Journal of Pathology and Translational Medicine

Volume 29(5); October 1995

In Vitro

The Effects of Excitatory Amino Acids and Their Receptors on Neuronal Damage of Rat Brain in Hypoxic-Ischemic Encephalopathy.: Heasoo Koo; Korean J Pathol. 1995;29(5):545-562.

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Abstract: Since the role of excitatory amino acids such as glutamate and aspartate and their receptors mediating cellular injury through various mechanisms were known in hypoxic-ischemic injury and associated diseases of central nervous system, blocking agents for transmitter release or receptors have been tried to reduce the cellular damages and subsequent sequelae experimentally. Several in vitro studies suggested two kinds of glutamate neurotoxicity: (1) rapid toxicity due to influx of sodium or chloride with resultant cellular edema and consequent damage, which is associated with N-methyl-D-Aspartate(NMDA) as well as non-NMDA receptors, (2) calcium mediated delayed toxicity associated mainly with NMDA receptor. This study was conducted to investigate the role of rapid toxicity in hypoxic-ischemic injury. Early lesions of 30 minutes to 24 hours after hypoxic-ischemic insult were examined by autoradiography with radiolabelled glutamate and kainitic acid (KA) as well as light and electron microscopy. Late changes were evaluated on formaldehyde-acetic acid-methanol(FAM) fixed brain 1 week after the insult. Cornus ammonis(CA) l of hippocampus showed the highest density of NMDA receptors, which was decreased constantly from 2 hours to 24 hours. In contrast, CA3 of hippocampus showed the highest density of KA receptors, which was the lowest at 6 hour and increased thereafter. Light microscopic examination showed the worst changes during 30 minutes to 6 hours. After 1 week, most of the cases showed degeneration of neurons and CAI and CA3 did not show the difference. Electron microscopic examination showed marked degenerative changes of neurons as well as neuropils starting from 30 minutes after the insult. In conclusion, rapid toxicity mediated by non-NMDA(KA) receptor seen in CA3 lead to permanent damage in 1 week old lesion.

Original Articles

Immunohistochemical Study of IL-4, IL-6, and TNF Expression in Cardiac Myxoma: Emphasis on Constitutional Symptoms of the Myxoma Patients.: Min Sun Cho, Soo Yeon Cho, Mi Jung Kim, Sung Sook Kim, Jeong Wook Seo, Woon Sup Han; Korean J Pathol. 1995;29(5):563-571.

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Abstract: It is well documented that cardiac myxomas are associated with immunologic features that can simulate systemic autoimmune diseases. Recently, it was reported that cardiac myxomas produce IL-6 constitutively, which could partly explain the immunologic features observed in these patients. However, only a few investigators have studied cytokines in regards to symptoms they may cause in patients with cardiac myxoma. Also there is very little information in the literature on the immunohistochmical localization of IL-6. We performed immunobistochemical stains for IL-4, TNF, and IL-6 on paraffm embbeded tissue of cardiac myxoma tissue. A bioassay of IL-6 activity in patient's serum and in cultured cells from fresh myxoma tissue was performed to ascertain the role of these cytokines in myxomas. In this study, we demonstrated inununohistochemically that there was a local overproduction of IL-4, TNF, and IL-6 in cytoplasm of the tumor cells in about half cases. Bioassays of the serum and cultured tumor cells revealed elevated IL-6 activities. Also these findings correlate to production of patient's constitutional symptoms with statistical significance (P<0.05). In conclusion, these results are of considerable importance in understanding the role of IL-4, TNF, and IL-6 in cardiac myxoma patient with constitutional symptoms, and have an impact on strategies for diagnosis and therapy of cardiac myxoma.

The Effects of Immunosuppressant and Immunostimulant on the Splenic Cell Subset of Rats Having Undergone Experimentally Induced Septal Fibrosis of Liver.: Mee Young Sol, Joon Yeon Kim, Sun Kyoung Lee; Korean J Pathol. 1995;29(5):572-583.

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Abstract: Although there have been many reports about the importance of the spleen's role in hepatic fibrogenesis, the exact mechanism is still uncertain. The author designed this study to evaluate splenic function on hepatic fibrogenesis. The degree of hepatic fibrosis and the population of splenocyte subsets were studied in the experimental animal model with fibrosis produced by injecting normal swine serum intra-peritoneally into Sprague-Dawley rats. The animals were divided into three groups; group A was subjected to injection of swine serum only, group B swine serum and complete Freund's adjuvant and group C swine serum and cyclosporin A. The experimental hepatic fibrogenesis by swine serum was augumented by coinjection with the adjuvant and inhibited by cyclosporin A. The study of the splenocyte subset revealed increased percentages of spienic B cell and CD4+ cell and a decreased percentage of CD8+ cell, and these changes of splenocyte subset were also augumented by the adjuvant and inhibited by cyclosporin A. The percent of monocytes was not significantly altered, although a tendancy of early decrease by the adjuvant was noted.

Nature of Stromal Cells in Cerebellar Capillary Hemangioblastoma: Immunohistochemical analysis.: Soon Won Hong, Tai Seung Kim, Ji Young Han; Korean J Pathol. 1995;29(5):584-589.

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Abstract: The origin of the stromal cell of cerebellar hemangioblastoma has long been studied electron microscopically and immunohistochemically. The results and theories about the stromal cell origin are variable and plentiful. However, the exact origin of the stromal cell remains controversial. The present study is aimed to elucidate the nature of the stromal cell of cerebellar hemangioblastoma. Ten cases of hemangioblastoma in Severance Hospital were used for immunohistochemical analysis of the stromal cell. The immunohistochemical staining of GFAP, S-100 protein, NSE, alpha-l-antichymottypsin, cytokeratin, CD 68, factor VIII related antigen, and synaptophysin were performed. The results were as follows; GFAP and S-100 protein were stained mainly but weakly in bellar capillary spindle cell and cellular process. NSE was stained mainly in foam cells, and 6 cases among them revealed strong reaction. ct-l-antichymotrypsin was stained in a few foam cells of 5 cases. Cytokeratin, CD 68, factor VIII related antigen, and synaptophysin showed negative reaction. Based on these results, it is considered that the origin of the stromal cell is histiocytic or neurogenic rather than glial. The weak positivity of GFAP and S-100 protein may support the neurogeriic origin but ct-l-antichymotrypsin positivity does not support the possibility. The positivity of GAP and S-100 protein supports the phagocytic action of histiocytic cell and suggests histiocytic origin rather than neurogenic.

Pathologic Characteristics of Colorectal Cancers with DNA Replication Errors.: Hoguen Kim, Yoon Mi Jeen, Jeong Yeon Shim, Chanil Park; Korean J Pathol. 1995;29(5):590-595.

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Abstract: Unstable microsatellite repeat sequences or DNA replication errors(RER) due to defective mismatch repair genes have been reported in a subset of sporadic colorectal tumors and in most tumors of patients of hereditary nonpolyposis colorectal carcinoma(HNPCC). To elucidate the clinicopathological correlation of these RER-positive cancers, we examined 16 cases of colorectal carcinoma of different histologic subtypes(6 cases of carcinoma with no gland formation, 5 cases of mucinous carcinoma and 5 cases of gland forming carcinoma). We detected RER in five cases. The patients with RER-positive cancers had a marked preponderance of carcinoma with no gland formations out of 6 carcinomas with no gland formation were RER-positive cancers) and of cancers proximal to splenic flexure(all of the RER-positive cases were proximal colon carcinomas). We conclude that RER-positive cancers have wiique pathologic features that may be useful for the screening and counselling of patients with hereditary colon cancers.

Expression of p53 Protein and c-erbB-2 Oncoprotein in Breast Carcinoma.: Eun Hee Lee, Dong Sug Kim, Tae Sook Lee, Soo Jung Lee; Korean J Pathol. 1995;29(5):596-606.

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Abstract: This study was conducted to evaluate the expression of p53 and c-erbB-2 using immuno-histochemical methods in 145 primary breast carcinomas and to correlate it with other histo-pathological prognostic factors. Invasive ductal carcinoma represented 129 of the cases. Expression of p53 protein and c-erbB-2 oncoprotein was present in 48% (62/129) and 30% (39/129) of invasive ductal carcinomas, respectively. The expression of p53 protein was stongly associated with a high score of degree of differentiation (p<0.05), nuclear pleomorphism (p<0.05), mitotic index (p<0.05), SBR grade (p<0.05) and MSBR grade (p<0.05), but it was not associated with patient's age, size of tumor or axillary node metastasis. The overexpression of c-erbB-2 C-erbB-2 oncoprotein was strongly associated with a high score of nuclear pleomorphism and a high SBR grade (p<0.05), but not associated with patient's age, size of tumor, axillary node metastasis, degree of differentiation, mitotic index or MSBR grade. An inverse relationship between the expression of p53 protein and estrogen receptor status was found, but the expression of c-erbB-2 was not associated with estrogen receptor status. It is concluded that p53 protein and c-erbB-2 oncoprotein are important prognostic factors in breast cancers, and that the aberrant expression of p53 protein is the most useful prognostic factor becausd of strong association of known histopathological prognostic factors and negative estrogen receptor status.

The Aberrant Expression of p53 Protein in Liver Cell Carcinoma.: Woo Young Jang, Dong Sug Kim, Ki Kwon Kim, Tae Sook Lee, Chang Yoon Kim, Hong Jin Kim; Korean J Pathol. 1995;29(5):607-614.

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Abstract: This study was carried out to evaluate the aberrant expression of p53 protein using immunobistochemical method in 54 surgically resected liver cell carcinomas and to correlate it with clinical and pathological findings. Twenty five out of 54 cases(46%) showed positive reaction in the nucleus of liver cell carcinoma and negative reaction in associated 30 cases of cirrhosis, one case of adenoma and two cases of adenomatous hyperplasia. The p53 protein expression was associated with alpha-FP level(p<0.05), but not associated with HBsAg positivity. It was significantly associated with WHO classification, Edmondson-Steiner grade and nuclear grade p53(p<0.05), but not associated with tumor size, capsule formation, portal vein invasion, cirrhosis in surrounding tissue, Eggel classification, special cell type and mitosis. In conclusion, our results suggest that the aberrant expression of p53 protein can be an advisory factor, at least, for prognosis evaluation.

HgCl2 Toxicity on Cultured Renal Tubular Cells of Rabbit.: Jung Young Lee, Seong Beom Lee, Suk Hyung Lee, Won Sang Park, Nam Jin Yoo, Sang Ho Kim, Choo Soung Kim; Korean J Pathol. 1995;29(5):615-623.

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Abstract: To understand the mechanism of cell injury when exposed to HgCl2, monitoring of cytosolic ionized free Ca2+([Ca2+]i), viability test, measurement of the amount of ATP, and Ca-ATPase activity were evaluated in cultured rabbit renal tubular cells(RTC) exposed to HgCl2. The results were as follows: 1) HgCl2 was cytotoxic to rabbit RTC at all doses except 10 uM and the rate of killing displayed a dose- and time-dependent relationship. 2) The absence of extracellular Ca provided partial protection from irreversible injury induced by HgCl2. 3) The increasing pattem of [Ca2+]i varied according to the concentrations of HgCl2. At the low concentrations of HgCl2 (2.5-10 microM), the level of [Ca2+]i increased slowly over the flat 2-3 min and then achieved plateau-state. In contrast, at the high concentrations of HgCl2 (25-100 microM) the level of [Ca2+]i achieved peak within 1 min and then decreased to a plateau state under normal concentrations. 4) The level of ATP was decreased to 27.5% of that of normal control cells within 3 min by using a treatment of 100 microM HgCl2. 5) HgCl2 did not affect the Ca2+ ATPase activity by enzyme histochemical observation. These findings suggest that the elevation of [Ca2+]i in response to the HgCl2-induced injury is an important event in accelerating injury that ultimately leads to cell death. But other possibilities such as HgCl2 might have direct deleterious effects on the also should be considered.

Morphometric Analysis of Preeciamptic Nephropathy with Focal Segmental Glomerulosclerosis.: Tae Sook Kim, Hyun Soon Lee; Korean J Pathol. 1995;29(5):624-633.

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Abstract: To evaluate the structural characteristics that might be related to the clinical features noted in preeclamptic patients with focal segmental glomerulosclerosis(FSGS), we analyzed post-partum renal biopsies of 8 preeclamptic patients with FSGS (group 1) by morphometry and glomerular studied the structural-functional relationships. These findings were also compared with those from three postpartum cases with minimal change lesion(group 2) and normal age-matched women(group 3). Mean glomerular volume (MGV) in group 1 and group 2 was (2.64 +/- 0.49) x 10(6) micrometer3 and (2.56+/-0.25)x 10(6) micrometer3, respectively. MGV in both groups was significantly increased compared with that of the control group [(1.11+/-0.22)x10(6) micrometer3](p<0.0005). The volume density of the mesangium/glomerulus [Vv(mes/glom)] in the group 1 patients was significantly increased (p<0.0001) when compared with that of the group 2 and the control group patients. The increment of Vv(mes/glom) was related to both the mesangial cell proliferation and expansion of mesangial matrix. The volume density of the capillary lumen/glomerulus [Vv(cap/glom)] in group I was significantly decreased(p<0.0001) when compared with that of group 2 and the control group. Vv(cap/glom) was directly related to Ccr in group l(r=0.70, p=0.05). These results suggest that reduced capillary luminal area caused by mesangial interposition is related to the decreased glomerular filtration rate in preeclamptic FSGS.

Cell Mediated Immunity in Tubulointerstitial Nephritis of Rats.: Hyeon Joo Jeong; Korean J Pathol. 1995;29(5):634-643.

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Abstract: To investigate the tubular major histocompatibility complex(MHC) expression and inflammatory phenotypes in tubulointerstitial nephritis, Lewis rats were inununized with azobenzen-earsonate-tyrosine in complete Freund adjuvant and challenged either foot pad or kidney, either by subcapsular injection or by ex vivo perfusion. The rats were sacrificed 2, 3, 5, 10 and 15 days after antigenic challenge. Foot pad swelling was significant at the antigenic challenge site (151.8 vs 6.8 x 10(-2) mm) at 24 hours. Tubulointerstitial nephritis was induced by both methods and the inflammatory infiltrate which first appeared on day 2, became prominent at day 5, then gradually subsided in ex vivo perfused rats, while inflannnation started on day 3 in subcapsular injected rats. The major site of inflammation was in the cortex and outer stripe of the outer medulla, with predominance of mononuclear cells throughout the course. The inflammatory cells showed mainly OX8 and ED1 positivity with OX19, W3/25 and CD5 positivity in minority. RT1B expression was diffuse in the cytoplasm of proximal tubules at day 2 and 5. These results suggest the involvement of cell mediated immunity in this experimental model, and the possibility that tubular epidielial cells process antigen and then become targets in immune injury.

A Scanning Electron Microscopic Study on Microvascular Changes in the Monocrotaline-induced Rat Lung by Corrosion Casting Method.: Na Hye Myong, Eui Keun Ham; Korean J Pathol. 1995;29(5):644-659.

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Abstract: To investigate the microvascular changes in primary pulmonary hypertension, the lungs of 24 Sprague-Dawley rats were treated by an intraperitoneal injection of 2% monocrotaline(MCT) solution and then examined with scanning electron microscopy(SEM) after microvascular corrosion casting. Histologic examination revealed significant medial thickening in the small to medium-sized pulmonary arteries. Scanning electron microscopic findings of the normal lungs showed two kinds of microvascular structures. One showed a well-fortned three-dimensional basket structure of uniform flat-tubular alveolar capillaries, which were connected to each other in a T or Y shape or at right angles. The other revealed a two-dimensional reticular sheet of round tubular branches mainly in the bronchial artery-supplying regions. The MCT-treated groups(remodelling) showed apparent changes in both kinds of microvasculatures in comparison to the normal group but the more prominent change was found in Lbe bronchial artery microvasculature showing the dense thick encasement around large pulmonary arteries. Alveolar microvasculature of the pulmonary artery revealed individually enlarged angular appearance, with generally deformed alveolar architecture. Quantitatively, the significant enlargement of diameter and intercapillary distance appeared in both microvasculatures of MCT-induced rat lungs, but the density was increased only in the bronchial artery microvasculature. In conclusion, our three-dimensional microvascular study of the MCT-treated rat lungs demonstrates a new morphologic finding of vascular remodeling in primary puhnonary hypertension, which is thought to play an important vascular role in the pathogenesis in addition to interstitial fibrosis.

Association of Ubiquitin-Positive Neuritic Threads in the CA 2-3 Region of the Hippocampus with Cortical Lewy Bodies.: Ki Hwa Yang, Ki Seung Yang, Choong Gu Kang, Joo Ho Sung; Korean J Pathol. 1995;29(5):660-668.

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Abstract: Ubiquitin-positive neuritic threads (UNTS) in the hippocampal CA 2-3 region are reported to occur exclusively in association with so-called diffuse Lewy body disease (DLBD). hi order to assess the association between the occurrence of Lewy bodies (LBs) and that of the UNTs, an immunohistochemical study on the hippocampus including the parahippocampal gyrus with antiubiquitin antibody (Chemicon Co., California, U.S.A.) was perfon-ned in four groups of patient's brains. All brains were selected from the large pool of brains referred to the Neuropadiology Laboratory of the University of Minnesota for studies of Alzheimer's disease. Group 1 consisted of 34 cases (20 male and 14 female) with LBs widespread in the frontal and temporal cortex and the brain stem nuclei (basal nucleus, substantia nigra, locus ceruleus and dorsal vagal nucleus) associated with varying degrees of degeneration of the substantia nigra; 21 (11 male and 10 female) combined with and 13 (9 male and 4 female) without Alzheimer's disease (AD) pathology. Group 2 included 12 cases (9 male and 3 female) in which LBs were observed only in the brain stem nuclei; 7 with and 4 without AD pathology. Group 3 consisted of 30 cases (9 male and 21 female) without LBs but with AD pathology and degeneration of the substantia nigra. Group 4 included 23 cases (11 male and 12 female) with neither LBs nor AD pathology but with degeneration of the substantia nigra. Ages of the patients varied among the groups. In the 46 cases in the first two groups with LBs. The mean age in the 28 cases with AD pathology was 77.3+/-8.9 and in the 18 cases without AD pathology it was 71.6+/-8.8 (P<0.05). In the 53 cases in groups 3 and 4 without LBs, which served as the controls, the mean ages were 80.8+/-7.7 and 74.0+/-9.7 respectively. UNTs were encountered in all (100%) of 34 cases of group I with widespread LBs which met the histological criteria of DLBD regardless of combined AD pathology. In 12 cases of group 2 with LBs confined to the brain stem, UNTs occurred in 3 (25%), 1 with and 2 without AD pathology. hi the group 3 cases with only AD pathology, UNTs occurred in 4 (13%) of 30 cases, while no UNTs were encountered in the 23 cases of group 4 without AD pathology or LBs. In conclusion, UNTs in the hippocampal CA 2-3 region occur invariably but not exclusively in association with widespread LBs, frequently when LBs are confined to the brain stem, and infrequently with AD pathology. It seems, therefore, that the UNTs are closely related to LBs and increase in ftequency as LBs spread beyond the brain stem with time, but the pathogenesis of the UNTs is little understood.

The Expression Rate and Pattern of HBcAg and HBsAg in the Hepatocytes According to the Histologic Activity of Cirrhosis.: Yoon Mi Jeen, Chanil Park; Korean J Pathol. 1995;29(5):669-677.

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Abstract: Since the discovery of hepatitis B virus as one of the causes of hepatitis, liver and hepatocellular carcinoma, many hepatitis B viral markers that appear in infected individuals have been discovered and many efforts to understand the relationship between the emergence of viral markers and the progression of hepatitis have been performed. Gudat (1975) compared the expression of HBcAg and HBsAg in various conditions and stages of hepatitis but the pattern of expression of viral markers and its significance have not been understood. Recently it was found by mierocytotoxicity assay that HBcAg might be the target of T lymphocytes. This study attempted to identify any correlation of the tissue expression rate and pattern of HBcAg and HBsAg with the histologic activity of 46 cases of liver cirrhosis using immunohistochemical staining. The expression rate and pattern of HBcAg and HBsAg in relation to the nodular size and positivity of serum HBeAg were also compared. The results were as follows; 1) The expression rate of HBcAg in the liver was 41.3% (19/46). and that of HBsAg was 67.4% (31/46). 2) The histologic activity of liver cirrhosis appeared to be correlated with the expression of HBcAg, especially cytoplasmic HBcAg. 3) The positivity of serum HBeAg was significantly higher in active liver cirrhosis. 4) There was no relationship between the tissue expression of HBsAg and the histologic activity of liver cirrhosis. relationship existed between the nodular size and expression rate and pattern of HBcAg and HBsAg. This study suggests that the tissue HBcAg, especially the cytoplasmic HBcAg is the most likely factor determining the histologic activity of liver cirrhosis, and that the cytoplasmic HBcAg may be the ultimate cause and target of most host immune response.

Case Reports

VATER Association: Three autopsy case reports with imusual defects.: Mi Ja Lee, Myeong Cherl Kook, Je G Chi; Korean J Pathol. 1995;29(5):678-683.

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Abstract: VATER association represents vertebral defects, anal atresia, tracheo-esophageal fistula with without esophageal atresia, renal defects and radial limb dysplasia. The probability of the simultaneous occurrence of any three of these defects is so unlikely that it suggests a sporadic non-random association. This non-random association appears to be related to some chromosomal anomalies, the caudal regression syndrome, mesodermal defects in early developmental period or the matemal use of sex hormones during embryogenesis. We report three autopsy cases of the VATER association that showed most of the known major and minor defects as well as an unusual concurrence of other defects, i.e., scoliosis, talipes varus, absent penis, urethral agenesis and stenosis, rectourethral fistula, rib anomaly, single umbilical artery, Meckel's diverticulum, diaphragmatic hemia, absent rectum, short neck, simian crease, low set ear, and hypoplastic lung.

Leiomyoma of the Female Urethra: A case report.: Kyoung Mee Kim, Anhi Lee, Sang In Shim; Korean J Pathol. 1995;29(5):684-686.

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Abstract: Leiomyomas are benign tumors of smooth muscle origin and are very rare in the female genital tract. To date, approximately 35 cases of urethral leiomyoma have been reported in the literature. A 34-year-old woman presented with a 3-year history of a mass at the urethral meatus. Physical examination showed 2 x 1.5 cm lump at the urethral meatus, posterior lip. Histologically the tumor was mainly composed of benign cigar shaped smooth muscle izells which were arranged in interlacing fascicles without cellular atypia or mitosis. Immunohistochemistry confirmed leiomyoma with positive staining for vimentin, desmin and muscle specific actin.

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