• 대한치과교정학회지
• /
• 제23권1호
• /
• pp.89-100
• /
• 1993

The purpose of this study was to evaluate the effect of orthodontic force on periodontal cellular activity by immunoperoxidase stain of epidermal growth factor, one of the tissue hormone. And supplementarily, to investigate of the changes of periodontal structures, periodontium was stained by H-E, Masson's Trichrome, P. A. S. stain after orthodontic force application. The experimental animals were four young adult dogs of average 8 month old. The fixed orthodontic appliance was cemented on mandibular right 4th premolar and 1st molar of each animal as experimental site. Mandibular left 4th premolar area of the same animal was used as control. The appliance consist of two silver crown soldered with 0.030' tube, $0.018\times0.022'$ S.S. sectional arch wire, and 0.009' open coil spring for manifestating of orthodontic force for bodily tooth movement of mandibular 4th premolar toward mesial direction. Experimental group was sacrificed at 1, 2, 3, 5 weeks from beginning of the experiment, and was investigated immunohistochemically and bistochemically by several staining methods. Findings were as follows: 1. The degree of EGF staining in control group was highest in epithelium of periodontium, and osteoclasts, osteoblasts and fibroblasts around the capillary were stained at higher level in periodontium. Generally, control group shows positive distribution of EGF all around the periodontal area. 2. The degree of EGF staining in control and 5 week group were similar, and did not show the significant different level between tension and pressure side. 3. All of 1, 2, 3 week group showed the same staining degree and distribution of EGF, and the tension side was more positive reaction of EGF stain than the pressure side. 4. The features of collagen fiber and periodontal fiber arrangement observed by H-E, Masson's Trichrome and P. A. S. stain revealed that oblique periodontal fibers were strectched in tension side, compressed in pressure side of all experimental group. Some fiber group in pressure side of 5 week group recovered the regular arrangement along the capillaries.

• Restorative Dentistry and Endodontics
• /
• 제11권1호
• /
• pp.63-75
• /
• 1985

This study was designed to identify lymphocytes and to compare the lymphocyte distribution in endoodontically treated periapical lesions with that in endodontically untreated periapical lesions by way of immunohistochemical staining. Twenty-one human dental periapical lesions were obtained, frozened, serially sectioned to $4-5{\mu}$, and stained using the three-stage indirect immunoperoxidase technique and monoclonal antibodies for detecting the presence of B,T lymphocyte and T suppressor cell. Following results were obtained; 1. All of the examined periapical lesions had positive staining for B,T lymphocyte and T suppressor cell. 2. The concentration of T lymphocytes in 18 lesions diagnosed as periapical cyst and granuloma in both groups was greater than that of B lymphocytes and 2 periapical lesions identified as abscess in treated lesions had more positive B lymphocytes than positive T lymphocytes. 3. The average numbers of T,B lymphocytes and T suppressor cells in Endodontically treated lesions were lower than those of untreated lesions, but no statistically significant difference was noted. 4. When the distribution ratios of T lymphocytes to B lymphocytes and T suppressor cells to T lymphocytes were compared in Endodontically treated lesions by the histological aspects of the lesions and at the intervals of the duration after Endodontic treatment, a statistically significant change was not found. 5. The mean values of T lymphocytes, B lymphocytes and T suppressor cells in Endodontically treated lesions were markedly decreased in the specimens obtained at 3 month after Endodontic treatment, but no statistically significant difference was found.

• 대한수의학회지
• /
• 제29권1호
• /
• pp.27-35
• /
• 1989

To develop more specific and sensitive diagnostic methods for infectious bovine rhinotracheitis, 7-C-2 monoclonal antibody specific to polypeptides of infectious bovine rhinotracheitis virus (IBRV) was applied in indirect immunofluorescence antibody assay (IFA), indirect immunoperoxidase assay(IPA) and radial immunodiffusion enzyme assay (RIDEA). It was found that IBRV infected in MDBK cells could be detected as early as 8 hours post infection by IFA, and that IFA was more rapid and specific to identify IBRV antigen than IPA. The diagnostic efficacy of RIDEA and SN test was studied with 88 bovine sera. It was evident that RIDEA could eliminate the false positive reaction encountered in serum neutralization(SN) test, being more rapid and sensitive than the latter. Highly significant correlation coefficiency (r=0.76, p<0.01) was evaluated between the titers of sera and the diameters of RIDEA. Tracheal membranes and sera collected from 96 slaughtered cattle with lesions in respiratory organs were examined to detect IBRV antigen and antibody by IFA, RIDEA and SN test. It was presented that positive rates were 32.3% in IFA, 20.8% in RIDEA and 21.9% in SN test, and that coincidence rate between RIDEA and SN test were 100% in positive sera and 98.7% in negative sera. In conclusion, it was assumed that application of monoclonal antibody could improve the diagnostic efficacy of IBR by enhancing sensitivity and specificity of IPA, IFA and RIDEA.

• 대한미생물학회지
• /
• 제22권2호
• /
• pp.167-174
• /
• 1987

N-myc, a DNA sequence related to the oncogene c-myc, was found to be amplified in untreated primary neuroblastomas and the amplification appeared to be associated with advanced disease at diagnosis and rapid tumor progression. Synthetic peptides have been useful immunogens for generating antisera and monoclonal antibodies to a number of native proteins. In order to identify myc-related protein in the tumor cells, an antiserum against a synthetic hexapeptide (-Glu-Asp-Ile-Trp-Lys-Lys-), whose sequence corresponds to a part of the exon 2 of oncogene N-myc, was prepared by immunizing a rabbit with BSA-conjugated peptide. After ammonium sulfate precipitation and affinity column chromatography, it appeared to be specific to the peptide. Strong nuclear staining in immunoperoxidase method using this serum was observed in both human promyeloid leukemic cell line, HL-60(containing high c-myc copy number), and human neuroblastoma cell line, LA-N-5 (containing high N-myc copy number), whereas LA351 (human lymphoid cell line) cells did not react with the serum. This reaction was completely abrogated by incubating the antiserum with soluble excess peptide. These data suggest that the protein encoded by N-myc could be localized in the nucleus as c-myc protein and this antiserum can be used to detect myc-related tumor cells in clinical samples and to determine if the N-myc expression correlates with genomic amplification in cell lines, untreated primary tumors, and untreated metastases.

• 대한세포병리학회지
• /
• 제7권1호
• /
• pp.1-11
• /
• 1996

19세기말과 20세기초에 각각 비르효와 파파니콜로에 의해 명료하게 된 세포 병리학과 탈락세포학의 개념에서 오늘날의 암진단의 일차적인 방법이 발전해 왔다. 파파니콜로의 탈락세포학의 개념에서 1960년대 초반에 세침흡인 세포검사가 개발되었다. 이 세침흡인 세포검사는 주된 진단방법이 되어져, 절개생검을 감소하게 하고 의료비용의 효과적인 이용에 공헌하였다. 1980년대에는 면역생화학적 기술들이 암 진단에 보충역활을 하게 되었다. 단 클론 항체를 이용하는 면역과산화효소법이 먼저 암의 본성을 밝히는 보조적인 방법으로 쓰여졌다. 특정 단클론 항체들이 이용가능하게 되어 세포산물이나 표면 표지자들을 인지하는 것을 훨씬 용이하게 하였다. 예를 들면 중간세사에 대한 항체들이 분화가 나쁜 종양의 조직기원을 결정하는데 가치가 있는 것이 증명되었다. 종양표지자들은 종양존재의 생화학적 표시자로 이용될 수도 있는데 이러한 종양표지자들은 혈장이나 다른 체액들에서 검출할 수 있다. 이 종양표지자들을 농출한 것을 진단적 검사에 이용하여 이미 진단된 암의 임상 경과를 추적하고 암 발생의 위험이 있는 집단에서 특정 종양을 발견해 내기 위한 선별검사로써 이용할 수 있다. 유세포 검사는 백혈병이나 림프종 세포들의 면역표현형을 알아내고, 종양세포들의 DNA함유량을 알아내며, 세포증식율을 알아내는 등의 몇가지의 세포의 특성을 분류해내는데 유용한 도구이다. 분자생물학적 방법들은, 암 환자를 진료하는데 있어 진단, 예후평가 및 치료 등의 분야에서 일보 전진하게 하였다. 핵산교잡법이 Southern blots, Nothern blot, Dot blot 및 in situ hybridization으로 이용된다. 분자생물학 및 그 기술이 암종 생물학을 이해하고 유전자 조작을 기초로한 치료법을 계획하는데 밝은 새로운 지평선을 열어줄 수 있을 것이다.

• 대한핵의학회지
• /
• 제26권2호
• /
• pp.371-379
• /
• 1992

악성종양의 진단 및 치료에 있어서 특정 종양에 대한 항체를 이용하는 연구가 활발히 진행되고 있다. 단세포군항체를 이용한 방사면역신티그라피로 암의 조기 진단 및 영상화가 가능하고 나아가 방사면역치료는 암의 선택적 치료에 도움이 될 수 있다. 위암은 우리나라에서 가장 흔한 악성종양으로 방사면역신티그라피와 방사면역치료법이 새로운 방법으로 모색되고 있다. 이러한 진단 및 치료법의 성공여부를 결정하는 중요한 인자의 하나가 종양조직내에서 종양관련 항원의 농도와 분포이다. 따라서 본 연구에서는 단세포군 항체를 이용한 방사면역학적 방법의 임상 이용을 위한 기초 연구의 일환으로 in vitro quantitative autoradiography를 이용하여 종양 관련항원인 TAG-72와 CEA의 위암조직내 농도 및 분포를 측정하였다. 33예의 위암조직에서 얻은 동결절편을 $1.3\sim83.3$ nmol/liter의 $^{125}I$ 표지 항 TAG-72 단세포군 항체 B-72.3과 항 CEA 단세포군 항체 CEA-79로 반응시킨 후 이 표본들의 자가방사법 디지탈 영상을 H & E 염색과 immunoperoxidase염색 표본과 비교하였으며, 특정 단세포군 항체의 결합에 대한 컴퓨터 분석으로 조직내 항원의 농도와 분포를 측정하였다. TAG-72는 25예(75.7%)의 조직에서 검출되었으며 그 농도는 $8.4\sim525.3$ pmol/gram이었고, CEA는 32예 (96.9%)에서 검출되었으며 그 농도는 $8.8\sim592.9$ pmol/gram 이었다. CEA의 위암 조직내 발현농도는 중앙치가 101.7 pmol/gram 으로 TAG-72의 중앙치인 27.9 pmol/gram 보다 높았다. TAG-72의 조직내 분포는 41.4%에서 병변 부위의 암세포 분포와 일치하였고, CEA의 분포는 병변 부위의 80.5%에서 암세포와 일치하는 소견을 나타내었다. TAG-72의 농도는 점액성 선종(mucinous adenocarcinoma)과 점액함유 선종(mucin containing adenocarcinoma)에서 다른 선종보다 더 높았다. CEA의 농도는 위암의 병리학적 종류에 따른 유의한 차이가 없었다. 이상의 결과로 위암조직에서 TAG-72와 CEA항원이 다양하게 발현됨을 알 수 있었고 CEA는 TAG-72 보다 더 빈번하게 균일한 분포로 발현하는 것으로 나타났다.

• 대한미생물학회지
• /
• 제21권2호
• /
• pp.211-226
• /
• 1986

To understand the pathogenesis of anicteric leptospirosis with severe pulmonary hemorrhage occured in Korea, the microbiological and pathological features were observed in the experimentally induced leptospirosis in guinea pigs infected with a virulent strain of Leptospira interrogans isolated from the patient at Wonju, Korea, and the results are summarized as follows. 1. The main pathological features were widespread hemorrhages, especially affecting lung, skeletal muscles, retroperitoneal and perirenal adipose tissues. The hemorrhages accompanied inflammatory process especially of vasculitic pattern as well as occasional coagulation necrosis in the liver, skeletal muscle, and myocardium. The main inflammatory cells were of plasma cell even in the fairly early stage of the infection. 2. Those pathologic changes were more exaggerated in the inoculation site. 3. Within 144 hours of infection, the longer the infection time, the more antigens were observed in the tissues, and the severer the pathologic changes. 4. Leptospiral antigens were detected at first by indirect immunofluorescent and immunoperoxidase technics. As the infection time extended, the antigens were observed in all of the tissues examined except in the skeletal muscle. The shape of the antigens was spiral or thread-like within 72 hours of infection. As the infection progressed, they became fragmented and granular. 5. Leptospires were detected in the blood within 144 hours of infection by darkfield microscopic examination. Thereafter, none was observed. 6. Antibody to leptospires were detected as early as 72 hours of infection. In summary, the virulent strain of L. interrogans used in this experiment induced widespread hemorrhages with inflammatory reaction especially in lung, skeletal muscles, and retroperitoneal adipose tissue. With these findings, it is suggested that the direct toxic effect of leptospires might playa great role in the pathogenesis of this infection.

• 한국동물위생학회지
• /
• 제35권1호
• /
• pp.9-17
• /
• 2012

The objective of the present study was to isolate and identify infectious bursal disease viruses (IBDVs) from broiler and layer chickens of outbreaks of infectious bursal disease (IBD) in three districts of Bangladesh. A total of 70 bursal samples were collected from dead broiler (n=40) and layer (n=30) chickens showing specific lesions of IBD from seven commercial poultry farms of three different districts (Mymensingh, Chittagong and Tangail) of Bangladesh during the year 2007. Five representative bursal samples from each farm were used for the isolation of IBDVs using 9-day-old embryonated eggs of seronegative flock of layer birds and for identification the samples were subjected to agar gel immunodiffusion test (AGIDT), immunohistochemistry (IHC) and reverse transcription-polymerase chain reaction (RT-PCR). Out of 35 bursal samples, IBDVs were successfully isolated from 28 (80%) samples. By AGIDT, 32 (91.4%) samples were found positive for IBDV antigen. Results of AGIDT clearly indicated that IBDVs detected in 29 bursal samples of six affected farms were identical to each other but not to IBDVs present in the remaining three samples of another farm. Indirect immunoperoxidase staining of the bursal sections revealed the presence of IBDV antigen in 32 (91.4%) samples and the IBDV antigen was detected mainly in the cortex of the lymphoid follicles of the bursal tissues. In histopathology, cell depletion, atrophy and necrosis were observed in many bursal follicles with severe edema of interfollicular septa. Of the 35 bursal samples, 34 (97.1%) samples generated 254 bp product by RT-PCR. In conclusion, the results of virus isolation and identification by AGIDT, IHC and the analysis of viral genome by RT-PCR confirmed the outbreaks of acute IBD in commercial poultry of Bangladesh. Moreover, histopathological findings and results of AGIDT gave a clear indication that the isolates from six outbreaks were different from classical strain and it seems to be of very virulent strain. On the other hand, the isolates from the other outbreak were similar to the classical strain.

• 대한수의학회지
• /
• 제34권1호
• /
• pp.77-83
• /
• 1994

유사산 태아의 폐, 청색증을 나타내는 자돈으로부터 돼지생식기 및 호흡기증후군(PRRS)의 원인체로 추정되는 바이러스주(KPRRSV) 들을 분리하였다. 분리된 바이러스주는 돼지콜레라, 돼지오제스키병, 돼지뇌심근염바이러스에 대한 형광항체반응에서는 음성이었으며 기니픽혈구에 대한 혈구응집 능력을 나타내지 않았다. 그리고 포유 마우스의 뇌내 접종시 이상을 나타내지 않았으나 돼지생식기 및 호흡기증후군에 대한 형광항체검사시 양성반응을 나타내었다. 분리된 바이러스는 돼지폐포탐식세포(porcine alveola macrophages)에서 세포변성효과(cytopathic effect)를 나타내었으며 세포변성효과를 나타내었던 바이러스주중 일주(KPRRSV-1)를 돼지폐포탐식세포에서 7대 연속 계대하여 돼지에 접종한 후 혈청을 분리하여 미국 및 유럽지역에서 분리된 돼지유행성 유사산 및 호흡기증후군의 바이러스를 탐식세포에 감염시켜 효소면역방법 (immunoperoxidase monolayer assay)으로 분석한 결과 분리된 바이러스는 미국형 돼지호흡기 및 유사산증후군에 가까운 항원형으로서 판명되었다.

• Restorative Dentistry and Endodontics
• /
• 제24권1호
• /
• pp.100-115
• /
• 1999

The purpose of this study was to investigate the distribution of calcitonin gene-related peptide containing nerve fibers in rat pulp after dentinl injury by means of immunohistochemistry and confocal laser scanning microscope. The Spague-Dawley rats weighing about 250-300gm were used. The animals were devided into normal control and experimental groups. Experimental animals were sacrified 1, 2, 4, 7, 10, 21days after dentinal injury (dentin cutting, and then acid etching with 35% phosphoric acid) on the maxillary molar teeth. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), then were decalcified with 15% formic acid for 10 days. Serial frozen $50{\mu}m$ thick sections were cut on a cryostat. The rabbit CGRP antibody was used as a primary antibody with a dilution of 1:2000 in 0.01M PB. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated antirabbit Ig G as a secondary anti body with dilution of 1:200 in 0.01M PB and incubated in ABC(avidin-biotin complex). The peroxidase reaction was visualized by incubating the sections in 0.05% 3,3 diaminobenzidine tetrahydrochloride containing 0.02% $H_2O_2$. For the confocal laser scanning microscopic examination, Primary antibody reaction was same as immunoperoxidase stainning, but fluorescein isothiocyanate(FITC)-conjugate antirabbit IgG as a secondary antibody was used. The confocal laser scanning microscope was used for the examination. A series of images of optical sections was collected with a 20x objective at $3{\mu}m$ intervals throughout the depth of specimen. FITC fluerescence was registrated through a 488nm and 568nm excitation filter, and images were saved on optical disk. The stereoscopic images and three dimentionnal images were reconstructed by computer software, and then were analyzed. The results were as follows : 1. In normal control group, CGRP containing nerve fibers were coursed through the root with very little branching, and then formed a dense network of terminals in coronal pulp. 2. A slight increase in CGRP containing nerve fibers at 1 and 2day postinjury was noted subjacent to the injury site. In the 4day group, there were an extensive increase in the number of reactive fibers, followed by a partial return toward normal levels at 7~10 day postinjury, and return by 21days. 3. The sprouting of the CGRP containing nerve fibers was evident within 2day after dentinal injury, and by 4days there was a maximal increased, but was decreased at 7days and returned to normal 10~21 day postinjury. 4. In confocal laser scanning microscopic exammination, the distinct distribution pattern and sprouting reaction of CGRP containing nerve fibers were observed in stereoscopic images and three dimentional images. These results suggest that CGRP containing nerve fiber can be important role in the response to dental injury and pain regulation.