• Title/Summary/Keyword: Tissue preparation

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Improved Procedure for Large-scale Isolation of Mitochondrial DNA from Mammalian Tissues

  • Hong, Sung-Soo;Lee, Chung-Choo
    • Animal cells and systems
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    • v.3 no.1
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    • pp.73-78
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    • 1999
  • Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

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Lactosaminated N-Succinyl-chitosan: Preparation and Biodistribution into the Intestine, Bone, Lymph Nodes and Male Genital Organs after I.v. Administration

  • Kato, Yoshinori;Onishi, Hiraku;Machida, Yoshiharu
    • Macromolecular Research
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    • v.11 no.5
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    • pp.382-386
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    • 2003
  • Reductive amination of N-succinyl-chitosan (1) and lactose using sodium cyanoborohydride in 1/15 M phosphate buffer (pH 6.0) for 6 d was suitable for the preparation of lactosaminated N-succinyl-chitosan (2). At 8, 24 and 48 h after i.v. administration of fluorescently labeled 1 (1') or 2 (2'), Peyer's patch, mesenteric lymph nodes, testes, prostate, preputial grand, intestine (small intestine plus cecum), femoral muscle, backbone and peritoneum were taken. Peyer's patch and mesenteric lymph nodes were put together as lymph nodes. Over 10% of dose/g tissue was distributed to the prostate and lymph nodes at 48 h post-administration in both l' and 2'.2' was easily distributed into not only the liver but also prostate, intestine, preputial gland and lymph nodes. Although galactose receptors are known to exist not only on the liver parenchymal cells but also on prostate and testes, the selective distribution of 2' into the prostate and the testes were not observed clearly. This study suggested that 1 and 2 should have possibilities for both the prevention and cure of lymph node metastasis as drug carriers.

Comparison of apical extrusion of intracanal bacteria by various glide-path establishing systems: an in vitro study

  • Dagna, Alberto;El Abed, Rashid;Hussain, Sameeha;Abu-Tahun, Ibrahim H;Visai, Livia;Bertoglio, Federico;Bosco, Floriana;Beltrami, Riccardo;Poggio, Claudio;Kim, Hyeon-Cheol
    • Restorative Dentistry and Endodontics
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    • v.42 no.4
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    • pp.316-323
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    • 2017
  • Objectives: This study compared the amount of apically extruded bacteria during the glide-path preparation by using multi-file and single-file glide-path establishing nickel-titanium (NiTi) rotary systems. Materials and Methods: Sixty mandibular first molar teeth were used to prepare the test apparatus. They were decoronated, blocked into glass vials, sterilized in ethylene oxide gas, infected with a pure culture of Enterococcus faecalis, randomly assigned to 5 experimental groups, and then prepared using manual stainless-steel files (group KF) and glide-path establishing NiTi rotary files (group PF with PathFiles, group GF with G-Files, group PG with ProGlider, and group OG with One G). At the end of canal preparation, 0.01 mL NaCl solution was taken from the experimental vials. The suspension was plated on brain heart infusion agar and colonies of bacteria were counted, and the results were given as number of colony-forming units (CFU). Results: The manual instrumentation technique tested in group KF extruded the highest number of bacteria compared to the other 4 groups (p < 0.05). The 4 groups using rotary glide-path establishing instruments extruded similar amounts of bacteria. Conclusions: All glide-path establishment instrument systems tested caused a measurable apical extrusion of bacteria. The manual glide-path preparation showed the highest number of bacteria extruded compared to the other NiTi glide-path establishing instruments.

SIMS Protein imaging with nanoparticle tagged antibody for simultaneous omic imaging

  • Lee, Seon-Yeong;Mun, Dae-Won
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.230.1-230.1
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    • 2015
  • One of the major problems of biological ToF-SIMS imaging is the lack of protein and peptide imaging. Most of biological story telling is mianly based on proteins. The biological implication of lipid ToF-SIMS imaging would be much higher if protein imaging is provided together. Utilizing high secondary ion yields of metals, proteins can be ToF-SIMS imaged with nanoparticle tagged proteins. Nanoparticles such as Fe3O4, SiO2, PbS were used for imaing NeuN, MCH, Orexin A, ${\alpha}$ synucline, TH(Tryosine Hydroxylase) in mouse tissues with a spatial resolution of ${\sim}2{\mu}m$ using a TOF-SIMS. Lipids and neurotransmitters images obtained simultaneously with protein images were overlayed for more deeper understanding of neurobiology, which is not allowed by any other bioimaging technqiues. The protein images from TOF-SIMS were compared with confocal fluorescence microscopy and NanoSIMS images. A new sample preparation method for imaging single cell membranes in a tissue using the vibrotome technique to prepare a tissue slice without any fixation and freeze drying will be also presented briefly for Hippocampus and Hypothalamus tissues.

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Imprint Cytologic Feature of Extraskeletal Osteosarcoma - A Case Report - (골외골육종의 압착도말 소견 - 1예 보고 -)

  • Gu, Mi-Jin;Bae, Young-Kyung;Kim, Mi-Jin;Choi, Joon-Hyuk;Choi, Won-Hee
    • The Korean Journal of Cytopathology
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    • v.11 no.1
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    • pp.59-63
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    • 2000
  • Extraskeletal osteosarcoma is an uncommon tumor originated from soft tissue without evidence of skeletal involvement. It usually affects adults and its common locations are extremity, buttock, and retroperitoneum. Although the histologic feature of this tumor is well known, there have been few reports on the fine needle aspiration cytologic findings. We report the imprint cytologic feature of extraskeletal osteosarcoma. The patient was a 49-year-old man with a mass of the left anterior chest for 2 years. On the Imprint preparation, the smears showed malignant round, polygonal or spindle cells with coarsely clumped chromatin and occasionally prominent nucleoli. The malignant cells occur singly, in clusters, or associated with amorphous eosinophilic osteoid. Mitotic figures are also seen.

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Fine Needle Aspiration Cytology of Pulmonary Lesions in Wegener's Granulomatosis - A Case Report - (Wegener's Granulomatosis 폐병변의 세침흡인 세포학적 소견 - 1예 보고 -)

  • Seo, Eun-Joo;Kwon, Hi-Jeong;Min, Ki-Ouk
    • The Korean Journal of Cytopathology
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    • v.9 no.1
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    • pp.85-88
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    • 1998
  • We described the findings of fine needle aspiration cytology of the lung from a patient with Wegener's granulomatosis. Early diagnosis and prompt treatment of the patients with Wegener's granulomatosis is essential for a better prognosis. However, the variety of clinical presentations and nonspecific radiologic infiltrates of Wegener's granulomatosis frequently make the diagnosis difficult. Although an open lung biopsy is required for a firm diagnosis, fine needle aspiration cytology & biopsy preparation can also provide an adequate tissue sample, when the findings of fine needle aspiration are considered with clinical manifestations and ANCA value in the serum. The cytologic smears showed scattered necrotic tissue fragments entrapping many neutrophils and occasional epithelioid cells. Multinucleated giant cells were infrequently observed. Ziehl-Neelsen stain for acid fast bacilli was negative. All the cytologic features recapitulated the histopathologic findings of purulent and necrotizing granulomatous inflammation seen in Wegener's granulomatosis.

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Preparation of a Porous Chitosan/Fibroin-Hydroxyapatite Composite Matrix for Tissue Engineering

  • Kim, Hong-Sung;Kim, Jong-Tae;Jung, Young-Jin;Ryu, Su-Chak;Son, Hong-Joo;Kim, Yong-Gyun
    • Macromolecular Research
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    • v.15 no.1
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    • pp.65-73
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    • 2007
  • Chitosan, fibroin, and hydroxyapatite are natural biopolymers and bioceramics that are biocompatible, biodegradable, and resorb able for biomedical applications. The highly porous, chitosan-based, bioceramic hybrid composite, chitosanlfibroin-hydroxyapatite composite, was prepared by a novel method using thermally induced phase separation. The composite had a porosity of more than 94% and exhibited two continuous and different morphologies: an irregularly isotropic pore structure on the surface and a regularly anisotropic multilayered structure in the interior. In addition, the composite was composed of an interconnected open pore structure with a pore size below a few hundred microns. The chemical composition, pore morphology, microstructure, fluid absorptivity, protein permeability, and mechanical strength were investigated according to the composition rate of bioceramics to biopolymers for use in tissue engineering. The incorporation of hydroxyapatite improved the fluid absorptivity, protein permeability, and tenacity of the composite while maintaining high porosity and a suitable microstructure.

The effect of the material properties for the commercially available cervical braces on the skin tissue (수종 시판 경추보조기의 물성이 피부조직에 미치는 영향)

  • Kim, K.T.;Park, J.C.;Choi, J.B.;Choi, K.;Mun, M.S.;Suh, H.
    • Proceedings of the KOSOMBE Conference
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    • v.1997 no.11
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    • pp.501-506
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    • 1997
  • This is to investigate the physical properties of the commercially available Soft, Thomas, Minerva, and Philadelphia cervical braces which are widely used in orthopedics, neurosurgery, and rehabilitation medicine clinics as assisting devices or physical stabilization of cervical vertebrates, to use as a basic data or designing new type brace. Tensile strengths were observed by universal mechanical measuring device and Thomas brace required the highest stress to break by tensile stress. Durabilities against continuous frictional forces were also determined, and Minerva brace demonstrated the longest frictional time until being perforated. and Thomas/minerva braces are superior to the other braces in the effect of the material property on the skin tissue, as the animal test. According to these results, polyethylene is recommendable as a frame and preparation of pores in the material is favorable to provide ventilation to skin.

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Detection of Virus in Fruit and Seed of Vegetables Using RT-PCR (RT-PCR에 의한 과채류 열매 및 종자의 바이러스 검정)

  • 최장경;김혜자;윤주연;박선정;김두욱;이상용
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.630-635
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    • 1998
  • Tobacco mosaic tobamovirus (TMV), cucumber mosaic cucumovirus (CMV), cucumber green mottle mosaic tobamovirus (CGMMV) and zucchini yellow mosaic potyvirus (ZYMV) from individual fruits and seeds of hot pepper and cucumber were detected by the reverse transcription-polymerase chain reaction (RT-PCR). The dilution end-points for RT-PCR in curde sap from TMV. and CMV - infected hot pepper leaves and CMV - and CGMMV-infected cucumber leaves were 10-5. However, the amount of PCR product obtained from preparation of ZYMV-infected cucumber leaf was 10-fold lower than those of CMV or CGMMV-infected cucumber leaves. In hot pepper, both TMV and CMV were detected in all parts of the fruit wall tissue, but the yields of PCR products in the fruit stalk and its surrounding tissues were higher than those of the end parts of the fruit. On the other hand, in cucumber fruit infected with CMV, CGMMV or ZYMV, the fruit wall tissue and seed located in both stalk and end parts showed higher yields of PCR products than those of intermediate parts. Of five viruses that were analysed, only TMV in hot pepper seed, and CGMMV and CMV in cucumber seed were detected in testa parts.

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A Case of Urethral Reconstruction Using a Superficial Circumflex Iliac Artery

  • Yoo, Kun-Woon;Shin, Hyun-Woo;Lee, Hye-Kyung
    • Archives of Plastic Surgery
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    • v.39 no.3
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    • pp.253-256
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    • 2012
  • A radial forearm free flap has been conventionally used for urethral reconstruction. However, aesthetic and functional complications occur frequently at the donor site. The use of a superficial circumflex iliac artery perforator (SCIP) flap can resolve these disadvantages. Here, we report our case with a review of literature. A 69-year-old man visited our hospital with multiple contusions of the abdomen and genital amputation. After necrotic tissue debridement, the length of the residual corpus carvernosum was 1.5 cm and that of the corpus spongiosum and urethra was 1 cm. For the reconstruction of the penis, a SCIP flap and anterolateral thigh free flap was performed. The primary closure was performed at the donor site. Three weeks postoperatively, the patient had a urethral foley catheter removed. The neourethra was functioning well without stricture. Four months postoperatively, the patient had no complications such as urethral stricture. A good recovery was also achieved with no aesthetic deficits at the donor site. SCIP flap is appropriate for urethral reconstruction. Because of its proximity to the recipient sites, it makes surgical preparation easier and the primary closure at the donor site available. It is also advantageous in that its location is almost unnoticeable.