fatty acid profile, and nitrogen retention, when given lucerne (Medicago sativa) or rhodes grass (Chloris guyana) hays. Entire stags ate 9% more dry matter (DM) than castrates (p<0.05). Both castrated and entire stags exhibited seasonal changes in DM intake, these being higher (p<0.05) in winter (62.3 g/kg $W^{0.75}$) than spring (56.9 g/kg $W^{0.75}$) or summer (55.3 g/kg $W^{0.75}$). Intakes of lucerne hay (211 g protein/kg DM) were significantly higher than of rhodes grass hay (49 g protein/kg DM) in all seasons (p<0.05). Digestibilities of DM (0.58), neutral detergent fibre (0.59) and acid detergent fibre (0.47) were similar between castrates and entires. DM digestibility was higher (p<0.0001) for lucerne (0.66) than rhodes grass (0.55), and was higher (p<0.05) in winter (0.60) than spring (0.58) or summer (0.57). The ruminal acetate concentrations were higher (p<0.001) in spring than summer (78 and 73 molar % respectively). Apparent digestibility of protein was significantly less (p<0.0001) for rhodes grass (0.37) than lucerne (0.75). N retention was positive for lucerne (15.2 g/d) but negative for rhodes grass (-2.8 g/d) (p<0.0001), and was higher (p<0.001) in summer (12.0 g/d) than spring (4.3 g/d) or winter (2.4 g/d). The tropical rusa deer exhibits seasonal variations in feed intake, food constituent digestibilities, VFA profile and N retention. Castration did not alter these traits. The results do not support the view that rusa deer can thrive on low-quality pastures. The productivity and commercial exploitation of rusa deer could be optimised if they are given high-protein feed during spring and summer.

Studies on the High Temperature Induced Stress on the Biochemical Profile and Fecundity of Daba and Laria Ecoraces of Tropical Tasar Silkworm Antheraea mylitta Drury (Lepidoptera: Saturniidae)

  • Lokesh, G.;Kar, P.K.;Srivastava, A.K.;Swaroopa, Saloni;Sinha, M.K.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.24 no.2
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    • pp.69-74
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    • 2012
  • Tropical tasar silkworm Antheraea mylitta Drury experiences extreme temperature stress conditions during its life cycle particularly during diapauses and first crop. The present study witnessed the impact of high temperature on some biochemical profiles and egg production (fecundity) of semi-domesticated Daba and Shorea robusta (Sal) based wild ecorace Laria during seed cocoon (pupa) preservation. Cocoons of Daba and Laria were treated with high temperature at $40^{\circ}C$ for 10 days in a BOD incubator. The protein profile and carbohydrate content in the hemolymph and fat body and total haemocyte count (THC) in the hemolymph of pupa were investigated. Further, the fecundity and fertility of egg was assessed. Significant increase in the protein concentration was observed in the hemolymph with reduction in the fat body (p<0.05). The difference in protein concentration was highly significant between the semidomesticated Daba and wild ecorace Laria (p<0.05). High pupal mortality (20%) and reduced fecundity (10-15%) in Daba was noticed compared to wild Laria. Also an increased THC (>28000) was recorded in Laria. The study infers the potentials of wild ecoraces in sustaining the extreme temperature conditions and need of adopting suitable package of practices for the preservation of diapause seed cocoons during extreme summer conditions. There is possibility to introgression thermal stress resistant traits in the semi-domesticated races of tasar silkworm by resorting to conventional breeding plans with wild races and keeping the thermal stress induced response as markers.

Effect of Gossypol from Cottonseed Meal Diets on Some Clinico-biochemical Parameters and Humoral Immune Response of Crossbred Calves Fed Barley or Sorghum

  • Pattanaik, A.K.;Sastry, V.R.B.;Singh, D.K.;Goswami, T.K.;Mohanty, D.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.9
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    • pp.1291-1296
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    • 2003
  • Effects of feeding of 9.95 mg free gossypol/kg live weight through cottonseed meal (CSM) were studied in 20 intact male calves fed barley or sorghum as source of cereal during the experimental duration of 210 days. Serum concentration of total protein, albumin, globulin and their ratio did not vary because of protein (gossypol) or cereal sources. Serum level of cholesterol and urea were lower (p<0.05) in sorghum than barley fed calves. Feeding of gossypol through CSM enhanced (p<0.05) serum cholesterol. An interaction between protein and period was observed with respect to serum concentrations of urea, creatinine and alanine transferase. The levels of serum creatinine and alanine transferase increased (p<0.05) following 120 days of experimental feeding in calves fed CSM diets compared to the control animals fed groundnut meal diets. No effect of feeding gossypol was, however, evident on the serum enzymes viz. alanine and aspartate transferases and alkaline phosphatase. Moreover, the source of cereal and protein did not appear to influence the metabolic profile of the calves. Humoral immune response, measured through antibody titre against Brucella abortus S99 innoculation, revealed a delayed and depressed seroreactivity indicative of immunocompromisation because of the phytotoxin gossypol. In conclusion, the feeding of gossypol at the designated levels, although had no deleterious clinico-biochemical manifestations, affected the humoral immune response of the calves.

Ecdysteroid Stimulates Virus Transmission in Larvae Infected with Bombyx mori Nucleopolyhedrovirus

  • Kang, Kyung-Don;Lee, Eun-Jung;Kamita, Shizuo George;Maeda, Susumu;Seong, Su-Il
    • BMB Reports
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    • v.33 no.1
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    • pp.63-68
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    • 2000
  • Most baculoviruses have an ecdysteroid UDP-glucosyltransferase (egt) gene, whose product inactivates ecdysteroid within the infected host. Bomhyx mori larvae infected with BmEGTZ, a mutant B. mori nucleopolyhedrovirus (BmNPV) in which the egt gene has been inactivated, die more rapidly compared to larvae infected with wild-type BmNPV. In this study, the profile of hemolymph proteins, and progression of virus infection in BmEGTZ- and BmNPV-infected B. mori larvae, was analyzed by SDS-PAGE and histochemically. These analyses showed that virus-encoded and virus-induced proteins were expressed quicker in BmEGTZ-infected larvae than in BmNPV-infected larvae. This suggests that the decrease in time to death, following BmEGTZ infection, results from the stimulation of virus-specific protein expression. In order to examine the effect of ecdysteroid on virus transmission, the profile of hemolymph proteins, and progression of virus infection, were analyzed following an ecdysteroid injection of BmEGTZ- or BmNPV-infected larvae. In the BmNPV-infected larvae, ecdysteroid treatment had no apparent effect on hemolymph protein expression. This suggests that the injected ecdysteroid was inactivated by the BmNPV-expressed ecdysteroid UDP-glucosyltransferase. An Ecdysteroid injection into BmEGTZ-infected larvae increased the speed of virus-specific protein expression and virus transmission. These results suggest that ecdysteroid stimulates protein expression, which in tum results in the stimulation of virus transmission.

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