• Korean Journal of Microbiology
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• v.30 no.6
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• pp.524-527
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• 1992

Synechococcus sp. cyanophage was isolated from Baekwoon reservoir located in KyonggiDo. The cyanophage was purified by employing ultrafiltration. differential centrifugation. and sucrose density gradient centrifugation. Electron microscopic observation indicated that the sizes of its isometric head and contractile tail are 89 nm and] II nm. respectively. which means that the isolated cyanophage is included in the group. Myoviridae. The cyanophage maintained the stability of more than 50 percent from $20^{\circ}C$ to $40^{\circ}C$ and from pH 5 to 8. and had the maximal infectivity at $30^{\circ}C$ and pH 9 implying its ecological significance.

• Parasites, Hosts and Diseases
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• v.32 no.4
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• pp.243-248
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• 1994

Plasma membrane proteins of a Korean isolate of Trichomonus vofinalis HY-1 were fractionated for antigen analysis. Homogenates of T. vaginalis were fractionated by the differential centrifugation using sucrose step-gradient method. The interface layer from the 25%/45% sucrose was collected as a plasma membrane fraction and its purity was examined by transmission electron microscopy. The antigenicity of plasma membrane fraction was analysed by enzyme-linked immunoelectrotransfer blot technique with immune rabbit serum and compared with surface antigen labelled with N. hydroxysuccinimide-biotin. The fluffy fraction of 25%/45% sucrose interface was homogeneous and membrane particles were present as extended sheet and concentric vesicles showing typical trilamellar appearance under transmission electron microscope. Seven fractions at 40, 50, 60, 110, 130, 140 and 150 kDa were identified as the antigenic membrane proteins in EITB with anti HY-1 rabbit serum. The common band at 60 kDa was detected both in antigenic fractions of plasma membrane and surface protein labelled with NHS-biotin. This result indicates that this protein is considered as a major surface antigen of T. vaginalis. The role of this surface antigen at 60 kDa should be studied further.

• Tuberculosis and Respiratory Diseases
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• v.45 no.1
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• pp.176-183
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• 1998

Background: The total and differential cell count of bronchoalveolar lavage(BAL) fluid are useful assessing activity, prognosis and response to therapy in diffuse interstitial lung disease. But controversy exist as to the appropriate method in processing BAL fluid. Therefore we investigated the effect of gauze filtration, centrifugation and different storage time of BAL fluid on the total and differential cell count. Methods: We obtained BAL fluid from 6 persons with no active lung lesion and divided pooled BAL fluid into several siliconized glass tubes and filtered through 0, 1, 2, 4 folds of cotton guaze(pore size: 1mm), and compared total cell count using hemocytometer after trypan blue staining and differential cell count after Wright-Giemsa staining of cytocentrifuged preparations. And we also counted total and differential cell count after centrifugation(400g for 30 min) and various storage time(2hr, 24hr, and 48hr). Results: There was no difference in total and differential cell count according to folds of gauze filtraion. But without gauze filtration, mucus threads that hampered total and differential cell count were found in 2 cases (33%). Centrifugation resulted in loss of total cell count($24{\pm}18%$) without change in differential cell count. There was no change in total cell count after 2hr storage but significant cell loss was found after 24hr storage time(24hr : $28{\pm}21%$, 48hr : $41{\pm}24%$). However there was no change in differential cell count with 48hr storage time. Conclusion: Total and differential cell count of BAL fluid may be best performed after cotton gauze filtration without centrifugation and within 2 hours.

• Korean Journal of Food Science and Technology
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• v.19 no.5
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• pp.441-445
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• 1987

The subcellular distribution of lipoxygenase in germinating soybean seeds (Glycine max[L.] AmSoy) was investigated by using differential centrifugation and sucrose density gradient fractionation. Most of lipoxygenase -1 and -2/3 activities was present in the supernatant fraction after differential centrifugation of homogenates prepared from three-day-old seedlings; only 1.5% of lipoxygenase activity remained in particulate fractions. The results of a sucrose density gradient fractionation (three-day-old) showed that the lipoxygenase activity coincided with acid phosphatase at the densities of 1.19, 1.23, $1.25g/cm^3$, even though most of lipoxygenase and acid phosphatase activities appeared in supernatant fractions. There was no indication that mitochondria contained any lipoxygenase activity, and it does not appear that glyoxysomes and ER contained any lipoxygenase activity either.

• The Korean Journal of Food And Nutrition
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• v.8 no.4
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• pp.307-317
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• 1995

The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

• Journal of agricultural medicine and community health
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• v.17 no.1
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• pp.46-55
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• 1992

A systematic study was conducted to identify and isolate a serologically pertinent antigen with high specific activity and low cross reactivity from P.westermani antigen. Differential centrifugation of the homogenate yield three particulate and one soluble fractions ; the $480{\times}G$ pellet(Pw1), the $7650{\times}G$ pellet(Pw2) treated with n-butanol(Pw3), and $100000{\times}G$ supernatnat(Pw4). Comparison of antigenicity of these antigens, based upon differential centrifugation, to that of saline extract of P. westermani worm(SEP) was performed by SDS-PAGE and immunoblot techniques. The results obtained were as follows : 1) The ratio of absorbance value of ELISA against paragonimiasis positive pool sera to that of negative sera was highest when using Pw3 as antigen and that was lowest using Pwl. 2) Silver stained and SDS-PAGE of each antigen showed 34 and 13Kd band as common antigen band, but Pw2 didn't show clear band. 3) By immunoblot 55 and 34Kd bands using SEP and Pw4 showed strong positive reaction without cross reaction with sera from other helmenthic infections. Using Pw3, 10Kd band was observed as specific band. In conclusion, Pw3($100000{\times}G$ pellet urea soluble, treated with n-butanol) and Pw4($100000{\times}G$ supernatant) were usable for ELISA and immunoblot technique.

• Parasites, Hosts and Diseases
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• v.21 no.1
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• pp.41-48
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• 1983

The activity and distribution of aspartate aminotransferase (EC 2.6. 1. 1) and alanine aminotransferase (EC 2.6.1.2) in adult Fascicle hepatica have been studied. Fasciola hepatica was fractionated by differential centrifugation into nuclear, mitochondrial and cytosolic fractions. The activity of GOT and GPT was measured by the method of Reitman and Frankel. Isozyme patterns of those enzyme were also examined by DEAE-cellulose column chromatography. The results obtained were as follows; 1. The activity of aspartate and alanine aminotransferase was about 0.55 unit and 0.92 unit per 1g of Fascicle hepatica, respectively. 2. The activity of those enzymes was relatively low compared with those in mammalian tissues. 3. The distribution of aspartate aminotransferase in the subcellular organelles showed that 71% of the activity was in cytosolic, 24% in mitochondrial and 5% was in nuclear fraction. 4. About 22% of the total alanine aminotransferase activity was found in the mitochondrial fratstion, about 66% in the cytosolic fraction. 5. Aspartate aminotransferase from cytosolic fraction was separated into two types of isozymes, whereas alanine aminotransferase from cytosolic fraction gave only one active peak on DEAE-cellulose column chromatography.

• YAKHAK HOEJI
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• v.38 no.1
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• pp.12-19
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• 1994

Capsaicin(8-methyl-N-vanillyl-6-nonenamide) is the principal pungent component of Capsicum fruits. This work is directed to the capsaicin-hydrolyzing enzyme playing a key role in the rate limiting and critical step of capsaicin metabolism. In order to get precise information on the enzyme's subcellular location, rat liver homogenate was divided into six subcellular fractions by differential centrifugation technique: crude nuclear pellet, PNS(post nuclear supernatant) fraction, lysosomal pellet, cytosol, Tris wash fraction, micrisomes. Capsaicin-hydrolysing enzyme activity was analysed by high performance liquid chromatography(HPLC). This enzyme was found at the highest specific activity in the microsomal fraction and co-distributed with marker enzymes of the endoplasmic reticulum, NADPH-cytochrome c reductase and nucleoside diphosphatase. This is compatible with the result of ninhydrin color reaction of vanillylamine, primary metabolite of capsaicin hydrolysis, on thin layer chromatography(TLC). This enzyme is most active at pH $8.0{\sim}9.0$. Definite subcellular location of this enzyme will make it easy to proceed with further study.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.6
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• pp.885-892
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• 2009

To investigate host defense mechanisms for protecting the mammary gland from mastitis infection, the membrane fraction of mammary tissues from Holstein cows was purified by differential velocity centrifugation, and then the sodium dodecyl sulfate-polyacrylamid gel electrophoresis (SDS-PAGE) separated proteins were identified by ion trap mass spectrometer equipped with a Surveyor high performance liquid chromatography (HPLC) system. A total of 183 proteins were identified. Bioinformatics software was applied to analyse physicochemical characteristics of the identified proteins and to predict biochemical function. These data may provide valuable information to investigate the mechanisms of mammary gland milk secretion and infectious disease, and enable a clear identification of proteins and potential protein targets for therapies.

• Applied Biological Chemistry
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• v.20 no.2
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• pp.175-181
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• 1977

Lysosomal enzyme latency was demonstrated for hydrolases from porcine leukocyte by suspending sediment sfrom differential centrifugation in 0.125 to 0.250 M sucrose. Specific activities pH optima and activation energies were determined for hydrolases distributed in various sedimentation fractions and for enzymes solubilized by n-butyl alcohol extraction. Specific activities of the hydrolases revealed the heterogeneity of the Iysosomal fractions relative to enzyme content. pH optima identified the enzyme as acid hydrolases with optima for cathepsin D and aryl sulfatase also at pH 6.8. Activation energies of some hydrolases were low revealing that these enzymes could function efficiently during low temperature aging of meat.