• Title/Summary/Keyword: Cell Extraction

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Purification and Biochemical Characterization of Lectin from Viscum album (겨우살이 Lectin의 정제 및 생화학적 특성)

  • Jang, Cheol-Su;O, Mi-Jeong;No, Gwang-Su
    • KSBB Journal
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    • v.14 no.5
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    • pp.578-584
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    • 1999
  • The lectin was purified through 0.15 M NaCl extraction, ammonium sulfate precipitation, sepharose 4B affinity chromatography and gel filtration using sephadex G-150 from the leaves of Visum album collected in Mt. Duk Yu. The final gel filtration step resulted in 11.64 folds purification with 0.14% of recovery yield. We also performed biochemical characterization of the purified Visum album lectin. HPLC analysis of lectin purified by gel filtration revealed a singel peak. The analysis of the purified lectin by SDS-PAGE showed a tetramer composed of two identical subunits with molecular weights of 32 and 30 kDa. The lectin was a glycoprotein containing 14.4% carbohydrate, which consist of glucose, fructose, arabinose and xylose, and the amino acids such as phenylalanine, lysine and tyrosine. The purified lectin agglutinated human red blood cell types with similar potency, but when tested against red blood cells from mouse, bovine, rabbit, chicken and porcine, significant difference in potency were observed. Hemaggluting activity was inhibited by D-galactose, D-mannose, D-lactose and D-raffinose, but not by D-glucose, D-glucosamine, D-mannosamine, L-fructose, D-xylose, D-arabinose, D-galacturonic acid, D-fructose, L-rhamnose and N-acetyl-D-galactosamine. The optimal pH and thermal stability of the purified lectin were pH 4.0-7.0 and 20-5$0^{\circ}C$, respectively.

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Immunomodulating Activity of Laminaria japonica Polysaccharides (참다시마 다당체의 면역 증강 활성)

  • Ryu, Deok-Seon;Oh, Seung-Min;Kim, Ki-Hoon;Kim, Soo-Hwan;Choi, Hyun-Ju;Lee, Dong-Seok
    • Korean Journal of Food Science and Technology
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    • v.42 no.3
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    • pp.350-354
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    • 2010
  • Laminaria japonica polysaccharides (LP) were prepared from L. japonica through hot water extraction, ultrafiltration and gel chromatography. In this study, we investigated the immunomodulating activity of LP (0.25-1 mg/mL) on the mitogen/alloantigen reactive proliferation and killing activity of the Balb/c mouse splenocytes. The LP directly induced the proliferation of splenocytes that was stimulated with mitogen or alloantigen in a dose-dependent manner. The killing activity of cytotoxic T lymphocytes (CTLs) and lymphokine activated killer cells (LAKs) were enhanced significantly in the LP treated cells. Also, the treatment of splenocytes with LP increased production of interleukin-2 (IL-2). These results suggest that polysaccharides from L. japonica show a substantial immunomodulating activity in mouse immune cells.

Effect of Anti-oxidant, Anti-inflammatory and Anti-invasive of PMA-induced Matrix Metalloproteinase (MMP-2) and MMP-9 Activities of Water Extract and Solvent Fractions of Saururus Chinensis (삼백초 물 추출물과 유기용매 분획물의 항산화, 항염증 및 PMA에 의해 유도된 MMP-2 및 MMP-9활성 침윤 억제 효과)

  • Kim, Jun-Ho;Kim, Eun-Jung
    • Journal of Life Science
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    • v.26 no.5
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    • pp.584-591
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    • 2016
  • Saururus chinensis is a perennial plants, its flavonoid compound is known to exhibit anti-oxidative activity. This study was aimed to investigate the effect of Water Extract and Solvent Fractions of Saururus chinensis on antioxidant, anti-inflammatory and anti-invasive of Phorbol 12-myristate 13-acetate (PMA)-induced matrix metalloproteinase (MMP-2) and MMP-9 activities. Plant samples were fractionated into hexane, CHCl3, ethyl acetate, butanol, and water fractions, and each of these was assayed individually. The water fraction showed the highest extraction yield at 9.25%(w/w). Anti-oxidative activity was analyzed by DPPH assay. Cell viability was detected by the MTS assay. Anti-inflammatory activity was assayed by the nitric oxide (NO) production in mouse macrophage Raw 264.7 cells. The activity and mRNA expression of MMP-2 and MMP-9 in human oral squamous carcinoma YD-10B cells were examined by zymography and RT-PCR. As results, MMP-2/-9 activation was increased in PMA induced YD-10B cells. In PMA-treated YD-10B cells, the increased mRNA expression and protein activation of MMP-2/-9 were significantly inhibited in the ethyl acetate fraction. The ethyl acetate fraction showed the highest anti-oxidative activity at 73.38%. The ethyl acetate fraction at non-cytotoxic concentrations significantly exhibited the anti-inflammatory activity of Raw 264.7 cells in dose-dependent manner. In conclusion, these findings demonstrate that the ethyl acetate fraction obtained from a chinensis water extract potentiates a promising therapeutic anti-invasive agent and, therefore, as an anti-cancer drug for cancer prevention and therapy in oral cancer.

Comparison of Biological Activities of Epimedium koreanum Nakai Produced in Korea and China (국내산과 중국산 삼지구엽초의 생리활성 비교)

  • Noh, Joon-Hyun;Kim, Young-Jin;Kim, Se-Won;Lee, Jin-Ha;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.3
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    • pp.195-200
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    • 2003
  • This study was carried out to compare the biological activities of Epjmedium koreanum Nakai grown wild in Korea and China. The antioxidative effect of E. koreanum Nakai-extracts grown wild in Gangwondo was 78%, which was higher than that in China as 71%. The inhibition ratio of growing cancer cells was estimated as 89, 91 and 86% for human liver, lung and breast cancer cell lines, respectively in adding 0.1 g/l of ethanol extracts of E. koreanum grown in Korea. The hepatoprotective effect and Angiotensin Converting Enzyme (ACE) inhibiting effect of Korean one were also observed as 124 and 87%, respectively, which were also higher than those produced in China. There was not much difference between Korean and Chinese one in inhibiting ${\alpha}-glucosidase$ activities in all ranges of supplement. It was proved that most effective extraction solvent was mixed type as water and ethanol (1:1, v/v) to have higher biological activities from both Korean and Chinese ones.

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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Production and Action of Microbial Piscicidal Substance (미생물에 의한 살어성물질의 생성 및 그 작용)

  • 도재호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.6 no.1
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    • pp.41-46
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    • 1978
  • Piscicidal substance produced by Streptomyces sp. isolated from soil was toxic against various kinds of fish. After extraction with CH$Cl_3$ from the culture medium, the substance was purified by avicel column chromatography. In order to test toxicity, various kinds of fish were subjected to the acqueous solution of 100 us of the substance per liter of water. Generally, the substance was toxic to most fish, but Macropodus chinenes and Misgurnus mizolepis are resistant to the substance than Gobius similis and Pseudorasbora parva. The substance was stable at pH range, 3.0 to 7.0, but labile at alkaline pH, and to heat as well. Succinic dehydrogenase on most of tissue cell of Cyprinus carpio was inhibited by this substance strongly, but spinal cord was not inhibited. By addition of Cu and Pb salts to the culture medium, piscicidal substance producibility was activated.

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Production of Biodiesel and Nutrient Removal of Municipal Wastewater using a Small Scale Raceway Pond (미세조류 옥외 배양시스템을 이용한 바이오디젤 생산 및 도시하수 영양 염류 제거)

  • Kang, Zion;Kim, Byung-Hyuk;Oh, Hee-Mock;Kim, Hee-Sik
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.207-214
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    • 2013
  • A concerted effort to develop alternative forms of energy is underway due to fossil fuel shortages and its deleterious effects. Recently, bioenergy from microalgae has gained prominence and the use of municipal wastewater as a low cost alternative for a nutrient source has significant advantages. In this study, we have employed municipal wastewater directly after primary treatment (primary settling basin) in a small scale raceway pond (SSRP) for microalgal growth. Indigenous microalgae in the wastewater were encouraged to grow in the SSRP under optimal conditions. The mean removal efficiencies of TN, TP, and $NH_3-N$ after 6 days were 77.77%, 63.55%, and 89.02%, respectively. The average lipid content of the microalgae was 19.51% of dry cell weight, and linolenate and linoleate (18:n) were the predominant fatty acids. The 18S rRNA gene analysis and microscopic observations of the indigenous microalgae community revealed the presence of Chlorella vulgaris and Scenedesmus obliquus as the dominant microalgae. These results indicate that untreated municipal wastewater, serving as an excellent nitrogen and phosphate source for microalgal growth, could be treated using microalgae in open raceway ponds. Moreover, microalgal biomass could be further profitable by the extraction of biodiesel.

Effect of dietary supplementation with Spirulina on the expressions of AANAT, ADRB3, BTG2 and FASN genes in the subcutaneous adipose and Longissimus dorsi muscle tissues of purebred and crossbred Australian sheep

  • Kashani, Arash;Holman, Benjamin William Behrens;Nichols, Peter David;Malau-Aduli, Aduli Enoch Othniel
    • Journal of Animal Science and Technology
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    • v.57 no.3
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    • pp.8.1-8.8
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    • 2015
  • Background: The demand for healthy, lean and consistent meat products containing low saturated fatty acid content and high quality polyunsaturated fatty acids (PUFA), especially long-chain (${\geq}C_{20}$) omega-3 PUFA, has increased in recent times. Fat deposition is altered by both the genetic background and dietary supplements, and this study aimed to assess the effect of dietary Spirulina supplementation levels on the mRNA expression patterns of genes controlling lipid metabolism in the subcutaneous adipose tissue (SAT) and Longissimus dorsi (ld) muscle of Australian crossbred sheep. Methods: Twenty-four weaned lambs belonging to four breeds under the same management conditions were maintained on ryegrass pasture and fed three levels of Spirulina supplement (control, low and high). In terms of nutrient composition, Spirulina is a nutrient-rich supplement that contains all essential amino acids, vitamins and minerals. It also is a rich source of carotenoids and fatty acids, especially gamma-linolenic acid (GLA) that infer health benefits. After slaughter, subcutaneous adipose tissue (SAT) and ld samples were subjected to mRNA extraction and reverse transcription using quantitative polymerase chain reaction (RT-qPCR) to assess the mRNA expression levels of the Aralkylamine N-acetyltransferase (AANAT), Adrenergic beta-3 receptor (ADRB3), B-cell translocation gene 2 (BTG2) and Fatty acid synthase (FASN) genes, which are associated with lipid metabolism. Results: Both low and high Spirulina supplementation levels strongly up-regulated the transcription of all the selected genes in both SAT and ld tissues (mostly in the subcutaneous adipose), but sheep breed and sex did not influence the gene expression patterns in these tissues. Conclusions: The evidence indicates that high Spirulina supplementation level resulted in a decrease in intramuscular fat content in Australian purebred and crossbred sheep due to the enhanced production of melatonin in sheep muscle tissues and strong up-regulation of mRNA expression of BTG2 in SAT which negatively affected fat deposition. In contrast, low Spirulina supplementation level strongly up-regulated the ADRB3 and FASN genes responsible for fat production. These findings are consistent with the observed phenotypic data suggesting that low Spirulina supplementation level can increase lamb production, with higher long-chain PUFA content.

Changes of Reserved Substance and the Peroxidase Activity in Tomato Fruits during the Storage of Sub-atmosphere Pressure (감압저장중 도마도 과실의 저장물질과 Peroxidase 활성의 변화)

  • Sohn, Tae Hwa;Choi, Jong Uck;Cheon, Seong Ho
    • Current Research on Agriculture and Life Sciences
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    • v.3
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    • pp.92-98
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    • 1985
  • The experiments were conducted to investigate the activity changes of peroxidase, existence of isoenzyme and the changes of reserved substances in tomato under subatmospheric pressure storage condition. The results obtained were as follows: 1. Soluble fraction showed the highest peroxidase activity and followed by cell wall fraction, mitochondria fraction and ribosome fraction in that order. 2. Peroxidase activity was decreased during the ripening and senescence period in tomato. Especially, peroxidase activity in tomato was higher at a room temperature ($25^{\circ}C$) than at a low temperature ($15^{\circ}C$). The decreasing inclination was similar in both treatment. The peroxidase activity was higher in 380 Torr, than in 570 Torr. 3. At least, two isoperoxidases(Soluble or solubilized) were identified from different extraction procedures. Three of four isoenzymes were recognized from a vertical slab of polyacrylamide gel electrophoresis. 4. The changes of components in tomato under SAP were generally affected by temperature and pressure. Especially, quality of tomato stored at a low temperature ($15^{\circ}C$) and SAP (380 Torr.) was best during storage.

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Synthesis and Small Animal Brain PET Study of a Serotonin Receptor Radiotracer, 18F-Mefway (세로토닌 5-HT1A수용체 방사성 추적자 18F-Mefway의 합성과 소동물 뇌 PET 연구)

  • Ahn, Sung-Min;Hong, Tae-Kee;Ryu, Young-Hoon;Choi, Jae-Yong;Kim, Sung-Chul
    • The Journal of the Korea Contents Association
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    • v.9 no.11
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    • pp.262-270
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    • 2009
  • $^{18}F$-mefway has been developed as radioligand for serotonin receptor 5-$HT_{1A}$. The object of this study was to obtain the mefway precursor with the higher yield than previous method and to identify whether $^{18}F$-mefway can bind to 5-$HT_{1A}$ or not. from microPET imaging of small animal brain. Precursor was prepared by a modification of the reported procedure then [$^{18}F$] labeling was performed by adding $^{18}F$ ion at $130^{\circ}C$ in the hot cell for 30min. After purification of reaction mixture using alumina Sep-pak and HPLC, microPET images of small animal brain were determined. The chemical yield of precursor was increased from 9% to 34% using oxalyl chloride and LAH/diethylether. We synthesized a precursor which was successfully labeled with no-carrier-added $^{18}F$-by new synthetic route. This research suggest that $^{18}F$-mefway will be used a radiopharmaceutical for evaluation of central nerve system disorder as imaging a gent for 5-$HT_{1A}$ receptor.