• 대한세포병리학회지
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• 제9권1호
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• pp.15-20
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• 1998

Mutant form of the p53 gene product is abnormally accumulated in the nuclei of the tumor cells due to prolonged half life, and readily detected by immunohistochemical methods. To determine the positivity rate of p53 in body cavity fluid according the primary site and histological types of tumors and the utility of p53 immunostaining as an adjunct in the diagnosis of malignancy, we reviewed 69 effusions, including pleural effusion, ascitic fluid, and pericardial fluid, that were diagnosed as overt malignancy and 21 effusions of suspicious malignancy, immunohistochemistry was performed on paraffin-embedded cell blocks using a monoclonal antibody to p53 supressor gene product(Clone DO7) and a standard avidin-biotin complex technique with a citrate buffer antigen retrieval solution. The results were as follows; of the 46 pleural effusions with overt malignancy, 22 were immunopositive for p53 protein; of the 21 ascitic fluids with overt malignancy, 5 were positive for p53. Positivity rates according to the primary sites of tumors were 18 of 34(52.9%), 8 of 21(38.1%), 1 of 9(11.1%) cases of the tumors of the lung, GI tract, and ovary, respectively. According to the histologic types of lung cancer, 11 cases(61.6%) were positive out of 18 adenocarcinomas, 2 of 5 large cell undifferentiated carcinomas, and 1 of 2 small cell undifferentiated carcinomas. Of 21 cases of suspicious malignancy, 6 were positive for p53 and all of them(6/6) were confirmed as adenocarcinoma of the lung or GI tract. These findings indicate that p53 immunostaining using paraffin embedded cell block is useful diagnostic and prognostic marker in body fluid cytology although negative immunostaining does not exclude malignancy.

• Restorative Dentistry and Endodontics
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• 제21권1호
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• pp.35-53
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• 1996

The development and repair requires the formation of new tissues comprised of various extracellular matrix components. The present study investigated the formation and distribution of the major ECM components such as type I collagen, type III collagen, fibronection, bone sialoprotein, and osteonection during development and repair. For developing observation. Sprague-Dawley rats weighing $27{\pm}1gm$ were sacrificed. For repair observation, Sprague-Dawley rats weighing $110{\pm}5gm$ were used. The pulp perforation were prepared on mesial surface of the maxillary first molar by using 1/2round bur. At 5 days after perforation, rats were sacrificed by perfusion with 3 % paroformaldehyde. The maxillary first molar region were cut, demineralized, dehydrated and embedded in paraffin. Immunostaining the ECM components was achieved by the avidin-biotin complex method. The results as follows : 1. Bright immunoreaction for fibronectin was present in the basement membrane at the inner epithelial-mesenchymal interface, especially concentrated in the blood vessel walls, cell membrane of odontoblasts, and initial predentin. 2. Type I and III collagen was observed in the newly formed pulp tissue, predentin, and its intensity increased as more of these components during repair. 3. Strong immunostaining for bone sialoprotein and osteonectin was found in dentin while no or weaker staining was observed loose connective tissue of the pulp. 4. These results suggest that develpment and repair is achieved through a series of cell differentiation and attachment by the specific ECM components.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권5호
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• pp.400-405
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• 2004

Angiogenesis inhibition is major concern to cancer chemotherapy and many studies about compound inhibiting angiogenesis is in progression. The long-known preventive effect of plant-based diet on tumorigenesis and other chronic diseases is well documented. Especially soy extract, genistein, is known to be potent angiogenesis inhibitor and prevent development and progression of tumor. In the present study, the effect of angiogenesis on tumorigenesis and chemopreventive effect of genistein by angiogenesis inhibition in hamster buccal pouch oral carcinigenesis model induced by 7.12-dimethylbenza(a)nthracene (DMBA) was studied. Forty eight Syrian Golden young adult hamsters (150-200 gm) were divided into two groups. In control group, 0.5% DMBA in heavy mineral oil was applied to hamster buccal pouch three times a week and in experimental group, 0.1 mg of genistein is administered orally everyday in addition to DMBA application. The animals were euthanized from 2 weeks to 16 weeks with interval of 2 week. H&E staining and immunohistochemistry was performed to evaluate microvessel density by using factor VIII-related antigen and avidin-biotin technique. Microvessels per area was quantified and compared between control and experimental group statistically. The results were as follows. 1. Microvessel density was increased time dependently in both groups and especially the increase was significant from 12 weeks to 16 weeks. 2. When comparing both group, the experimental group showed significantly low microvessel density than control group in 12 weeks (p=0.043), 14 weeks (p=0.050), 16 weeks (p=0.037). Based on these results, it was concluded that genistein influenced oral carcinogenesis by angiogenesis inhibition.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.321-340
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• 1995

The regeneration of destructed periodontal tissues is one of the ultimate objectives of periodontal therapy. Guided tissue regeneration technique was developed for the ideal regeneration of periodontal tissues. In order to investigate the role of fibronectin, laminin and tenascin in the regenerating process of periodontal tissues, the expanded PTFE barrier membranes(Gore Associates, USA) removed from the patients who had been treated by guided tissue regeneration(GTR) and guided bone regeneration(GBR) techniques were fixed in neutral formalin for 6-24 hours, embedded with paraffin, sectioned at $4-6{\mu}m$ in thickness, and immunohistochemically processed by Avidin-Biotin peroxidase complex method for detecting fibronectin, laminin and tenascin. Monoclonal mouse anti-human fibronectin antibody(Oncogene Science, USA., 1:100), monoclonal mouse anti-human laminin antibody(Oncogene Science, USA., 1:50) and mouse anti-human tenascin antibody(Oncogene Science, USA, 1:10) were used as primary antibodies. The light microscopic findings were as follows: (1) The distribution of fibronectin, laminin and tenascin was various according to the area of barrier membranes. (2) The distribution of fibronectin in case of GBR was extensive in the tissue on the outer surface of barrier membranes, and rare in the intervening space and on the inner surface. In case of GTR it was extensive on the outer surface and in the intervening space, and rare on the inner surface. (3) The distribution of laminin was rare in the tissue on the outer, the inner surface and intervening space of barrier membranes, regardless of GBR or GTR. (4) In case 'of GBR rare distribution of tenascin was observed on the outer surface only, except the inner surface and the intervening space of barrier membranes. In case of GTR the distribution of tenascin was extensive in the tissue on the outer surface, rare in intervening space and the inner surface. The results suggest that fibronectin, laminin and tenascin may play a important role in the regenerating process of periodontal tissue, and they may affect the outcome of healing.

• 대한수의학회지
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• 제35권2호
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• pp.317-326
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• 1995

The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제15권3호
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• pp.217-228
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• 1993

The purpose or this study was to observe the histological alteration of Langerhans cells on wound healing process in applying low energy laser irradiation. For this study, 50 Spraque-Dewly rats, weighing 150Gm or more were devided into control, experimental control group(0), 47.5Hz(1), 190Hz(3), 380Hz(5), 760Hz(7), lased group. All the experimental animals were made excision wound on buccal mucosa, 2mm depth, and lased with stoma laser (904nm, semconductor type ASGaAI, Sedalac France) 47.5Hz, 380Hz, 960Hz, 3minutes one time respectively except experimental control group. After the experiment, experimental animals were sacrificed after 24hours, 48hours, 72hours on each. Taken specimens were embedden in paraffin, sectioned 6-8u in thickness. And the langerhans cell were detected using ant S-100 protein antibody, and histochemically processed with Avidin Biotin complex method. All the Langerhan cells were calculated under light microspe in 400 multiplication field and standard deviation, probability test between each group were evaluated using statistical analysis system(S.A.S)program. Following results were obtained. 1. Langerhan cells were increased in experimental control group compared to that in control group(P<0.01). 2. 24hour after experiments, Langerhans cell were decreased compare to that in control group and control experimental group 5, 1, 3. Probability test shows significance between control experimental and 5, 1, 3 group on a =0.05 range. 3. 48our after experiment, Langerhans cells were decreased compare to that on experimental control group, and probability test shows significance between control experimental and 3, 7, 5 group an a=0.05 range. 4. 72hour after experiments, Langerhans cells were decreased compare to that on experimental control group and probability test on group comparison shows significance between control experimental and 1, 5 and 1 between 3, 7 between 3, and 5, between 7, respeilively on a=0.05 range. 5. Langerhans cells number in experimental group were decreased compare to that on experimental control group in applying laser irradiation.

• Restorative Dentistry and Endodontics
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• 제19권2호
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• pp.372-383
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• 1994

Saliva plays an important role in modulating the oral microbial ecology. And it is suggested to influence the initiation and progression of the dental caries. To evaluate the correlations between the salivary antimicrobial agents and the caries susceptibility, the 51 subjects were divided into 3 groups according to caries experience ; caries resistant group, medium caries susceptible group, and high caries susceptible group. Stimulated whole saliva was collected, and the salivary levels were measured for lysozyme, lactoferrin, and secretory-IgA to Streptococcus mutans. The lysozyme level was estimated using Micrococcus diffusion plate, lactoferrin level was determined with a non-competitive avidin-biotin enzyme immunoassay, and the titer of secretory IgA to Streptococcus mutans was assayed with ELISA. The results were as follows: 1. Lysozyme levels of each group showed no significant difference statistically (p>0.05). 2. The caries resistant group and the medium caries susceptible group had significantly higher levels of lactoferrin than the high caries susceptible group (p<0.05). But no clear difference was observed between the caries resistant group and the medium caries susceptible group(p>0.05). 3. The caries resistant group and the medium caries susceptible group showed relatively higher levels of the secretory IgA to Streptococcus mutans than the pigh caries susceptible group, but no significant difference was observed statistically (p>0.05).

• Restorative Dentistry and Endodontics
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• 제15권2호
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• pp.58-76
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• 1990

This study was designed to elucidate the distribution of the immunoglobulins in the experimentally induced rat periapical lesions. The pulp exposure was performed in 80 molars from 40 rats and the animals were sacrificed at 15, 30, 60 and 90 days after the operation and examined and radiographed. Of the 80 samples, 56 samples were routinely sectioned ($4-6{\mu}$ in thickness) and stained with Hematoxylin-Eosin for the light microscopic examination and 50 samples were stained with toluidin blue for mast cells and 50 samples were stained using the Avidin-Biotin horseradish peroxidase for detecting the presence of Ig A, Ig E, Ig M and Ig G containing cells. The following results were obtained : 1. The periapical lesions could be observed in all of 80 teeth by radiogragh (100%) and the periapical lesions were detected in 50 samples of 51 samples by light microscopy (98%). The size of lesions increased with time lapse both by radiograph and by light microscopy(p<0.05). 2. Of the 50 samples, 19 samples were diagnosed as periapical abscesses, 18 as periapical granulomas, 10 as fibrous scar tissues and 3 cysts. 3. After pulp exposure, periapical granulomas were developed mostly in the 15 day group, with time lapse periapical abscesses and fibrous scar tissues increased. 4. In the 50 periapical lesions, the numbers of Ig G containing cell (57.2%) were prominent and the percentage of Ig A, Ig E and Ig M containing cells were 16.4%, 14.7% and 11.8% respectively. The numbers of all classes of immunoglobulin containing cell were highest in the periapical granulomas and lowest in the cysts(p<0.05). 5. The number of the mast cell and immunoglobulin containing cells decreased generally with time lapse after the pulp exposure and Ig A, Ig E, Ig M and Ig G containing cells and mast cells had the high correlation one another(>0.6).

• 한국가축번식학회지
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• 제22권1호
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• pp.19-27
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• 1998

The present study was performed to investigate the patterns of a, pp.arance of lectin in uterus of fetuses of 90 and 120 days old and neonates of Korean native goat. The carbohydrate markers were used in histochemistry for the determination of the lectin by staining of avidin-biotin-per-oxidase complex(ABC), rincinus communis agglutinin(RCA-I), ulexeuropalus communis agglutinin(UEA) and wheat germ agglutinin(WGA). 1. The effects of this study were as follows; 1. The binding reactions for Con-A were weak on the mucosal epithelium of endometrium in 90 and 120 days old fetuses, and neonates and moderate at the free surface of mucosal epithelia. 2. The binding reactions for DBA was partially moderate on the mucosal epithelium of endometrium and partially strong at the free surface of mucosal epithelia in 120 days old fetuses. In neonates, the reactions were strong on the mucosal epithelium and gland primordium of endometrium, and the secretions at the free surface showed strong reactions for DBA. But, in 90 days old fetuses, the reaction was negative. 3. The binding reactions for RCA-I were moderate on the mucosal epithelium of endometrium and at the free surface of mucosal epithelia in 90 days old fetuses. In 120 days old fetuses, the reactions were weak on the mucosal epithelium of endometrium and moderate at the free surface of mucosal epithelia. In neonates, the reactions were moderate on the mucosal epithelium of endometrium and strong at the free surface of mucosal epithelia and also strong in the uterine gland. 4. The binding reactions for UEA were negative in 90 and 120 days old fetuses and neonates. 5. In 90 days old fetuses, the binding reactions for WGA were generally weak on the mucosal epithelium of endometrium, but several epithelial cells showed moderate reaction for WGA. In 120 days old fetuses and neonates, the reactions were moderate on the mucosal epithelium and blood vessels of endometrium and strong at the free surface of mucosal epithelia.

• 한국동물위생학회지
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• 제41권1호
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• pp.57-61
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• 2018

Canine peripheral nerve sheath tumors (PNSTs) are spindle cell tumors that arise from Schwann cells, perineural cells, fibroblasts or all of them. Based on the morphology and biologic behavior, PNSTs are divided into benign PNST (BPNST) and malignant PNST (MPNST) forms. The aim of this study is to diagnose the two cases of neoplastic tissue samples with features of PNSTs by the histopathology and immunohistochemistry. The study was performed using two specimens from small animal clinic. The first case, A was a mass, 3~4 cm in diameter, extruded from vaginal mucosa of 10-year-old spayed female mixed-breed dog. And the second case, B was a subcutaneous mass, 1.5 cm in diameter, which is originated from right hind leg of 9-year-old castrated male mixed-breed dog. Two cases were stained with hematoxylin and eosin (H&E) for histopathological examination. And also immunohistochemistry (IHC) was performed by the avidin-biotin peroxidase complex (ABC) method with antibodies specific for the following proteins: S-100 protein, smooth muscle actin (SMA) and epidermal growth factor receptor (EGFR). In results, Antoni B schwannoma pattern characterized by pleomorphic, round and fusiform polygonal cells was seen in A. In B, Antoni A pattern, densely packed spindle cells arranged in interlacing bundles was seen in addition to Antoni B pattern. In IHC, cytoplasms of neoplastic cells were diffusely labeled for S-100 expression in A and B. For SMA, both A and B show negative expression. And for EGFR, A shows negative expression but B shows partially positive expression in areas of Antoni B schwannoma pattern. The histopathologic features of two cases coupled with the S-100 immunoreactivity led to a diagnosis of PNST. For SMA, both A and B show negative expression. The diagnosis of A will be a BPNST with the negative result and B will be a MPNST with the positive result for EGFR.