• Korean Journal of Veterinary Research
• /
• v.36 no.2
• /
• pp.277-281
• /
• 1996

A technique to improve the analysis of micronuclei(MN) in lymphocytes as a cytogenetic indicator is reported. For the purpose of diminishing the variation of the result from individual reader and making it easier to distinguish accurately a cytokinesis blicked(CB) lymphocyte and micronuclei, we tried a modified one-step silver staining technique as a method for detection of the argyrophilic nucleolar organizer region(AgNOR) with or without conventional Giemsa stain in the slide from CB method. Compared with the conventional Giemsa stain, the preparation processed with this method are especially useful for the accurate analysis of MN of cultured lymphocyte with cytochalasin B. This method will be a useful technique for automated calculation of MN.

• Tuberculosis and Respiratory Diseases
• /
• v.42 no.4
• /
• pp.513-521
• /
• 1995

Background: Nucleolar organizer regions(NORs) are chromosomal segments encoding for ribosomal RNA and associated with argyrophilic nonhistone protein. Ribosomal RNA genes ultimately direct ribosome and protein synthesis, and it has been suggested the numbers of NORs detected in the cell may reflect nuclear and cellular activity. This study was performed to evaluate the applicability of AgNORs to the diagnosis of squamous cell carcinoma of the lung. Method: The one step silver methods(AgNORs) was used to stain NORs in the routinely processed, formalin fixed, paraffin embedded sections of 36 cases of squamous cell carcinoma of the lung obtained by surgical resection of primary tumor. In each specimen, 100 tumor cells and 100 normal cells adjacent to the tumor chosen at random were examined under an oil immersion lens at a magnification of ${\times}1000$. The mean number of AgNORs per nucleus was calculated for each specimen. Results: The mean number of AgNORs per nucleus(mAgNORs) of normal bronchial epithelium and squamous cell carcinoma of the lung was $1.74{\pm}0.25$ and $4.05{\pm}0.80$, respectively. The difference of mAgNOR between normal and tumor tissue was statistically significant(p<0.001). There was no statistical difference among tumors of different stages. The difference of mAgNOR between normal and tumor tissue was statistically significant in each TNM stage(p<0.05). Conclusion: Mean AgNOR count may be used as a useful marker for the differential diagnosis of benignancy and malignancy, and proliferative activity of the cell in squamous cell carcinoma of the lung. But there was no statistical difference in mean AgNOR count among tumors of different surgical stages. Further studies for the application of mAgNORs to the diagnosis of other histologic types and cytologic specimens of the lung cancer are needed.

• Toxicological Research
• /
• v.14 no.4
• /
• pp.489-494
• /
• 1998

In the present study, we have evaluated cytotoxic effects of Taraxaci Herba extract on human skin melanoma cells. The light microscopic study showed morphological changes AG-NOR (argyrophylic nucleolar organizer region) by silver chloride stain, and glycoprotein by PAS reaction of the treated cells. Disruptions in cell organelles were determined by SRB assay.

• Biomedical Science Letters
• /
• v.10 no.4
• /
• pp.361-366
• /
• 2004

The purpose of this study was to determine the effect of high voltage pulsed current (HVPC) stimulation on proliferative activities of basal keratinocytes by measured nucleolar organizer region (NOR) expression and thickness of spinous layer in aged rat skin. Fifty-one weeks old twelve male Sprague-Dawley rats (300∼350g) were divided into control and HVPC stimulation groups. Each animal's hair on the back were removed. The HVPC stimulation group received an negative monophasic twin peak pulsed current stimulation with 50 V, while the control group was given the same treatment without electricity. The rats were sacrificed after 3 weeks. The biopsy specimens were fixed in formalin, embedded in paraffin and stained with hematoxyline-eosin and silver nitrate. The thickness of basal to granular layer of the epidennis were measured using a light microscope and computerized image analysis system. The number of argyrophilic nucleolar organizer region (AgNOR) were counted using a light microscope and computerized image analysis system and calculated as the mean number of AgNOR per nucleus in the basal keratinocyte. By using a Student's t-test, an increase in the thickness of basal-spinous layer (P<0.001) of epidermis can be observed in HVPC stimulation rats as compared with the control rats, whereas the thickness of the granular layer is not affected. A Student's t-test showed a significantly higher mean NOR number per nucleus of the basal keratinocyte in the HVPC stimulation rats than control rats (P<0.001). There was significantly positive correlation between the NOR number and the thickness of basal-spinous layer (r=0.80, P<0.05). These results suggest that the HVPC stimulation may increase the thickness of spinous layer in the epidennis due to increased proliferative activities of basal keratinocytes in epidennis in aged rat skin.

• The Journal of Korean Physical Therapy
• /
• v.12 no.1
• /
• pp.1-7
• /
• 2000

The purpose of this study was to determine the effect of microamperage electrical stimulation on the number of argyrophilic nucleolar organizer region (AgNOR) in rat skin. Twenty four male Sprague-Dawley rats were divided into electrical stimulation and control group. Bach animals hair on the back was removed. The electrical stimulation group received an positive rectangular positive electrical stimulation with $500{\mu}A$, while the control group was given the same treatment without electricity. The rats were sacrificed at 4 and 7 day of stimulation, respectively. The biopsy specimens were fixed in formalin, embedded in paraffin and stained with silver nitrate. The AgNOR were counted using a light microscope and computerized image analysis system and calculated as the mean number of AgNOR per nucleus in the epidermal keratinocyte. In control skin, the mean AgNOR count of epidermal keratinocyte at 4 and 7 day were 1.67 and 1.72, whereas electrical stimulated rat had mean AgNOR counts of 2.0 and 2.14, respectively. A Student's t-test showed a significantly higher mean AgNOR number at 4 ana 7 day in the electrical stimulated rats than control rats (p<0.05). The microamperage electric current stimulation increased the epidermal AgMOR expression in incisional wound skin. These results suggest that the microamperage electrical stimulation may promote migration and proliferative activity of epidermal keratinocyte in surgical wound.

• Environmental Mutagens and Carcinogens
• /
• v.18 no.2
• /
• pp.104-108
• /
• 1998

In the present study, we have evaluated cytotoxic effects of Taraxaci Herba extract in human oral epitheloid carcinoma cells. An antitumor activity was measured by colorimetric assays using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and sulforhodamine protein B (SRB). The light microscopic study showed morphological changes, Ag-NOR (argyrophylic nucleolar organizer region) number and PAS positive reaction or the treated cells. These results obtained are as follows : MTT and SRB quantities were significantly decreased in cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations. The number of Ag-NORs were significantly decreased in cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations and the rate of Ag-NORs was shifted to left side (one Ag-Nounucleus was increased and five Ag-NORs/nucleus were decreased) by the high concentration. PAS reaction of cultured KB cells treated with 10$^{-2}$ /mg/ml and 10$^{-3}$ /mg/ml concentrations was negative. These results suggest that Taraxaci Herba retains a potential antitumor activity.

• YAKHAK HOEJI
• /
• v.43 no.2
• /
• pp.173-179
• /
• 1999

In the present study, we have evaluated cytotoxic effects, antitumor activities and metastasis inhibitory effects of Palsun brewing water in NIH 3T3 cells, human epitheloid carcinoma cells, and human skin melanoma cells. The light microscopic study showed morphological changes, AG-NOR (argyrophylic nucleolar organizer region) by silver chloride stain, and glycoprotein by PAS (periodic acid stain) reaction of the treated cells. Disruptions in cell organelles were determined by colorimetric methods; MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide) and SRB (sulforhodamine B protein) assays. These results suggest that Palsun Brewing Water retains no cytotoxic effects in NIH 3T3 cells and a growth-inhibitory activity in cancer cell lines.

• Korean Journal of Veterinary Research
• /
• v.30 no.4
• /
• pp.459-464
• /
• 1990

Inhibitory effect of caffeine on NIH3T3 cell proliferation was studied by using [$^3H$]thymidine incorporation and a modified one-step silver staining technique. The latter technique shows argyrophilic nucleolar organizer region-associated proteins (Ag-NORs), which are seen in nuclei as black dots. The result was compared with the counts of [$^3H$] thymidine incorporation. The results were summarized as follows; 1. The Ag-NORs numbers of NIH3T3 cells were $6.81{\pm}1.38$ at 24 hrs, $7.13{\pm}1.26$ at 48 hrs, $8.50{\pm}2.04$ at 72 hrs after incubation. 2. The numbers of Ag-NORs were significantly decrease in caffeine treated groups (p<0.01). 3. The counts of [$^3H$] thymidine incorporation were similar to the result of using Ag-NORs staining technique. It is concluded that Ag-NOR is a rapid, simple and compatible method to quantitate cell proliferation as compared with [$^3H$]thymidine incorporation.