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Expression of Recombinant Human Bone morphogenetic protein 2 (hBMP2) in Insect cells

  • Kim, Seong-Wan;Kim, Seong-Ryul;Park, Seung Won;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.34 no.1
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    • pp.1-5
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    • 2017
  • Bone morphogenetic protein 2 (BMP2) plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. It is involved also in cardiac cell differentiation and epithelial to mesenchymal transition. In this study, We expressed human BMP2 (hBMP2) recombinant protein using Baculovirus Expression Vector System (BEVS) in Sf9 insect cells. The hBMP2 cDNA was cloned into baculovirus transfer vector, pBacgus-4x-1 and recombinant baculovirus was screened out through X-gal and GUS-fusions assay. Western blot analysis shown that molecular weight of hBMP2 recombinant protein was about 44.71 kDa.

Effects of Mud Flat Bacteria Origin Protease Supplementation by Crude Protein Level on Growth Performance, Nutrient Digestibility, Total Protein and BUN Concentration in Broiler (조단백질 수준에 따른 갯벌 미생물 유래 단백질 분해 효소제의 급여가 육계의 생산성, 영양소 소화율 및 혈액 내 총 단백질과 혈중 요소태 질소 함량에 미치는 영향)

  • Kim, H.J.;Cho, J.H.;Chen, Y.J.;Yoo, J.S.;Min, B.J.;Jang, J.S.;Kang, K.R.;Kim, I.H.
    • Korean Journal of Poultry Science
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    • v.34 no.3
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    • pp.217-222
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    • 2007
  • This study was conducted to investigate the effects of mud flat bacteria origin protease supplementation by crude protein level on growth performance, nutrient digestibility, total protein and BUN (blood urea nitrogen) concentration in broilers. A total of four hundred eighty broilers were randomly allocated into four treatments with six replications for five weeks. Dietary treatments included 1) high crude protein diet, 2) high crude protein diet + 0.1% protease, 3) low crude protein diet and 4) low crude protein diet + 0.1% protease. During the entire experimental period, weight gain and feed/gain were improved in treatments of high crude protein diets and low crude protein diet added protease compared with treatment of low crude protein diet without protease (P<0.05). Similarly, DM digestibility was also improved in treatments of high crude protein diets and low crude protein diet added protease compared with treatment of low crude protein diet without protease (P<0.05). N digestibility was improved in treatment of high crude protein diet added protease compared with low crude protein diet without protease (P<0.05). Total protein concentration in blood was increased in treatment of high crude protein diet without protease compared with other treatments (P<0.05). In conclusion, mud flat origin protease was effective in improving weight gain, feed/gain and nutrient digestibility, and influenced blood total protein in broilers.

Oxidative Stress in Extrahepatic Tissues of Rats Co-Exposed to Aflatoxin B1 and Low Protein Diet

  • Rotimi, Oluwakemi A.;Rotimi, Solomon O.;Oluwafemi, Flora;Ademuyiwa, Oladipo;Balogun, Elizabeth A.
    • Toxicological Research
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    • v.34 no.3
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    • pp.211-220
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    • 2018
  • Early life exposure to aflatoxin B1 (AFB1) and low protein diet through complementary foods during weaning is common in parts of Africa and Asia. This study evaluated the effect of co-exposure to AFB1 and low protein diet on the extrahepatic tissues of rats. Twenty-four three-week old weanling male albino rats were used for this study and were randomly assigned into four groups: group 1 served as control and was fed normal protein diet (20% protein), group 2 was fed low protein diet (5% protein), group 3 was fed normal protein diet + 40 ppb AFB1 while group 4 received low protein diet + 40 ppb AFB1, all for eight weeks. Afterward, biomarkers of anemia (packed cell volume (PCV), hemoglobin) and kidney function (urea, uric acid, and creatinine) were determined in the blood while biomarkers of oxidative stress were determined in the tissues spectrophotometrically. Co-exposure to AFB1 and low protein diet significantly (p < 0.05) decreased body weight gain and PCV, increased biomarkers of kidney functions and induced oxidative stress in the tissues studied. There was significant (p < 0.05) reduction in glutathione concentration while TBARS was significantly increased in the tissues. Co-exposure to AFB1 and low protein diet had additive effects on decreasing the weight gain and potentiation effect of kidney dysfunction in the rats. The co-exposure also decreased antioxidant enzymes and increased oxidant status in the tissues. Our results demonstrate that this co-exposure has deleterious health effects on extrahepatic tissues and should be a public health concern especially in developing countries where AFB1 contamination is common.

Identification and Characterization of pH-Regulated Genes in Saccharomyces cerevisiae

  • Hong, Sung-Ki;Choi, Eui-Yul
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.327-333
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    • 1996
  • Yeast, like many other microbes, encounters large variations in ambient pH in their natural environments. Microorganisms capable of growing over a wide pH range require a versatile, efficient pH homeostatic mechanism protecting intracellular processes against extremes of pH. In several organisms, fusions to the bacterial lacZ gene have been extremely useful for the identification of genes expressed at different time during the life cycle or under different growth conditions. In this study, using the lacZ gene screening system, we surveyed a large number of yeast strains with lacZ insertion to identify genes regulated by pH. A yeast genomic library was constructed and inserted with lacZ by a shuttle mutagenesis procedure. The yeast transformants were individually picked up with a toothpick, replica-plated, and grown in alkaline pH medium. Among the 35,000 colonies screened, 10 candidate strains were identified initially by the $\beta$-gal assay. We finally confirmed two yeast strains carrying the genes whose expression are strictly dependent on pH of growth medium. One of the fusions showing a 10-fold induction in expression level in response to alkali pH was selected and further characterized. The pH-regulated gene was cloned by inverse PCR and a partial sequence of the gene was determined. Identification and characterization of the gene is currently under investigation.

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Nitrogen metabolism and mammary gland amino acid utilization in lactating dairy cows with different residual feed intake

  • Xie, Yunyi;Miao, Chao;Lu, Yi;Sun, Huizeng;Liu, Jianxin
    • Animal Bioscience
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    • v.34 no.10
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    • pp.1600-1606
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    • 2021
  • Objective: This study was conducted to enhance our understanding of nitrogen (N) metabolism and mammary amino acid (AA) utilization in lactating cows with divergent phenotypes of residual feed intake (RFI). Methods: Fifty-three multiparous mid-lactation Holstein dairy cows were selected for RFI measurements over a 50-d experimental period. The 26 cows with the most extreme RFI values were classified into the high RFI (n = 13) and low RFI (n = 13) groups, respectively, for analysis of N metabolism and AA utilization. Results: Compared with the high RFI cows, the low RFI animals had lower dry matter intake (p<0.01) with no difference observed in milk yield between the two groups (p>0.10). However, higher ratios of milk yield to dry matter intake (p<0.01) were found in the low RFI cows than in the high RFI cows. The low RFI cows had significant greater ratios of milk protein to metabolizable protein (p = 0.02) and milk protein to crude protein intake than the high RFI cows (p = 0.01). The arterial concentration and mammary uptake of essential AA (p<0.10), branched-chain AA (p<0.10), and total AA (p<0.10) tended to be lower in the low RFI cows. Additionally, the low RFI cows tended to have a lower ratio of AA uptake to milk output for essential AA (p = 0.08), branched-chain AA (p = 0.07) and total AA (p = 0.09) than the high RFI cows. Conclusion: In summary, both utilization of metabolizable protein for milk protein and mammary AA utilization are more efficient in cows with lower RFI than in the high RFI cows. Our results provide new insight into the protein metabolic processes (related to N and AA) involved in feed efficiency.

Responses of Labeo rohita to Dietary Lathyrus sativus Seeds

  • Barse, A.V.;Jadhao, S.B.;Sahu, N.P.;Srivastava, P.P.;Jain, K.K.;Pal, A.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.1
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    • pp.127-130
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    • 2004
  • Lathyrus sativus, locally known as Khesari, is a leguminous pulse crop grown in many parts of the world for food (used by poor people) and animal feed/fodder. Its seeds are rich in protein and energy but contains anti-nutritional factors prominent among which is ${\beta}$-N-Oxalylamino-L-Alanine (BOAA), a neurotoxin causing lathyrism in humans due to prolong consumption. Keeping in view the chemical characteristics of this toxin and literary facts on L. sativus feeding in terrestrial animals, it was hypothesized that aquatic species may better utilise this ingredient in mixed extruded diets. Diets were prepared with varying levels (0, 10, 18, 26 and 34%) of L. sativus seeds and fed for 60 days to study growth, body composition and digestibility of nutrients. Final body weight, specific growth rate and feed and protein conversion ratio did not differ (p>0.05) between treatments. Crude protein digestibility was reduced (p<0.01) beyond 26% inclusion level of L. sativus. Final carcass composition with regard to protein, lipid and ash did not differ (p>0.05) among the treatments. Comparable craniosomatic, viscerosomatic, renosomatic and hepatic indices and no mortality implies no apparent adverse effect on the vital organs and fish health. It was concluded that L. sativus can be a promising feed ingredient that can be used up to 34% or possibly higher level in fingerlings diet. To our knowledge, it appears to be the first report of its kind under laboratory conditions.

Stage-Specific Changes and Regulation of Endogenous Protein Phosphorylation in Allomyces macrogynus

  • Park, Young-Shik;Oh, Keun-Hee;Lee, Soo-Woong;Seong, Chang-Soo;Park, I-Ha;Yim, Jeong-Bin
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.374-378
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    • 1996
  • In the aquatic fungus Allomyces macrogynus the effects of $Ca^{2+}$ and cAMP on the intracellular signal transduction of zoospore germination were studied using in vitro protein phosphorylation assay system. An endogenously phosphorylated protein (p50) having molecular weight of 50 kDa on SDS-PAGE was found in soluble fractions of both zoospore and mycelium. In zoospore extract, the endogenous phosphorylation of p50 was weak without any effectors, but was enhanced by $Ca^{2+}$ and even more by cAMP. Phosphorylation of the same protein in mycelial extract was high only in the absence of cAMP. Irrespective of the presence of $Ca^{2+}$ and cAMP, its phosphorylation was antagonistically suppressed in assay of combined zoospore and mycelial extracts. These results suggest that p50 is interconvertible in phosphorylation/dephosphorylation as a novel protein involved in germination of A. macrogynus. The antagonistic effect of cAMP to the phosphorylation of p50s from different developmental stages may be important in the regulation of cellular differentiation.

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Changes of Plasminogen Activator Activity under Heat Stress Condition in Porcine Endometrium

  • Hwangbo, Yong;Cheong, Hee-Tae;Park, Choon-Keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.240-246
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    • 2019
  • The aim of this study was to investigate effect of heat stress on expression levels of plasminogen activators (PAs) related mRNAs and proteins, and changes of PAs activity in porcine endometrial explants. The endometrial explants (200 ± 50 mg) were isolated from middle part of uterine horn at follicular phase (Day 19-21) and were pre-incubated in serum-free culture medium at 38.5℃ in 5% CO2 for 18 h. Then, the tissues were transferred into fresh medium and were cultured at different temperature (38.5, 39.5, 40.5 or 41.5℃) for 24 h. The expression level of urokinase-type PA (uPA), type-1 PA inhibitor (PAI-1), type-2 PAI (PAI-2), and heat shock protein-90 (HSP-90) mRNA were analysis by reverse-transcription PCR and proteins were measured by western blotting. The supernatant were used for measurement of PAs activity. In results, mRNA and protein levels of HSP-90 was higher in 41.5℃ treatment groups than other treatment groups (p < 0.05). The expression of uPA, PAI-1, and PAI-2 mRNA were slightly increased by heat stress, however, there were no significant difference. Heat stress condition suppressed expression of active uPA and PAI-2 proteins (p < 0.05), whereas PAI-1 protein was increased (p < 0.01). Although PAI-1 protein was increased and active uPA was decreased, PAs activity was greatly enhanced by exposure of heat stress (p < 0.05). These results suggest that heat stress condition could change intrauterine microenvironment through regulation of PAs activity and other factors regarding with activation of PAs might be regulate by heat stress. Therefore, more studies regarding with regulatory mechanism of PAs activation are needed.

Effect of Oyster Shell Calcium Powder on the Quality of Restructured Pork Ham

  • Choi, Jung-Seok;Lee, Hyun-Jin;Jin, Sang-Keun;Lee, Hyun-Joo;Choi, Yang-Il
    • Food Science of Animal Resources
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    • v.34 no.3
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    • pp.372-377
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    • 2014
  • This study was conducted to evaluate the effects of oyster shell calcium powder (OSCP) as a substitute for phosphates in curing agent, on the quality of restructured pork ham. Restructured pork ham was processed under six treatment conditions: T1 (no additives), T2 (0.3% sodium tripolyphosphate), T3 (1.5% NaCl+0.5% whey protein), T4 (1.5% NaCl+0.5% whey protein+0.15% OSCP), T5 (1.5% NaCl+0.5% whey protein+0.3% OSCP), and T6 (1.5% NaCl+0.5% whey protein+0.5% OSCP). Addition of OSCP significantly increased the ash content and pH of restructured pork ham (p<0.05), but did not affect the cooking loss and water holding capacity values of restructured pork ham. Addition of OSCP had no effect on Hunter a and b surface color values of restructured pork ham, but did decrease the Hunter L surface color value (p<0.05). The addition of 0.5% OSCP showed significantly higher chewiness and springiness values of restructured pork ham, compared with the addition of phosphates (p<0.05). In conclusion, the addition of OSCP combined with low NaCl and 0.5% whey protein can be considered a viable substitute for phosphates in the curing agent, when processing restructured pork ham.

Himalayan dock (Rumex nepalensis): the flip side of obnoxious weed

  • Wangchuk, Kesang
    • Journal of Animal Science and Technology
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    • v.57 no.11
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    • pp.34.1-34.5
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    • 2015
  • Himalayan dock (Rumex nepalensis) was evaluated for forage value and antinutrients under three, five and seven weeks cutting intervals in the temperate environment. Dry matter (DM) content was measured for each cutting interval. Forage quality parameters such as Crude Protein (CP), Acid Detergent fiber (ADF), Neutral Detergent Fiber (NDF), Calcium (Ca) and Phosphorus (P) were analyzed. Plants with seven weeks cutting interval gave higher DM yield. CP and P content were significantly higher for three weeks cutting intervals. Average CP contents were 31.38 %, 30.73 % and 27.32 % and average P content 0.58 %, 0.52 % and 0.51 % for three, five and seven weeks cutting intervals, respectively. Ca content did not differ significantly between cutting intervals. The average Ca content were 0.91 %, 0.90 % and 90 %, for three, five and seven weeks cutting intervals, respectively. Tannin and mimosine contents were not significantly different between cutting intervals. Average tannin contents were 1.32 %, 1.27 % and 1.26 % and mimosine 0.38 %, 0.30 % and 0.28 % for three, five and seven weeks cutting intervals, respectively. The study concluded that R. nepalensis could be a potential source of protein for livestock. The study also suggests seven weeks harvesting interval to provide plants with high dry matter yield, high forage quality and very low levels of anti-nutrients.