• Title/Summary/Keyword: Trypsin Digestion

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Effects of Heat-oxidized Soy Protein Isolate on Growth Performance and Digestive Function of Broiler Chickens at Early Age

  • Chen, X.;Chen, Y.P.;Wu, D.W.;Wen, C.;Zhou, Y.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.4
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    • pp.544-550
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    • 2015
  • This study was conducted to investigate effects of heat-oxidized soy protein isolate (HSPI) on growth performance, serum biochemical indices, apparent nutrient digestibility and digestive function of broiler chickens. A total of 320 1-day-old Arbor Acres chicks were randomly divided into 4 groups with 8 replicates of 10 birds, receiving diets containing soy protein isolate (SPI, control group) or the same amount of SPI heated in an oven at $100^{\circ}C$ for 1, 4, or 8 h, for 21 days, respectively. The results indicated that compared with the control group, body weight gain and feed intake of birds fed diet containing SPI heated for 8 h were significantly lower (p<0.05). Serum urea nitrogen concentration was higher in the broilers fed diet containing SPI heated for 4 or 8 h at d 21 (p<0.05). In contrast, serum glucose content was decreased by HSPI substitution at d 21 (p<0.05). The relative pancreas weight in HSPI groups was higher than that in the control group at d 21 (p<0.05). Meanwhile, the opposite effect was observed for relative weight of anterior intestine and ileum in broilers fed a diet containing SPI heated for 8 h (p<0.05). Birds fed diets containing SPI heated for 4 or 8 h had a decreased lipase activity in anterior intestinal content at d 14 and 21 (p<0.05), respectively. In addition, the same effect was also noted in broilers given diets containing SPI heated for 1 h at d 21 (p<0.05). Similarly, amylase, protease and trypsin activity in anterior intestinal content were lower in broilers fed diets containing SPI heated for 8 h at d 21 (p<0.05). The apparent digestibility of dry matter (DM) from d 8 to 10 and DM, crude protein (CP), and ether extract from d 15 to 17 were lower in broilers fed diets containing SPI heated for 8 h (p<0.05). Besides, birds given diets containing SPI heated for 4 h also exhibited lower CP apparent digestibility from d 15 to 17 (p<0.05). It was concluded that HSPI inclusion can exert a negative influence on the growth performance of broilers, which was likely to result from the simultaneously compromised digestive function.

Expression of receptors of Vitamin D and cytokines in osteoclasts differentiated by M-CSF and ODF (Macrophage Colony-Stimulating Factor와 Osteoclast Differentiation Factor로 분화 유도된 생쥐 파골세포에서 Vitamin D 및 수종의 싸이토카인 수용체의 발현)

  • Seong, Soo-Mi;Um, Heung-Sik;Ko, Sung-Hee;Woo, Kyung-Mi;Chang, Beom-Seok
    • Journal of Periodontal and Implant Science
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    • v.32 no.4
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    • pp.865-873
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    • 2002
  • The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.

Generation of Transgenic Plant (Nicotiana tabacum var. Petit Havana SR1) harboring Bacillus thuringiensis Insecticidal Crystal Protein Gene, cry II A (Bacillus thuringiensis 살충성 결정단백질 유전자(cry II A)의 형질전환 식물 제작)

  • 이정민;류종석;권무식
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.305-311
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    • 1997
  • Bacillus thuringiensis, a gram-positive soil bacterium, is characterized by its ability to produce crystalline inclusions during sporulation. The crystal proteins exhibit a highly specific insecticidal activity. An insecticidal crystal protein (ICP), Cry II A, is specifically toxic to both lepidopteran and dipteran insects. In this study, tobacco plants transformed by the cry II A gene have been generated. The Cry II A crystal protein was purified from E. coli JM103 harboring cry II A gene by differential solubility. The activated Cry II A was prepared by tryptic digestion. The purified protoxin (70 kDa) and the activated toxin (50 kDa) were analyzed by SDS-PAGE. To generate the transgenic tobacco having cry II A gene, the cry II A gene was subcloned to a plant expression vector, pSRL2, having two CaMV 35S promoters. The recombinant plasmid was transformed into tobacco (N. tabacum var. Petit Havana SR1) by Agrobacterium-mediated leaf disc transformation. Through the regeneration, six putative transgenic tobacco plants were obtained and three transformants were confirmed by Southern blot analysis. It has been found that one plant had single copy of cry II A gene, another had two copies of the gene, and the third had a truncated gene. After the immunochemical confirmation of cry II A expression in plants, the transgenic tobacco plants will be used to study the genetics of future generation with the insecticidal crystal protein gene cry II A.

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Antimicrobial Activity of Lactobacillus reuteri Against Major Food-Borne Pathogens (Lactobacillus reuteri의 주요 식품 위해 미생물에 대한 항균 효과)

  • Kwon, Nam-Hoon;Kim, So-Hyun;Bae, W.K.;Kim, J.Y.;Lim, J.Y.;Noh, K.M.;Kim, J.M.;Ahn, J.S.;Hur, J.;Park, Y.H.
    • Journal of Food Hygiene and Safety
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    • v.16 no.4
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    • pp.264-273
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    • 2001
  • Antimicrobial activities of ave different probiotics (Lactobacillus reuteri, L. acidophillus, L. bulgaricus, L. casei and Bifidobacterium longum) against 8 bacterial pathogens were determined on the Mueller Hinton Agar containing supernatant of probiotics obtained from 3 different growth conditions (MRS without glycerol, MRS with 0.5 M glycerol or 0.25 M glycerol solution). Though antimicrobial activity of L. reuteri in the first two conditions was not better than the others', the activity was significantly higher than that of others in 0.25 M glycerol solution. This prominent effect might be attributable to reuterin, produced by L. reuteri using glycerol. We could detect the presence of reuterin in the supernatant of 0.25 M glycerol solution with 500 MHz Nuclear Magnetic Resonance (NMR). The result of minimum bactericidal concentration (dilution fold) has revealed that reuterin showed pan-bactericidal effects against 8 major food-borne pathogens. To examine any changes of antimicrobial activities of the probiotics, the probiotics were treated with different pH conditions, pepsin or trypsin digestion. Antimicrobial activity of reuterin was not entirely affected by any of these treatments, while the activities of the other probiotics were significantly decreased.

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Effect of Environmental Factors on the Growth of Rabbit Oral Keratinocytes (토끼 구강점막 상피세포 성장에 미치는 환경인자의 영향)

  • Yoon, Moon-Young;Park, Hee-Jung;Lee, Doo-Hoon;Jang, In-Keun;Park, Jung-Keug;Kim, Woo-Sik
    • Korean Chemical Engineering Research
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    • v.43 no.1
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    • pp.103-109
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    • 2005
  • Isolation and primary culture technique of rabbit oral keratinocytes, and the study for effect of environmental factors on the cell growth were carried out in T75-flask. $1.92{\pm}0.59{\times}10^6$ viable cells were isolated by trypsin enzymatic digestion method from $0.25cm^2$ biopsy of rabbit oral mucosa. Primary culture with 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 0.15 mM $Ca^{2+}$ showed confluence after 8 days and doubling time was 2.54 days. Effect of medium types, medium volume and supplement types on the cell growth was investigated after the cultured keratinocytes had been harvested from primary confluence. Serum addition showed adverse effect and the increase of serum concentration didn't have an effect on the cell growth. The increase of medium volume decreased the cell growth. The increase of calcium concentration increased the cell growth and 2.0 mM was optimum value. In conclusion, when rabbit oral keratinocytes was cultured in T75-flask, the most effective conditions was to use 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 2.0 mM $Ca^{2+}$, and doubling time was 1.32 days. This study can provide the useful informations to develop a process and design a bioreactor for the culture of keratinocytes in human body like skin and cornea, as well as mucosa.