• Proceedings of the Korean Society of Toxicology Conference
• /
• 2003.05a
• /
• pp.88-88
• /
• 2003

n view of the role of haptoglobin as a candidate for physical performance, we investigated the protein polymorphism of the haptoglobin in elite Korean male athletes. The serum sample was collected from 120 Korean male eliteathletes. The haptoglobin phenotypes were determined by polyacrylamide gel electrophoresis, followed by peroxidase staining. (omitted)

• Journal of Periodontal and Implant Science
• /
• v.24 no.1
• /
• pp.178-184
• /
• 1994

This study was undertaken to examine the protein content of GCF and serum from noraml population in order to standardize the sample loading on SDS/PAGE gels. The resulats were as follows ; 1. The protein concentration of serum was not different between normal group and diseased group. 2. In GCF, the bands of lower molecular weight than albumin were heavily stained, but in serum, the protein bands of higher molecalar weight were found. 3. The profile of protein in normal GCF was characterized by heavily staining bands at 77, 66, 55, 26 KDa corresponding to the positions of transferrin, albumin, heavy and light chains of Ig G. Also 47, 37 KDa nonplasma proteins were found.

• Journal of Oral Medicine and Pain
• /
• v.15 no.1
• /
• pp.91-104
• /
• 1991

After alkaline slab polyacrylamide gel electrophoresis and 3.3'-DMB staining of parotid saliva from 48 peoples in Ullung-do and 35 peoples in Jawall-do, the author have got following conclusions. 1. The gene frequencies of Proline-rich protein in Ullung-do were Pr1 =0.719, Pr2=0.281 2. The gene frequencies of Proline-rich protein in Jawall-do were Pr1=0.671, Pr2=0.329 3. The gene frequencies of Double band protein in Ullung-do were Db+=0.087, Db-=0.913 4. The gene frequencies of Double band protein in Jawall-do were Db+=0.0.106, Db-=0.0.894 5. The gene frequencies of Pa protein in Ullung-do were Pa+=0.179, Pa-=0.821 6. The gene frequencies of Pa protein in Jawall-do were Pa+=0.167, Pa-=0.833 7. The gene frequencies of Pr1 of Ullung-do and jawall-do were lower than those of Pr1 in Seoul and Onyang, but similar to those of Pr1 in kangnung and Cheju. 8. The gene frequencies of Db- of Ullung-do and Jawall-do were much higher than those of Db+ in Japanese, Chinese and other populations in Korean.

• Animal Bioscience
• /
• v.36 no.7
• /
• pp.1022-1033
• /
• 2023

Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.

• Journal of Korean Ophthalmic Optics Society
• /
• v.8 no.1
• /
• pp.65-71
• /
• 2003

The corneal epithelium is constantly being shed. The mechanism of corneal desquamation is not fully understood. Apoptosis, programmed cell death, may play a role. Apoptosis can be induced by a number of factors and different mechanisms. The study was performed to examine the apoptotic index induced in human corneal epithelial cells maintained in tissue culture by various apoptotic inducers. Various inducers, recombinant human cytokines($INF{\gamma}$, $TNF{\alpha}$, FASAb), actinomycin D. camptothecin, cycloheximide, dexamethasone and etoposide, were purchased from commercial suppliers. Inducers at manufacturer-recommended concentration were added to the corneal epithelial cells for 48 hours. Cell viability was measured using MTT assay. The cells were then assessed for the level of apoptosis. Morphologic changes and quantification of apoptotic cells were determined and counted under fluorescence microscope after inducers-treated human corneal epithelial (HCE) cells for 48 hours with Hoechst 33342 staining. Annexin V-FITC/PI staining and DePsipher assay. The expression of Fas protein was studied by immunocytochemistry. All inducers induced apoptosis in HCE cells in a dose dependent manner. Actinomycin D. camptothecin and etoposide induced apoptosis at lower than manufacturer-recommended concentration, while cytokines, cycloheximide and dexamethasone induced apoptosis at higher concentrations at the end of 48 hours. All inducers elicited typical apoptotic morphologic changes (chromatin condensation, nucleus fragmentations non-orange-red colored mitochondria) and expresses Fas protein highly. Apoptotic index of HCE cells by these inducers was different from the other cell lines. RNA synthesis inhibitor and topoisomerase inhibitors induced apoptosis at lower concentration than manufacturer-recommended concentration. Cytokines, cycloheximide and dexamethasone were able to produce apoptosis at 10 times higher concentrations. RNA synthesis inhibitor and topoisomerase inhibitors are more sensitive than intracellular receptor-activators in apoptotic induction of HCE cells.

• Journal of Life Science
• /
• v.23 no.12
• /
• pp.1509-1515
• /
• 2013

The term lipotoxicity has been used to describe how excess lipid accumulation leads to cellular dysfunction and death in non-adipose tissues, including skeletal muscle. While lipotoxicity has been found in cultured skeletal muscle cells with high-fat feeding, the consequences of lipotoxicity in vivo are still unknown, particularly in Type-I muscle, which is metabolically affected by lipotoxicity. The aim of this study was to investigate the effects of a high-fat diet on changes in the morphology and apoptotic protein expression of Type-I muscle loss in rats. The rats were fed either a high-fat diet or a normal diet for six weeks, and then lipid accumulation, inflammation response, and nucleus infiltration were measured, and PARP protein expression was cleaved by Oil Red O staining, H & E staining, and Western blot, respectively. Lipid accumulation, inflammation response, nucleus infiltration, and cleaved PARP protein expression were significantly (p<0.05) higher in the high-fat diet group than they were in the normal diet group. The weight of Type-I muscle tended to be lower in the high-fat diet group compared to the normal diet group, but the difference was not statistically significant. These results indicate that a high-fat diet triggers cell death in Type-I muscle via lipotoxicity, which suggests that a high-fat diet may be associated with sarcopenia.

• Journal of Periodontal and Implant Science
• /
• v.32 no.1
• /
• pp.1-12
• /
• 2002

The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.11 no.1
• /
• pp.203-220
• /
• 1989

Ever since the expression of new tumor-specific antigens was reported during malignant transformation, studies on separation, purification and characterization of these proteins have been so activated recently. Following experiment was performed to observe tumor-specific antigens by implanting DMBA pellet into submaxillary gland of rat for inducing salivary gland tumor. After dividing 280 rats into 2 groups, in control group, sham operation was performed on right submaxillary gland and, in experimental group, DMBA pellet (5mg) was implanted into right submaxillary gland. Then proteins from excised submaxillary gland by killing 10 rats every two weeks for 28 weeks were extracted with 3M KCl, and SDS-PAGE and PAS-staining were carried out for biochemical examination. The obtained results were summarized as follows; 1) At 12th week since implantation of DMBA pellet, tumor mass formation was inspected. And dysplasia at 6th week and invasive epidermoid carcinoma at 10th week were observed by microscope. 2) In control group, the weight ratio of both submaxillary glands had no any change, however, in experimental group, the ratio was increased remarkably. And at 28th week after DMBA implantation, there was more than 15 times of differences in weight between control and experimental group. 3) There was no DMBA remnant after 22nd experimental week. 4) In the SDS-PAGE, high molecular protein bands (more than 100 kd) were appeared much, and new prominent protein bands (66, 48, 41.5, 39, 37, 37.5 kd) were appeared after 4th week since DMBA implantation. However, 38, 27, 22kd protein bands were disappeared. 5) In PAS-staining, high molecular proteins were proteins were all glycoproteins and 37.5kd protein was proved as to be glycoprotein. And 38kd glycoprotein was disappeared after 4th week since DMBA implantation.

• Korean Journal of Clinical Laboratory Science
• /
• v.39 no.3
• /
• pp.217-222
• /
• 2007

In the last 5 years the Epidermal Growth Factor Receptor (EGFR) has emerged as one of the most important targets for drug development in oncology. Monoclonal antibodies targeting the external domain of EGFR have been shown to have clinical benefits in colorectal and head and neck cancer when combined with chemotherapy and/or radiation. Also the targeting of the epithelial growth factor receptor (EGFR) kinase domain using the closely related inhibitors gefitinib and erlotinib has generally been ineffective against solid tumors, many of which over express the receptor. We found that there were some differential expressions according to primary antibodies of the EGFR protein which being used as one of the histological tumor markers for non-small cell lung cancer (NSCLC). We also found that there are some differential expressions according to antibodies, the pH of the antigen retrieval (AR) buffer solutions and kinds of enzymes. There were some differential expressions according to the secondary antibodies and the detection systems. We analyzed the correlations between the immunohistochemical expressions of the EGFR protein and the gene mutations of the EGFR. The differences between automatic stainers and manual staining methods were also evaluated.

• The Korean Journal of Physiology and Pharmacology
• /
• v.20 no.2
• /
• pp.147-152
• /
• 2016

Present study aimed to investigate the effect of curcumin-pretreatment on intestinal I/R injury and on intestinal mucosa barrier. Thirty Wistar rats were randomly divided into: sham, I/R, and curcumin groups (n=10). Animals in curcumin group were pretreated with curcumin by gastric gavage (200 mg/kg) for 2 days before I/R. Small intestine tissues were prepared for Haematoxylin & Eosin (H&E) staining. Serum diamine oxidase (DAO) and tumor necrosis factor (TNF)-${\alpha}$ levels were measured. Expression of intestinal TNF-${\alpha}$ and tight junction protein (ZO-1) proteins was detected by Western blot and/or immunohistochemistry. Serum DAO level and serum and intestinal TNF-${\alpha}$ leves were significantly increased after I/R, and the values were markedly reduced by curcumin pretreatment although still higher than that of sham group (p<0.05 or p<0.001). H&E staining showed the significant injury to intestinal mucosa following I/R, and curcumin pretreatment significantly improved the histological structure of intestinal mucosa. I/R insult also induced significantly down-regulated expression of ZO-1, and the effect was dramatically attenuated by curcumin-pretreatment. Curcumin may protect the intestine from I/R injury through restoration of the epithelial structure, promotion of the recovery of intestinal permeability, as well as enhancement of ZO-1 protein expression, and this effect may be partly attributed to the TNF-${\alpha}$ related pathway.