• Title/Summary/Keyword: Heparin extraction

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Characteristic Properties of Fucoidan Sulfate Purified from Gompi, Ecklonia stolonifera (곰피에서 정제한 Fucoidan Sulfate의 특성)

  • Lee, Hong-Soo;Jin, Sung-Hyun;Kim, Hee-Sook;Ryu, Byung-Ho
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.716-723
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    • 1995
  • The fucoidan purified from Korean brown seaweed, Ecklonia stolonifera was characterized on molecular structure and blood anticoagulant activities. Extraction was conducted at $100^{\circ}C$ with water and repeated twice. The crude fucodian was 151.1g out of 20.0 kg of Ecklonia stolonifera. The Fucoidan-1, which was purified from crude fucoidan using calcium chloride and cetyl pyridium chloride (CPC), was 35.2% against crude fucoidan. Fucoidan-5 was obtained approximately 28.1% from Fucoidan-1 through DEAE-Toyopearl 650 M ion-exchange column chromatography and showed one band by cellulose acetate electrophoresis. The molecular weight of Fucoidan-5 was estimated to be about 21,000∼23,000 dalton by Sephacryl S-300 gel filtration chromatography. Fucoidan-5 consists of 35.7% of fucose and 4.3% of galactose and the molar ratio of fucose and sulfate was about one to one. IR spectrum of Fucoidan-5 showed absorption at $1240\;cm^{-1}\;and\;850\;cm^{-1}$ and specific rotation value, $[\alpha]$, was $[\alpha]$. These results suggests that the sulfate maybe bind at $C_{4}$ carbon on ${\alpha}-L-fucose$. Gas chromatograph of methyl alditol acetate revealed that Fucoidan-5 is a fucose containing sulfated polysaccharide with $({\alpha}l-2)\;or\;({\alpha}l-2)$ glycosidic linkage. Anti-thrombin activity of the Fucoidan-5 was estimated as 1.4 time stronger than heparin. From above results, the purification methods using CPC and ion exchange chromatography is effective tools for obtaining highly purified fucoidan from Gompi, Ecklonia stolonifera.

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Mastitis Diagnostics by Near-infrared Spectra of Cows milk, Blood and Urine Using SIMCA Classification

  • Tsenkova, Roumiana;Atanassova, Stefka
    • Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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    • 2001.06a
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    • pp.1247-1247
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    • 2001
  • Constituents of animal biofluids such as milk, blood and urine contain information specifically related to metabolic and health status of the ruminant animals. Some changes in composition of biofluids can be attributed to disease response of the animals. Mastitis is a major problem for the global dairy industry and causes substantial economic losses from decreasing milk production and reducing milk quality. The purpose of this study was to investigate potential of NIRS combined with multivariate analysis for cow's mastitis diagnosis based on NIR spectra of milk, blood and urine. A total of 112 bulk milk, urine and blood samples from 4 Holstein cows were analyzed. The milk samples were collected from morning milking. The urine samples were collected before morning milking and stored at -35$^{\circ}C$ until spectral analysis. The blood samples were collected before morning milking using a catheter inserted into the carotid vein. Heparin was added to blood samples to prevent coagulation. All milk samples were analyzed for somatic cell count (SCC). The SCC content in milk was used as indicator of mastitis and as quantitative parameter for respective urine and blood samples collected at same time. NIR spectra of blood and milk samples were obtained by InfraAlyzer 500 spectrophotometer, using a transflectance mode. NIR spectra of urine samples were obtained by NIR System 6500 spectrophotometer, using 1 mm sample thickness. All samples were divided into calibration set and test set. Class variable was assigned for each sample as follow: healthy (class 1) and mastitic (class 2), based on milk SCC content. SIMCA was implemented to create models of the respective classes based on NIR spectra of milk, blood or urine. For the calibration set of samples, SIMCA models (model for samples from healthy cows and model for samples from mastitic cows), correctly classified from 97.33 to 98.67% of milk samples, from 97.33 to 98.61% of urine samples and from 96.00 to 94.67% of blood samples. From samples in the test set, the percent of correctly classified samples varied from 70.27 to 89.19, depending mainly on spectral data pretreatment. The best results for all data sets were obtained when first derivative spectral data pretreatment was used. The incorrect classified samples were 5 from milk samples,5 and 4 from urine and blood samples, respectively. The analysis of changes in the loading of first PC factor for group of samples from healthy cows and group of samples from mastitic cows showed, that separation between classes was indirect and based on influence of mastitis on the milk, blood and urine components. Results from the present investigation showed that the changes that occur when a cow gets mastitis influence her milk, urine and blood spectra in a specific way. SIMCA allowed extraction of available spectral information from the milk, urine and blood spectra connected with mastitis. The obtained results could be used for development of a new method for mastitis detection.

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AN EXPERIMENTAL STUDY ON THE BONE INDUCTION CAPACITY OF THE PORCINE BONE MATRIX-DERIVED BONE MORPHOGENETIC PROTEIN (돼지의 골기질유도 골형성단백질의 골유도능에 관한 실험적 연구)

  • Park, Young-Wook;Lee, Jong-Ho;Kim, Soo-Kyeong
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.19 no.3
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    • pp.265-286
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    • 1997
  • Bone morphogenetic proteins(BMPs) are a group of transforming growth factor beta(TGF-${\beta}$)-related factors and multifunctional proteins, especially the only known biologic factors capable of inducing endochondral bone formation at an extraskeletal site. This study was performed to investigate the effect of the partially purified porcine BMP(pBMP) at an ectopic site. PBMP was partially purified from porcine bone matrix and its activity was monitored by an in vivo bioassay. The purification method utilized extraction of the bone-inducing activity with 4M guanidine, followed by chromatography on heparin-Sepharose. Active fractions were assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. And the fractions were reconstituted with inactive insoluble collagenous bone matrix from rats, acid soluble type I collagen from rat tail and chondroitin-6-sulfate sodium salt and implanted into the pectroralis muscle pouches of Sprague-Dawley rats. And the carrier complex was implanted on the opposite side as control. The rats were sacrificed at the day of 1st, 3rd, 5th, 7th, 11th, 14th and 21st after implantation and examined histologically, radiologically and biochemically. And alkaline phosphatase activity and calcium content were used as indices of bone formation. The results were as follows ; 1. Active fractions were localized in a zone between 31 and 40 KDa on SDS-PAGE. 2. The implanted 3.0mg of the partially purified pBMP induced cartilage and bone in the muscle tissue of rats through an endochondral ossification process. 3. Inactive insoluble bone matrix, type I collagen and chondroitin-6-sulfate have functioned as carriers for pBMP, but revealed some foreign body reactions. 4. Soft X-ray didn't reveal significant change between the experimental and the control group. 5. The alkaline phosphatase activities in the experimental group of 5th, 7th, 11th, 14th and 21st were increased significantly compared with control (p<0.01) with the peak in the group of 11th day. 6. With time, the calcium content of the experimental group increased. And the calcium contents in the experimental group of 11th, 14th and 21st were increased significantly compared with control (p<0.01).

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