• Journal of Korean Neurosurgical Society
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• v.30 no.4
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• pp.443-450
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• 2001

Objective : The cyclin-dependent kinase inhibitor $p27^{kip1}$ protein is a negative regulator of the cell cycle, and its degradation is required for entry into the S phase. Loss of $p27^{kip1}$ expression has been reported to be associated with aggressive behavior in a variety of tumors of epithelial and lymphoid origin. However, its association with various astrocytic tumors has not been clearly demonstrated. We studied to investigate the relationship of $p27^{kip1}$ expression with the biological behavior of astrocytic tumors in addition to study on the role of $p27^{kip1}$ in the tumorigenesis of these tumors. Patients and Methods : From 1990 to 1998, a total of 29 astrocytic tumor of all grades obtained by operative resection were included for evaluation. We studied the expression of $p27^{kip1}$ protein immunohistochemical assay in astrocytic tumors and compared the findings with the clinicopathologic parameters. Immunohistochemical staining was performed on formalin-fixed paraffin-embedded sections by the avidin-biotin-peroxidase complex method. According to WHO classification, all cases were divided into astrocytomas(4 cases), anaplastic astrocytomas(9 cases), and glioblastomas(16 cases) by 3 pathologists. Clinical information was obtained from medical records, and others such as location and size of tumors from imaging studies. Results : Mean $p27^{kip1}$ protein labeling indexes(LI, mean${\pm}$standard deviation) of astrocytomas, anaplastic astrocytomas, and glioblastomas were $80.6{\pm}9.1$, $63.6{\pm}21.0$, and $28.9{\pm}18.7$, respectively, and were inversely correlated with grade of glial tumors(p<0.0001). Mean $p27^{kip1}$ protein LI in the recurrent group was lower than that in the nonrecurrent group, but there was no significant difference statistically(p=0.464). Additionally, $p27^{kip1}$ protein expression did not show any significant relationship to other prognostic factors such as age(p=0.1643), tumor size(p=0.8), or location(p=0.8). Conclusion : These results suggested that reduced expression of $p27^{kip1}$ protein may play a important role in the malignant transformation process of astrocytic tumor cells.

• Journal of Korean Neurosurgical Society
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• v.30 no.4
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• pp.430-436
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• 2001

Objective : The proliferative potential of intracranial glioma affects the histological malignancy and prognosis of patients with these tumors. In this study, we present the relationship between MIB-1 labeling index(LI) and clinical variables which might play the major role in determining the prognosis of patient with astrocytic tumors. Patients and Methods : Excised tumor specimens from a total of 52 patients were stained to detect monoclonal MIB-1-Ki-67 antibody by avidin-biotin complex immunohistochemistry. The MIB-1 LI was evaluated with histological grades, demograpghic data, and survival time. The statistical significance of their correlation was analyzed by Pearson correlation test. Results : The 52 patients included 30 male patients and 22 female patients. The tumors according to the criteria of the World Health Organization(WHO) classification were verified as pleomorphic xanthoastrocytoma in one, pilocytic astrocytomas 4, astrocytomas 1, anaplastic astrocytomas 3, and glioblastomas 31. MIB-1 LI in astrocytic ttumors showed no correlation with age and gender. However, the patients under 10 years had the longest survival time, whereas short survival time was observed in the older patients. The mean MIB-1 LI of different tumor grades were as follows : pleomorphic xanthoastrocytoma, $4.40{\pm}0.00$ ; pilocytic astrocytoma, $4.53{\pm}3.09$ ; astrocytoma, $5.50{\pm}6.03$ ; anaplastic astrocytoma, $12.68{\pm}12.50$ ; Glioblastoma, $21.31{\pm}19.63$. Although the levels of MIB-1 LI were varied in individual tumors, the MIB-1 LI was increased in parallel with the histological grades. Glioblstomas showed significantly higher MIB-1 LI compared with that of anaplastic astrocytomas and low grade astrocytomas (p = 0.001). The mean survival time of entire group of patients was also well correlated with MIB-1 LI in astrocytic tumors(p = 0.015). Moreover, the mean survival time of the entire group of patients with Lis < 10 was $125.33{\pm}113.57weeks$, and the mean survival of those with $Lis{\geq}10$ was $60.71{\pm}62.58weeks$. This difference was also statistically significant(p = 0.004). Conclusion : The results of this study suggest that MIB-1 LI correlates with histological grades and might play a significant role in predicting the survival of patients with astrocytic tumors.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.121-130
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• 1994

Expression of gonadotropin releasing hormone(GnRH) has been described in the rat ovary. It remains, however, unkown whether GnRH is synthesized as a prohormone. Therefore, this study was performed to verify the expression of pro-GnRH by in situ hybridization and further to investigate the effect of gonadotropin on GnRH or GnRH mRNA in rat ovary by immunohistochemical and in situ hybridization techniques. Adult female Sprague-Dawely rats were used and the estrous cycle was synchronized by intraperitoneal injection of pregnant mare's serum gonadotropin(PMSG). Ovaries were fixed with 4% paraformaldehyde and embedded with G.C.T. compound and cut by cryostat. For immunohistochemistry, avidin-biotin peroxidase complex(ABS) method was employed and for in situ hybridization, $^{35}S$-end labeled oligonucleotide was used and followed by autoradiography. By in situ hybridization using GnRH oligomer and GAP(GnRH associated protein) oligomer, GnRH mRNA and GAP mRNA were co-localized in the fullicular cells, luteal cells, interstitial cells and theca cells. GnRH or GnRH mRNA signals in the ovary increased by human chorionic gonadotropin(hCG) injection. At the 3 and 6 hrs after hCG injection, the number of GnRH and GnRH mRNA containing cells increased rapidly and the density of GnRH and GnRH mRHA culminated at 9 hrs after heG injection. With the follicular development, the high expression of GnRH and GnRH mRNA was also observed within the follicles. After ovulation, the density of GnRH or GnRH mRNA decreased in the follicles but increased in the corpus lutea.

• Proceedings of the Korean Society for Bioinformatics Conference
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• 2001.10a
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• pp.61-86
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• 2001

All cancers are caused by abnormalities in DNA sequence. Throughout life, the DNA in human cells is exposed to mutagens and suffers mistakes in replication, resulting in progressive, subtle changes in the DNA sequence in each cell. Since the development of conventional and molecular cytogenetic methods to the analysis of chromosomal aberrations in cancers, more than 1,800 recurring chromosomal breakpoints have been identified. These breakpoints and regions of nonrandom copy number changes typically point to the location of genes involved in cancer initiation and progression. With the introduction of molecular cytogenetic methodologies based on fluorescence in situ hybridization (FISH), namely, comparative genomic hybridization (CGH) and multicolor FISH (m-FISH) in carcinomas become susceptible to analysis. Conventional CGH has been widely applied for the detection of genomic imbalances in tumor cells, and used normal metaphase chromosomes as targets for the mapping of copy number changes. However, this limits the mapping of such imbalances to the resolution limit of metaphase chromosomes (usually 10 to 20 Mb). Efforts to increase this resolution have led to the "new"concept of genomic DNA chip (1 to 2 Mb), whereby the chromosomal target is replaced with cloned DNA immobilized on such as glass slides. The resulting resolution then depends on the size of the immobilized DNA fragments. We have completed the first draft of its Korean Genome Project. The project proceeded by end sequencing inserts from a library of 96,768 bacterial artificial chromosomes (BACs) containing genomic DNA fragments from Korean ethnicity. The sequenced BAC ends were then compared to the Human Genome Project′s publicly available sequence database and aligned according to known cancer gene sequences. These BAC clones were biotinylated by nick translation, hybridized to cytogenetic preparations of metaphase cells, and detected with fluorescein-conjugated avidin. Only locations of unique or low-copy Portions of the clone are identified, because high-copy interspersed repetitive sequences in the probe were suppressed by the addition of unlabelled Cotl DNA. Banding patterns were produced using DAPI. By this means, every BAC fragment has been matched to its appropriate chromosomal location. We have placed 86 (156 BAC clones) cytogenetically defined landmarks to help with the characterization of known cancer genes. Microarray techniques would be applied in CGH by replacement of metaphase chromosome to arrayed BAC confirming in oncogene and tumor suppressor gene: and an array BAC clones from the collection is used to perform a genome-wide scan for segmental aneuploidy by array-CGH. Therefore, the genomic DNA chip (arrayed BAC) will be undoubtedly provide accurate diagnosis of deletions, duplication, insertions and rearrangements of genomic material related to various human phenotypes, including neoplasias. And our tumor markers based on genetic abnormalities of cancer would be identified and contribute to the screening of the stage of cancers and/or hereditary diseases

• The Korean Journal of Malacology
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• v.15 no.1
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• pp.1-11
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• 1999

An immunohisochemical study on the cerebral ganglion of the African giant snail, Achatina fulica. was conducted by applying the AB/AY staining and the avidin-bovine-peroxidase complex staining methods. The followings are the results obtained throughout the study. The cerebral ganglion of Achatina fulica is an ellipsoidal body of 2 x 1 mm in size, which is connected by the cerebral commissure of 1 mm in diameter. The cross-section through the cerebral ganglion, shaped like a butterfly, is divided into the medio-dorsal parts, the latero-dorsal parts, the caudo-dorsal parts, and the lateral lobes. In the medio-dorsal and latero-dorsal parts, the LG cells and the DG cells are found mixed, although the LG cells are dominant. In lateral lobe, however, the Y cells are quite dominant, while the LG cells and the DG cells are seldom found. The LG cells are 20-70 $\mu\textrm{m}$ in sizes and circular or ellipsoidal in shapes. They are stained light green with the AB/AY. 1 - 3 nucleoli are found in karyolymph, where granular chromantins are evenly distributed. In cytoplasm, it is found that the secretory granules are evenly developed.

• Journal of Periodontal and Implant Science
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• v.23 no.2
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• pp.300-314
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• 1993

Periodontal disease research has been focused on understanding the immunopathologic mechanisms which may operate in the development and maintenance of peiodontal inflammatory changes. Immunologic and inflammatory responses may relate to the etiology and pathogenesis of periodontal disease. In order to research immunopathology of periodontal disease, previous investigators have spent much time on the distribution of lymphocyte subpopulations and NK cells but they have spent less time on the changes of those cells to the periodontal disease severity. The purpose of study was performed to investigate the changes of the distribution of T lymphocytes, B lymphocytes, T lymphocyte subsets, and Natural Killer cells in the gingival epithelium and connective tissue of the periodontal disease with the various clinical parameters including Gingival Index, Sulcular Bleeding Index, and pocket depth. Gingival tissues were obtained from 25 patients with different severity of periodontal disease. Serial cryostat sections displaying a cross section of gingiva were labelled with monoclonal antibody for pan T cells, T cytotoxic/suppressor cells, T helper/inducer cells, pan B cells, and NK cells were develped using an avidin-biotin-peroxidase system. Lymphocyte populations were enumerated in repeatable fields from gingival section. 1. T cells were more increased at grade 1 and 3 than at grade 0 of gingival index (p<0.05). Helper T cells and NK cells were significantly increased at grade 1, 2, 3 than at grade 0(p<0.05). 2. T cells were more decreased at grade 3 and 4 than at grade 1 of sulcular bleeding index (p<0.01, p<0.05). Especially, Natural Killer cells were significantly increased at grade 1, 2, 3, 4 than at grade 0 (p<0.05, p<0.001). 3. The ratios of helper T/suppressor T cells were more decreased at grade 4 than at grade 0 and at grade 4 than at grade 2 of sulcular bleeding index (p<0.05, p<0.05). 4. Helper T cells were significantly decreased at grade II and III than at grade I, however the Natural Killer cells showed a increasing tendency with the increase of the pocket depth, there were no significant differences between each grade of pocket depth. 5. The ratios of helper T/suppressor T cells were tended to be decreased with the increase of the pocket depth, there were no significant differences between each grades of pocket depth. There was a very weak change in the distribution of T lymphocytes, B lymphocytes, T lymphocyte subsets, and Natural Killer cells in the gingival epithelium and connective tissue of the periodontal lesion with the various clinical parameters including gingial index, sulcular bleeding index, and pocket depth. But, the number of T lymphocytes and Natural Killer cells were significantly changed in gingival index and sulcular bleeding index.

• Parasites, Hosts and Diseases
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• v.31 no.1
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• pp.49-56
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• 1993

Specific antibody test in serum and cerebrosinal fluid (CSF) is still the main mode of serological diagnosis of cystiercosis. Of different techniques of artibody test, enzyme-linked immunosorbent assay (micro-ELISA) has widely been applied. This study was undertaken to observe whether diagnostic capability can be Improved by applying more sensitive techniques such as Protein A-ELISA and avidin biotin complex ELISA (ABC-ELISA). When evaluated using 115 sera of human cysticercosis, the antibody positive rates were not significantly improved in Protein A-ELISA (82.6%) and in ABC-ELISA (86.1%) than in micro-ELISA (81.7%). The specificities, evaluated in 165 sera from other diseases and normal controls, were significantly improved (88.5% by micro-ELISA, 93.3% by Protein A-ELISA and 93.8% by ABC-ELISA). Antibody levels (absorbance, abs.) in individual serum were correlated well (r : 0.83∼0.86) each other. An actual benefit of Protein A-ELISA and ABC-ELISA was that they needed smaller amount of test sample.

• Korean Journal of Clinical Laboratory Science
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• v.39 no.2
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• pp.128-135
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• 2007

This study was designed to investigate the correlation between the expression rate of p53 and p21 proteins by immunohistochemical staining and tumor prognostic factors including the tumor size, histological differentiation and Dukes' stage of tumor prognostic factors in colon cancer, and to acquire necessary data for the presumption of diagnosis, treatment and prognosis of colon cancer patients. From January 2000 to January 2003 at Hanyang University Guri Hospital, the paraffin blocks of 35 patients diagnosed with colon cancer whose pathologic reports were possible to review were selected. Harris hematoxylin & eosin (H&E) staining and immunohistochemical staining by ABC (Avidin Biotin Conjugate) method were performed. The histological differentiation grade and stage were classified according to the classification of the World Health Organization (WHO) and modified Dukes's stage from H&E staining. The expression rate of p53 and p21 proteins were analyzed by immunohistochemical staining. The results was analyzed statistically by SPSS (Windows version 8.0). As a result, the expression rate of p53 protein was 11.4% (4 cases) in clear differentiation, 48.6% (17 cases) in moderate differentiation, and 17.1% (6 cases) in poor differentiation. In other words, the poorer the differentiation, the higher the expression rate of p53 protein (p<0.05). The expression rate of p21 was 17.1% (6 cases) in clear differentiation, 40.0%(14 cases) in moderate differentiation, and 8.6% (3 cases) in poor differentiation, According to the progression of histological malignant degeneration, the expression rate of p21 protein decreased distinctively (p<0.05). However, the correlation between the two above mentioned proteins and the tumor-size and Dukes' stage was not of statistical significance. In the comparison of the expression rate of p53 protein with that of p21 protein, in 10 cases, p53 protein expression was positive while p21 protein expression was negative, and in 6 cases, p53 protein expression was negative whereas p21 protein expression was positive. Consequently a statistically significant inverse correlation between the expression rate of p53 protein and that of p21 protein was observed (p<0.05). In conclusion, we found a significant correlation between histological differentiation and the expression rate of p53 and p21 proteins (p<0.05), and a significant inverse correlation between the expression rate of p53 protein and that of p21 protein (p<0.05). Also, it could be confirmed that the over expression of p53 and p21 proteins is closely associated with the occurrence of colon cancer and its progress. Therefore, it is thought that this study may be greatly beneficial to the presumption of diagnosis, treatment and prognosis of colon cancer patients.

• Journal of Food Hygiene and Safety
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• v.6 no.1
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• pp.27-40
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• 1991

;ABSTRACT-The effects of gastric ulcer induced by restraint and water-immersion stress on gastric carcinogenesis in Wistar male rats treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were examined. Rats of group 1 were 2iven stress for 8 hours before they were received MNNG (100 mg/l) in drinking water for 20 weeks. Rats of group 2 were received MNNG first for 2 weeks and then were given stress once a week from 3rd to 12th weeks, with simultaneous MNNG adminitration and followed by MNNG only until 20th weeks. Rats of group 3 were received MNNG only as a positive control and rats of group 4 were not treated with carcinogen. All groups were sacrificed in 20 weeks. Sections of the pyloric mucosa were stained by avidin-biotin peroxidase complex (ABC) immuno-histochemical method. PAPG (pepsinogen isozyme 1 altered pyloric gland), body weight change, gross lesions and histopathological changes were examined. The results obtained from these studies were summarized as follows: 1. The mean body weight gains of the rats fed with carcinogens (group 1, 2, 3) were significantly lower than that of group 4 (control group, without carcinogen. p<0.05). However, the differences of the mean body weight of rats treated with carcinogen were not significant. 2. Stress treatment (group 1 and 2) increased the appearance of the numbers of PAPG (Pepsinogen 1 Altered Pyloric Gland) induced by carcinogen significantly compared with that of group 3 (carcinogen only, p<0.01). 3. The incidence rate of mucosal hyperplasia in pylorus was significantly increased in group 2 compared with group 3 (p<0.05).0.05).

• Korean Journal of Veterinary Research
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• v.39 no.3
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• pp.575-582
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• 1999

Morphometrical analysis of chicken Cryptosporidium baileyi in various stages of life cycle in the bursa of Fabricius were carried out by electron microscope to establish a differential point for identification of C baileyi. By avidin-biotin complex method, protozoans of the bursa of Fabricius were identified Cryptosporidium spp. The size and area on each developmental stages of C baileyi, as measured by Morphomat 10 attached to electron microscope were as follows. Trophozoites' size with range of $3.21{\pm}0.70{\times}2.49{\pm}0.59{\mu}m$, area with range of $118.82{\pm}41.92{\mu}m^2$; meronts' size $3.99{\pm}1.07{\times}2.96{\pm}0.52{\mu}m$, area $210.11{\pm}57.11{\mu}m^2$; merozoites' size $1.98{\pm}0.43{\times}0.60{\pm}0.18{\mu}m$, area $24.10{\pm}5.97{\mu}m^2$; microgametes' size $1.36{\pm}0.83{\times}0.50{\pm}0.23{\mu}m$, area $20.23{\pm}6.73{\mu}m^2$; macrogametes' size $4.57{\pm}0.65{\times}4.02{\pm}0.55{\mu}m$, area $258.37{\pm}51.83{\mu}m^2$; oocytes' size $4.39{\pm}0.56{\times}3.44{\pm}0.50{\mu}m$, area $187.21{\pm}62.68{\mu}m^2$. In conclusion, the size and area on each developmental stages of Cryptosporidium baileyi is different from that of other Gryptosporidia spp. It suggests, with considering tissue tropism and life cycle, morphometrical analysis can be quite a good way to identify C baileyi.