OBJECTIVE: The discovery of new biomarkers for ovarian cancer is clearly necessary for the detection and monitoring of the disease. Experion(TM) automated electrophoresis system can be employed in the identification of differentially expressed proteins in cancer cells. The objective of this study was to discover potential diagnostic serological biomarkers for ovarian cancer. METHODS: We performed protein expression difference analyses for 14 healthy women and 28 ovarian cancer patients with stage I, III and IV using Experion(TM) system. And then we checked the protein expression as silver staining after loading at 8~16% gradient gel for comparison with Experion(TM) gel image. The candidate biomarkers were purified and determined using MALDI-TOF mass spectrometer. RESULTS: The distinctive polypeptide peaks were detected at 115.40, 15.96, 14.8, 11.66, and 10.69 kDa and these five peaks were identified as ceruloplasmin, hemoglobin beta chain, hemoglobin sigma chain, serum amyloid A4, and amyloid related serum protein SAA, respectively. These proteins were significantly different between the sera of normal healthy women and ovarian cancer patients. CONCLUSIONS: Five proteins were found to be significantly different between the sera of normal healthy women and ovarian cancer patients. In addition, Experion(TM) assay system can provide high performance for analysis of ovarian cancer-related proteins by increasing the throughput while maintaining a high level of accuracy.
Amyloid ; Biomarkers* ; Ceruloplasmin ; Electrophoresis ; Female ; Humans ; Ovarian Neoplasms* ; Silver Staining

BACKGROUND AND OBJECTIVES: The Nucleolar organizer regions (NORs) are one of markers of cellular proliferation. Because the NORs can be visualized by a silver staining technique, the NORs are called the argyrophilic nucleolar organizer regions (AgNORs). The expression patterns of proliferative markers have been reported in the cholesteatoma, but the AgNORs have not been studied in the cholesteatoma. We investigated the proliferative activities of the cholesteatoma by the AgNORs and the usefulness of the AgNORs as a proliferative index in the cholesteatoma. MATERIALS AND METHODS: We assessed 5 postauricular skin samples and 20 cholesteatoma specimens by the numbers of the total AgNORs and the large AgNORs (large AgNOR means a diameter of over 6 nm) in high power fields and each cell. And the total areas of the AgNORs in high power fields (HPF) were calculated. RESULTS: The numbers of the large AgNORs in HPF, the numbers of AgNORs in each cell and the total areas of the AgNORs in HPF of the cholesteatoma were higher than those of the controls (p<0.05). In the cholesteatoma, the numbers of the large AgNORs and the total areas of the AgNORs in HPF were the highest in the keratinizing squamous epithelium of thick portion followed by the non-keratinizing squamous epithelium, and the keratinizing epithelium of thin portion. The numbers of the large AgNORs in each cell of the basal and superficial layers were the highest in the thick keratinizing squamous epithelium. In the suprabasal layer, the non-keratinizing squamous epithelium showed higher numbers of the large AgNORs but showed no statistical significance. CONCLUSION: 1) The proliferative capacity of the epithelium of cholesteatoma is reactive proliferative status. 2) The proliferative activity is varied with the differentiation status of the squamous epithelium in the cholesteatoma.
Cell Proliferation ; Cholesteatoma* ; Epithelium* ; Nucleolus Organizer Region* ; Silver Staining ; Skin

OBJECTIVE: To assess the value of Ploton silver staining and phalloidin-iFlour 488 staining in observation of the morphology of osteocyte dendrites of mice at different developmental stages. METHODS: The humerus and femurs were harvested from mice at 0 (P0), 5 (P5), 15 (P15), 21 (P21), 28 (P28), and 35 days (P35) after birth to prepare cryo-sections and paraffin sections. HE staining of P35 mouse femur sections served as a reference for observing osteocytes in the trabecular bone and cortical bone. The humeral sections at different developmental stages were stained with Ploton silver staining to observe the morphology of osteocytes and canaliculi, and the canalicular lengths in the cortical and trabecular bones of the humerus of the mice in each developmental stage were recorded. The cryo-sections of the humerus from P10 and P15 mice were stained with phalloidin iFlour-488 to observe the morphology of osteocytes and measurement of the length of osteocyte dendrites in the cortical bone. RESULTS: In the trabecular bone of the humerus of P0-P15 mice, Ploton silver staining only visualized the outline of the osteocytes, and the morphology of the canaliculi was poorly defined. In P21 or older mice, Ploton silver staining revealed the morphology of the trabecular bone osteocytes and the canaliculi, which were neatly arranged and whose lengths increased significantly with age (P21 CONCLUSIONS Mouse osteocyte dendrites elongate progressively and their arrangement gradually becomes regular with age. Ploton silver staining can clearly visualize the morphology of the osteocytes and the canaliculi in adult mice but not in mice in early stages of development. Phalloidin iFlour-488 staining for labeling the cytoskeleton can be applied for mouse osteocytes at all developmental stages and allows morphological observation of mouse osteocytes in early developmental stages.
Animals ; Bone and Bones ; Dendrites ; Mice ; Osteocytes ; Phalloidine ; Silver Staining

silver nitrate staining techniques. Five, 10-mm holes were placed in each rabbit mandible to simulate defective regions with the use of a low speed trephine bur. In the experimental group, the previously cited defects were grafted with both activated osteoblastic and autogenous bone. The control group, however, was only grafted with autogenous bone. Both groups were then analyzed at 2, 4, and 8-week intervals using bone histomorphometric analysis.RESULTS: According to histomorphologic analysis, the rates of new bone formation at the 2, 4, and 8-week intervals were 36%, 51%, and 23% for the control group, respectively; 52%, 39%, and 28%, for the experimental group, respectively. The experimental group showed higher rates of new bone formation compared to the control group at both the 2-week and 8-week interval.CONCLUSION: Bone marrow-derived osteoblasts seems to be a promising bone graft material.]]>
Dexamethasone ; Durapatite ; Mandible ; Osteoblasts ; Osteogenesis ; Rabbits ; Silver Staining ; Transplants

Nanogold-silver staining is a pre-embedding immunocytochemical technique, making it possible to label antigens with particulate markers at the electron microscopic level. This technique is based on the advancement of gold technology and on the development of silver staining or enhancement. In the present study, the method of Nanogoldsilver staining that includes the use of high concentration of glutaraldehyde and gold toning as well, was used to localize gp100 or a membrane bound antigen in melanosome of the cultured melanocytes. With the HMB45 antibody against gp100, biotinylated secondary antibody and streptavidin-nanogold followed by silver enhancement and gold toning led to highly specific labeling of gold particle over the melanosome compartments. The specificity of labelings obtained with this protocol was confirmed by the control experiment. Omission of the primary antibody led to very low background of labeling. The specific signal that appeared as a collection of gold particles was localized mainly to the peripheral part of melanosome. This finding provides the more detailed nature of gp100 localization in melanosome, which has not yet been shown by the previous immunocytochemical study such as the post-embedding immunoelectron microscopy.
Glutaral ; Melanocytes* ; Melanosomes ; Membranes ; Microscopy, Immunoelectron ; Sensitivity and Specificity ; Silver ; Silver Staining

We examined a series of changes that occur in the trabecular meshwork fibers of human eyes during fetal development at 12-30 weeks of gestation. At 12 and 15 weeks, the uveal meshwork was stained black with silver impregnation (indicating the predominance of collagen types III and IV) in the endomysium of the ciliary muscle. At 20 weeks, in combination with Schlemm's canal, a dense fibrous tissue mass corresponding to the trabecular meshwork anlage appeared and was colored black. The anlage was continuous with the corneal endothelium rather than with the ciliary muscle. Until 25 weeks, the trabecular meshwork was identifiable as fragmented fiber bundles that stained red-black, suggesting a mixture of collagen types I, III, and IV. At 30 weeks, half of the ciliary muscle fibers were inserted into the scleral spur and not into the meshwork. Therefore, any contribution of ciliary muscle contraction to the differentiation of the trabecular meshwork would appear to be limited. We hypothesize that an uneven distribution of mechanical stresses in the area of the cornea-sclera junction causes a tear thereby creating Schlemm's canal and is accompanied by a change in the collagen fiber types comprising the meshwork.
Collagen ; Endothelium, Corneal ; Eye ; Fetal Development ; Humans ; Muscle Contraction ; Muscles ; Pregnancy ; Silver ; Silver Staining ; Stress, Mechanical ; Trabecular Meshwork

PURPOSE: FSGS do not respond well to any kind of therapy and gradually progress to end-stage renal disease. This study was conducted to investigate the difference of protein expression between MCNS and FSGS as a preliminary study for understanding the pathophysiology of FSGS. METHODS: Renal biopsy samples of MCNS and FSGS were obtained, which was diagnosed by one pathologist. They were solubilized with a conventional extraction buffer for protein extraction. The solution was applied on immobilized linear gradient strip gel (pH 4-7) using IPGphor system. Silver staining was carried out according to standard method. Protein identification was done by searching NCBI database using MASCOT Peptide Mass Fingerprint software. RESULTS: The differences in protein expressions between MCNS and FSGS were shown by increased or decreased protein spots. Most prominently expressed spot among several spots in FSGS was isolated and analyzed, one of which was glutathione S-transferase (GST) P1-1, whereas it was not found in MCNS. So GSTP1-1 was considered as the one of the key biomarkers in pathogenesis of FSGS. CONCLUSION: This result would be helpful in diagnosing FSGS and researching FSGS. Further studies for glutathione S-transferase P1-1 might be necessary to elucidate the mechanisms regarding FSGS.
Biomarkers ; Biopsy ; Dermatoglyphics ; Glutathione Transferase ; Kidney Failure, Chronic ; Nephrotic Syndrome ; Proteomics ; Silver Staining

The validation study for two STR loci on X-chromosome, DXS7132 and GATA31D10, was done including allelic distribution and frequency of each allele to use these results for individual identification and paternity testing. For 496 unrelated Koreans, above two STR loci were amplified simultaneously using duplex PCR amplification method. The amplified products were analyzed by polyacrylamide gel electrophoresis followed by silver staining. In male DXS7132 locus revealed 7 different alleles ranging from 276bp to 300bp. The largest allele was consisted of 14 repetition of [TCTA] unit and took 0.3417. The allele 15 followed next as 0.3165 and allele 13 as 0.1726. In female general distribution was same except one allele, allele 18 was found additionally. The heterozygosity was 0.7706 and 23 different genotypes were found. Polymorphism information content(PIC) was 0.727. Two cases of mutation were noted in DXS7132 locusIn both male and female 7 different alleles were noted in GATA31D10 locus and the alleles ranged from 195bp to 231bp. The allele 15(199bp) took the majority of all as 0.825. The other alleles showed rather relatively low frequency. The heterozygosity was 0.2385 and 11 different genotypes were found. PIC was 0.2521, and no mutation was noted in GATA31D10 locus. Considering these two loci together, 22 different halpotype were noted.
Alleles ; Electrophoresis, Polyacrylamide Gel ; Female ; Genotype ; Haplotypes ; Humans ; Male ; Paternity ; Polymerase Chain Reaction ; Silver Staining

Two quadruplex PCR reactions were designed for 7 tetrameric (D3S2406, D4S2368, D5S818, D7S821, D9S925, D13S317, D19S253) and one trimeric (D6S1043) short tandem repeats loci to study the allele frequency and the applicability of genetic variation in these loci in forensic case works. For 310 unrelated Koreans DNA was isolated from peripheral blood using phenol/chloroform method. Quadruplex I was consisted of D4S2368, D6S1043, D7S821, D9S925 and quadruplex II D3S2406, D5S818, D13S317, D19S253. The amplified products were analyzed by 5%polyacrylamide gel electrophoresis followed by silver staining. The heterozygosity in each loci ranged 92.91-66.13%, and PD(Power of Discrimination) was above 0.85 in each loci. Every loci except D6S1043 followed hardy-Weinberg expectation. The cumulative PI was low as 1.65x10-10. Two mutations were noted, one in D19S253 and the other in D9S925 among 234 gametes. With these results above eight STR loci studied here proved to be highly polymorphic enough to be used in forensic field. This study provides valuable population data in these loci for Korean.
DNA ; Electrophoresis ; Gene Frequency ; Genetic Variation ; Germ Cells ; Microsatellite Repeats ; Polymerase Chain Reaction* ; Silver Staining

Two quadruplex PCR reactions were designed for 7 tetrameric (D3S2406, D4S2368, D5S818, D7S821, D9S925, D13S317, D19S253) and one trimeric (D6S1043) short tandem repeats loci to study the allele frequency and the applicability of genetic variation in these loci in forensic case works. For 310 unrelated Koreans DNA was isolated from peripheral blood using phenol/chloroform method. Quadruplex I was consisted of D4S2368, D6S1043, D7S821, D9S925 and quadruplex II D3S2406, D5S818, D13S317, D19S253. The amplified products were analyzed by 5%polyacrylamide gel electrophoresis followed by silver staining. The heterozygosity in each loci ranged 92.91-66.13%, and PD(Power of Discrimination) was above 0.85 in each loci. Every loci except D6S1043 followed hardy-Weinberg expectation. The cumulative PI was low as 1.65x10-10. Two mutations were noted, one in D19S253 and the other in D9S925 among 234 gametes. With these results above eight STR loci studied here proved to be highly polymorphic enough to be used in forensic field. This study provides valuable population data in these loci for Korean.
DNA ; Electrophoresis ; Gene Frequency ; Genetic Variation ; Germ Cells ; Microsatellite Repeats ; Polymerase Chain Reaction* ; Silver Staining