• Molecular & Cellular Toxicology
• /
• v.4 no.3
• /
• pp.253-259
• /
• 2008

Human follicular fluid (HFF) is the in vivo microenvironment for oocyte maturation and includes a variety of proteins that could be involved in oocyte development and fertilization. We therefore used a proteomic approach to identify new HFF proteins. HFF from mature human follicles was obtained from five women following oocyte collection for in vitro fertilization (IVF). Ethanol-precipitated HFF run on two-dimensional gel electrophoresis (2DE) produced approximately 250 Coomassie brilliant blue-stained spots, 64 of which were identified using matrix-assisted laser desorption/ionization-mass spectrometry (MALDIMS). In this study, several proteins including complement factor H, inter-${\alpha}$ (globulin) inhibitor H4, inter-${\alpha}$-trypsin inhibitor heavy chain H4 precursor, human zinc-${\alpha}$-2-glycoprotein chain B, PRO2619, PRO02044, and complex-forming glycoprotein HC were new proteins that have not been previously reported in HFF using proteomic methods. Additionally, we identified alloalbumin venezia for the first time from trichloroacetic acid (TCA)-precipitated HFF. These HFF proteins could serve as new biomarkers for important human reproductive processes.

• Molecules and Cells
• /
• v.20 no.2
• /
• pp.271-279
• /
• 2005

Although macrophages are an important first line of cellular defense, they are unable to effectively kill phagocytosed C. albicans. To determine the physiological basis of this inability, we investigated the alterations of macrophage proteins caused by C. albicans infection. Since the formation of C. albicans hyphae caused cell death, proteins were prepared 3 h after infection and examined by two-dimensional gel electrophoresis (2-DE). The most prominent changes were in glycolytic enzymes, which could have caused energy depletion of the infected cells. Also changed were proteins involved in maintenance of cellular integrity and NO production. Treatment of the macrophages with either cytochalasin D or taxol did not alter their inability to kill C. albicans. Our results indicate that multiple factors contribute to cell death as the pathogenic form of C. albicans becomes fully active inside macrophage cells.

• 한국생물공학회:학술대회논문집
• /
• 2005.10a
• /
• pp.674-679
• /
• 2005

With addition of uracil during fermentation, the production rate of $1,3-{\beta}-glucan(curdlan)$ was enhanced. Since uracil was used as precursor of UDP-Glucose, the UDP-glucose level was increased and further glucan synthesis rate was increased.

• Proceedings of the Korean Society of Embryo Transfer Conference
• /
• 2006.10a
• /
• pp.58-58
• /
• 2006

• Korean Journal of Veterinary Research
• /
• v.42 no.2
• /
• pp.231-239
• /
• 2002

Proteomics is an emerging powerful tool in studying protein expression and function. At present study, proteomics was employed to evaluate the antigenicity among Escherichia coli O157:H7 strains using 2-dimensional gel electrophoresis (2-DE) and immunoblotting, SDS-PAGE and immunblotting analysis revealed no big differences among E coli O157:H7 strains. 2-DE analysis, however, revealed common antigens as well as specific antigens. The immunoblotting analysis revealed 20 common antigenic spots among E coli O157:H7 strains. In addition, there were 3 and 13 spots as common antigens between ATCC 43894 and KSC 109, and between ATCC 43894 and ACH 5, respectively. Antigenic spots specific for individual strain were also identified as 15, 8 and 22 for ATCC 43894, ACH 5 and KSC 109, respectively. The common antigens would be useful by employing either vaccine development or diagnosis marker, or both, whereas the specific antigens of individual strains would be applicable for epidemiological study. This study suggest that proteome analysis, representative as 2-DE, is valuable tool in exploring the E. coli antigenicity.

• Food Science and Biotechnology
• /
• v.18 no.1
• /
• pp.249-252
• /
• 2009

Two-dimensional gel electrophoresis (2-DE) was employed to assess the thermo-tolerance characteristics of Bifrdobacterium infantis ATCC 27920 to mild heat adaptation. When exposed to various heat levels, pH, and hydrogen peroxide ($H_2O_2$) stress conditions, B. infantis ATCC 27920 exhibited high level of stress resistance. Under mild-heat treatment ($46^{\circ}C$), no significant change in viability level was observed after 2 hr. Interestingly, improved viability was observed in mild-heat adapted ($46^{\circ}C$ for 1 hr) cultures exposed to $55^{\circ}C$, in comparison to control experiments. Viability was not affected by pH, bile, and $H_2O_2$ stress conditions. 2-DE analysis revealed those mild-heat adaptation up-regulated 4 proteins and down-regulated 3 proteins. Among these protein spots, isopropyhnalate dehydratase (leuD), glycosyltransferase (glgA), and ribosomal protein L5 (rp1E) were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALD1-TOF/MS).

• Journal of Magnetics
• /
• v.20 no.3
• /
• pp.246-251
• /
• 2015

This study was aimed to study the effect of Zn content on the hyperfine parameters and the structural variation of $Ni_{1-x}Zn_xFe_2O_4$ for x = 0, 0.2, 0.4, 0.6, and 0.8. To achieve this, a sol-gel route was used for the preparation of samples and the obtained ferrites were investigated by X-ray diffraction, scanning electron microscopy, and $M{\ddot{o}}ssbauer$ spectroscopy. The formation of spinel phase without any impurity peak was identified by X-ray diffraction of all the samples. Moreover, the estimated crystallite size by X-ray line broadening indicates a decrease with increasing Zn content. This result was in agreement with the scanning electron microscopy result, indicating the reduction in grain growth with further zinc substitution. The room-temperature $M{\ddot{o}}ssbauer$ spectra show that the hyperfine fields at both the A and B sites decreased with increasing Zn content; however, the rate of reduction is not the same for different sites. Moreover, the best fit parameter showed that the quadrupole splitting values of B site increased from the pure nickel ferrite to the sample with x = 0.8.

• Reproductive and Developmental Biology
• /
• v.33 no.4
• /
• pp.243-248
• /
• 2009

To examine the differential expression of proteins during the cycling (70~80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0~10.0, were analyzed by 2-DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2.D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up-regulated in the cycling cell and 5 were up-regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide-isomerase A3, microsomal protease ER-60, alpha-actinin-2, and heat-shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.

• Korean Journal of Agricultural Science
• /
• v.38 no.1
• /
• pp.65-70
• /
• 2011

Development of mouse fetus brains can be defined morphologically and functionally by three developmental stages, embryo day (ED) 16, postnatal stage one week and eight weeks. These defined stages of brain development may be closely associated with differential gene expression rates due to limited cellular resources such as energy, space, and free water. Complex patterns of expressed genes and proteins during brain development suggests the changes in relative concentrations of proteins rather than the increase in numbers of new gene products. This study was designed to evaluate early protein expression pattern in mouse fetus brain. The mouse brain proteome of fetus at ED 15.5, and 19.5 was obtained using 2-dimensional gel electrophoresis (DE). Analysis of the 2-DE gels in pH 3-10 range revealed the presence of 15 differentially expressed spots, of which 11 spots were identified to be known proteins following MALDI-TOF analysis; 3 spots were up-regulated and 8 spots were down-regulated in the mouse fetus brain at ED 15.5. UP-regulated proteins were identified as MCG18238, isoform M2 of pyruvate kinase isozymes M1/M2, isoform 2 of heterogeneous nuclear ribonucleoprotein K, heterogeneous nuclear ribonucleoprotein H2, creatine kinase B-type, 40S ribosomal protein SA and hemoglobin subunit beta-H1. Down-regulated proteins were putative uncharacterized protein, lactoylglutathione lyase and secreted acidic cysteine rich glycoprotein. Our results revealed composite profiles of mouse fetus brain proteins related to mouse fetus development by 2-DE analysis implying possible roles of these proteins in neural differentiation.

• Restorative Dentistry and Endodontics
• /
• v.38 no.4
• /
• pp.210-214
• /
• 2013

Objectives: This study aimed to evaluate the capacity of 2% chlorhexidine gel associated with 8% papain gel in comparison with 5.25% sodium hypochlorite in bovine pulp tissue dissolution. Materials and Methods: Ninety bovine pulps of standardized sizes were used and fragmented into 5-mm sizes. The fragments were removed from the root middle third region. They were divided into 6 experimental groups (n = 15), 1) 8% papain; 2) 2% chlorhexidine; 3) 2% chlorhexidine associated with 8% papain; 4) 0.9% saline solution; 5) 2.5% sodium hypochlorite; and 6) 5.25% sodium hypochlorite. The pulp fragments were weighed and put into immobile test tubes for dissolution for time intervals of 30, 60, 90, and 120 min. Results: The 5.25% sodium hypochlorite had greater dissolution potential than the pure papain, and when associated with chlorhexidine, both promoted greater dissolution than did the saline solution and 2% chlorhexidine groups (p < 0.05). The 2.5% sodium hypochlorite promoted dissolution to a lesser extent than the groups with papain within a period of 30 min (p < 0.05), but, was comparable to the saline solution and chlorhexidine. After 120 min, the 2.5% and 5.25% sodium hypochlorite promoted dissolution of 100% of the pulp fragments, and papain, 61%, while chlorhexidine associated with papain and chlorhexidine alone dissolved only 55% and 3%, respectively. Conclusions: The 8% papain in gel, both alone and in association with chlorhexidine, was able to dissolve bovine pulp tissue, but to a lesser extent than did 5.25% sodium hypochlorite.