• Title/Summary/Keyword: Tissue preparation

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Affinity Purification of Human Alpha-Fetoprotein (吸着 크로마토그라피法에 의한 사람 ALpha-Fetoprotein의 分離)

  • Kang, Shin-Sung;Bang, Ok-Sun;Park, Tae-Kyu
    • The Korean Journal of Zoology
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    • v.29 no.4
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    • pp.283-293
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    • 1986
  • For the preliminary step to make and characterize the monoclonal antibodies of human alpha-fetoprotein (AFP) was purified from 534g of human fetal tissues through the procedures of tissue extraction, DEAE-cellulose, concanavalin A-Sepharose, Cibacron Blue F3GA-agarose and immunoadsorbent affinity chromatography. The isolated AFP preparation showed a single band on polyacrylamide gel electrophoresis and a single precipitin are against rabbit anti-human cord serum and anti-human AFP on immunoelectrophoresis. Our AFP also displayed a single band on SDS-polyacrylamide gel electrophoresis. The recovery of AFP was 8.76mg total.

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Preparation of Biodegradable Thermo-responsive Polyaspartamides with N-Isopropylamine Pendent Groups (I)

  • Moon, Jong-Rok;Kim, Ji-Heung
    • Bulletin of the Korean Chemical Society
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    • v.27 no.12
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    • pp.1981-1984
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    • 2006
  • Novel amphiphilic, thermo-responsive polyaspartamides which showed both LCST (lower critical solution temperature), and sol-gel transition were prepared and characterized. The polyaspartamide derivatives were synthesized from polysuccinimide, the polycondensate of aspartic acid monomer, via successive nucleophilic ring-opening reaction by using dodecylamine and N-isopropylethylenediamine (NIPEDA). At the intermediate composition ranges, the dilute aqueous solution exhibited a thermally responsive phase separation due to the presence of LCST. The phase transition temperature was controllable by changing the content of pendent groups. In addition, a physical gelation, i.e. the sol-gel transition was observed from the concentrated solutions, which was elucidated by dynamic viscoelastic measurements. These novel injectable and thermo-responsive hydrogels have potential for various biomedical applications such as tissue engineering and current drug delivery system.

A Blood-brain Barrier Permeable Derivative of 5-Fluorouracil: Preparation, Intracellular Localization, and Mouse Tissue Distribution

  • Im, Jung-Kyun;Biswas, Goutam;Kim, Wan-Il;Kim, Kyong-Tai;Chung, Sung-Kee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.3
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    • pp.873-879
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    • 2011
  • 5-Fluorouracil (5-FU), an anticancer agent was covalently attached to the recently developed sorbitol-based G8 transporter, and the conjugate (7) with FITC was found to have an affinity toward mitochondria and to readily cross BBB to gain an entry into mouse brain. Measured by $IC_{50}$, the conjugate (9) without the fluorophore showed enhanced cytotoxic activity toward two types of multidrug-resistant cell lines. These results strongly suggest that the sorbitol-based G8 transporter can be utilized as a good CNS delivery vector.

Effects of Forskolin on Endogenous Dopamine and Acetylcholine Release in Rat Neostriatal Slices

  • Kim, Hwa-Jung
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.520-528
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    • 1996
  • The involvement of the cyclic AMP (cAMP) effector system in the release of endogenous dopamine and acetylcholine from the rat neostriatum was assessed. Forskolin, an activator of adenylate cyclase, was used to enhance cAMP production, and the consequence of this enhancement on the spontaneous and potassium stimulated release of dopamine and acetylcholine was evaluated. Neostriatal slices were prepared from Fischer 344 rats and after a preincubation period the release of each endogenous neurotransmitter was measured from the same slice preparation. To measure acetylcholine release the slice acetylcholinesterase (AChE) activity was inhibited with physostigmine, but the release from slices with intact AChE activity was also determined (choline, instead of acetylcholine was detected in the medium). Under both conditions forskolin induced a significant dose-dependent increase in the potassium-evoked release of dopamine. In the same tissue preparations the release of neither acetylcholine (AChE inhibited) nor choline (AChE intact) was affected by forskolin. The results indicate that the CAMP second messenger system might be involved in neuronal mechanisms that enhance neostriatal dopamine release, but stimulation of this second messenger by forskolin does not further enhance neostriatal acetylcholine release.

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Time-dependent effects of EDTA for smear layer removal on microhardness of root canal dentine

  • Song, Yoon-Kyoung;Cho, Yong-Bum
    • Proceedings of the KACD Conference
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    • 2003.11a
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    • pp.587-589
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    • 2003
  • I. Objectives The purpose of this study was to evaluate the effects of EDTA on microhardness of roor canal dentine according to different working time. II. Materials and Methods Forty recently extracted, intact single root teeth were used. Debris and soft tissue remnants on the root surface were cleaned with scaler and stored in saline at $4^{\circ}C$ until used. All selected roots were similar in size and shape. 1. Preparation of teeth. The crown of the teeth were removed at the level of the CEJ, using a water-cooled diamond bur.(omitted)

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Use of stem cells in bone regeneration in cleft palate patients: review and recommendations

  • Amiri, Mohammad Amin;Lavaee, Fatemeh;Danesteh, Hossein
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.48 no.2
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    • pp.71-78
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    • 2022
  • This study was conducted to review the efficacy of different sources of stem cells in bone regeneration of cleft palate patients. The majority of previous studies focused on the transplantation of bone marrow mesenchymal stem cells. However, other sources of stem cells have also gained considerable attention, and dental stem cells have shown especially favorable outcomes. Additionally, approaches that apply the co-culture and co-transplantation of stem cells have shown promising results. The use of different types of stem cells, based on their accessibility and efficacy in bone regeneration, is a promising method in cleft palate bone regeneration. In this regard, dental stem cells may be an ideal choice due to their efficacy and accessibility. In conclusion, stem cells, despite the lengthy procedures required for culture and preparation, are a suitable alternative to conventional bone grafting techniques.

Preparation and Characterization of Ipriflavone-Loaded Poly(L-lactide-co-glycolide) Scaffold for Tissue Engineered Bone (조직공학적 골을 위한 애프리플라본을 함유한 다공성 지지체의 제조 및 그 특성)

  • Jang, Ji-Wook;Lee, Bong;Han, Chang-Whan;Lee, Il-Woo;Lee, Hai-Bang;Khang, Gil-Son
    • Polymer(Korea)
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    • v.27 no.3
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    • pp.226-234
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    • 2003
  • Ipriflavone (IP), a non-hormonal isoflavone derivative, has been shown to interfere with bone remodeling by inhibiting bone resorption and stimulating bone formation. IP consistently increased the amount of Ca incorporated into the cell layer by mesenchymal stem cells (MSCs). In this study, we developed the novel IP loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds for the possibility of the application of the tissue engineered bone. IP/PLGA scaffo1ds were prepared by solvent casting/salt leaching method and were characterized by porosimeter, scanning electron microscopy, determination of residual salt amount, differential scanning calorimetry, and X-ray diffractometer, respectively. IP/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of IP on the osteoinduction compared with control PLGA scaffo1ds. Thin sections were cut from paraffin embedded tissues and histological sections were stained H&E, von Kossa, and immunohistochemical staining for Type I collagen and osteocalcin. It can be observed that the porosity was above 91.7% and the pore size was above 101 $\mu\textrm{m}$. Control scaffo1d and IP/PLGA scaffo1ds of 50% IP were implanted on the back of athymic nude mouse to observe the effect of IP on the induction of cells proliferation for 9 weeks. The evidence of calcification, osteoblast, and osteoid from the undifferentiated stem cells in the subcutaneous sites and other soft connective tissue sites having a preponderance of stem cells has been observed. From these results, it seems that IP plays an important role for bone induction in IP/PLCA scaffolds.

Preparation and Characterization of Biodegradable Hydrogels for Tissue Expander Application (조직 확장기용 생분해성 하이드로젤의 제조 및 특성분석)

  • Yuk, Kun-Young;Kim, Ye-Tae;Im, Su-Jin;Garner, John;Fu, Yourong;Park, Ki-Nam;Park, Jeong-Sook;Huh, Kang-Moo
    • Polymer(Korea)
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    • v.34 no.3
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    • pp.253-260
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    • 2010
  • In this study, we prepared and evaluated a series of biocompatible and biodegradable block copolymer hydrogels with a delayed swelling property for tissue expander application. The hydrogels were synthesized via a radical crosslinking reaction of poly(ethylene glycol) (PEG) diacrylate and poly(D,L-lactide-co-glycolide)-poly(ethylene glycol)-poly(D,L-lactide-co-glycolide)(PLGA-PEG-PLGA) triblock copolymer diacrylate as a swelling/degradation controller (SDC). For the synthesis of various SDCs that can lead to different degradation and swelling properties, various PLGA-PEG-PLGA triblock copolymers with different LA/GA ratios and different PLGA block lengths were synthesized and modified to have terminal acrylate groups. The resultant hydrogels were flexible and elastic even in the dry state. The in vitro degradation tests showed that the delayed swelling properties of the hydrogels could be modulated by varying the chemical composition of the biodegradable crosslinker (SDC) and the block ratio of SDC/PEG. The histopathologic observation after implantation of hydrogels in mice was performed and evaluated by macrography and microscopy. Any significant inflammation or necrosis was not observed in the implanted tissues. Due to their biocompatibility, elasticity, sufficient swelling pressure, delayed swelling and controllable degradability, the hydrogels could be useful for tissue expansion and other biomedical applications.

Isolation of Protoplasts from Tomato Root by Two-step Osmotic Treatment (토마토 뿌리조직으로부터 두 단계 삼투압 처리에 의한 원형질체의 분리)

  • Shin, Dae-Seop;Han, Min-Woo;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.192-196
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    • 2004
  • In order to measure cellular physiological activity including ion channel activity, protoplasts were isolated from the root tissue of tomato plant. The general methods recommended were not efficient enough to make protoplasts from the root tissue. Among various conditions tested, we found that a two-step treatment of osmosis is very efficient for the isolation of protoplasts. In this procedure, root tissues were preincubated in a solution containing 300 mM sorbitol for 30 min. Then, they moved to the reaction solution containing 700 mM sorbitol as well as cell wall-digesting enzymes. The formation of protoplast was greatly increased by this method. In order to find the optimal condition of the two-step method, various conditions of pH, osmotic pressure, incubation time, and the concentrations of cell wall-digesting enzymes were tested. The yield of protoplast isolation was maximal at pH 5.0 after 2 hr incubation. Mixed enzymes of 3% cellulase, 1 % macerozyme, and 0.1 % pectolyase showed maximal protoplast isolation. The physiological activity of isolated protoplast evaluated by measuring the cellular ATPase activity was as high as that measured from the preparation of root tissue. The protoplasts isolated by this method were remained healthy up to 4 hrs which is enough time to measure the cellular physiological activity. These results show that the two-step treatment of osmotic pressure was successful to obtain high yield of healthy protoplast from tomato root tissue.

Possibility of Local Recurrence Caused by Surgical Instruments in the Mouse Skin Cancer Model (백서 모델에서 수술 기구를 통한 피부악성종양의 국소 재발 가능성)

  • Kim, Gook-Jin;Lee, Hyoung-Suk;Kim, Nam-Gyun;Lee, Kyung-Suk;Kim, Jun-Sik;Park, Sang-Woo
    • Archives of Plastic Surgery
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    • v.38 no.4
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    • pp.339-344
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    • 2011
  • Purpose: The goal of cancer surgery is complete removal of cancer tissue and prevention of recurrence. Surgeons can change the surgical instruments after total resection of the cancer mass. The purpose of this procedure is to prevent dissemination of the cancer cells attached to the surgical instruments. Authors hypothesize the possibility of local recurrence caused by the cancer cells attached to the surgical instruments in the skin cancer cases. Methods: Skin cancers were induced by using DMBA-TPA two-stage carcinogenesis model in 10 of Balb/c mice. In 2-weeks, skin cancer was developed in all 10 mice. cancer cell attached surgical instruments were made by pinching the removed cancer tissue using Adson tissue forcep 10, 20, 30 times each. To count number of cancer cells in each forcep with different number of pinching was done, the forceps were washed in 30 mL of the normal saline and Cytospin preparation was done. To make recurrence models from cancer cell attached surgical instrument, three incisions were made in normal skin of each mouse, and local seeding was done by pinching subcutaneous tissue in 10, 20, 30 times each by using Adson teeth forceps mentioned above as cancer cell attached surgical instrument. Results: All skin cancers were squamous cell carcinoma. Local recurrences were developed in 7 mice (3 in 10 times forceping site, 2 in 20 times forceping and 3 in 30 times forceping). In the cytospin test, the mean number of squamous cells in 100 microscope was 28.6 in 10 times, 47.2 in 20 times, 93.6 in 30 times, respectively. P value was 0.002 in Wilcoxon-Sign test. Conclusion: The number of cell count was significantly increased as number of pinching was increased. And these cells are able to induce local recurrence by local seeding. Considering this result, authors are able to confirm that the minimal handling in cancer surgery is important factor to prevent local recurrence.