• Title/Summary/Keyword: Staining solutions

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Differential Expression of EGFR Protein by Immunohistochemical Staining Methods and the Relationship Between the Degree of EGFR Protein Expression and EGFR Gene Mutation (면역조직화학적 염색 방법에 따른 상피세포 성장 수용체 단백(EGFR)의 발현정도의 차이 및 EGFR의 발현정도와 EGFR 유전자의 돌연변이와의 상관관계에 대한 고찰)

  • Yoon, In-Sook;Kim, Keuk-Jun;Lee, Eun-Hwa;Seok, Sang-Hee;Kim, Sang-Hee;Kim, Hyun-Yong;Song, Ho-Jung;Lee, Tae-Jong
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.217-222
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    • 2007
  • In the last 5 years the Epidermal Growth Factor Receptor (EGFR) has emerged as one of the most important targets for drug development in oncology. Monoclonal antibodies targeting the external domain of EGFR have been shown to have clinical benefits in colorectal and head and neck cancer when combined with chemotherapy and/or radiation. Also the targeting of the epithelial growth factor receptor (EGFR) kinase domain using the closely related inhibitors gefitinib and erlotinib has generally been ineffective against solid tumors, many of which over express the receptor. We found that there were some differential expressions according to primary antibodies of the EGFR protein which being used as one of the histological tumor markers for non-small cell lung cancer (NSCLC). We also found that there are some differential expressions according to antibodies, the pH of the antigen retrieval (AR) buffer solutions and kinds of enzymes. There were some differential expressions according to the secondary antibodies and the detection systems. We analyzed the correlations between the immunohistochemical expressions of the EGFR protein and the gene mutations of the EGFR. The differences between automatic stainers and manual staining methods were also evaluated.

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Effect of high concentrations of carboxymethyl chitosan on preventing formation of peritoneal adhesion in rats (랫드에서 고농도의 Carboxymethyl Chitosan이 복벽 유착 예방에 미치는 영향)

  • Kwon, Young-Sam;Jang, Hwan-Soo;Jeong, Jae-Hoon;Bae, Jae-Sung;Rhee, Jeong-Woo;Kang, Han-Saem;Yang, Jeong-Hoon;Li, Wen-Xue;Jang, Kwang-Ho
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.151-156
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    • 2003
  • The objective of this study was to determine the effective concentration of carboxymethyl chitoaan(CMC) solution on preventing intraperitoneal adhesions. In this study, 60 rats were divided into four groups : an untreated control group and three experimental groups that were treated with 3 ml of 3, 5, and 7% CMC solution. Adhesions were induced by suturing both the ileal serosa and peritoneum scraped until petechial bleeding occurred. CMC solutions were instilled intraperitoneally immediately before closure of the peritoneomuscular layer. The tensile strength of formed adhesions was measured by using a computerized tensiometer. Histopathological changes of adhesions were evaluated by hematoxylin-eosin for light microscopic examination and Massons's trichrom staining for collagen fibers. All of CMC solutions reduced significantly the tensile strength of the adhesions. CMC inhibited the increase of inflammatory cell infiltration and damage of adhesion spot, and accelerated the collagen synthesis and neovascuiarization. No significant differences were observed among all of CMC concentration at inhibiting adhesion formation. Therefore, we could conclude that 3% CMC, the lowest concentration adopted in this study, was appropriate to prevent the formation of peritoneal adhesion.

Effects of supplementation with antifreeze proteins on the follicular integrity of vitrified-warmed mouse ovaries: Comparison of two types of antifreeze proteins alone and in combination

  • Kim, Min Kyung;Kong, Hyun Sun;Youm, Hye Won;Jee, Byung Chul
    • Clinical and Experimental Reproductive Medicine
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    • v.44 no.1
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    • pp.8-14
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    • 2017
  • Objective: The aim of this study was to analyze the effect of supplementing vitrification and warming solutions with two types of antifreeze proteins (AFPs) and the combination thereof on the follicular integrity of vitrified-warmed mouse ovaries. Methods: Ovaries (n=154) were obtained from 5-week-old BDF1 female mice (n=77) and vitrified using ethylene glycol and dimethyl sulfoxide with the supplementation of 10 mg/mL of Flavobacterium frigoris ice-binding protein (FfIBP), 10 mg/mL of type III AFP, or the combination thereof. Ovarian sections were examined by light microscopy after hematoxylin and eosin staining, and follicular intactness was assessed as a whole and according to the type of follicle. Apoptosis within the follicles as a whole was detected by a terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling assay. Results: The proportion of overall intact follicles was significantly higher in the type III AFP-supplemented group (60.5%) and the combination group (62.9%) than in the non-supplemented controls (43.8%, p<0.05 for each). The proportion of intact primordial follicles was significantly higher in the FfIBP-supplemented (90.0%), type III AFP-supplemented (92.3%), and combination (89.7%) groups than in the non-supplemented control group (46.2%, p<0.05 for each). The proportions of non-apoptotic follicles were similar across the four groups. Conclusion: Supplementation of the vitrification and warming solutions with FfIBP, type III AFP, or the combination thereof was equally beneficial for the preservation of primordial follicles in vitrified mouse ovaries.

Effect of Platelet-rich Plasma (PRP) on Regeneration of Rat Sciatic Nerve in a Silicone Chamber

  • Minn, Kyung-Won;Jeong, Eui-Cheol;Chang, Hak;Kwon, Sung-Tack;Kim, Suk-Wha;Baek, Rong-Min
    • Archives of Plastic Surgery
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    • v.37 no.2
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    • pp.105-109
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    • 2010
  • Purpose: The purpose of this study is to determine the effect of platelet-rich plasma (PRP) on rat sciatic nerve regeneration in a 10 mm silicone chamber. Methods: A total of 6 inbred Lewis rats were used in this study. Bilateral sciatic neurectomy was performed on each rat. On one side, silicone chambers containing PRP solutions were implanted; on the contralateral side, the chambers without PRP were implanted as a control. In 12 weeks post-implantation, chambers were retrieved and both gastrocnemius muscles were excised. Nerves biopsy samples were examined under a light microscope after Masson trichrome staining. Results: Cross sections of the midpoints of PRP treated nerves were significantly larger and appeared more mature than those of controls. Conclusion: Based on morphological evidence, PRP has a positive effect on neural regeneration, and it may therefore be useful for treating peripheral nerve injuries.

Cryopreservation of Sporothalli of the Genus Porphyra (Bangiales, Rhodophyta) from Korea

  • Jo, Young-Hyun;Kang, Sung-Pil;Seo, Tae-Ho;Choi, Sung-Je;Kho, Kang-Hee;Kuwano, Kazuyoshi;Saga, Naotsune;Kim, Min-Yong;Shin, Jong-Ahm
    • ALGAE
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    • v.18 no.4
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    • pp.321-331
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    • 2003
  • Cryopreservation of sporothalli of the red alga Porphyra (P. seriata, P. yezoensis, P. tenera, P. pseudolinearis and P. dentata) was performed by the two-step cooling method in liquid nitrogen. The algal samples were suspended in various cryoprotective solutions, and slowly cooled to -40$^{\circ}C$ in 4 hours using a programmed freezer. After the first slow cooling the suspensions with cryoprotectants were immediately immersed in liquid nitrogen. The suspension from the programmed freezer was thawed quickly by agitation of the vial in a water bath at 40°C. When ice in the suspension of cryogenic vial was mostly melted, the vial was transferred to an ice bath for complete melting of the residual ice. The cryoprotectants in the vial were washed off by gradual dilution with seawater. The viability of the cell was assessed with neutral red staining. The viability of Porphyra samples ranged 54.6-70.9% when the mixed suspension of 10% dimethylsulfoxide and 0.5 M sorbitol in 50% seawater used as a cryoprotectant.

COLOR STABILITY OF NEW SILORANE-BASED COMPOSITE RESIN: AN IN VITRO SPECTROPHOTOMETRIC STUDY (Silorane-based 복합레진의 색안정성에 관한 연구)

  • Son, Yu-Jin;Hyun, Hong-Keun;Kim, Young-Jae;Kim, Jung-Wook;Lee, Sang-Hoon;Kim, Chong-Chul;Hahn, Se-Hyun;Jang, Ki-Taeg
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.1
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    • pp.73-81
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    • 2010
  • The objective of this study was to evaluate the color stability of new silorane-based composite resin compared with methacrylate-based composite resins after immersion in 3 staining solutions. One silorane-based composite(Filtek P90) and 3 methacrylate -based composites(Filtek Z250, Filtek Z350, Tetric Ceram) were evaluated. Twenty disk-shaped specimens($10{\times}2mm$) of each of 4 composite resins were prepared. The specimens were then divided into 4 groups of 5 specimens each and immersed in 3 staining solutions( coffee, red wine, curry solution) or distilled water(control) for 28-day test period. Color of the specimens was measured with a spectrophotometer(Color Eye 7000A) using CIE $L^*a^*b^*$ color space relative to CIE standard illuminant D65 at baseline, 1day, 3days, 7days, 14days, 21days and 28days after staining. The color differences( ${\Delta}{E^*}_{ab}$) were calculated. The results were obtained as follows ; 1. The mean color changes(${\Delta}{E^*}_{ab}$) of the composites were greatest in curry solution, then red wine, coffee and distilled water, in decreasing order. 2. The mean color changes(${\Delta}{E^*}_{ab}$) of the composites were greatest in Z350, Z250, Tetric Ceram and P90, in decreasing order.

Highly sensitive and selective detection of cyanide in aqueous solutions using a surface acoustic wave chemical sensor (표면음향파 화학센서를 이용한 수용액 중 시안화이온의 선택적인 고감도 검출)

  • Lee, Soo Suk
    • The Journal of the Acoustical Society of Korea
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    • v.35 no.6
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    • pp.473-479
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    • 2016
  • We report a highly selective and sensitive 200 MHz Surface Acoustic Wave (SAW) sensor that can detect cyanide ion in aqueous solution using surface immobilized thioester molecules in combination with gold nanoparticles (AuNPs). To construct the sensor device, a monolayer of thioester compound was immobilized on the SAW sensor surface. At the sensor surface, hydrolysis of thioester group by nucleophilic addition of cyanide occurred and the resulting free thiol unit bound to AuNP to form thiol-AuNP conjugate. For the signal enhancement, gold staining signal amplification process was introduced subsequently with gold (III) chloride trihydrate and reducing agent, hydroxylamine hydrochloride. The SAW sensor showed a detection ability of $17.7{\mu}M$ for cyanide in aqueous solution and demonstrated a saturation behavior between the frequency shift and the concentration of cyanide ion. On the other hand, our SAW sensor had no activities for other anions such as fluoride ion, acetate ion and sulfate ion, moreover, no significant interference observed by other anions. Finally, all the experiments were carried out in-house developed sensor and fluidics modules to obtain highly reproducible results.

An Experimental Study on the Effect of the Coronary Vasodilator-mixed Cardioplegic Solution on Myocardial Protection during Prolonged Aortic Cross-Clamping (심혈관 확장제를 첨가한 심정지액의 심근보호에 미치는 효과)

  • Jo, Gyu-Do;Kim, Se-Hwa;Lee, Hong-Gyun
    • Journal of Chest Surgery
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    • v.18 no.4
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    • pp.732-739
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    • 1985
  • This study was experimentally undertaken to evaluate the effect of the coronary vasodilator-mixed cardioplegic solution on myocardial protection during prolonged aortic cross-clamping. The dogs were divided into two groups: control group A[received hypothermic cardioplegic solution without any additive coronary vasodilator], and comparing group 8[received hypothermic cardioplegic solution, mixed with various coronary vasodilators and Inderal]. Group A further was divided into two subgroups: subgroup A-1[ischemic time, 90 minutes], and subgroup A-2 [ischemic time, 240 minutes]. Group B further was divided into five subgroups: subgroup B-1 [received papaverine mixed hypothermic cardioplegic solution], subgroup B-2[received nitroglycerin mixed hypothermic cardioplegic solution], subgroup B-3 [received nitroprusside mixed hypothermic cardioplegic solution, subgroup B-4[received hydralazine mixed hypothermic cardioplegic solution], and subgroup B-5 [received inderal mixed hypothermic cardioplegic solution]. The specimens from all of the subgroups were studied by electron microscopic examination. The specimens of subgroups [B-l, B-2 8-3, and B-4], received coronary vasodilators mixed hypothermic cardioplegic solutions, were also compared by methylene blue induced staining of the myocardium and coronary vessels. The results obtained were as followings: l. On electron microscopic examination, all of the specimens, including subgroup A-2, showed no irreversible change of the myocardium. But the best result was obtained from the subgroup B-l, treated by papaverine mixed hypothermic cardioplegic solution. The subgroup B-2, treated by nitroglycerin, was next. And the subgroup B-5, treated by Inderal, was agreeable, comparing the electron microscopic finding with control group in the effect of myocardial protection. 2. The distribution in the myocardium of cardioplegic solution was demonstrated with the aid of methylene blue staining in the subgroups of B-l, B-2, B-3, and B-4, and they were the groups treated by papaverine, nitroglycerin, nitroprusside, and hydralazine in their grouping order. The best result was obtained from the subgroup B-1 [papaverine]. The subgroup B-2 [nitroglycerin] was next. The subgroup B-3 [nitroprusside] was moderate in finding of the colorization. The subgroup B-4 [hydralazine] was the poorest in the distribution of the cardioplegic solution in the myocardium. From these results, it appeared that myocardial protection during ischemic arrest for open heart surgery could be enhanced considerably when coronary dilatation was assured.

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A Study on the Morphological Analysis of Sperm (정자의 형태학적 특성 분석에 관한 연구)

  • Paick, Jae-Seung;Jeon, Seong-Soo;Kim, Soo-Woong;Yi, Won-Jin;Park, Kwang-Suk
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.2
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    • pp.153-165
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    • 1997
  • In male reproducible health, fertility and IVF (in-vitro fertilization), semen analysis has been most important. Semen analysis can be divided into concentration, motional and morphological analysis of sperm. The existing method which was developed earlier to analyze semen concentrated on the sperm motility analysis. To provide more useful and precise solutions for clinical problems such as infertility, semen analysis must include sperm morphological analysis. But the traditional tools for semen analysis are subjective, imprecise, inaccurate, difficult to standardize, and difficult to reproduce. Therefore, with the help of development of microcomputers and image processing techniques, we developed a new sperm morphology analyzer to overcome these problems. In this study the agreement on percent normal morphology was studied between different observers and a computerized sperm morphology analyzer on a slide-by-slide basis using strict criteria. Slides from 30 different patients from the SNUH andrology laboratory were selected randomly. Microscopic fields and sperm cells were chosen randomly and percent normal morphology was recorded. The ability of sperm morphology analyzer to repeat the same reading for normal and abnormal cells was studied. The results showed that there was no significant bias between two experienced observers. The limits of agreement were 4.1%${\sim}$-3.8%. The Pearson correlation coefficient between readers was 0.79. Between the manual and sperm morphology analyzer, the same findings were reported. In this experiments the slides were stained by two different methods, PAP and Diff-Quik staining methods. The limits of agreement were 7.2%${\sim}$-5.7% and 6.0%${\sim}$-6.3%, respectively. The Pearson correlation coefficients ware 0.76 and 0.91, respectively. The limits of agreement was tighter below 20% normal forms. In the experiments of repeatability, 52 cells stained by PAP and Diff-Quik staining methods were analyzed three times in succession. Estimating pairwise agreement, the kappa statistic for the pairs were 0.76, 0.81, 0.86, and 0.75, 0.88, 0.88 respectively. In this study it was shown that there was good agreement between manual and computerized assessment of normal and abnormal cells. The repeatability and agreement per slide of computerized sperm morphology analyzer was excellent. The computer's ability to classify normal morphology per slide is promising. Based on results obtained, this system can be of clinical value both in andrology laboratories and IVF units.

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Development of FISH Methods for Detection of Genus Bacillus (Bacillus 속 세균을 검출하기 위한 Fluorescent In Situ Hybridization 방법의 개발)

  • 홍선희;김옥선;송홍규;이동훈;안태석
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.204-208
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    • 2001
  • A technique for detection of Bacillus in soil and waste water treatment system was developed. Mixed col- tured solutions of 22 Bacillus strains were applied for selection of probe and pretreatment method for FISHAmong the three probes known as useful tool for FISH method,S-G-Bacill-0597-a-A-22 was best for detec-tion of Bacillus. Ninety five percent of DAPl count was observed with FISH method with S-G-Bacill-0597-a-A-22 probe. For increasing the permeability to Bacillus cell walls, pretreatment with lysozyme was betterthan that with lysozyme and SDS/DTT (dithiothreitol). Bacillus spore was not detected with FISH. So, Bacil-lus detection in ecosystem requires FISH with pretreatment of lysozyme and spore staining.

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