• Title/Summary/Keyword: 조직염색법

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Expression of Calponin 3 in the Striatum Following 3-Nitropropionic Acid-induced Neurotoxicity (선조체에서 3-nitropropionic acid 투여 후 calponin 3의 발현 연구)

  • Choi, Yun-Sik
    • Journal of Life Science
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    • v.23 no.1
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    • pp.125-130
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    • 2013
  • Calponin 3 is an F-actin-binding protein and plays a key role in regulating spine plasticity and synaptic activity in neurons. Unlike the other subtypes, calponin 1 and 2, which are expressed in smooth and cardiac muscle cells, calponin 3 is highly expressed in the brain. The goal of this study was to elucidate the spatiotemporal expression pattern of calponin 3 following repeated administration of 3-nitropropionic acid in mice. The repeated administration of 3-nitropropionic acid generated necrotic neuronal cell death in the striatum. Calponin 3 was up-regulated in the neuroprotective penimbral region from 1.5 days after the last injection and thereafter. Double immunofluorescence study revealed that calponin 3 was induced in GFAP-positive astrocytes. These results suggest that calponin 3 induction in the neuroprotective penumbral area following 3-nitropropionic acid intoxication may play a key role in reactive astrogliosis in the striatum.

Spinal Syringomyelia Secondary to an Intracranial Microcystic Meningioma in a Dog (개에서 척수공동증을 동반한 소낭성 뇌수막종 1례)

  • Won, Sangcheol;Kim, Jae-Hoon
    • Journal of Veterinary Clinics
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    • v.31 no.5
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    • pp.439-444
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    • 2014
  • A 5-year-old, 2.7 kg female maltese dog was presented a local veterinary clinic with one week history of hindlimb lameness associated with patellar luxation. Reduction of bilateral medial patellar luxation was operated using trochlear resection and lateral reinforcement technique. Three weeks after the surgery, the dog showed bending spine with pain in thoracic and lumbar region, continuous ataxia and intermittent convulsion. Magnetic resonance imaging scanning revealed a hyperintense mass in right frontal lobe of brain and abnormal cavitation from cervical cord to third lumbar cord. Histopathologically, neoplasm in brain composed of meningothelial cells showed loosely reticular or lace-like morphology with numerous extracellular cystic spaces of variable size and shape. Neoplastic cells were positive for vimentin and negative for neuron specific enolase and glial fibrillary acidic protein. Irregular shaped enlarge central canal-like cavity was existed in cervical and lumbar cords. In our best knowledge, this report described the clinical findings, imaging and histopathologic characteristics of unusual intracranial microcystic meningioma with secondary syringomyelia in a dog.

EVALUATION OF PERIODONTAL LIGAMENT CELL VIABILITY IN RAT TEETH ACCORDING TO VARIOUS EXTRA-ORAL DRY STORAGE TIMES USING MTT ASSAY (구강 외 노출시간에 따른 흰쥐 치아 치주인대세포 활성도의 MTT 검색법을 이용한 평가)

  • Jeon, In-Soo;Kim, Eui-Seong;Kim, Jin;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.31 no.5
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    • pp.398-408
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    • 2006
  • The purpose of this study was to verify the usefulness of MTT analysis as a tool of measurement of the periodontal ligament cell viability from the extracted rat molar. A total of 80 Sprague-Dawley white female rat of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted under Ketamine anesthesia. Twenty-four teeth of each group (divided as five groups depending upon the time-lapse after extraction such as Immediate, 10, 20, 40 and 60 minutes) were immersed in $200{\mu}l$ of MTT solution (0.5 mg/ml) and processed for optical density measurements. Another 10 teeth of each group were treated as same as above and sectioned at $10{\mu}m$ for microscopic examination. All measurements values were divided by the value of hematoxylin-eosin staining which represented the volume of each corresponding samples. Immediate and 10 minute groups showed highest MTT values followed by 20, 40, and 60 minutes consecutively. Statistical significance (p<0.05) existed between all groups except in immediate versus 10 minute groups and 40 versus 60 minutes. Histological findings also showed similar findings with MTT results in crystal shape and crystal numbers between the experimental groups. These data indicate that in vivo MTT analysis nay be of value for evaluation of the periodontal ligament cell viability without time- consuming cell culturing processes.

A Study for the Mechanical Properties in Scouring and Heat Treatment of PLA Woven Fabric (PLA직물의 정련 및 열처리 시 물성변화에 대한 연구)

  • Kim, Ji-Yeon;Park, Sung-Min;Kang, Suk-Hee;Park, Jeong-Young;Yeom, Jeong-Hyeon
    • Proceedings of the Korean Society of Dyers and Finishers Conference
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    • 2011.03a
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    • pp.108-108
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    • 2011
  • PLA 즉 폴리유산섬유는 옥수수를 발효하여 글루코오스(포도당)상태를 만든 후 젖산(유산, Lactic acid)으로 만들고 이것을 탈수, 축합반응시켜 polylactic acid로 만든 것이다. 생분해성이 있으므로 저탄소, 녹색성장의 모토를 대변하는 소재라는 이점이 있다. 구조는 에스테르기의 반복단위를 가지는 소수성 섬유로 벤젠환은 없으나 그 외 구조는 폴리에스테르와 비슷하며, 에스테르기가 존재하므로 분산염료와 수소결합하여 염착된다. 그러나 PLA는 융점이 $170^{\circ}C$, Tg $57^{\circ}C$로 내열성이 낮아서 염색온도, 열처리온도, 다림질에 제약이 있으며, 알칼리에 약한 단점이 있다. 따라서 PLA섬유는 낮은 염착량, 내알칼리성, 염착온도 때문에 염색 및 후가공 단계에 많은 사전 실험을 통한 조건 설정이 필요한 까다로운 섬유이다. 본 연구에서는 (주)휴비스의 PLA원사로 제직한 직물(경사:DTY 75/72SD, 위사:DTY 100/72SD, 조직:DOBBY) 생지에 대하여 열처리 시 장력의 유무, 온도, 시간에 따른 폭의 변화를 측정하여 수축률을 알아보았다. 또한, PLA직물을 온도별로 정련한 후 열처리하여 인열강도 측정을 통해 최적 전처리 조건을 조사하였다. 실험결과, PLA생지를 무장력 상태에서 열처리 시 수축이 심하게 일어나고, 장력이 주어져도 열처리 온도에 따라 수축의 정도에 차이가 나타났다. 열처리 시간은 30, 60, 90, 120초로 주었으나 큰 편차는 없었고, 경사가 위사보다 수축 정도가 더 컸으며, $130^{\circ}C$에서는 전체적으로 수축이 심하였다. 생지의 정련에는 인산에스테르계 정련제와 약알칼리인 탄산나트륨으로 조액하여 60, 70, 80, $90^{\circ}C$에서 10분간 처리한 후, Lab. tenter(Mathis, LTE)를 이용하여 110, 120, $130^{\circ}C$에서 30, 60, 90, 120초간 열처리한 다음, KS K 0535 펜듈럼법에 의거하여 인열강도를 측정하였다. 그 결과, 상기 정련온도에서는 인열강도에 영향을 주지 않았으나, 열처리 온도가 $130^{\circ}C$일 때 현저한 강도의 저하를 나타내었다. 실험조건 하에서 가장 적절한 열처리 조건은 $110^{\circ}C$, 60초로 사료된다. 따라서 PLA의 약한 내열성과 내알칼리성 실험결과, 강도나 수축 등 물성변화가 일어나지 않도록 열처리 온도의 제어에 주의가 필요함을 확인할 수 있었다. 실제 섬유가공 작업현장에서는 일반적으로 열처리기가 $180^{\circ}C$이상의 고온으로 고정된 경우가 많은데, 작업자들에게 PLA소재에 대한 사전주의 및 공정변경에 대한 주지가 요구된다.

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Immunohistochemieal study on the antigenicity of body compartments of Payugonimus westermani (폐흡충 충체 부위별 항원성에 대한 면역 조직화학적 연구)

  • Lee, Sun-Hyeong;Seong, Suk-Hwan;Chae, Jong-Il
    • Parasites, Hosts and Diseases
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    • v.27 no.2
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    • pp.109-118
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    • 1989
  • Production of circulating specific antibodies to the lung fluke (Paragenimus westermani) by its host is well known and used in various kinds of immunodiagnostic methods, However, it has not been well documented which compartments (or structures) of the lung fluke are most responsible for the production of specific antibodies. The present immunohistochemical study was undertaken to demonstrate the antigenicity of each body compartment of p. westermani such as suckers, tegument, spines, vitelline glands, intestine, reproductive organs(male and female), and eggs. Indiret immunoperoxidase(IP) stain technique was applied, using formalin-fked, paraffin- embedded lung tissues of P westermani-infected cats sectioned in 4 Um thickness as the antigen and cat antisera (11~20 weeks of infection) as the primary antibody. Peroxidase-conjugated goat anti-cat IgG was used as the secondary antibody and diaminobensidine(DAB) as the coloring agent. Strong yellow or yellowish brown staining was regarded positive. The primary and secondary antibody dilutions were made at 1 : 500~1 : 2, 000 and 1 : 200~1 : 500 respectively, and IP stain was repeated 10 times for each dilution. A consistent result obtained was that the intestinal epithelial border, intestinal content, vitelline glands, and eggs scattered around the worm capsule showed strong positive staining, while uterine eggs and some parenchymal portions showed weak positive reaction. On the other hand, the suckers, tegument, spines, subtegumental cells, cytoplasm of intestinal epithelial cells, male reproductive organs, and ovary revealed negative staining. The body compartments showing higher antigenicity were, in the decreasing order, the intestinal epithelial border, intestinal content, eggs in the worm capsule, vitelline glands, uterine eggs, and parenchymatous portions. The intestinal epithelial border and luminal contents revealed positive staining even at a few concentration of 1 : 4, 000 primary antibody(secondary ab., 1 : 200) whereas the parenchymatous portion showed positive reaction only at higher concentrations than 1'500 (secondary ab., 1 : 200). The results suggest that the specific antibody responses of the host to p. westermani occur most strongly upon the excretes from the intestinal epithelium of the worm and e99s Produced around the worm capsule,

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Analysis of disease mechanism of subacute necrotizing lymphadenitis in children (소아 아급성 괴사성 림프절염의 임상적, 방사선학적, 면역조직화학적 소견)

  • Kim, Hyun Jung;Yeom, Jung Suk;Park, Ji Suk;Park, Eun Sil;Seo, Ji Hyun;Lim, Jae Young;Park, Chan Hoo;Woo, Hyang Ok;Cho, Jae Min;Lee, Jeong Hee;Youn, Hee Shang
    • Clinical and Experimental Pediatrics
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    • v.51 no.11
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    • pp.1198-1204
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    • 2008
  • Purpose : The cause of subacute necrotizing lymphadenitis, a rare disease in children, has not been completely clarified. This study was aimed to investigate the disease mechanism by examining clinical, radiologic, and immunohistochemical findings in children diagnosed with subacute necrotizing lymphadenitis after an excisional biopsy. Methods : We examined 19 lymph node tissue specimens from 17 children diagnosed with subacute necrotizing lymphadenitis at Gyeongsang National University Hospital from March, 1998 to July, 2006. A retrospective survey of the medical records was performed. CT findings were analyzed. Immunohistochemical staining was done on tissues obtained by excisional biopsy from all patients. Results : The patient's age ranged from 5 to 19 years (average age :11.8 years). The main symptoms included a neck mass (17/19), pain in the mass (6/17), and fever (12/19). The palpable lymph nodes were mostly cervical in location; the maximum diameter, which was measured radiologically, was less than 3 cm in all 10 cases. The masses were pathologically divided into proliferative, necrotic, and xanthomatous types. With immunohistochemical staining the masses were divided into lesion (L), perilesion (PL), and necrosis (N). The CD8 staining was stronger than the CD4 staining for all regions in three types. The CD4 staining intensity was mainly increased in the perilesion, and CD8 was mainly increased in the lesion. Conclusion : We compared the radiologic findings, clinical symptoms, and pathology to help understand the cause of disease in patients with subacute necrotizing lymphadenitis.

Injury of Neurons by Oxygen-Glucose Deprivation in Organotypic Hippocampal Slice Culture (뇌 해마조직 절편 배양에서 산소와 당 박탈에 의한 뇌신경세포 손상)

  • Chung, David Chanwook;Hong, Kyung Sik;Kang, Jihui;Chang, Young Pyo
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1112-1117
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    • 2008
  • Purpose : We intended to observe cell death and apoptotic changes in neurons in organotypic hippocampal slice cultures following oxygen-glucose deprivation (OGD), using propidium iodide (PI) uptake, Fluoro-Jade (FJ) staining, TUNEL staining and immunofluorescent staining for caspase-3. Methods : The hippocampus of 7-day-old rats was cut into $350{\mu}m$ slices. The slices were cultured for 10 d (date in vitro, DIV 10) and and exposed to OGD for 60 min at DIV 10. They were then incubated for reperfusion under normoxic conditions for an additional 48 h. Fluorescence of PI uptake was observed at predetermined intervals, and the cell death percentage was recorded. At 24 h following OGD, the slices were Cryo-cut into $15{\mu}m$ thicknesses, and Fluoro-Jade staining, TUNEL staining, and immunofluorescence staining for caspase-3 were performed. Results : 1) PI uptake was restricted to the pyramidal cell layer and DG in the slices after OGD. The fluorescent intensities of PI increased from 6 to 48 h during the reperfusion stage. The cell death percentage significantly increased time-dependently in CA1 and DG following OGD (P<0.05). 2) At 24 h after OGD, many FJ positive cells were detected in CA1 and DG. Some neurons had distinct nuclei and processes while others had fragmented nuclei and disrupted processes in CA1. TUNEL and immunofluorescent staining for caspase-3 showed increased expression of TUNEL labeling and caspase-3 in CA1 and DG at 24 h after OGD. Conclusion : The numerous dead cells in the slice cultures after OGD tended to display apoptotic changes mediated by the activation of caspase-3.

A study on the mechanism of NO-induced apoptosis in human gingival fibroblast (사람 치은 섬유아세포에서 산화질소 유도 세포고사에 대한 연구)

  • Kim, Kang-Moon;Chung, Hyun-Ju;Kim, Won-Jae
    • Journal of Periodontal and Implant Science
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    • v.34 no.4
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    • pp.807-818
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    • 2004
  • 산화질소는 생리적 농도에서 세포내 신호전달자로 작용하지만 높은 농도에서는 세포독성을 일으킨다. 최근 치은 섬유아세포와 치주인대 섬유아세포는 산화질소 합성효소를 가지고 있고 세균의 lipopolysaccharide나 cytokine에 의해 대량의 높은 농도의 산화질소가 합성된다는 보고가 있음에도 지금까지 치은 조직에서 산화질소의 세포독성에 대한 연구는 아직 이루어 지지않고 있다. 본 연구는 사람의 치은 섬유아세포에서, 산화질소유도세포 고사기전을 밝히는데 목적이 있다. 세포 생장력은 MTT 방법으로 측정하였고, 세포의 형태적 변화는 Diff-Quick 염색법으로 조사하였다. Bcl-2 famly와 Fas 발현 정도는 RT-PCR 방법에 의해 확인하였으며, caspase-3, -8 와 -9의 활성은 spectrophotometer로 reactive oxygen species (ROS)는 형광분광계에 의해 측정되었다. 미토콘드리아에서 세포질로 분비된 cytochrome c는 western blot으로 조사하였다. 산화질소 유리제인 sodium nitroprusside (SNP) 처리는 사람 섬유아세포의 생존률을 시간과 농도 의존적으로 감소시켰고, 세포용적축소, 염색사 용축, DNA 절편화를 일으켰다. 또한, SNP 처리로 미토콘드리아에서 세포질로 유리되는 cytochrome c 양이 증가되었고, caspase-9 과 caspase-3 의 활성이 증가되었다. 한편, SNP 처리에 의해 death receptor 구성요소인 Fas 발현이 증가되었고, caspase-8의 활성이 증가되었다. Bcl-2 family 에 대한 RT-PCR 분석결과, 세포고사를 억제하는 Bcl-2 발현은 감소되었으나 세포고사를 자극하는 Bax와 Bid의 발현은 증가되었다. Soluble guanylate cyclase 억제제인 ODQ는 SNP에 의한 세포 생존율 감소를 차단하지 못했다. 따라서, 본 실험의 결과들은 사람 섬유아세포에서 산화질소유도 세포고사에 Bcl-2 family나 ROS가 매개하는 미토콘드리아 의존 및 death receptor 의존 세포고사기전이 관여함을 시사하였다.

The nervous system of Fibricola seoulensis by acetylcholinesterase histochemistry (Acetylcholinesterase 조직화학법으로 관찰한 Fibricola seoulensis 신경계의 분포)

  • 전은우;김창환
    • Parasites, Hosts and Diseases
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    • v.31 no.4
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    • pp.321-330
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    • 1993
  • F. seoulensis were obtained from artificially infected albino rats at 3, 4, 5, 6, 7 days after infection. The worms and metacercariae were washed in physiological saline solution, and fled with 10% neutral formalin. The acetylcholinesterase (AchE) stained by one histochemistry using acetylthiocholine iodide as substrate. Eserine, ism-OMPA and BW284C51 were used as inhibitors of AchE. The nervous system consists of three pairs longitudinal nerve trunks interconnected with excretory plexus in posterior half, and phinmc and oral sucker in anterior half of metacercariae and adults. The longitudinal nerve trunks are interconnected with transverse commissures and numerous circular commissures. Considerable numbers of circular commissures are interconnected with longitudinal nerve trunks tying on the surface of the worms. At each stage of juvenile worms, AchE and nonspecific cholinesterase activites were observed in the oral sucker, ventral sucker, pharynx and nerve system. Isoxymes of AchE in f seoderuts were separated into the two bands, 69 kDa and 132 kDa. The major band was 69 kDa.

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The Effects of Polyampholyte on Vitrification Process for cryopreservation of Bovine Oviduct Epithelial Cell (Polyampholyte가 소난관상피세포의 초자화 동결방법에 미치는 영향)

  • Kim, Sung Woo;Lee, Jae-Yeong;Kim, Chan-Lan;Yu, Yeonhee;Lee, Sung Soo;Ko, Yeoung-Gyu
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.6
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    • pp.527-535
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    • 2020
  • The purpose of this study was to establish a simple vitrification protocols to preserve animal cell lines derived from tissues of livestock that could be recultured. Bovine oviduct epithelial cells (BOEC) were used for the vitrification process using a 0.25 ml straw to increase cryopreservation efficiency. BOEC was cultured from the oviduct of 3.5-day estrus state, and the commercially available polyampholyte StemCell KeepTM was used as a cryoprotective agent. Using different concentrations, the viability rates of BOEC in 5, 10, 25, 50, 75, and 100% in freezing media were investigated. Survivability was determined using a differential staining technique using a trypan blue test and a CYTO-13/PI staining protocol. The viability rates of BOEC in the trypan blue test were 5.6±11.8, 12.5±7.2, 53.0±2.7, 85.1±6.9, 79.8±0.6, and 60.7±6.7% with a respective concentration of StemCell KeepTM. The viability rates in CYTO-13/PI staining were 4.6±2.5, 30.8±12.1, 58.4±2.5, 85.5±1.2, 79.8±0.6, and 71.2±1.2%, respectively. These results indicate that BOEC could be preserved with StemCell KeepTM without toxicity in a 0.25-ml straw. The optimal concentration of vitrification solution with StemCell KeepTM was determined to be 50% and can be considered as a proper preservation method for cryobanking.