• ALGAE
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• v.25 no.4
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• pp.197-204
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• 2010

Cytoskeletal changes were observed during cell division of the green alga Zygnema cruciatum using flourescein isothiocynate (FITC)-conjugated phallacidin for F-actin staining and FITC-anti-$\alpha$-tubulin for microtubule staining. Z. cruciatum was uninucleate with two star-shaped chloroplasts. Nuclear division and cell plate formation occurred prior to chloroplast division. Actin filaments appeared on the chromosome and nuclear surface during prophase, and the F-actin ring appeared as the cleavage furrow developed. FITC-phallacidin revealed that actin filaments were attached to the chromosomes during metaphase. The F-actin ring disappeared at late metaphase. At telophase, FITC-phallacidin staining of actin filaments disappeared. FITC-anti-$\alpha$-tubulin staining revealed that microtubules were arranged beneath the protoplasm during interphase and then localized on the nuclear region at prophase, and that the mitotic spindle was formed during metaphase. The microtubules appeared between dividing chloroplasts. The results indicate that a coordination of actin filaments and microtubules might be necessary for nuclear division and chromosome movement in Z. cruciatum.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.2
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• pp.60-66
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• 2019

Objective: To compare black rice (Oryza sativa L) extract with three different staining methods for human sperm head assessment. Methods: Semen samples were collected from 34 volunteers. Four smears of each ejaculate were prepared for staining using the rapid Papanicolaou (PAP) stain, SpermBlue, DipQuick, and black rice extract. The percentage of defective sperm heads (mean${\pm}$standard deviation) was compared. Results: Black glutinous rice extract, a natural dye, was used instead of hematoxylin to stain the nuclei of the sperm heads. The percentage of defective sperm heads showed a significant difference between black rice extract and DipQuick (p= 0.000). In contrast, black rice extract and rapid PAP showed no statistically significant difference (p= 0.974). A strong correlation (r = 0.761) was found between the findings obtained using rapid PAP and black rice extract. In contrast, a weak correlation (r = 0.248) was obtained between DipQuick and black rice extract for the percentage of defective sperm heads. Conclusion: The results showed good agreement and a strong correlation between the rapid PAP and black rice extract stains. The advantages of black rice extract as a novel substitute for hematoxylin for nuclear staining include ease of preparation, local availability, and favorable nuclear staining properties. Further studies could also focus on comparing staining techniques in clinical samples.

• Proceedings of the Korean Nuclear Society Conference
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• 2002.05a
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• pp.388-388
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• 2002

• Journal of Radiation Protection and Research
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• v.29 no.1
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• pp.9-15
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• 2004

Firstly, we compared the two staining techniques, Giemsa and Acridine orange, to determine micronuclei on samples of cultures of five healthy human peripheral blood lymphocytes after ${\gamma}-irradiation\;(^{137}Cs)$ in dose ranges of 0 to 800cGy. It was found that the Acridine orange staining method gives more reliable results than the usual Giemsa staining method in micronucleus tests. Moreover, the frequency of micronuclei in cytokinesis-blocked human B-lymphocytes was studied after in vitro irradiation in dose ranges of 0 to 50cGy. After setting and separating the B-lymphocytes, the frequency of radiation-induced micronuclei were observed as the end-point markers for the low-dose radiation dosimetry after staining with Giemsa and Acridine orange dyes. The micronuclei frequency in B-lymphocytes was significantly elevated from 10 to 30cGy ${\gamma}-irradiation$. The determination of micronuclei in B-lymphocytes after staining with Acridine orange was higher than that of Giemsa. The frequency of micronuclei in B-lymphocytes was observed to be at least two times higher than those of T-lymphocytes Giemsa in dose increasing. Therefore, the determination of low-dose radiation-induced micronuclei in B-lymphocytes after staining with Acridine orange is likely to have the greatest potential in the estimation of low dose radiation exposure.

• The Korean Journal of Nuclear Medicine
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• v.38 no.6
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• pp.511-515
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• 2004

Purpose: The sodium-iodide symporter (NIS) expression is an important factor in determining the sensitivity of radioiodine therapy in well-differentiated thyroid cancers. Several previous studies for the expression of NIS in thyroid tissues show diverse results. To investigate whether there is difference between methods in determining the expression of NIS in thyroid tissues of patients with thyroid nodules, we measured the expression ot NIS using two different methods (RT-PCR and immunoshistochemical staining) and compared the results. Materials & Methods: We measured the expression of NIS by reverse transcriptase-polymerase chain reaction (RT-PCR) and also by immunohistochemical staining using anti-NIS antibody in thyroid cancers and other benign thyroid diseases. We compared the results of each method. We included 19 papillary carcinomas, 1 follicular carcinoma, 7 medullary carcinoma, 4 adenomas and 7 nodular hyperplasias. Results: By RT-PCR analysis, 10 of 19 papillary carcinomas expressed NIS, but 1 follicular cancer didn't express NIS. By immunohistochemical staining, 15 of 19 papaillary carcinomas express NIS, but 1 follicular lancer didn't express NIS. There was a significant correlation between the semiquautitative results of RT-PCR and immunohistochemical staining of NIS expression. (p<0.01) Conclusion: Our data demonstrated that the expression of NIS in thyroid cancers and other benign diseases investigated by RT-PCR and immunohistochemical staining correlated well each other. However, by immunohistochemical staining, more NIS expression was found.

• Journal of Korean Physical Therapy Science
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• v.10 no.1
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• pp.198-205
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• 2003

We evaluated the effect of low power GaAsAl laser on re-epithelization in full-thickness excisional wound of rat skin. Two full-thickness excisions were made on the back of the experimental animals. Low power laser applications with 10mW intensity were treated experimental animals twice a day for 7 days. On the seventh postoperative day the quantitative analysis of re-epithelization was performed using immunohistochemical staining for proliferating cell nuclear antigen (PCNA). The majority of PCNA immunoreactive cells was observed at epithelial cells in the margin of full thickness excisional wound. The low power laser treatments significantly increased the number of PCNA immunoreactive cell as compared to that of non treated animal group (p<0.01). The shape of PCNA immunoreactive cell appeared as small dark, round to ovoid structures. Most PCNA immunoreactive cells exhibited a high intensity of staining that contrasted sharply with the surrounding background. In conclusion, these findings suggest that GaAlAs laser treatments effectively enhance the epithelial wound healing by the stimulating cell proliferation. Furthermore, the majority of cell proliferation occurred in the margin of full thickness excisional wound.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.9
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• pp.3959-3963
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• 2014

The tumour suppressor genes, p53 and pRb, are known to play important roles in neoplastic transformation. While molecular routes to the uncontrolled growth of hepatocytes, leading to primary liver cancer have generated considerable interest, the roles of p53 and pRb mutations in hepatocellular carcinoma (HCC) and hepatoblastoma (HB) remain to be clarified. We examined the immunohistochemical expression of p53 and pRb gene products in 26 HCC and 9 HB, sampled into tissue microarray blocks. 10 (38%) of 26 HCC showed > 10% tumour nuclear staining for p53 protein, 3 of these also being HbsAg positive. Conversely, none of 9 HB expressed nuclear p53 immunopositivity. Some 24 (92%) HCC and 8 (89%) HB showed loss of pRb nuclear expression. Two of the 26 HCC and one of the 9 HB showed >10% tumour nuclear staining for pRb protein. Our results suggest that p53 does not have an important role in the development of HB but may contribute in HCC. There is also loss of pRb expression in the majority of HCC and HB, supporting loss of pRb gene function in the hepatocarcinogenesis pathway. However, a comparison of the staining profiles of p53 and pRb proteins in HCC and HB did not reveal a consistent pattern to differentiate between the two types of tumours immunohistochemically. Hence the use of p53 and pRB protein expression has no contribution in the situation where there is a diagnostic difficulty in deciding between HCC and HB.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.8
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• pp.3627-3630
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• 2012

The epithelial cell adhesion molecule (EpCAM) is a pan-epithelial differentiation antigen that is expressed on almost all carcinomas. However, a role in rat liver carcinogenesis has never been reported previously. Thus, its expression was investigated herein in rat liver tumors induced by diethylnitrosamine (DEN). Twenty male 5-week-old F344 rats were used in this experiment. Mini-osmotic pumps containing doses of 47.5 mg of DEN were inserted into the abdominal cavity of each animal to initiate liver carcinogenesis. All animals were sacrificed at 26 weeks after DEN treatment. At necropsy, hepatic masses were processed for histopathological examination, which revealed forty-four hepatocellular adenomas (HCAs) and twenty hepatocellular carcinomas (HCC). Tumors were immunohistochemically analyzed for EpCAM, proliferating cell nuclear antigen (PCNA) and co-localization of the two. EpCAM expression was mainly detected in hepatic tumor cells, showing a cytoplasmic staining pattern. However, expression was also slightly observed in normally-appearing surrounding hepatic cells. PCNA expression was highly detected in tumor cells, showing nuclear staining. Double staining of EpCAM and PCNA in tumors showed many cells with co-localization. Taken together, EpCAM and PCNA expression were increased in DEN-induced tumors and many tumor cells showed co-expression. It is suggested that EpCAM may increase during DEN-induced tumors, possibly associated with cell proliferation.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.407-414
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• 2011

The development of embryos reconstructed by somatic cell nuclear transfer (SCNT) is dependent upon numerous factors. Central to development is the quality and developmental competence of the recipient cytoplast and the type of the donor nucleus. Typically metaphase of the second meiotic division (MII) has become the cytoplast of choice. Production of a cytoplast requires removal of the recipient genetic material, however, it may remove proteins which are essential for development or reduce the levels of cytoplasmic proteins to influence subsequent reprogramming of the donor nucleus. In this study, enucleation at MII did not affect the activities of either MPF or MAPK kinases. Immunocytochemical staining showed that both Cyclin B1 (MPF) and Erk1/2 (MAPK) were associated with the meiotic spindle of AI/TI oocytes with little staining in the cytoplasm, however, at MII association of both proteins with the spindle had reduced and a greater degree of cytoplasmic distribution was observed. The analysis of oocyte proteins removed during enucleation is a difficult approach to the identification of factors which may be depleted in the cytoplast. This is primarily due to the large numbers of aspirated karyoplasts which would be required for the analysis.

• 대한핵의학회:학술대회논문집
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• 2004.11a
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• pp.408.2-408.2
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• 2004