Journal of Pathology and Translational Medicine

Volume 34(11); November 2000

Original Articles

Distribution of Free Radicals in Reperfusion Injury after Transient Brain Ischemia.: Eunkyoung Kwak, Hyungho Suh, Jiyoung Park, Yunsup Kum, Taein Park, Jungwan Kim, Yoonkyung Sohn; Korean J Pathol. 2000;34(11):893-900.

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Abstract PDF: Free radicals are known as an important factor which may act on reperfusion injury after transient or permanent brain ischemia. Numerous studies about cytotoxic function of free radical have been done. Most of these studies demonstrate the function of free radical in reperfusion injury by using radical scavenger or antioxidant as inhibitor of radicals. We used a modification of Karnovsky's Mn2 /diaminobenzidine (DAB) technique to demonstrate intravascular free radicals following transient occlusion and reperfusion of one middle cerebral artery in Sprague-Dawley rats. The MCA was occluded for 2 hours using an intraluminal suture method. The reperfusion time after transient ischemia was 1 hour, 6 hours, and 24 hours, respectively. Animals were perfused transcardially with solution containing Mn2 and DAB. After DAB perfusion, the brains were removed promptly, sectioned in frozen, and stained with methylene blue for light microscopic examination. Upon light microscopic examination, free radicals were confined within intravascular lumen and the amount of deposits increased according to the duration of reperfusion. Upon electron microscopic examination, free radicals were located in nuclear membrane and membrane of mitochondria and RER, and demonstrated as electron dense deposits. In addition, cell processes of the neuron revealed an electron dense deposits beneath the inner side of the membrane. In conclusion, free radicals demonstrated in the reperfusion injury area indicate that free radical acts as an important cytotoxic factor. Intracellular localization of free radicals may explain the relationship between free radical and delayed neuronal injury.

Expressions of MAGE-3, PCNA, p21, and p53 Proteins in the Hypopharyngeal Squamous Cell Carcinoma Cell Line (PNUH-12) Analysed by Bivariate Flow Cytometry.: Hee Kyung Chang, Deok Jun Kim, Hwan Jung Roh, Bang Hur, Kang Dae Lee, Spagnoli Spagnoli; Korean J Pathol. 2000;34(11):901-908.

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Abstract PDF: MAGE (melanoma antigen gene) is a tumor specific shared antigen, presented by HLA class I molecules, which is recognized by cytotoxic T lymphocytes. MAGE proteins are expressed in malignant tumor cells, in contrast to no expression in normal or benign tissues except for testis and placenta. MAGE might be a potential target for immunotherapy of malignant tumors. However, its biological aspects associated with cell cycle are not yet described. The flow cytometry is a useful tool for objective and quantitative analyses of heterogenous tumor cell population. To understand the status of MAGE related to cell cycle and its relationship with p53 as the G1 checkpoint regulator, p21, and PCNA as a proliferative index, we investigated expression of MAGE-3 protein, mutant p53, p21, and PCNA by flow cytometry and immunohistochemical stain. In addition, double stains for MAGE-3/p53, p53/PCNA, and p53/p21 were analysed with bivariate flow cytometry. DNA histograms using MAGE-3/PI (DNA) and p53/PI (DNA) were also analysed. The cell line (PNUH- 12) used for this study originated from a hypopharyngeal squamous cell carcinoma, which has point mutation (exon 7, C-->G) of p53. The expression rate of MAGE-3 was 83%, PCNA 85%, and p53 81%. No expression for p21 was identified. MAGE-3 was expressed in cytoplasm, while both PCNA and p53 were expressed in nuclei of tumor cells. With bivariate analyses, coexpression rates of MAGE-3/p53 and p53/PCNA were 0.96 and 0.97, respectively. Both MAGE-3 and p53 showed constantly high level throughout the cell cycle. These results suggest that expression of MAGE-3 and mutant p53 is not dependent on the cell cycle. p21 seems to be inactivated.

A Clinicopathologic Study of 53 Gastrointestinal Mesenchymal Tumors.: Young Kyung Bae, Dong Sug Kim, Mi Jin Gu, Joon Hyuk Choi, Mi Jin Kim, Young Jin Kim, Won Hee Choi, Sun Kyo Song, Koing Bo Kwun; Korean J Pathol. 2000;34(11):909-918.

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Abstract PDF: The gastrointestinal mesenchymal tumors (GIMTs) form a heterogenous group with controversy centering on both the cell of origin and the prediction of clinical behavior. They include a small group of tumors with mature smooth muscle or Schwann cell differentiation and a larger group with inconsistent or no evidence of differentiation. Tumors in the latter are now referred to as gastrointestinal stromal tumors (GISTs). A clinicopathologic and immunohistochemical study was performed on 53 cases of GIMTs to identify cellular differentiation and predictors of clinical behavior. Fifty three cases of GIMTs could be histologically and immunophenotypically divided into three categories, 6 leiomyomas (11.3%), 4 schwannomas (7.6%), and 43 GISTs (81.1%). All leiomyomas (SMA desmin ) and schwannomas (S-100 ) were located in stomach and negative for CD34 and CD117. Thirty nine cases of GISTs were either CD34 (n=26) or CD117 (n=23) immunoreactive. Of these 39 GISTs, 26 were negative for myoid (SMA, desmin) and neural marker (S-100), 10 SMA desmin-S-100-, two SMA-desmin-S-100 , and one SMA desmin-S-100 . Two out of 4 GISTs, which were negative for CD34 and CD117, were immunohistochemically considered leiomyosarcoma (SMA desmin ). GISTs of small intestine had a tendency to be malignant than those of stomach. Pathologic grade of GISTs was not correlated with cellular differentiation. In 29 GISTs with clinical follow-up information, tumor size, mitotic counts, Ki-67 labelling index, tumor necrosis, mucosal invasion, and CD34 expression were significantly correlated with metastasis/recurrence.

Correlation between bcl-2 and Caspase-3 Expression and Proliferating Activity in Squamous Neoplasia of the Uterine Cervix.: Kyung Sun Park, Mi Seon Kang, Hye Kyoung Yoon; Korean J Pathol. 2000;34(11):919-926.

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Abstract PDF: Detailed mechanism of uterine cervical cancer progression still remains unclear. Altered programmed cell death (apoptosis) and cellular proliferation are associated with the development of neoplasia. The authors investigated the expressions of bcl-2, which inhibit apoptosis, and caspase-3, which is involved in the induction of apoptosis and has been considered to be correlated with apoptosis, and proliferating activity according to the degree of malignancy in the squamous neoplasia of the uterine cervix. Correlation between bcl-2 and caspase-3 expression and proliferating activity was done. The materials were low grade squamous intraepithelial lesions (LSIL, n=15), high-grade squamous intraepithelial lesions (HSIL, n=15), microinvasive squamous cell carcinoma (n=15), and squamous cell carcinoma (n=15). Immunohistochemical stainings for bcl-2, caspase-3, and MIB-1 were done. bcl-2 and MIB-1 expressions were progressively increased in accordance with the increasing degree of malignancy, but caspase-3 immunoreactivity was higher in LSIL than invasive cancers. There was an inverse relationship between bcl-2 and caspase-3 expression, but the difference did not reach statistical significance. No significant correlation between MIB-1, bcl-2, and caspase-3 expressions was observed. These results suggest that an inhibition of apoptosis and the augmentation of proliferating activity of tumor cells might be separately involved in the development of the cervical squamous neoplasia.

Telomerase Activity and Expression of MIB-1 and bcl-2 in Human Chorionic Villi from Early and Term Normal Pregnancy.: Jung Sook Cho, Young Soon Kang, In Gul Moon, Bum Chae Choi, Jong Pyo Lee, Hoon Taek Lee, Sung Ran Hong; Korean J Pathol. 2000;34(11):927-933.

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Abstract PDF: Telomerase is an enzyme that maintains telomeres and prevents telomere shortening, and may be linked with cellular proliferation or the aging process. The purpose was to examine telomerase activity in human chorionic villi from early and term normal pregnancies, and to analyze the correlation of telomerase activity (TA) with MIB-1 & bcl-2. A total of 37 placentae were obtained from 16 early and 21 term pregnancies. TA was assayed by telomeric repeat amplification protocol, and immunohistochemical staining was performed for MIB-1 & bcl-2 expression. TA & MIB-1 expression were strong in early placenta, but bcl-2 was highly expressed in term placentae. Thirteen (81.25%) of 16 early placentae showed TA, but only 2 (9.52%) of 21 term placentae expressed TA (p<0.01). MIB-1 was observed in nuclei of cytotrophoblast, and the expression rate was 16.09% in early placentae and 2.87% in term placentae (p<0.01). bcl-2 was observed only in the cytoplasm of syncytiotrophoblast. Term placenta demonstrated stronger expression of bcl-2 compared to early placentae (p<0.05). These findings suggest that TA, MIB-1 & bcl-2 expression are critically regulated over the course of gestation: cytotrophoblast, main cells of early chorionic villi, may be a common source of telomerase and proliferative activity. The TA showed good correlation with cellular proliferative activity. Syncytiotrophoblast, may be a main source of bcl-2 expression which is stronger in the term placentae.

Expression Patterns of S100A6 Gene in Human Thyroid Diseases.: Joo Heon Kim, Jae Wha Kim, Seon Young Yoon, Jong Hyuck Joo, In Seong Choi, Mee Ja Park; Korean J Pathol. 2000;34(11):934-940.

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Abstract PDF: S100A6 (calcyclin) is a member of the S100 family and has been originally isolated from the cDNA library of Syrian baby hamster kidney cells. The S100A6 gene expression is reported to remain high throughout the cell cycle following induction by serum or growth factors, suggesting that the gene may be required for cell cycle progression. Nevertheless, the role that S100A6 may play in tumor progression remains unknown. In this study, we have explored the expression patterns of S100A6 gene in human thyroid tissues by northern blot analysis. Using the S100A6 monoclonal antibody, we carried out the immunohistochemical staining to determine the distribution/localization of S100A6 protein within tumor or non-tumorous cells of the thyroid. To modulate the regulation of endogenously expressed S100A6 protein in the intracellular level, overexpressed or anti-sense treated transfectant was constructed by using the eukaryotic expression vector. As a result, immunohistochemistry for S100A6 showed a strong positivity in the malignant tumors of thyroid and a high expression level of S100A6 protein affected cell proliferation in the overexpressed transfectant. These findings suggest that S100A6 may be involved in the tumor pathogenesis and provides another parameter for the differentiation of malignant and benign lesions. A well defined monoclonal antibody against S100A6 protein is now available for the immunohistochemical studies of the various thyroid tissues.

Molecular Diagnosis of Cutaneous T Cell Lymphoproliferative Diseases.: Ji Young Park, Myung Hoon Lee, Eun Kyung Kwak, Dong Ja Kim, Tae In Park, Han Ik Bae; Korean J Pathol. 2000;34(11):941-949.

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Abstract PDF: It is often problematic to diagnose T-cell lymphoproliferative disorders of the skin because of the difficulty in establishing clonality in paraffin-embedded tissue. We used polymerase chain reaction single strand conformational polymorphism (PCR-SSCP) and heteroduplex analysis in paraffin embedded tissue to detect clonal rearrangement of T-cell receptor gamma (TCRgamma) gene in 17 T-cell lymphoproliferative disorders and 6 atypical lymphoproliferative diseases. We used polymerase chain reaction to detect TCR beta gene rearrangement in 8 of 17 cases which did not show TCRgamma gene rearrangement. Jurkat cell lines were used as monoclonal controls. DNA was extracted from 5 biopsies of T-cell lymphomas, 10 biopsies of mycosis fungoides, 2 biopsies of lymphomatoid papulosis, and 6 biopsies of atypical lymphoproliferative lesions. We detected monoclonality in 5 of 5 T-cell lymphoma cases, 2 of 2 lymphomatoid papulosis cases, 6 of 10 mycosis fungoides cases, and 2 of 6 atypical lymphoproliferative disease cases. We conclude that nonradioactive PCR-SSCP for TCR gene rearrangement analysis is a useful adjunct to routine histological and immunophenotypic methods in the diagnosis of cutaneous T cell lymphoproliferative disorders in paraffin embedded tissue.

Case Reports

Dendriform Pulmonary Ossification: A case report.: Chan Kwon Jung, Kyo Young Lee, Chang Suk Kang, Sang In Shim, Byung Kee Kim; Korean J Pathol. 2000;34(11):950-952.

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Abstract PDF: The dendriform pulmonary ossification (DPO) is a rare condition of unknown origin in which branching mature bony spicules, usually containing marrow, are found within the alveolar septa. DPO manifests slow progression over many years or may remain unchanged; spontaneous regression has not been recorded. Most patients have no symptoms directly attributed to the ossification, although they may have symptoms due to the underlying fibrotic process. We experienced a case of DPO in 38 year-old-man who presented with cough and sputum for a month. The chest X-ray showed marked coarsened interstitial lung markings in both lungs, especially in the lower lobes. Open lung biopsy was done. Grossly, there were significant dendriform osseous structures. Histologically, branching arrays of mature bone were found in the interstitium and occasionally in alveolar spaces. Some bony trabeculae contained fatty or cellular marrow. The alveolar septa showed fibrous thickening with chronic inflammation. The transition between fibrosis and bone tissue was observed. Our case suggests that dendriform pulmonary ossification may be a rare special manifestation of chronic fibrosing interstitial inflammation of the lung. Osseous structures seem to derive from metaplastic bone formation in the vicinity of undergoing fibrous process.

Epithelioid Angiomyolipoma of the Kidney: A case report.: You Kyung Kim, Jong Sil Lee, Ky Hyun Chung, Sun Hoo Park, Gyung Hyuck Ko; Korean J Pathol. 2000;34(11):953-956.

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Abstract PDF: Angiomyolipoma is considered by many authors to be a hamartoma, occurring in a sporadic form or in association with tuberous sclerosis. This lesion consists of thick walled blood vessels, smooth muscle, and mature adipose tissue in varying amounts. We have experienced a case of the angiomyolipoma composed of monotypic epithelioid cells. The patient was a 69-year-old female. Clinically, there was no evidence of tuberous sclerosis. Microscopically, the tumor was composed of polygonal cells with abundant eosinophilic granular or clear cytoplasm, pleomorphic nuclei, prominent nucleoli, and multinucleated giant cells. The tumor cells showed positive reaction for HMB45, CD68, smooth muscle actin, and S-100, and negative reaction for epithelial membrane antigen, cytokeratin, vimentin, desmin, CD34, estrogen receptor, and progesterone receptor. Ultrastructual analysis showed the presence of glycogen, mitochondria, and other microorganelles in neoplastic cells. Melanosome or premelanosome was not identified.

Trichofolliculoma in Korean Patients: A report of six cases.: Kyoung Mee Kim, Jai Kyung Ko, Anhi Lee, Chang Suck Kang, Byung Kee Kim, Seok Jin Kang; Korean J Pathol. 2000;34(11):957-960.

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Abstract PDF: Trichogenic tumors are a very rare group of primary cutaneous neoplasms and a few tumors have been recognized. Trichofolliculoma is a benign, papular, organoid adnexal tumor of the skin and presents as a solitary skin-colored nodule. We report six cases of trichofolliculoma occurred in Korean patients consisting of three male and three female patients. The duration of disease was variable from 3 months to 5 years. The site of tumor was limited to the face and included eyelid in two cases, nose in two cases, cheek and posterior aspect of ear in each one case. The size of the mass varied from 4 to 8 mm in diameter. Microscopically, all the cases showed large central cystic hair follicle with numerous secondary follicular buds, producing a radiating pattern. The stroma was fibrotic and well demarcated from the adjacent dermis.

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