• Title/Summary/Keyword: Immunoreaction

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Differential expression of glycoconjugates, estrogen receptor-α, c-fos and c-jun in the vagina of normal and ovariectomized rat (흰쥐 발정주기와 난소절제에 따른 질상피의 glycoconjugates, estrogen receptor-α, c-fos 및 c-jun 분포변화)

  • 최병태;길영기;김강련;김순옥;최영현;이준혁
    • Journal of Life Science
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    • v.12 no.3
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    • pp.317-324
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    • 2002
  • We investigated the compositional changes of glycoconjugates (GCs) and expression of estrogen receptor (ER)-$\alpha$, c-fos and c-jun in the vagina of normal and ovariectomized rats by histochemical and immunohistochemical methods. The mucinous transformation of the superficial layer that occurred from late diestrus to proestrus was accompanied with extensive enrichment of GCs. According to the cyclic changes of the vagina, distinct reactivity patterns such as SBA affinity in the diestrus and Con A affinity in the diestrus and estrus phase was observed. However, weak staining for GCs was detected in the atrophied vaginal epithelium of ovariectomized rats. ER-$\alpha$ immunoreaction was mainly demonstrated in the basal layer of epithelium and estrus cycle-related variation in the number of ER-$\alpha$ immunoreaction were not pronounced. But the stromal cells showing ER-$\alpha$ immunoreaction were abundantly observed from diestrus to estrus phase. The most numerous c-fos immunoreactive cells were observed in the basal and intermediate layer of epithelium and stromal fells from the proestrus to estrus phase and c-jun in the basal layer of epithelium during estrus phase. The c-jun immunoreaction of stromal cells expressed only in the estrus phase. In the ovariectomized rats, a few of ER-$\alpha$, c-fos and c-jun immunoreactive cells were observed in the vaginal epithelium and no immunoreaction were found in the stromal cells. ER-$\alpha$ and c-fos immunoreaction fully expressed in the proestrus coincident with the cell proliferation, mutinous transformation and cornification of vaginal epithelium. These data indicate that vagina epithelium and stromal reals express multiple protein such as ER-$\alpha$, c-fos and c-iun by estrogen that may function in process of cells proliferation and differentiation of vagina epithelium.

Effects of Electroacupuncture on the Expression of Cyclooxygenase in the Spinal Cord of Carrageenan-injected Rat (전침이 carrageenan유발 동통모델의 척수배각내 cyclooxygenase 발현에 미치는 영향)

  • Choi, Yung-Hyun;Lee, Yong-Tae;Choi, Byung-Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.3
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    • pp.749-752
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    • 2005
  • We investigated the effects of electroacupuncture (EA) on the expression of cyclooxygenase in the spinal cord of acute inflammatory pain model. Inflammation was induced by an intraplantar injection of 1% carrageenan into the right hind paw of Sprague-Dawley. Bilateral 2 Hz EA stimulation with 0.5 mA, 1 mA and 3 mA were delivered at those acupoints corresponding to Zusanli and Sanyinjiao in man via the needles in carrageenan-injected rats. Three hours after carrageenan injection, effects of EA on cyclooxygenase (COX) expression were observed in the dorsal horn of the spinal cord using immunohistochemical method. The immunoreaction of COX-1 tended to increase in the superficial laminae and the neck of the dorsal horn as compared with normal. The COX-2 immunoreaction in the carrageenan-injected rat was also significantly increased in the all regions of the dorsal horn as compared with normal one. However, COX-1 immunoreaction in carrageenan-injected rat were decreased in the superficial laminae and neck of the dorsal horn by low intensity of EA stimulation. Except high intensity of EA stimulation in the superficial laminae, COX-2 expression was attenuated in all regions of the dorsal horn by all types of EA treatment. It is concluded that EA treatment may attenuate inflammatory pain in carrageenan-injected rat through modulating expression of COX-2 in the dorsal horn of the spinal cord.

Hapten Synthesis and Influence of Coating Ligands on Enzyme-linked Immunoreaction of DDT

  • Hong, Ji- Youn;Kim, Jong-Hyun;Choi, Myung-Ja
    • Bulletin of the Korean Chemical Society
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    • v.23 no.10
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    • pp.1413-1431
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    • 2002
  • For the development of immunodetection method of 4,4'-dichlorodipheny-2,2,2-trichloroethane (p,p'-DDT), a persistent and broad toxic organochlorine insecticide, various DDT derivatives were synthesized and characterized for the use of immunogens and the coating ligands for the antibody evaluation. The appropriate lengths of linkers were introduced to investigate more efficient DDT derivatives. Among these hapten derivatives, 2,2-Bis(4-chlorophenyl)acetic acid (DDA), 5,5-Bis(4-chlorophenyl)-5-hydroxypentanoic acid (DDHP) and 5,5-Bis(4-chlorophenyl)-5-chloropentanoic acid (DDCP) were conjugated with keyhole limpet hemocyanin (KLH) for the use of immunogen to produce antibodies. 6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoic acid (DDHH) and 3-[6,6-Bis(4-chlorophenyl)-6-hydroxyhexanoylamino]propanoic acid (DDHHAP) in addition to above hapten derivatives were conjugated to ovualbumin (OVA) and bovine serum albumin (BSA) for the use of coating ligands to measure the titration level of antibody and the displacement of free analytes. Three matching pairs of antibodies and coating ligands were selected for the simultaneous detection of p,p'-DDT and its related compounds of DDA and 2,2-bis(4-chlorophenyl)-1,1-dichloroethylene (p,p'-DDE) by investingating the displacement of free analytes in an indirect ELISA. These were PAb #1 and coating ligand DDCP-OVA, PAb #1 and DDHHAP-OVA, and PAb #3 and DDHHAP-OVA. The most useful immunoreaction for DDT analytes were obtained using PAb #3 and coating ligand DDHHAP-OVA showing 3.4 ng/mL of lower limit of detection. These results indicated that titration level and free analytes displacement were greatly influenced by hapten derivatized and carrier proteins conjugated.

Effect of Garlic Extract on the Activation Pattern of MAPK Signaling in the Rat Heart After a Bout Exercise (마늘추출물이 운동부하 흰쥐의 심장내 MAPK signaling 활성에 미치는 영향)

  • Lee, Jun-Hyuk;Chung, Kyung-Tae;Lee, Yang-Tae;Choi, Yung-Hyun;Choi, Byung-Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.5
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    • pp.1299-1303
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    • 2008
  • Since exercise training induces mechanical stress to the heart, we examined the activation pattern of mitogen-activated protein kinase(MAPK)s signaling pathway by immunohistochemistry. The immunoreactions of MAPKs signaling with c-fos and Schiff's reaction were increased in the cardiac muscle of exercised rat compared to normal one except immunoreaction for MEK1/2 and ERK1/2 and p38. However, the immunoreaction of phospho-JNK and phospho-p38 with early gene c-fos were arrested markedly in water extract of Alliium sativum (WEAS) treated rat compared to exercised one. Since MAPKs signaling does play a protective role in response to pathological stimulus in the heart, results in the present study suggest that WEAS may act as a alleviating agent for exercise-induced stress to. heart through regulating MAPKs signaling activation.

Primary Culture and Characteristics of Blood-Brain Barrier Endothelial Cells from Rat Brain Microvessel (쥐의 뇌 미세혈관 내피세포를 이용한 뇌혈관장벽 내피세포의 배양과 특성)

  • Lee, Hee-Sang;Kim, Seok-Jung;Kim, Dae-Jin;Chung, Yoon-Hee;Kim, Sung-Su;Lee, Won-Bok;Kim, Kyung-Yong
    • Applied Microscopy
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    • v.35 no.4
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    • pp.55-63
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    • 2005
  • The characteristics of primary cultured rat brain microvessel endothelial cells (RBMECs) were studied using microscopy, immunohistochemistry and measuring of transendothelial electrical resistance (TER). The RBMECs formed a monolayer by $5{\sim}6$ days after plating and showed characteristics of whirling appearance. The TER increased until day 5 and decreased then. There was few immunoreaction with anti-GFAP, anti-GalC, anti-neurofilament 160/200 kD antibodies. So the contamination of astrocyte, oligodendrocyte, and neuron. could be ruled out.. Immunoreaction to vWF antigen was widespread througout the cytoplasm as Weibel-Palade granule. Immunoreaction to tight junction proteins, i.e. occludin, ZO-1, and ZO-2 was seen at cell contact. In summary, RBMECs isolated and cultured showed morphological, immunohistochemical and electrical characteristics of blood-brain barrier (BBB). The in vitro BBB model can be used in studying characteristics of in vivo BBB.

Immunohistochemical Studies on S-100 Protein and Lactoferrin in Salivary Gland Tumors (타액선 종양에 있어서 S-100 단백과 Lactoferrin에 관한 면역조직화학적 연구)

  • Choi Dae-Sik;Kim Sang-Hyo
    • Korean Journal of Head & Neck Oncology
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    • v.9 no.1
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    • pp.74-87
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    • 1993
  • Immunohistochemical studies on S-100 protein and lactoferrin were carried out to evaluate the existence and distribution pattern of S-100 protein and lactoferrin positive cells in salivary gland tumors. The specimens used were 25 cases of pleomorphic adenoma, 2 cases of monomorphic adenoma, 2 cases of mucoepidermoid tumor, 2 cases of acinic cell tumor, 3 cases of adenoid cystic carcinoma and 2 cases of adenocarcinoma occured in parotid and submandibular salivary gland. ABC kits(Dako corp. Copenhagen. Denmark) for S-100 protein and lactoferrin were used. The results obtained were summarized as follows: In the normal salivary gland. positive immunoreaction for S-100 protein was observed in myoepithelial cells of acini and intercalated ducts. Positive immunoreaction for lactoferrin was observed in serous acinic cells, epithelial cells of intercalated ducts, and excretory material in the ductal lumina. In the pleomorphic and monomorphic adenomas. most of tumor cells were positive for S-100 protein, while luminal tumor cells in gland-like or duct-like structures were rarely positive for lactoferrin. In mucoepidermoid tumor, most of squamous cells and a few of intermediate cells were positive for S-100 protein, but all of tumor cells were negative for lactoferrin. In acinic cell tumor, most of tumor cells were positive for lactoferrin, but all of tumor cells were negative for S-100 protein. In adenoid cystic carcinoma, basaloid tumor cells in trabecular structure were focally positive for S-100 protein. and in adenocarcinoma, many of tumor cells were posivive for both S-100 protein and lactoferrin. Thus, according to the embryonic stage of the development of the tumor cell origin, it was possible to classify the salivary gland tumor as followings: mucoepidermoid carcinoma which originated from the earliest stage, acinic cell tumor which originated from the end stage. Between these two extremes, there were pleomorphic adenoma, adenoid cystic carcinoma and adenocarcinoma which originated in the middle stage of the development of .the salivary glands. Based on the above results, it can be stated that S-100 protein is demonstrated in tumor cells orginated from myoepithelial cells and lactoferrin in glandular differentiated tumor cells.

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Study of Effects of Electroacupuncture on the Hippocampal Cholinergic Neuronal Activity

  • Lee Kwang Ro;Kang Sei Young;Yoon Ji Won;Cho Chang Hyun;Sung Kang Keyng
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1850-1855
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    • 2004
  • The purpose of this report is to investigate the electroacupuncture effect on the cholinergic neuronal activation in the hippocampal CA1 section. The electroacupuncture was performed on S36 of white rats and its consequencies were investigated by immunohistochemical method. Hippocampal CA1 sections of Sprague Dawley white male rats electroacupunctured on S36 at 20㎐ and 100㎐ are stained by cresyl violet to show that the values of 100㎐ and 20㎐ group increased significantly compared to sham group's one. Especially, 100㎐ group shows stronger neuronal activation compared to 20㎐ group. Induction of AChE, a neurotransmitter, in hippocampal CA1 is increased significantly in 100㎐ and 20㎐ group compared to sham group. Especially 20㎐ group shows higher AChE immunoreaction than 100㎐ does, although it wasn't significant enough. Induction of NGF(Nerve Growth Factor) in hippocampal CA1 sections was observed higher in 20㎐ and 100㎐ group than in sham group. Especially, 20㎐ group shows higher NGF immunoreaction compared to 100㎐. The facts above indicate that the eletroacupucture is effect to the cholinergic neuronal activation of hippocampus induced by focal ischemia.

Ultra-sensitive Determination of Salinomycin in Serum Using ICP-MS with Nanoparticles

  • Cho, H.K.;Lim, H.B.
    • Bulletin of the Korean Chemical Society
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    • v.35 no.11
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    • pp.3195-3198
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    • 2014
  • An ultra-sensitive detection method for small molecules such as antibiotics was developed using ICP-MS with magnetic and $TiO_2$ nanoparticles. Since most of the antibiotics are too small to employ a sandwich-type extraction through an immunoreaction, a non-specific platform was employed, in which the target was extracted by magnetic separation, followed by tagging with $TiO_2$ nanoparticles of 11.2 nm for ICP-MS measurement. The detection limit for salinomycin obtained from spiked serum samples was $0.4ag\;mL^{-1}$ (${\pm}10.3%$), which was about $1.5{\times}10^6$ times lower than that of LC-MS/MS and about $1.2{\times}10^{11}$ times better than that of ELISA. Such an excellent sensitivity enabled us to study the toxicity of antibiotics exposed to human beings by determining them in serum.

Bacillus cellulyticus K-12 Crystalline Cellulose-Degrading Avicelase Gene and Expression in Eschterichia coli

  • Cheorl-Ho Kim;Woo
    • The Korean Journal of Food And Nutrition
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    • v.6 no.4
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    • pp.314-321
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    • 1993
  • We have cloned the Bacillus cellulyticus K-12 avicelase (Avi, E.C.3.2.1.4) gene (ace A) In E. coli. This was accompanied by using the vector PT7T3U 19 and Hind W -Hind m libraries of Bacillus cellulyticus K-12 chromosomal inserts created in 5.cofi. The Libraries were screened for the expression of avicelase by monitoring the immunoreaction of the anti-avicelase (immunoscreening). Positive clones (Ac-3, Ac-5, and Ac-7) contained the identical 3.5kb Hind III fragment as determined by restriction mapping and Southern hybridization, and expressed avicelase efficiently and constituvely using its own promoter in the heterologous host. From the immunoblotting analysis, a polypeptide which showed a CMCase activity with an Mr of 54000 was detected.

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DISTRIBUTION IN FIBRONECTIN OF THE RABBIT TEMPOROMANDIBULAR JOINT TISSUES FOLLOWING SURGICAL INDUCTION OF ANTERIOR DISK DISPLACEMENT : IMMUNOHISTOCHEMICAL STUDY (악관절원판의 인위적 전방변위술시행후 악관절구성조직에서 Fibronectin의 분포변화)

  • Kim, Uk-Kyu;Chung, In-Kyo;Park, Bong-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.25 no.4
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    • pp.337-349
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    • 1999
  • The extracellular matrix(ECM) is a complex network of different combination of collagens, glycosaminoglycans, laminin, fibronectin, and many other glycoproteins including proteolytic enzymes. The composition and organization of the ECM contributes to the uniques physical or biomechanical properties of a tissue. Fibronectins(FN) are dimeric glycoproteins located on cell surfaces, in the matrix of connective tissue, and in blood. Fibronectins mediate cell attachment to collagen substratum and have been implicated in a variety of important biological processes, including embryogenesis and cell differentiation. The purpose of this study was to determine the effects of surgical induction of anterior disk displacement(ADD) on distribution of fibronectin in the rabbit temporomandibular joint(TMJ) tissues included the articular cartilage, disc, retrodiscal tissue, articular eminence using an immunohistochemical technique. The left TMJ was exposed surgically, and all discal attachments were severed except for the posterior attachment. The disk was then repositioned anteriorly and sutured to the zygomatic arch. The right TMJ served as a shamoperated control. Normal joints were used as a nonoperated control. Fourty-five rabbits were used for experiments in total. For fibronectin immunohistochemical study, eighteen rabbits (one normal group and 5 experimental groups, each group consists of 3 rabbits) were used. The experimental rabbits were sacrified after operation period of 2, 3, 4, 6 and 8 weeks on fibronectin. The obtained results were as follows ; 1. Fibronectin immunoreaction on all TMJ tissues(mandibular condyle, articular disc, retrodiscal tissue, articular eminence) in the normal rabbit was observed. Especially the reverse cell layer and proliferation zone of articular cartilage of condyle show strong positive reaction. 2. Depletion of fibronectin in the all TMJ tissues except hypertrophic zone of articular cartilage occurred at 2 weeks following induction of ADD. 3. The restoration of immunoreaction at 4 weeks was observed and a progressive increasing reaction at 6 weeks, 8 weeks also was found. Our study generally showed degenerative changes in TMJ tissues after ADD although TMJ tissues adapted or degenerated to abnormal loads and stress distribution according to the remodeling capacity of TMJ tissues.

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