• Title/Summary/Keyword: In vitro Technique

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Effect of Crude Protein Levels in Concentrate and Concentrate Levels in Diet on In vitro Fermentation

  • Dung, Dinh Van;Shang, Weiwei;Yao, Wen
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.6
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    • pp.797-805
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    • 2014
  • The effect of concentrate mixtures with crude protein (CP) levels 10%, 13%, 16%, and 19% and diets with roughage to concentrate ratios 80:20, 60:40, 40:60, and 20:80 (w/w) were determined on dry matter (DM) and organic matter (OM) digestibility, and fermentation metabolites using an in vitro fermentation technique. In vitro fermented attributes were measured after 4, 24, and 48 h of incubation respectively. The digestibility of DM and OM, and total volatile fatty acid (VFA) increased whereas pH decreased with the increased amount of concentrate in the diet (p<0.001), however CP levels of concentrate did not have any influence on these attributes. Gas production reduced with increased CP levels, while it increased with increasing concentrate levels. Ammonia nitrogen ($NH_3$-N) concentration and microbial CP production increased significantly (p<0.05) by increasing CP levels and with increasing concentrate levels in diet as well, however, no significant difference was found between 16% and 19% CP levels. Therefore, 16% CP in concentrate and increasing proportion of concentrate up to 80% in diet all had improved digestibility of DM and organic matter, and higher microbial protein production, with improved fermentation characteristics.

In-vitro Study on Hemorheological Behaviors of Blood Flow Through a Micro Tube (미세튜브 내부를 흐르는 혈액유동의 유변학적 특성에 대한 in-vitro 연구)

  • Kang, Myung-Jin;Ji, Ho-Seong
    • Journal of Biomedical Engineering Research
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    • v.31 no.2
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    • pp.99-105
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    • 2010
  • In order to obtain velocity profile of blood flow with high spatial resolution, a micro PIV technique consisted of a fluorescent microscope, double-pulsed YAG laser, cooled CCD camera was applied to in-vitro blood flow experiment through a micro round tube of a diameter $100{\mu}m$. Velocity distributions of blood flow for rabbit were obtained. The viscosity profiles for shear rate were found at flowing condition. To provide hemorheological characteristics of blood flow, the viscosities for shear rate were evaluated. The viscosity of blood also steeply increase by decreasing shear rate resulting in Non-Newtonian flow, especially in low shear rate region caused by RBC rheological properties. The results show typical characteristics of Non-Newtonian characteristics from the results of velocity profile and viscosity for blood flow. From the inflection points, cell free layer and two-phase flow consisted with plasma and suspensions including RBCs can be separated.

Effect of Heat Treatment on the In Vitro Protein Digestibility and Trypsin Indigestible Substrate (TIS) Contents in Some Seafoods (수산단백질(水産蛋白質) 소화화(消化華)에 미치는 가열처리(加熱處理)의 영향(影響))

  • Ryu, Hong-Soo;Lee, Kang-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.1
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    • pp.1-12
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    • 1985
  • In an attempt todetermine the optimum heat treatment, the changes in TIS content and in vitro protein digestibility of squid, shrimp, oysterand pollock under various heating conditions were studied. The effect of drying method and cold storage on the in vitro digestibility and TIS content were also studied. Optimal boiling conditions were 1 min, for squid, 0.5min. for oyster(eviscerated), 1 min. for whole oyster, and 5 min. for pollock. Steaming times that yieled products with the highest in vitro digestibility value were: 1 min. at $100^{\circ}C$ for squid, 1 min, at $88^{\circ}C$ for oyster and $1{\sim}2.5min$. at $100^{\circ}C$ for pollock. All of freeze dried samples showed the highest in vitro digestibility value and sundried one were comparble to freeze dried samples except high fat level or noneviscerated samples. Fat content was the nain inhivbitory factor of the seafood enzymic digestion during processing and storage. The multi-enzyme assay, used to predict the quality change of dried seafoods stored in a cold room for long periods of raw seafoods treated with various heating methods, offers many advantages over the convetional methods of determining protein quality.

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Diets with Different Forage/Concentrate Ratios for the Mediterranean Italian Buffalo: In vivo and In vitro Digestibility

  • Fabio, Zicarelli;Calabro, Serena;Piccolo, Vincenzo;D'Urso, Simona;Tudisco, Raffaella;Bovera, Fulvia;Cutrignelli, Monica I.;Infascelli, Federico
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.1
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    • pp.75-82
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    • 2008
  • In vivo and in vitro digestibility of 6 diets with a forage to concentrate ratio (F/C) ranging from 100 to 50:50 (diet 1: all hay, diet 2: 90:10, diet 3: 80:20, diet 4: 70:30, diet 5: 60:40, diet 6: 50:50) were investigated using 6 buffaloes in a $6{\times}6$ Latin square design. For the in vivo trial, the individual faeces of buffaloes were collected 3 times per day for 7 days. Individual pooled faeces and samples of each diet were analysed for chemical composition and insoluble acid ash (AIA) contents in order to estimate the coefficient of apparent digestibility (ADC). On the last day of the in vivo trial a sample of faeces was collected from each animal and used as inoculum for the in vitro test, using the gas production technique (IVGPT). The in vivo organic matter digestibility (ADC) rose as the percentage of concentrate increased up to the 70:30 (F/C) diet (67.01, 73.03, 78.06 and 79.05, respectively for diets 1, 2, 3 and 4); the other two diets (60:40 and 50:50 F/C) unexpectedly did not follow this trend (75.11 and 79.06, respectively for diet 5 and 6). However, these data agree with the results of the in vitro trial. The ADC was positively correlated with the dOM (p<0.001), but not with the gas production at different times; cumulative gas production recorded at the end of incubation (OMCV) showed an irregular trend and was not closely correlated to degraded OM. Estimation of in vivo digestibility from in vitro fermentation data was acceptable, despite leaving room for improvement.

Predicting the Nutritional Value of Seafood Proteins as Measured by Newer In Vitro Model -1. C-PER and DC-PER of Shellfish Proteins- (수산식품단백질 품질평가를 위한 새로운 모델 설정 -1. 패류의 C-PER 및 DC-PER-)

  • Ryu, Hong-Soo;Lee, Kang-Ho;Kim, Jang-Yang;Choi, Byeong-Dae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.3
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    • pp.265-273
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    • 1985
  • To predict the nutritional quality of seafood proteins using a newer in virto model, 10 species of shellfish protein samples were used in determining the extent of in vitro digestibility, trypsin indigestible substrate (TIS), computed protein efficiency ratio (C-PER), discriminant computed protein efficiency ratio (DC-PER) and predicted digestibility which calculated solely from amino acid profile. The content of TIS in eviscerated samples were ranged from 1.10 to 5.09 mg/g solid, whereas the whole samples were ranged from 1.26 to 7.30 mg/g solid expressed quantitatively as mg of soybean trypsin inhibitor. The in vitro digestibility showed $82{\sim}86%$ for eviscerated samples in contrast with $78{\sim}84%$ for whole ones. Therefore, the results suggested that in vitro digestibility of shellfish was influenced by the present of viscera. The lysine content of Mya arenaria, Saxidomus purpuratus, Anadara subcrenata, and Anadara broughronii were lower than that of ANRC casein, but Corbicula fluminea, Cyclina sinensis, and eviscerated Mytilus edulis, were showed the value about 10.0 g/16g N. In all samples, the content of tryptophan and cystein were more higher than those of ANRC casein. The C-PER of whole samples showed the value below 2.0 while the values above 2.5 noted in the eviscerated samples. DC-PER of most samples were greater than those of C-PER and a greater discrepancies were revealed in whole shellfish which possesses the lower in vitro digestibility. The shellfish sample showed a high in vitro digestibility and a low TIS content such as eviscerated ones may need the DC-PER and predicted digestibility procedures rather than C-PER and four-enzyme in vitro digestibility procedure could offer more advantages in predicting the protein quality of whole shellfish samples which have poor in vitro digestibility and high TIS content.

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In Vitro Transcription Analyses of Autographa californica Nuclear Polyhedrosis Virus Genes

  • Huh, Nam-Eung
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.183-190
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    • 1994
  • Cell-free extracts prepared from cultured insect cells, Spodoptera. frugiperda, were analyzed for activation of early gene transcription of an insect baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV). The template DNA used for in vitro transcription assays contained promoter sites for the baculovirus genes that have been classified as immediate early ($\alpha$) or early genes. These genes are located in the HindIII-K/Q region of the AcNPV genome. Nuclei isolated from the AcNPV-infected Spodoptera frugiperda cells were also used for in vitro transcription analysis by RNase-mapping the labeled RNA synthesized from in vitro run-on reaction in the isolated nuclei. The genes studied by this technique were p26 and pl0 genes which were classified as delayed early and late gene, respectively. We found that transcription of the genes from the HindIII-K region was accurately initiated and unique in the whole cell extract obtained from uninfected cells, although abundance of the in vitro transcripts was reverse to that of in vivo RNA. With isolated nuclei transcription of the p26 gene was inhibited by $\alpha$-amanitin suggesting that the p26 gene was transcribed by host RNA polymerase II. However, transcription of the pl0 gene in isolated nuclei was not inhibited by $\alpha$-amanitin, but rather stimulated by the inhibitor. We also found that the synthesis of $\alpha$-amanitin-resistant RNA polymerase was begun before 6 hr p.i., the time point at which the onset of viral DNA replication as well as the appearance of a-amanitin-resistant viral transcripts were detected. These studies give us strong evidence to support the previous data that early genes of AcNPV were transcribed by host RNA polymerease III, while transcription of late genes was mediated at least by a novel $\alpha$-amanitin-resistant RNA polymerase.

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Improved Detection and Purification of Grapevine Leafroll-associated 3 Closterovirus Using Tissue Culture (포도 조직배양에 의한 Grapevine Leafroll-associated 3 Closterovirus의 증식과 검출효율 증대)

  • 김현란;정재동;정봉남;이봉춘;박진우;최용문
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.335-339
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    • 2001
  • Grapevine leafroll-associated 3 closterovirus (GLRaV-3) is phloem limited virus, and one of the most severe viral diseases found in Korea. However, nonhomogenous distribution and low concentration and seasonal variations of GLRaV-3 in grapevines remain as main problems which prevent the introduction and molecular biology or serology experiments. Virus-infected plantlet in vitro was obtained from node tissue cuttings, which was GLRaV-3 infected 'kyoho' vines. The amount of purified virus was highest in vitro plantlet. Moreover, viruses seem to be relatively homogeneously distributed in all organs including leaf, stem and callus derived from in vitro plantlets. RT-PCR detection using in vitro plantlet tissue as template was most effective. When comparing ELISA to RT-PCR, RT-PCR detection was 1,000 times as effective as ELISA. These results can be explained by improved quality such as tenderness or less tannins in plantlet in vitro. In conclusion, until infected herbaceous host will be available, tissue culture can be usefully adopted as a technique for a good source of GLRaV-3 closterovirus for further studies.

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Antibacterial potential of the extracts derived from leaves and in vitro raised calli of medicinal plants Pterocarpus marsupium Roxb., Clitoria ternatea L., and Sanseveiria cylindrica Bojer ex Hook

  • Shahid, M;Shahzad, A;Anis, M
    • Advances in Traditional Medicine
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    • v.9 no.2
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    • pp.174-181
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    • 2009
  • Pterocarpus marsupium, Clitoria ternatea, and Sanseveiria cylindrica are some of the important and endangered medicinal plant species of India. Despite of medicinal properties, antibacterial potential of the plants have not yet been explored. The present study was designed to optimize the in vitro technique for micropropagation and to screen the extracts from leaves and in vitro raised calli for antibacterial properties. Excised leaf-explants from the parent plants were surface sterilized and cultivated on Murashige & Skoog's (MS) medium containing $N^6$-benzyladenine (BA) in concentrations of 1, 2, 5, and $10{\mu}M$. Optimal growth of calli was noticed at a concentration of $5{\mu}M$, therefore the extracts from calli grown at this concentration were further studied for antibacterial activity. Both alcoholic and aqueous extracts from leaves of respective plants, and their in vitro raised calli were tested for antibacterial activity by agar well diffusion method against a range of Gram-positive and Gram-negative bacteria. Aqueous extracts showed antibacterial activity against limited number of bacterial species; notably the extracts of C. ternatea which showed antibacterial activity against Streptococcus pyogenes, Bacillus subtilis and Bacillus cereus. Alcoholic extracts of all three plants showed antibacterial activity against a wider range of bacteria. Among the Gram-positive bacteria, extracts from C. ternatea showed strong antibacterial activity against Bacillus spp., whereas the extracts of S. cylindrica showed good antibacterial potential for Staphylococcus aureus, S. epidermidis and S. pyogenes. The extracts from all three plants showed antibacterial activity against Gram-negative bacteria, including, Salmonella spp. and Shigella dysenteriae; organisms causing enteric fever and dysentery. In most of the cases, the extracts from respective calli showed comparable, and in some cases better, result in comparison to the extracts from parent leaves. To the best of our knowledge this is the first preliminary report on antibacterial potential, especially through calli extracts, of these plants; and in vitro cultivation of the explants may be used to obtain phytotherapeutic compounds.

A Study of OSIRIS Technique in in vivo Localized NMR Spectroscopy (생체내 체적선택 자기공명분광법 OSIRIS 기법의 연구)

  • Yi, Y.;Lim, T.H.;Mun, C.W.;Lee, T.K.;Oh, C.H.
    • Proceedings of the KOSOMBE Conference
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    • v.1993 no.11
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    • pp.63-66
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    • 1993
  • Localized NMR spectra were obtained from phantom by OSIRIS technique. The selected volume, which can be controlled by frequency and its bandwidth, was 0.125cc out of 25cc and free of contamination from outer volume. With this technique NMR spectrum of a living tissue can be obtained without biopsy. i.e. in vivo state in which the metabolism of tissue may be quite different from in in vitro state. It is expected of this technique to be useful in the study of metabolism of living tissue as well as in diagnosis of deseases.

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Interspecies Somatic Cell Nuclear Transfer Technique for Researching Dog Cloning and Embryonic Stem Cells

  • Sugimura, Satoshi;Sato, Eimei
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.1-8
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    • 2011
  • Large quantities of high-quality recipient oocytes with uniform cytoplasm are needed for research in the promising field of somatic cell nuclear transfer (SCNT) and embryonic stem cell research. In canines, however, it is difficult to obtain large quantities of oocytes because each donor produces a limited number of mature oocytes in vivo. Although in vitro maturation (IVM) is considered an alternative approach to oocyte production, this technique is still too rudimentary to be used for the production of highquality, uniform oocytes in large quantities. One technique for overcoming this difficulty is to use oocytes obtained from different species. This technique is known as interspecies SCNT (iSCNT). This review provides an overview of recent advances in canine - porcine interspecies SCNT.