• Title/Summary/Keyword: Positive stain

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Studies on antigenicity and production of monoclonal antibody for diagnosis of canine heartworm(Dirofilaria immitis) (개 심장사상충(Dirofilaria immitis) 진단을 위한 항원성 조사 및 단크론항체 생산)

  • Lee, Cheol-soon;Jee, Cha-ho
    • Korean Journal of Veterinary Research
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    • v.40 no.1
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    • pp.130-137
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    • 2000
  • In order to diagnose canine heartworm infection by antigen capture ELISA, the crude somatic(S), partial somatic(below 45kDa) and excretory/secretory(E/S) antigen of adult heartworm were identified and the antigenicity was examined by silver stain, immunoblot and ELISA. Then, production of monoclonal antibody to specific antigen carried out in this experiment. The bands to S antigen and E/S antigen were recognized between 10 and 200kDa and common bands were recognized strongly 14, 18, 28, 43kDa by silver stain. By western blot analysis, fractions to S antigen were recognized 14, 16, 18, 20, 24, 28, 32, 43, 50, 55kDa, etc. and only a 14kDa to E/S antigen in positive sera which were positive in modified Knott's test and necropsy. In ELISA, the positive sera reacted to antigens(SA, $SA_{45}$, E/S) were significantly different from negative sera by Student's t-test(p<0.05). Four hybridoma cell lines(14, 16, 17, 32kDa) than produce specific monoclonal antibodies for these antigens were obtained by immunizing BALB/c mice with a partially purified somatic antigen (below 45kDa) preparation, by fusing spleen cells with SP2/O cell myeloma cells, and by screening cell culture supernatants for antibody. In these results, it was confirmed that partial somatic antigen(below 45kDa) or E/S antigen can be used for serologic diagnosis of heartworm infection and monoclonal antibody reacting with specific antigen(14kDa) can be used for antigen capture ELISA in prepatent period of canine heartworm infection.

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Studies on the ploidy distributions of the hepatocytes isolated in hepatocarcinogensis of rats (랫드의 간발암화과정에서 분리한 간세포의 ploidy 분포변화에 관한 연구)

  • Choi, Kyung-chul;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.32 no.4
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    • pp.649-661
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    • 1992
  • This study was performed to compare DNA content by flow cytometer (FCM) and glutathione S-transferase placental form (GST-P) positive foci for searching objective and accurated properties of tumor. Sprague-Dawley rats aged six weeks were divided into three groups and group 1 and 2 of rats were given an intraperitoneal injection of diethylnitrosamine at 200mg/kg body weight and group 3 of rats were given saline. Three weeks after beginning of the experiment, all groups were performed partial hepatectomy. Group 1 of rats were begun to feed on diets containing 0.02% 2-acetylaminofluorene as a promoter for six weeks, group 2 and 3 of rats were begun to feed on basal diets. At 4, 6, and 8 weeks after initiation, all groups of rats were killed, livers were extracted for H & E stain, immunohistochemical stain, and DNA ploidy analysis. In quantitative analysis for GST-P positive lesion number and area by using Image Analyzer, group 1 and 2 represented significant difference in comparison with group 3. In ploidy distribution, diploid cells of group 1 and 2 were increased significantly in comparison with those of group 3 at 4, 6, and 8 weeks after initiation, respectively tetraploid cells were reduced. But S-phase cells were not changed significantly. It is concluded that ploidy change by FCM is useful as objective data for early detection in hepatocarcinogenesis. Therefore, methodology and study of DNA content are carried out for more objective and accurate ploidy analysis in liver tumor.

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Distribution of positive cells by two monoclonal antibodies (MT1, MB2) in rat spleens (Rat 비장에서 MT1 과 MB2 항체의 양성반응세포 분포)

  • Kwak, Soo-dong;Koh, Phil-ok;Kim, Chong-sup
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.455-460
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    • 1998
  • This study was designed to investigate the distributions of the positive cells in rat spleens by two monoclonal antibodies of MT1 and MB2. The spleens of immature 10 rats (Sprague Duwely, approximately 200gm) were collected and paraffin-embedded sections of spleens were stained with immunohistochemical methods. Higher proportions of MT1-positive cell number in spleens were ordered as marginal zone(8.5~18.1%), red pulp(2.1~8.8%) and periarterial lymphoid sheath(0~1.6%) in white pulp, and those of MB2-positive cell number are ordered as the central area of the periarterial lymphoid sheath(100%), red pulp(29.1~45.0%), marginal zone(15.2~30.4%), and peripheral area of periarterial lymphoid sheath(2.3~3.5%). The positive cells by MB2 are more numerous in number than by MT1. The above results were concluded that the positive cells by above two monoclonal antibodies were scattered throughout the red pulp and marginal zone, but in the central area of the periarterial lymphoid sheath, the MB2-positive cells only were present.

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A Clinical Study on 61 Cases of Tuberculous Tracheobronchitis (기관 및 기관지 결핵 61예의 임상적 고찰)

  • Ahn, Chul-Min;Kim, Hyung-Jung;Hwang, Eai-Suk;Kim, Sung-Kyu;Lee, Won-Young;Kim, Sang-Jin
    • Tuberculosis and Respiratory Diseases
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    • v.38 no.4
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    • pp.340-346
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    • 1991
  • Tuberculous tracheobronchitis is defined as a specific inflammation of the trachea or major bronchi caused by the tubercle bacillus and recognized as one of the most common and serious complication of pulmonary tuberculosis. It had been a diagnostic challenge in prebronchoscopic era and since 1968, fiberoptic bronchoscopy has been accepted as a safe and valuable diagnostic procedure of tuberculous tracheobronchitis. Now, it remains a troublesome therapeutic problem due to its sequelae such as bronchostenosis, bronchiectasis and bronchial deformity. The authors analyzed the clinical features, radiological findings and bronchoscopic findings with pathologic and bacteriologic study on 61 cases of tuberculous tracheobronchitis and following results were obtained. 1) The peak incidence was in the fourth decade and male to female ratio was 1:3.4. 2) The most common symptom was cough (86.9%) and followed by sputum (49.2%), dyspnea (27.9%), fever (19.8%), weight loss (11.5%), hemoptysis (6.6%), hoarseness (6.6%) and chest discomfort (3.3%) and localized wheezing was heard in 18%. 3) In chest X-ray, consolidation with collapse was observed in 70.5%, and followed by consolidation only (18.0%), mediastinal node enlargement (8.2%), cavitary lesion (6.6%), suspicious hilar mass (3.3%) and miliary lesion (1.6%) and there was no abnormal findings in 4.9%. 4) Bronchoscopy showed hyperplastic lesion in 67.2%, mucosal lesion (18.0%), ulcerative lesion (9.8%) and stenotic lesion (4.9%). The most common site of bronchial lesion was right upper bronchus (36.1%) and followed by right main bronchus (34.4%), left main bronchus (29.5%), left upper bronchus (16.4%), right middle bronchus (8.2%), right lower bronchus (6.6%) and left lower bronchus (3.3%). 5) Chronic granulomatous inflammation with or without caseation necrosis on microscopic examination was confirmed in 69.7%, bronchial washing AFB stain was positive in 34.1%, prebronchoscopic sputum AFB stain was positive in 88.1% and postbronchoscopic sputum AFB stain was positive in 30.1%.

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THE BONE CHANGES OF THE MIDPALATAL SUTURE AFTER RAPID PALATAL EXPANSION IN YOUNG ADULT DOGS (유성견에서 상악골 급속확장 후 정중구개봉합부에서의 골조직 변화)

  • Park, Chang-Wook;Yoon, Young-Jooh;Kim, Kwang-Won
    • The korean journal of orthodontics
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    • v.27 no.3 s.62
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    • pp.391-399
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    • 1997
  • Rapid palatal expansion(RPE) is a method of inducing the new bone formation in the palate by separation of the midpalatal suture, which can be done conveniently by placing heavy force across the maxillary dental arch. This experiment was undertaken to examine the histologic changes after RPE and during retention period. Four young adult dogs(a control dog, three experimental dogs) aged 4 to 6 months old were used for this experiment. Expansion screw($Hyrax^{\circledR}$, Dentarum Inc.) was delevered to the palate and fumed 180 degrees every morning and evening for 8 days, giving a total expansion of 7.2mm. A control dog was sacrified at the starting point of this study without any treatment and three experimental dogs were sacrified after RPE, 14-day retention, and 28-day retention in each. Thereafter, those samples were observed with hematoxylin-eosin(H-E) stain, ground section(Villanueva stain), alkaline phosphatase(ALP) stain, tartrate-resistant acid phosphatase(TRA) stain. The results were as followings: 1. After RPE, collagen fiber bundles were stretched along the midpalatal suture and few osteoblasts were flattened-inactive state and also, a little osteoid tissues was observed. Few multinucleated osteoclasts which had TRAP-positive activity in their cytoplasm were seen in horizontal section, whereas a few osteoclasts were seen in frontal section, especially in the nasal floor side of palatal bone. 2. After 14-day retention, collagen fiber bundles were stretched along the midpalatal suture and few osteoblasts which had ALP-positive activity in their cytoplasm were seen. Few multinucleated osteoclasts which had TRAP-positive activity in their cytoplasm were seen in horizontal section, whereas a few osteoclasts were seen in frontal section, especially in the nasal floor side of palatal bone. 3. After 28-day retention, collagen fiber bundles were arranged like those of control dog and osteoblasts which showed a lot of immature bone formation were cuboidal shape and exhibited ALP-positive activity in their cytoplasm. Few multinucleated osteoclasts which had TRAP-positive activity in their cytoplasm were seen in horizontal section, whereas a few osteoclasts were seen in frontal section, especially in the nasal floor side of palatal bone. According to the above results, the new bone formation after rapid palatal expansion was examined after 14-day retention and significantly increased after 28-day retention.

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The Influences of Vascular Endothlelial Growth Factor and Endothelin-1 on Speramtogenesis in Testis (정자형성 과정에서 Vascular Endothelial Growth Factor 및 Endothelin-1 발현의 면역조직화학적 연구)

  • Park, Sung-Woo;Park, Hyun-Jun;Park, Nam-Cheol
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.4
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    • pp.235-244
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    • 2004
  • Objective: The effects on spermatogenesis by expression of vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) were investigated. Materials and Methods: Testicular specimens were obtained from 40 infertile males due to primary testicular failure and from 10 fertile males with other urologic problems. The specimens of infertile males were devided into 4 groups according to histologic findings; Sertoli cell only syndrome (A), maturation arrest (B), hypospermatogenesis (C) and sloughing and disorganization (D). VEGF and ET-1 expression were detected with immunohistochemical stain. Results: VEGF expression on Leydig cell was detected in all cases. But, VEGF expression rates on germ cell were significantly higher in infertile group B, C, D compared to that of the control group (p<0.05). ET-1 expression rates on Leydig cell was significantly lower in all infertile group compared to that of the control group (p<0.05). But, ET-1 expression rates on Sertoli cell was significantly higher in all infertile group compared to that of the control group (p>0.05). In germ cell of infertile group, LH, FSH and prolactin were significantly decreased, and estradiol is increased in positive stain group on ET-1 immunohistochemical stain (p<0.05). VEGF and ET-1 expression were not correlated mean seminiferous tubule diameter (p>0.05). Conclusions: Abnormal spermatogenesis would be reflected in VEGF expression in germ cell.

Screening for Resistance to Purple Seed Stain 'Cercospora kikuchii' by Seed Inoculation Technique. (종자접종법에 의한 대두 자반병 저항성 검정에 관한 연구)

  • Shin-Han Kwon;Jeong-Hae Oh
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.23 no.2
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    • pp.113-117
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    • 1978
  • An attempt was made to establish a mass-screening technique for resistance to purple seed stain .disease in soybean. Seeds sterilized in 1 : 10000 'mercuric chloride for 1 minute and transffered to Petridishes containing 20ml water agar plus 50ppm of the sodium salt of 2,4-D were inoculated by placing a small agar cube containing fungal mycelium on the seed coat of each seed. A positive correlation between natural infection of purple seed :stain and purple discoloration by seed inoculation technique was highly significant and by this technique, some native soybean collections and introduced varieties were tested for resistance to the disease. Most of the soybean varieties tested were susceptible except for the varieties Hill, Harosoy and Sac, resistant comparatively.

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Study on functional elevations of sperm-host glands in domestic hens 1. Histochemical and electron microscopical observations (닭의 정자선(精子腺) 기능(機能) 향상(向上)을 위한 연구(硏究) 1. 조직화학적(組織化學的) 및 전자현미경적(電子顯微鏡的) 관찰(觀察))

  • Kwak, Soo-dong;Woo, Ho-choon;Choe, Sang-yong
    • Korean Journal of Veterinary Research
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    • v.31 no.1
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    • pp.1-10
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    • 1991
  • The present investigation was focussed on histochemical and electron microscopical observations of utero-vaginal(U-V) glands in U-V junctions of domestic hens. In histochemical observations, fine granules by PAS technique for mucopolysaccharides and nile blue stain for acidic lipids were slightly stained on the cytoplasm of U-V glandular epithelium. Larger granules by Sudan black B stain for neutral fat and phospholipids and Sudan III stain for neutral fat were heavily stained on the perinuclear region of the U-V gland epithelial cells. These positive materials were heavily stained on the U-V glandular epithelium of lowfecundity hens and non-laying hens. In scanning electron microscopic findings of the U-V junction surface, the orifices of U-V glands are seen as the crater-like invagination. The neck of the U-V gland and the epithelium of U-V iunction were covered by ciliated epithelial cells. Aggregates of spermatozoa are observed often to be on the necks of the U-V gland. These spermatozoa heads are embedded in the glandular tubules and many spermatozoa tails are free on the epithelium of uterine surface. In transmission electron microscopic findings, the epithelial cells of the U-V glandular orifices were ciliated, columnar cells. The apical regions of these cells contained numerous electrondense, round secretory granules of uniformly size. The epithelial cells of the U-V glandular tubule were columnar or pyramid shape with round or oval nuclei. These epithelial cells have numerous microvilli and also contained electron-dense, round secretary granules of uniformly size and electron-lucent vesicles of various size. Spermatozoa are seen as the cross-sections of various regions of heads and tails in glandular tubules. Also spermatozoa arranged longitudinally parallel within the glandular tubules.

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A report of 38 unrecorded bacterial species in Korea within the classes Bacilli and Deinococci isolated from various sources

  • Kang, Heeyoung;Kim, Haneul;Bae, Jin-Woo;Lee, Soon Dong;Kim, Wonyong;Kim, Myung Kyum;Cha, Chang-Jun;Yi, Hana;Im, Wan-Taek;Kim, Seung Bum;Seong, Chi Nam;Joh, Kiseong
    • Journal of Species Research
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    • v.8 no.2
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    • pp.176-190
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    • 2019
  • A total of 38 bacterial strains within the classes Bacilli and Deinococci were isolated from various sources in Korea. Samples were collected from animal intestine, urine, soil, tidal flat mud, and kimchi. In the sequence comparison and phylogenetic analysis of 16S rRNA sequences, the 38 isolates were assigned to the classes Bacilli and Deinococci with sequence similarities more than 98.7%. Twenty-four strains and 13 strains were classified the order Bacillales and Lactobacillales in the class Bacilli, respectively. In the order Bacillales, there were nine species in the genus Bacillus, seven species in the genus Paenibacillus, and the remaining eight species in the genera Domibacillus, Halobacillus, Virgibacillus, Lysinibacillus, Paenisporosarcina, Planococcus, Savagea, and Staphylococcus. In the order Lactobacillales, there were four species in the genus Lactobacillus, three species in the genus Leuconostoc, three species in the genus Lactococcus, and the remaining three species in the genera Aerococcus, Enterococcus, and Streptococcus. One species was related to the genus Deinococcus of the order Deinococcales. Most of the isolated strains were Gram-stain-positive, but some were Gram-stain-variable or Gram-stain-negative. Cells were rod or cocci-shaped. Based on the results of 16S rRNA analysis, we report 38 strains as previously unrecorded species to Korea, and the basic characteristics of strains are described herein.

EFFECT OF COBALT-60 IRRADIATION ON THE DEVELOPING TOOTH GERM OF RAT (Cobalt-60이 발육치배조직에 미치는 영향에 관한 실험적 연구)

  • Lee Ki Sik
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.6 no.1
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    • pp.33-38
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    • 1976
  • The author observed the effects of the cobalt-60 irradiation on the amelogenesis and dentinogenesis of the albino rat fetuses by means of histological and histochemical methods. Females in oestrus were mated overnight and examined the next morning for evidence of copulation. The lower left abdomen of mothers were exposed to cobalt-60 irradiation on the 10th day of gestation, l00R 200R and 300R respectively. The fetuses were removed from the mothers on the 18th day of gestation. The employed histochemical methods were PAS reaction, colloidal iron reaction, aldehyde fuchsin stain, α-amino acid reaction, -SH radical reaction and methyl- green pyronin stain. The results were as follows; 1. The group irradiated by l00R made no histological differences in comparison with the control group. 2. Increasing the irradiation to 200R, abnormal dentin formation occured, and resulted in enamel hypoplasia and in atrophy and necrosis of odontoblasts. In dentinal papilla, the dilation and the degeneration of the blood vessels, excessive reticular atrophy and osteodentin were revealed. 3. With the more irradiation (200R-300R), the positive material of PAS, α-amino acid and aldehyde fuchsin tended to decrease in the ameloblast and the odontoblast. No significant changes appeared in DNA, the stainability of methylgreen pyronin.

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