• Journal of Embryo Transfer
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• v.13 no.3
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• pp.277-289
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• 1998

To establish a system for sperm swim-up separation through sucrose layer, indiscreet sperm migration should sufficiently to block but movement of sperm shouldn't inhibit. Thus, the effects of sucrose levels in sucrose layer, incubation times and types of sucrose layer on sperm separation were examined. And the results obtained were as follows; 1. Layer of 10mM sucrose inhibited sperm swim-up migration through sucrose layer. 2. Incubation for 25 minutes without sucrose layer significantly increased sperm swim-up migration. However, incubation for 10 minutes to induce swim-up through sucrose layer significantly stimulated sperm migration and maintained sperm movement. 3. There was no significant difference between Type I and Type II in barrier effect of sucrose layer. However, sucrose layer of Type II with shorter distance of barrier was efficient for sampling. To elucidate a function of follicular fluid on sperm chemotaxis using in vitro system of sucrose layer of Type II and incubation for 10 minutes, the effects of dilution, heat treatment, and protein and lipid extracts of follicular fluid on sperm swim-up separation were examined. And the results obtained were as follows; 4. Follicular fluid stimulated sperm migration and movement, and significantly-attracted capacitated-sperm at 10% level. 5. Follicular fluid heated at 55$^{\circ}C$ for 30 minutes maintained the effect of follicular fluid stimulating sperm migration and movement. 6. Follicular protein stimulated sperm movement that was reduced by filtration of the protein. 7. Follicular lipid didn't significantly stimulate sperm migration and movement. 8. Both of follicular protein and lipid reduced the effect of follicular fluid stimulating sperm migration and movement. In conclusion, sucrose layer could be used for a barrier against indiscreet sperm migration by swim-up. And follicular fluid stimulated migration and movement of sperm and attracted capacitated-sperm through sucrose layer. Especially, heat-resistant protein of follicular fluid stimulated sperm migration.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.60 no.1
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• pp.29-32
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• 2015

Two-dimensional electrophoresis (2-DE) was executed to separate the seed storage proteins from the buckwheat. The proteins extracted from the whole seed proteins were better separated and observed in the use of lysis buffer. Using this method, the highly reproducible isoelectric focusing (IEF) can be obtained from polyacrylamide gels, and IEF from the polyacrylamide gel at all the possible pH range (5.0-8.0) was more easily separated than IPG (immobilized pH gradient) gels. The polyacrylamide gels in the first dimension in 2-DE was used to separate and identify a number of whole seed proteins in the proteome analysis. In this new apparatus using 2-DE, 27cm in length of plate coated with polyacrylamide gel was used and the experiment was further investigated under the various conditions.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.4
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• pp.240-245
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• 2003

The use of microfabricated microfluidic devices offers significant advantages over current technologies including fast analysis time and small reagent requirements. In the context of proteomic research, the possibility of using affinity-based separations for prefractionation of samples using microfluidic devices has significant potential. We demonstrate the use of microscale devices to achieve affinity separations of proteins using a device fabricated from borosilicate glass wafers. Photolithography and wet etching are used to pattern individual glass wafers and the wafers are fusion bonded at 650$^{\circ}C$ to obtain enclosed channels. A polymer has been successfully polymerized in situ and used either as a frit for packing beads or, when derivatized with Cibacron Blue 3GA, as a separation matrix. Both of these technologies are based on in situ UV photopolymerization of glycidyl methacrylate (GMA) and trimethylolpropane trimethacrylate (TRIM) in channels.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.4
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• pp.601-604
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• 2006

The objective of this study was to investigate technical methods for extraction of mucopolysachharide-protein containing chondroitin sulfate from keel cartilage of chickens. The chemical composition of chicken keel cartilage was determined. For the preparation of mucopolysaccharide-protein from lyophilized chicken keel cartilage, hot water extraction and alcalase hydrolysis methods were examined. Results showed that the optimum condition of hot water extraction was incubation for 120 min with a yield of 40.09% and chondroitin sulfate content of 28.46%. For alcalase hydrolysis, the most effective condition was 2% alcalase in 10 volumes of distilled water for 120 min. The yield of hydrolysate was 75.87%, and chondroitin sulfate content was 26.61%. For further separation of chondroitin sulfate from the alcalase hydrolysate, which has a higher yield than that of hot water, 60% ethanol precipitation was performed. The yield of the ethanol precipitate was 21.41% and its chondroitin sulfate content was 46.31%. The hot water extract, alcalase hydrolysate and ethanol precipitate showed similar electrophoretic migration with standard chondroitin sulfate (chondroitin sulfate A), using cellulose acetate membrane electrophoresis. These results indicated that a significant amount of mucopolysaccharide-protein containing chondroitin sulfate could be acquired form chicken keel cartilage. Therefore, keel cartilage in chicken may provide an inexpensive source of chondroitin sulfate for commercial purposes.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.37 no.2
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• pp.85-90
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• 2004

The effect of the hydraulic residence time (HRT) for the removal of aquarium waste, such as protein, total suspended solids (TSS) and turbidity were investigated by using a foam separator Protein, TSS and turbidity removal efficiencies were increased with the increase of hydraulic residence time. The optimum hydraulic residence time was 0.5 min, and the highest protein and TSS removal rates were $14.4\;g/L{\cdot}day\;and\;38.9\;g/L{\cdot}day,$ respectively. The tendency of turbidity removal rate and efficiency was similar to that of protein.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.5
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• pp.1911-1915
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• 2012

Aim: New technologies for the early detection of pancreatic cancer (PC) are urgently needed. The aim of the present study was to screen for the potential protein biomarkers in serum using proteomic fingerprint technology. Methods: Magnetic beads combined with surface-enhanced laser desorption/ionization (SELDI) TOF MS were used to profile and compare the protein spectra of serum samples from 85 patients with pancreatic cancer, 50 patients with acute-on-chronic pancreatitis and 98 healthy blood donors. Proteomic patterns associated with pancreatic cancer were identified with Biomarker Patterns Software. Results: A total of 37 differential m/z peaks were identified that were related to PC (P < 0.01). A tree model of biomarkers was constructed with the software based on the three biomarkers (7762 Da, 8560 Da, 11654 Da), this showing excellent separation between pancreatic cancer and non-cancer., with a sensitivity of 93.3% and a specificity of 95.6%. Blind test data showed a sensitivity of 88% and a specificity of 91.4%. Conclusions: The results suggested that serum biomarkers for pancreatic cancer can be detected using SELDI-TOF-MS combined with magnetic beads. Application of combined biomarkers may provide a powerful and reliable diagnostic method for pancreatic cancer with a high sensitivity and specificity.

• YAKHAK HOEJI
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• v.55 no.3
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• pp.203-212
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• 2011

Adipocytes have been known to secrete a number of important proteins called adipokines with roles in energy metabolism, reproduction, cardiovascular function and immunity. In this study we have attempted to identify intensively secretory proteins from 3T3-L1 adipocytes. 3T3-L1 preadipocytes were differentiated into mature adipocytes and then the cells were left in serum-free medium. The supernatant was filtrated and dialyzed. Lyophilized secretome was fractionated by two different methods, 1-D SDS PAGE and RP-FPLC. The tryptic peptides from the gel slices and the FPLC fractions were analyzed by nanoLC/ESI-MS/MS. We identified a total of 303 identical proteins from two methods, 251 proteins from 1-D gel and 184 proteins from RP-FPLC. 86 of them were listed as a secretory protein Finally, we identified many known or unknown secreted proteins existed in the low level including adiponectin, angiotensinogen, bone morphogenetic protein-1 (BMP-1), macrophage migration inhibitory factor (MIF), insulin like growth factor-II (IGF-II), interleukin-6 (IL-6), follistatin-related protein-1, minecan, and resistin. The existence of some of secreted proteins has been confirmed in RNA level. This proteomic experiment is useful for the intensive screening of secretory proteins in many kinds of other cells.

• The korean journal of orthodontics
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• v.34 no.1 s.102
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• pp.71-81
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• 2004

In this study, we attempt to investigate the mechanisms by which PDL cells regulate osteoclast formation and also tc know whether PDL retained their characteristic phenotype during tooth eruption and interdental separation. Rats were prepared at developmental days 21 (pre-root formation), 27(toot development), 34(advanced root formation/eruption) and at later times(adult rats). To induce severe resorption state of alveolar bone and tooth root, interdental separation with brass wire was performed between the lower first and second molars for 2 weeks in adult rats. Rat mandibles were demineralized and embedded in paraffin, and horizontal and frontal section were prepared for immuno-histochemical analysis using PDL-specific protein 22 (PDLs22), receptor activator of NFKB ligand (RANKL) and osteoprotegerin (OPG) antibodies. 1. Root formation and eruption stage of tooth development. 1) PDLs22 immunolocalization was observed in tooth follicle/PDL cells and osteoblasts throught out the root formation and eruption stages of tooth development. 2) RANKL expression became stronger at eruption stage than root formation stage of tooth development. 3) Strong expression of OPG was detected in follice/PDL cells of toot formation stage but it was decreased with tooth eruption. 2. Interdental separation between lower first and second molar 1) Comparared to normal animal, multinucleated osteoclasts and odontoclasts were markedly induced in the alveolar bone and tooth root with PDL remodeling in hematoxylin-eosin section. 2) PDLs22 expression was decreased with interdental separation. 3) RANKL expression was Increased with interdental separation in PDL fibroblasts, osteoblasts, odontoclasts and it lacunae, resorting dentin, cementum and bone matrix. 4) OPG expression was slightly decreased in the PDL cells adjacent to the alveolar bone and root surface with interdental separation. These results suggested that during tooth eruption and tooth movement, RANKL and OPG in the periodontal tissues are important determinants regulating balanced alveolar bone and tooth root resorption. And it is also suggested that PDL cells retained their characteristic phenotype during tooth eruption and interdental separation except for the short period of PDL remodeling.

• Food Science and Preservation
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• v.15 no.6
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• pp.903-908
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• 2008

This study investigated protein changes in soymilk and whole soymilk due to enzymatic hydrolysis. The total free amino acid contents of low molecular weight soymilk (LSM) and low molecular weight whole soymilk (LWSM) were higher than soymilk (SM) and whole soymilk (WSM). The essential amino acid content was similar in SM and LSM, but was higher in LWSM than WSM. In SDS-PAGE performed to tendency of becoming low molecules, the soy protein molecular weights were 3372 kDa for SM and WSM, but 17 kDa or less for LSM and LWSM. Also, high molecular weight protein spots were evident in 2-D electrophoresis of SM and LSM, but only low molecular weight protein spots of various sizes were evident in WSM and LWSM. This suggests that the high molecular weight protein in SM and WSM is changed to low molecular weight protein by enzymatic hydrolysis. Further investigations of the separation and qualities of these proteins are required.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.2
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• pp.195-198
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• 1989

The water binding capacity (WBC) of hoe plasma protein was investigated. The centrifugal condition for optimal separation of plasma from hog blood was fixed at 1400 g-force. The WBC of 5%-plasma-protein-solution eel increased rapidly between pH 6 and 7 but gradually after pH 7 at $85^{\circ}C$ for 30 min. The higher heating temperature demonstrated the higher WBC of 5%-plasma-protein-solution gel at pH 7 within short period of time. The WBC of 5%-plasma-protein-solution gel increased rapidly at the beginning of heating. The WBC per gram of plasma protein at pH 7 and $85^{\circ}C$ for 30 min decreased as protein concentration of the plasma solution increased.