• 제목/요약/키워드: Protein profile

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Proteome Analysis of Recombinant CHO Cells Under Hyperosmotic Stress

  • 이문수;김경욱;김영환;이균민
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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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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아세트아미노펜에 의해 간손상이 유발된 랫드의 유전자 발현 분석 (Gene Expression Analysis of Acetaminophen-induced Liver Toxicity in Rat)

  • 정희경
    • Toxicological Research
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    • 제22권4호
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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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    • 제19권10호
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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.

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

  • 권오윤;김미연;손찬욱;류희문;김형진;윤원기;김환묵;김미리
    • 한국식품영양과학회지
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    • 제37권1호
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    • pp.117-123
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    • 2008
  • 국내에서 생산된 감자 중 세풍, 남서, 수미, 조풍 및 대서의 5가지 품종에 대하여 단백질 profile 및 아미노산 조성을 분석하였다. 총 질소함량은 $1.27{\sim}1.64%$이었으며, 남서가 높았고 수미가 낮게 나타났다. 아미노산 조성은 품종 간에 유의적인 차이가 있었다. 한편, 주요 감자 단백질은 papatin(40 kDa), trypsin inhibitor(20 kDa) 및 protease inhibitor(15 kDa)이었으며, 이들의 함량은 각각 $22.16{\sim}25.81%$, $25.22{\sim}20.91%$$14.12{\sim}25.23%$이었다. Papatin 함량은 조풍, 세풍, 수미감자가 높은 함량을 보인 반면, trypsin inhibitor는 조풍감자가 5.22%로 가장 낮은 함량을 보였다. Protease inhibitors인 20 kDa와 15 kDa를 합한 값은 $24.7{\sim}35.0%$이었으며, 세풍이 가장 적었고 조풍에 가장 많이 함유되어 있었다.

Control of Encapsulation Efficiency and Initial Burst in Polymeric Microparticle Systems

  • Yeo, Yeon;Park, Ki-Nam
    • Archives of Pharmacal Research
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    • 제27권1호
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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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    • 제4권2호
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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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    • 제17권8호
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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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    • 제24권2호
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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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    • 제16권9호
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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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    • 제33권1호
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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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