• Title/Summary/Keyword: in vitro digestion.

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Characteristics of Hemolysin in Mosquitocidal Bacillus thuringiensis strain 21-2 (모기 살충성 Bacillus thuringiensis 21-2균주의 용혈성 내독소 단백질의 특성)

  • 김광현;김위종;김영희;김병우
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.230-234
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    • 2002
  • To describe characteristics of a hemolysin in mosquitocidal Bacillus thuringiensis subsp. gyangiensis strain 21-2, Escherichia coli HB101 was transformed with a gene encoding hemolysin in the strain 21-2. Transformant 47 con-tained 2.5 kb DNA was selected by ELISA, immunoblot and DNA electrophoresis. Transformant 47-5 was recon-structed after digestion of the 2.5 kb DNA with Hind m. Transformant 47-5 contained 1.8 kb DNA and expressed 23 kDa Protein which had mosquitocidal activity to Aedes aegypti. The 23 kDa Protein itself in vitro didn't show hemolytic activity on human erythrocytes, but the protein had the activity after proteinase K treatment.

Protective Effects of Gamipalmi-hwan on Elastase-induced Apoptosis of A549 Cells (가미팔미환(加味八味丸)의 elastase 유도성 A549 세포사멸에 대한 보호효과)

  • Oh, Ji-Seok;Park, Yang-Chun
    • The Journal of Korean Medicine
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    • v.31 no.2
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    • pp.137-148
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    • 2010
  • Objective: This study aimed to evaluate the protective effects of Gamipalmi-hwan (GPH) on elastase-induced lung cell injury. Materials and Methods: As an in vitro model of emphysema, the current study was performed to investigate potential activity of GPH in regulating injury responses of A549 human type II cell line mediated by elastase treatment. Results: GPH treatment increased the number of A549 cells which was reduced by elastase digestion. Elastin protein level, which was reduced by elastase treatment, was increased by GPH treatment. Labeling intensity with caspase 3 protein in elastase-treated cells was reduced by GPH treatment. Both Erk1/2 and Cdc2 protein levels, which were decreased by elastase treatment, were increased to a level similar to that of the normal cells. mRNA levels encoding IL-$1{\beta}$ and TNF-$\alpha$ were increased by elastase and then down-regulated by GPH. Conclusion: The present data suggest that A549 cells are subjected to inflammatory damage by elastase and can be recovered by GPH treatment. Further studies examining the protective activity of GPH in elastase-treated lung tissue would be useful for therapeutic strategies of emphysema treatment.

Estimation of Rumen By-pass Rate of Chromium-methionine Chelates by Ruminal Bacteria Analysis (반추미생물 분석에 의한 Chromium-methionine Chelate의 반추위 By-pass율 추정)

  • Kim, C.H.;Park, B.K.;Park, J.G.;Kim, H.S.;Sung, K.I.;Shin, J.S.;Ohh, S.J.
    • Journal of Animal Science and Technology
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    • v.47 no.5
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    • pp.759-768
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    • 2005
  • The study was designed to estimate the in vitro rumen by-pass rate of both chromium methionine chelate as an organic supplement and $ClCl_3$ as an inorganic supplement. Rumen by-pass rates of the supplements were evaluted by comparing ruminal metabolites in rumen fluid and Cr and methionine contents in the body of ruminal microorganism. For in vitro digestion examination, basic nutrients for ruminal microbes were supplied with 7g(DM) of feed, 2g of rice straw, and 2g of corn silage per each incubation jar. Three treatments including Control(no supplementation of Cr), T1(1000ppb supplementation of $ClCl_3$) and T2(chromium methionine chelate supplementation equivalent to 1000ppb of Cr content) were prepared with five replications per each treatment. pH of T2 was lower than that of Control and T1 regardless of incubation time. Ammonia content was higher in T2 than in Control and T1 during first 6 hours of incubation. However, the ammonia content in Control was remained low after 6 hours. Total volatile fatty acids(VFA) content in control was increased constantly as incubation time was extended. Therefore, VFA content in T1 and T2 were significantly lower (P<0.05) than those of Control. Dry matter recovery rate by ruminal microorganism was the lowest in T1, however ruminal microbial population was increased most efficiently in T2 during 12 hours of in vitro incubation. Cr concentrations in the body of ruminal microbes were not different(P>0.05) between Control and T2, but it was significantly high in T1(P<0.05). Contents of methionine and cystine in ruminal microbes also were not different between Control and T2(P>0.05), but it was relatively low in T1. Based on the above results, the chromium methionine chelate was believed to by-pass rumen and could remain intact until it reaches small intestine compared to inorganic chromium. This results implies that chromium methionine chelate could be more effective to function in the small intestine of ruminant animals.

Deoxyribonucleic Acid Was Responsible for the Anticoagulatory Effect of an Earthworm, Lumbricus rubellus

  • Paik, Seung-R.;Woo, Jeong-Im;Kim, Gyoung-Mi;Cho, Jin-Mo;Yu, Kyoung-Hee;Chang, Chung-Soon
    • BMB Reports
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    • v.30 no.1
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    • pp.37-40
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    • 1997
  • Earthworm extracts are known for anti-inflammatory, analgesic. antipyretic, and anticancer effects but can also influence blood circulation. It was previously shown that an earthworm, Lumbricus rubelius. contained a water-extractable anticoagulant which was a heat- and acid-stable molecule with hydrophilic property. In order to uncover the biochemical nature of this molecule, the anticoagulant was processed with various hydrolases such as trypsin, DNase, RNase. and lysozome. When the digested samples were analyzed with an in vitro coagulation test measuring activated partial thromboplastin time (APTT) and agarose gel electrophoresis, the anticoagulant proved to be a relatively homogeneous DNA fragment with relative molecular size around 72 base pairs. Interestingly, the activity was further stimulated with a trypsin digestion. RNA. on the other hand, did not prolong the APTT. It was also demonstrated that the DNA accelerated the antithrombin III (AT-III) inhibition of thrombin from $IC_{50}$ of 0.34 to 0.16 unit determined with S-2238 as a substrate, whereas heparin, a popular anticoagulant. shifted the value to 0.05. Therefore, it is suggested that the DNA could be considered as an alternative antithrombotic agent to heparin, which would exhibits bleeding side effects.

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Effects of Supplementation of Ruminally Protected Amino Acids on In vitro Ruminal Parameters and Milk Yield and Milk Composition of Dairy Cows in Mid-lactation (보호아미노산의 추가 공급이 반추위 발효성상 및 비유중기 착유우의 유량 및 유성분에 미치는 영향)

  • Lee, Jong-Min;Nam, In-Sik;Ahn, Jong-Ho
    • Journal of Animal Science and Technology
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    • v.50 no.2
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    • pp.199-208
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    • 2008
  • This study was undertaken to investigate the effects of ruminally protected amino acids (Methionine and Lysine) on in vitro ruminal parameters, and in vivo milk yield and milk composition in mid-lactating cows. In the first in vitro experiment, there were no statistical significances between treatments in ruminal pH and dry matter digestibility during various incubation times. In the second in vivo experiment, milk yield decreased by 11.92% in control and 5.68% in the treatment respectively, but decrease rate of milk yield in the treatment was lower than control. Milk yields naturally decreased as time goes by since the DIMs(Days in milk) of the cows in experiment were in mid-lactation period. 4% FCM(Fat corrected milk) and milk protein yields also, respectively, decreased by 11.25% and 11.09% in control and 6.16% and 5.47% in the treatment as compared with the intial. Milk protein and milk fat production were higher in the treatment(0.90kg, 1.10kg) than those of control(0.66kg, 0.79kg). Milk fat content significantly increased with supplementing protected amino acids as compared to control(P<0.05). From the above results, protected amino acids were positively utilized in the performances of mid-lactating cows without inhibiting rumen fermentation. Further investigation is suggested for essential amino acid composition and intestinal digestion rate out of rumen bypass protein in dietary protein to be estimated.

The importance of NIR spectroscopy in the estimation of nutritional quality of grains for ruminants

  • Flinn, Peter C.
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1612-1612
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    • 2001
  • The production of grain for export and domestic use is one of Australia's most important agricultural industries, and the NIR technique has been used extensively over many years for the routine monitoring of grain quality, particularly moisture and protein content. Because most Australian grain is intended for human food production, the determinants of grain quality for livestock feed, apart from protein, have been largely ignored. However the increasing use of grain for feeding to pigs, poultry, beef cattle and dairy cows has led to an important national research project entitled “Premium Grains for Livestock”. Two of the objectives of this project are to determine the compositional and functional characteristics of grains which influence their nutritional quality for the various classes of livestock, and to adopt rapid and objective analytical tests for these quality criteria. NIR has been used in this project firstly to identify a set of grain samples from a large population of breeders' lines which showed a wide spectral variation, and hence a potentially wide variation in nutritional value. The selected samples were not only subjected to an extensive array of chemical, physical and in vitro analyses, but also were grown out to produce sufficient quantities of grain to feed to animals in vivo studies. Additional grains were also strategically selected from farms in order to include the effect of weather damage, such as rain, drought and frost. In this study to date, NIR calibrations have been derived or attempted, on both ground and whole grains, for in vivo dry matter digestibility (DMD), pepsin-cellulase dry matter disappearance, protein, fat, acid detergent fibre, neutral detergent fibre, starch, in sacco DMD and in vitro assays to simulate starch digestion in the lumen and small intestine. Results so far indicate high calibration accuracy for chemical components (SECV 0.3 to 2.6%) and very promising statistics for in vivo DMD (SECV 1.8, $R^2$ 0.93, SD 7.0, range 61.9 to 92.3, n=60). There appears to be some potential for NIR to estimate some in vitro properties, depending upon the accuracy of reference methods and appropriate sample populations. Current work is in progress to extend the range of grains with in vivo DMD values (a very laborious and expensive process) and to increase the robustness of the various NIR calibrations, with the aim of implementing uniform testing procedures for nutritional value of grains throughout Australia.

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The Effects of Processing Methods of Corn on In vitro DM Digestability and In sacco Degradability in Rumen (옥수수 가공방법이 In vitro 건물소화율 및 In sacco 건물 분해율에 미치는 영향)

  • Son, K.N.;Kim, Y.K. Kim;Lee, S.K.;Kim, H.S.
    • Journal of Animal Science and Technology
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    • v.45 no.3
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    • pp.433-442
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    • 2003
  • The objective of these studies were to examine the effects of processing methods of corn grains on in vitro dry matter digestability and in sacco degradability in the rumen by three ruminally cannulated dry Holstein cows. The corns for these experiments were untreated; whole corn L(density; 660 g/$\ell$), whole corn H(density; 740 g/$\ell$), and treated by four different types: Ground corn, 3.8 mm, 2.8 mm, and 1.5 mm flaked corns. The results obtained were summarized as follows: The DM degradabilities, after 48 hr incubation by in sacco method, were the highest(94.4, 88.0 and 87.0%, respectively) in 1.5 mm flaked corn, ground corn, and 2.8 mm flaked corn. The 3.8 mm flaked corn was degraded significantly lower than these. Until 12 hr incubation, whole corn L tended to be degraded little more than whole corn H, was not significantly different. However, after 24 hr incubation, the significant differences between whole corn L and whole corn H were shown(P<0.05). The DM digestabilities by in vitro digestion were the highest for 1.5 mm flaked corn and ground corn(92.3 and 91.2%, respectively)(P<0.05), followed by 2.8 mm and 3.8 mm flaked corn(83.9 and 83.4%, respectively), tended to be similar to those by in sacco method. Whole corn L was digested twice more rapidly than whole corn H. Summarizing the experimental data, compared with unprocessed corns, the flaked corns were significantly increased in the degradabilities of dry matter in the rumen. In addition, as increasing the flaking degree of corn, the degradabilities of dry matter were significantly improved. Referring to these kinds of physical characteristics of grain sources in the ruminal degradabilities, it is believed to be possible to optimize the environment of the fermentation in the rumen.

Evaluation of Forage Production and Feed Value of Sasa borealis in the Jeju Area (제주지역 조릿대의 사초생산성 및 사료가치평가)

  • Chung, Sang Uk;Seong, Hye Jin;Yun, Yeong Sik;Lee, Ga Eul;Oh, Young Kyoon;Baek, Youl Chang;Lee, Seul;Moon, Sang Ho
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.38 no.2
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    • pp.135-139
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    • 2018
  • This study was conducted to evaluate the forage production and feed value of Sasa borealis (S. borealis) in Jeju Island in order to improve the utilization of Sasa borealis and to help mitigate the problem of reduced plant species diversity caused by S. borealis in Hanlla Mountain. To investigate the forage production, three quadrat structures were installed in the S. borealis natural community in the middle part of Hanlla Mountain. From May to October 2017, S. borealis in quadrats was cut at a fixed time of each month, and then forage production and regenerated acidity per kg/ha were evaluated. For the evaluation of feed value, compositional analysis was performed on the monthly samples. In vitro digestion experiments were carried out using cannula mounted Hanwoo. In vitro neutral detergent fiber digestibility(IVNDFD) and in vitro acid detergent fiber digestibility(IVADFD) were measured after the experiment. Forage production of S. borealis showed relatively good regeneration ability in May and June, but the regeneration ability decreased as the cutting was repeated. In order to use S. borealis as a forage, it is considered efficient to feed black goats with good fiber decomposition or horses good palatability to S. borealis and relatively good digestibility.

Anthocyanin Stability and Silage Fermentation Quality of Colored Barley

  • Song, Tae Hwa;Han, Ouk kyu;Park, Tae Il;Kim, Dae Wook;Yoon, Chang;Kim, Kee Jong;Park, Ki Hun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.32 no.4
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    • pp.335-342
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    • 2012
  • This study was conducted to observe the fermentative quality and anthocyanin content in whole crop colored barley silage during storage periods and anthocyanin stability in in vitro ruminal fluid. Silages of colored barley cultivar "Boanchalbori" and normal barley cultivar "Yuyeonbori" were stored during 0, 2, 4, 6, and 12 months. The in vitro ruminal fluid was fermented for 0, 6, 12, 24, and 48 hrs. For the feed value, crude protein of colored barley silage was slightly increased in the silage compared to that of normal barley silage, and being increased up to 2 months after ensiling and thereafter maintained at the similar level. Neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents of both the barley significantly increased by prolonged storage of 2 months (p<0.05), but they were maintained at the constant level after 2 months of storing silage. Whereas TDN (total digestible nutrients) contents of them were decreased by the prolonged storage of 2 months (p<0.05), then maintained at the constant levels. The fermentative quality and pH values in both the barley silages were slightly decreased during the storage time. Lactic acid and acetic acid contents were increased during prolonged storage period, but not significantly different among treatments. Butyric acid was not detected. In the colored barley silage, pH value showed slightly lower compared to that of the normal barley silage but not significant, and lactic acid content was significantly higher than the normal barley silage (p<0.05). The total anthocyanin content in the whole crop colored barley silage decreased to 42% after 2 months of ensilage, however maintained at the constant level until 12 months of ensilage. In the case of anthocyanin stability on in vitro ruminal fluid digestion, the pH value of the ruminal fluid was slightly lower at 6, 12, 24, 48h incubation time and the content of anthocyanin was at similar levels. These results indicated that the colored barley showed higher fermentation quality, and total anthocyanin content was maintained stable at 42% level of the first value in storing silage. As the anthocyanin had higher stability in the ruminal fluid, the colored barley has a potential as functional feeds for Ruminants.

Changes in Nutritive Value and Digestion Kinetics of Canola Seed Due to Microwave Irradiation

  • Ebrahimi, S.R.;Nikkhah, A.;Sadeghi, A.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.3
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    • pp.347-354
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    • 2010
  • This study aimed to evaluate effects of 800 W microwave irradiation for 2, 4 and 6 min on chemical composition, antinutritional factors, ruminal dry matter (DM) and crude protein (CP) degradability, and in vitro CP digestibility of canola seed (CS). Nylon bags of untreated or irradiated CS were suspended in the rumen of three bulls from 0 to 48 h. Protein subfractions of untreated and microwave irradiated CS before and after incubation in the rumen were monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Microwave irradiation had no effect on chemical composition of CS (p>0.05). There was a linear decrease (p<0.001) in the phytic acid and glucosinolate contents of CS as irradiation time increased. Microwave irradiation for 2, 4 and 6 min decreased the phytic acid content of CS by 8.2, 27.6 and 48.6%, respectively. The total glucosinolate contents of CS microwave irradiated for 2, 4 and 6 min decreased by 41.5, 54.7 and 59.0% respectively, compared to untreated samples. The washout fractions of DM and CP and degradation rate of the b fraction of CP decreased linearly (p<0.001) as irradiation time increased. Microwave irradiation for 2, 4 and 6 min decreased effective degradability (ED) of CP at a ruminal outflow rate of 0.05 $h^{-1}$ by 4.7, 12.3 and 21.0%, respectively. Microwave irradiation increased linearly (p<0.001) in vitro CP digestibility of ruminally undegraded CS collected after 16 h incubation. Electrophoresis results showed that napin subunits of untreated CS disappeared completely within the zero incubation period, whereas cruciferin subunits were degraded in the middle of the incubation period (16 h incubation period). In 4 and 6 min microwave irradiated CS, napin subunits were degraded after 4 and 16 h incubation periods, respectively, and cruciferin subunits were not degraded untile 24 h of incubation. In conclusion, it seems that microwave irradiation not only protected CP of CS from ruminal degradation, but also increased in vitro digestibility of CP. Moreover, microwave irradiation was effective in reducing glucosinolate and phytic acid contents of CS.