• Title/Summary/Keyword: blue stain

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Comparative Analysis of Muscle Proteome from Porcine White and Red Muscles by Two-dimensional Electrophoresis (이차원전기영동법을 이용한 white muscle과 red muscle간의 단백질 발현양상의 비교분석)

  • Kim, N.K.;Joh, J.H.;Chu, K.S.;Park, H.R.;Park, B.Y.;Kim, O.H.;Lee, C.S.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.731-738
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    • 2003
  • The technique known as proteomics is useful for characterizing the protein expression pattern of a particular tissue or cell type as well as quantitatively identifying differences in the levels of individual proteins. In present study, we carried out the comparative expression patterns of white and red muscles. We used the two-dimensional electrophoresis(2-DE) for analyzing the protein expression. Proteins isolated from porcine white and red muscles were separated by 12% poly-acrylamide gel and then were detected by coomassie blue and silver staining. More than 600 protein spots were detected on each 2-DE gel. By visual analysis of the stained gel, five proteins were identified to be differentially expressed in the white vs red muscle. By database searching based on the molecular weights and pI(isoelectric point) of the five proteins, three of them were found to be most close to troponin I, T and myoglobin. However, further researche is needed for identification and functional analysis of the unidentified proteins. In conclusion, we found five proteins, which are differentially expressed in the white vs red muscle. The functional analysis of the differentially expressed proteins will provide valuable information on biochemical characteristics of the muscle type.

FORMATION OF BASEMENT MEMBRANE AND STRATIFICATION OF RABBIT ORAL KERATINOCYTES CULTURED ON HUMAN ACELLULAR DERMAL MATRIX (인간 무세포성 진피기질 위에 배양한 가토 구강각화상피세포의 중충화와 기저막 형성에 관한 연구)

  • Kim, Yong-Deok;Ahn, Kang-Min;Yum, Hak-Yeol;Chung, Hun-Jong;Kim, Soung-Min;Jang, Jeong-Won;Sung, Mi-Ae;Park, Hee-Jung;Hwang, Soon-Jung;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.6
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    • pp.510-522
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    • 2005
  • To assess the clinical applicability of bio-artificial mucosa which was made with autologous oral keratinocytes and human acellular dermal matrix, the formation of basement membrane and stratification of oral keratinocytes were evaluated. Six New Zealand white rabbits (around 2kg in weight) were anesthetized and its buccal mucosa was harvested (1.0 $\times$ 0.5cm size). Oral keratinicytes were extracted and cultured primarily with the feeder layer of pretreated NIH J2 3T3 fibroblast. These confluent cells were innoculated on the human acellular dermal matrix and cultured in multiple layer by air-rafting method. After 3, 5, 7, 10, 14 days of culture, each cultured bio-artificial mucosa was investigated the number of epthelial layer of by H&E stain and toluidine blue stain. The immuhohistochemical methods were used to evaluate the cell division capacity, the formation of basement membrane, and it's property of specific cells (PCNA, cytokeratin 14, laminin). Transmission electromicroscopy was used for the attachment between cells and matrix with the number of hemidesmosome. In result, the numbers of layer of stratified growth of oral keratinocyte cultured on the human acellular dermal matrix and the number of hemidesomal attachment between epithelial cells and human acellular dermal matrix were similar to the layers of normal oral mucosa after 10 days of culture. The cell division rate, basement membrane formation and proliferation rate increased as culture period increased. With these results, bio-artificial mucosa with autologous oral epithelial cells cultured on the acellular dermal matrix had clinically adaptable properties after 10 days' culture and this new bio-artificial mucosa model with relatively short culture time can be expected clinical applicability.

The Morphology Study of Organ Surface BongHan Ducts and Corpuscle (장기표면의 내외봉한관과 봉한소체의 형태학적 관찰)

  • Ahn, Seong-Hun;Kim, Min-Su;Lee, Sang-Hun;Kwon, O-Sang;Kim, Jae-Hyo;Soh, Kwang-Sup;Sohn, In-Chul
    • Korean Journal of Acupuncture
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    • v.26 no.1
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    • pp.79-84
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    • 2009
  • Objective : In 1960's Bonghan Kim's team found BongHan(BH) ducts which were presumed as acupuncture meridians and BH corpuscles. They asserted Bonghan theory and SanAl theory which was involved in cell division and cell restoration. However, many other experiments which had been operated to demonstrate and find the existence of BH ducts had failed because of the secret of blue stain drugs. During the last several years, BongHan theory has been revived through experimental researches to find the anatomical structures of BH ducts and corpuscles by Soh's Biomedical Physics Lab. Soh's research team used the staining with Janus Green B, Alcian blue, nanoparticles and Acridine Orange. We used DAPI staining to find the existence of BH ducts and the corpuscles and to observe nuclear arrangement. Methods : We used japan white rabbits as experimental animals. BH ducts and corpuscles were stained with DAPI. The nucleus configuration in BH ducts stained with DAPI were observed with microscope. Results : In this study, we found thread like structures in silver white color distinguished from the blood vessels, nerves and lymph vessels. These thread like vessels in the linear duct shape were connected to same colored mass in the ball shape. Thread like structures we found could be separated easily from the surrounding other organ mass. The nuclei of the thread like structure in DAPI staining, are about 10${\sim}$20${\mu}m$ length, in rod shape and linear arrangement. Conclusion : We concluded that the thread like structure we found was same vessel reported by Soh's research team, BongHan ducts and corpuscle.

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Effects of 0.1% Tacrolimus on Canine Skin Mast Cells and Eosinophils (개에서 0.1% FK-506 연고의 피부 비만세포와 호산구에 미치는 영향)

  • Jeong, A Young;Bae, Seulgi;Oh, Taeho
    • Journal of Veterinary Clinics
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    • v.31 no.2
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    • pp.95-101
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    • 2014
  • Five dogs were used to determine whether 0.1% tacrolimus ointment application for one day would inhibit IgE-mediated late-phase reactions (LPRs). It was consisted of three periods: one period without therapeutic administration (control) and two periods of treatment with either the tacrolimus ointment or vehicle. Induction of IgE-mediated LPRs was induced by intradermal injections of 0.05 ml (0.14 mg/ml) of solution of goat anti-canine IgE polyclonal antibodies. Each section for mast cells (MCs) and eosinophils (EPs) was stained with acidified toluidine blue, and Luna's stain, respectively. Assessment of anti-inflammatory effect of tacrolimus ointment composed of cell counts of MC and EP from lesions of induced LPR. In normal canine biopsies, the number of dermal MCs and EPs were $12.3{\pm}1.4cells/mm^2$ and $3.1{\pm}1.3cells/mm^2$, respectively. MC counts dramatically decreased at time dependent manner after anti-IgE administration. However, the number of MCs on 6 hours after challenge was significantly less decreased in the groups treated with the tacrolimus, as compared with control and vehicle group. The number of EPs on 24 hours after challenge was significantly lower in the group treated with the tacrolimus than in the control and vehicle groups. In conclusion, this study revealed that 0.1% tacrolimus ointment in dogs may exert a potent anti-inflammatory effect on inhibition of MC degranulation and also secondary prevention of EP infiltration during LPR.

The Efficacy of Conjugated Linoleic Acid and Carprofen in Progression of Early Stage of Experimentally Induced Osteoarthritis in Dogs (개에서 Conjugated Linoleic Acid와 Carprofen이 실험적 골관절염 초기 진행에 미치는 효능)

  • Park, Se-il;Bae, Jae-sung;Kwon, Young-sam;Jang, Hwan-soo;Li, Wen-xue;Lim, Jae-hyun;Eom, Ki-dong;Kim, Jung-eun;Jang, Kwang-ho
    • Journal of Veterinary Clinics
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    • v.20 no.3
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    • pp.278-285
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    • 2003
  • This study was performed to compare early stage changes following the administrations of Conjugated Linoleic Acid (CLA), Carprofen and their combinations in the experimental canine cranial cruciate ligament rupture. Twenty five mongrel dogs were divided into five groups; Groups 1, 2, and 3 received a sectioning the cranial cruciate ligament (CCL) of the right stifle joint, and were administered CLA (250 mg/kg/day/orally), carprofen (4.4 mg/kg/day/orally), and their combinations for 4 weeks beginning 4 weeks postsurgery, respectively. Group 4 received sectioning CCL and no treatment. Group 5 was composed of unoperated normal dogs. The macroscopic observation of cartilage erosions on both the condyles and plateaus were evaluated, and the severity of the cartilage lesions and synovial inflammation was examined histologically at eight weeks after surgery. Histological examinations including hematoxylin and eosin stain, standard toluidine blue method, PAS technique and Masson trichrome technique, hematologic and radiographic evaluation were perfonned after experimental surgery. Slight yellowish discoloration of the surface was found in some of the experimental dogs. However macroscopic findings showed no significant differences among the groups. In radiographic findings, cranial displacement of the proximal tibia relative to the femoral condyles in all groups was observed but no significant differences among the groups was seen. This study showed that oral administration of CLA, carprofen and their combinations revealed no early stage change in the canine stifle joint following experimental rupture of the cranial cruciate ligament.

Histochemical Studies of the Intestinal Mucosubstances in Agramus agramus, Inimicus japonicus, Epinephelus chlorostigma, and Helirolenus dactylopterus (노래미, 쑤기미, 구실우럭 및 홍감펭 장관 점액질에 대한 조직화학적 연구)

  • Jeong, Gil-Nam;Jo, Un-Bock
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1669-1674
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    • 2007
  • The conventional histochemical stains were used to study the properties of mucosubstances of the intestine in four teleostean species, i. e., Agramus agramus, Inimicus japonicus, Epinephelus chlorostigma, and Helicolenus dactylopterus, all belonging to the order Perciformes. The following methods were used; periodic acid Schiff (PAS), alcian blue (AB) pH at 2.5, AB pH at 1.0, AB pH at 2.5-PAS, and aldehyde fuchsin (AF) pH at 1.7-AB pH at 2.5 stain. The mucosal folds of intestines show differences in species and regions. Widely distributed in all portions studied, goblet cells situated between cylinderical epithelial cells are round or ovoid in shape. They were most densely distributed in distal intestine. In the middle and. distal intestine of Epinephelus chlorostigma and all regions of intestines of Agramus agramus, Inimicus japonicus, and Helicolenus dactylopterus, the presence of both acidic and neutral mucin was confirmed. The property of acidic mucosubstance was sialomucin. Neutral mucin was only encountered in the proximal intestine of Epinephelus chlorostigma. The amount and property of mucin showed differences in species and regions. In the distal intestine of Inimicus japonicus, the amount of acidic mucin is similar to that of neutral mucin. In all regions of intestine of Agramus agramus, proximal and middle intestine of lnimicus japonicus, middle and distal intestine of Epinephelus chlorostigma, and distal intestine of Helicolenus dactylopterus, acidic mucin occured more frequently than neutral mucin. The proximal and middle intestine of Helicolenus dactylopterus have more neutral mucin than acidic mucin.

Effect of irradiation on the acinar cells of submandibular gland in streptozotocin-induced diabetic rats (방사선조사가 당뇨 백서의 악하선 선포세포에 미치는 영향)

  • Lee Seung-Hyun;Hwang Eui-Hwan;Lee Sang-Rae
    • Imaging Science in Dentistry
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    • v.33 no.3
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    • pp.161-169
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    • 2003
  • Purpose: To observe the histologic changes and clusterin expression in the acinar cells of the submandibular gland in streptozotocin-induced diabetic rat following irradiation. Materials and Methods: Mature Sprague-Dawley rats were divided into three groups: control, diabetic, and diabetic-irradiated groups. Diabetes mellitus was induced in the Sprague-Dawley rats by injecting streptozotocin, while the control rats were injected with citrate buffer only. After 5 days, rats in diabetic-irradiated group were irradiated with single absorbed dose of 10 Gy to the head and neck region. The rats were killed at 1, 3, 7, 14,21, and 28 days after irradiation. The specimen including the submandibular gland were sectioned and observed using histologic and immunohistochemical methods. Results : Morphologic change of acinar cells was remarkable in the diabetic group, but was not observed in the diabetic-irradiated group. Necrotic tissues were observed in the diabetic-irradiated group. Coloring of toluidine blue stain was most increased at 14 days in the diabetic group, however there were no significant change throughout the period of the experiment in the diabetic-irradiated group. Expression of clusterin was most significant at 14 days in the diabetic group, but gradually decreased with time after 7 days in the diabetic-irradiated group. Degeneration of clusterin was observed in the diabetic-irradiated group. Conclusion : This experiment suggests that the acinar cells of submandibular gland in rats are physiologically apoptosed by the induction of diabetes, but that the apoptosis is inhibited and the acinar cells necrotized after irradiation.

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Peripheral Nerve Regeneration After Various Conditioned Side to Side Neurorrhaphy in Rats (말초신경 손상 후 측측문합을 이용한 신경이식시 신경이식의 수에 따른 신경재생 및 근육 기능 회복에 관한 비교 연구)

  • Kim, Sug-Won;Chung, Yoon-Kyu;Kang, Sang-Yoon;Cho, Pil-Dong
    • Archives of Reconstructive Microsurgery
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    • v.10 no.1
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    • pp.12-17
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    • 2001
  • Recovery of nerve injury is conditioned by various factors including physical state, injured site, cause of injury, and neurorrhaphy Many researchers have reported on regeneration of nerve using end to side neurorrhaphy. The purpose of this study was to examine regeneration of nerve in various conditioned side to side neurorrhaphy. Total of 25 male Sprague-Dawley rats weighing 220 to 250 gm were divided into five groups of five rats each. The group 1, sham group, composed of dissection only without nerve transaction. The group 2, control group, composed of nerve division only without neurorrhaphy or sural nerve graft. The group 3 composed of one segmental sural nerve graft between the tibial and peroneal nerve after division. Group 4 had two segment graft, and the group 5 with three segment graft, each segment being 6mm long and 5 mm apart. The side to side neurorrhaphy was performed between peroneal nerve and tibial nerve using segmental sural nerve graft in rats. We exposed the sciatic nerve, tibial nerve, peroneal nerve, and sural nerve on left side with prone position. The peroneal nerve was cut on the bifurcation site from tibial nerve and the side to side epineurial neurorrhaphy was performed between peroneal nerve and tibial nerve through 6 mm sural nerve segment graft with 11-0 nylon under operating microscope. The electromyography and the weight from ipsilateral tibialis anterior muscle was performed at one month after neurorrhaphy Peroneal and tibial nerve was examined at distal and proximal to the neurorrhaphy site by methylene blue stain under light microscope for histologic appearance. The number of nerve fibers were counted using the image analyzer. Statistically, both in electromyography and number of nerve fibers, the differences in values between the groups were significant.

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Flocculation Effect of Alkaline Electrolyzed Water (AEW) on Harvesting of Marine Microalga Tetraselmis sp.

  • Lee, Su-Jin;Choi, Woo-Seok;Park, Gun-Hoo;Kim, Tae-Ho;Oh, Chulhong;Heo, Soo-Jin;Kang, Do-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.3
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    • pp.432-438
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    • 2018
  • Microalgae hold promise as a renewable energy source for the production of biofuel, as they can convert light energy into chemical energy through photosynthesis. However, cost-efficient harvest of microalgae remains a major challenge to commercial-scale algal biofuel production. We first investigated the potential of electrolytic water as a flocculant for harvesting Tetraselmis sp. Alkaline electrolyzed water (AEW) is produced at the cathode through water electrolysis. It contains mineral ions such as $Na^+$, $K^+$, $Ca^{2+}$, and $Mg^{2+}$ that can act as flocculants. The flocculation activity with AEW was evaluated via culture density, AEW concentration, medium pH, settling time, and ionic strength analyses. The flocculation efficiency was 88.7% at 20% AEW (pH 8, 10 min) with a biomass concentration of 2 g/l. The initial biomass concentration and medium pH had significant influences on the flocculation activity of AEW. A viability test of flocculated microalgal cells was conducted using Evans blue stain, and the cells appeared intact. Furthermore, the growth rate of Tetraselmis sp. in recycled flocculation medium was similar to the growth rate in fresh F/2 medium. Our results suggested that AEW flocculation could be a very useful and affordable methodology for fresh biomass harvesting with environmentally friendly easy operation in part of the algal biofuel production process.

EXPERIMENTAL STUDY ON THE ANTERIORLY DISPLACED TEMPOROMANDIBULAR JOINT MENISCUS IN RABBIT (실험적으로 전방이동시킨 가토의 악관절원판에 관한 연구)

  • Choi, Nack-Jun;Chang, Young-Il
    • The korean journal of orthodontics
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    • v.21 no.1 s.33
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    • pp.53-76
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    • 1991
  • The study was designed to analyse the reorganization of the rabbit TMJ meniscus which was anteriorly displaced by surgery. The author compared the anteriorly displaced groups with control group. After surgical opening of the left rabbit TMJ space, cut the posterior attachment of the meniscus, and pushed it under the undercut area of the condyle head. Experimental groups were sacrificed by 1, 2, 4, 8 weeks after surgery. The samples were analysed with light microscope under T-B stain and electron microscope. The results were as follows: 1) The rabbit TMJ meniscus consisted of thick anterior and posterior band running different way, and comparative thin intermediate band runining antero-posteriorly. 2) Round oval shape chondrocyte-like cells were imbeded between the collagen fiber bundles and composed of proteoglycan granules, that showed metachromasia with toluidine blue, around the cell matrix. 3) Type II collagen fiber bundles in experimental group occured degenerative changes in organic patterns at 8 weeks, but those of type I collagen fiber bundles sustained longer, 4) The typical fibrocartilage of the rabbit TMJ meniscus was changed into fibrotic mode in process of time and showed the degenerative changes, which contained hyperplasia, calcification, resorption and hyalinization in the connective tissue. 5) The hyperplastic change of the synovial membrane in 4 week group and transitional change from fibrocyte to chondrocyte in cell type in 8 week group were observed. 6) The diameters of collagen fibers were diminished with the degenerative changes, the shape of the fibers became wavier and more nonorganic in running pattern and fiber bundle spaces widened.

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