• Title/Summary/Keyword: Fluorescence probe technique

Search Result 68, Processing Time 0.029 seconds

Determination of HER2 Gene Amplification in Breast Cancer using Dual-color Silver Enhanced in situ Hybridization (dc-SISH) and Comparison with Fluorescence ISH (FISH)

  • Unal, Betul;Karaveli, Fatma Seyda;Pestereli, Hadice Elif;Erdogan, Gulgun
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.10
    • /
    • pp.6131-6134
    • /
    • 2013
  • Background: The two basic methods that are currently accepted to identify the HER2 status are immunohistochemistry and flyorescence in situ hybridization (FISH). The aim of this study was to perform the dual-color silver in situ hybridization (dc-SISH) technique as an alternative to FISH. Materials and Methods: A total of 40 invasive breast carcinoma cases were assessed for HER2 gene amplification by FISH and dual-color SISH. Results: Significant correlation was found in the HER2 expression results obtained with the two approaches (p=0.001, p<0.05). The concordance rate was 92.3%. Conclusions: Foutine practical use of the dc-SISH method, which is much easier to apply, score, and evaluate, has many advantages. HER2 and CEN17 status can be evaluated simultaneously with the newly developed "Dual-Color Probe". All these specifications and the reliable results obtained support the widespread use of SISH technique in clinical practice.

Effects of Barbiturates on the Fluidity of Phosphatidylethanolamine Model Membranes (Barbiturates가 소의 신선한 대뇌피질 Synaptosomal Plasma Membrane Vesicles로 부터 추출하여 제제한 Phosphatidylethanolamine 인공세포막의 유동성에 미치는 영향)

  • Yun, Il;Kim, Hyung-Il;Hwang, Tae-Ho;Kim, Jong-Ryol;Kim, In-Se;Chung, Yong-Za;Shin, Yong-Hee;Jung, Hyun-Ok;Kang, Jung-Sook
    • The Korean Journal of Pharmacology
    • /
    • v.26 no.2
    • /
    • pp.209-217
    • /
    • 1990
  • Intramolecular excimer formation with 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization with 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effects of barbiturates on the bulk fluidity of the model membranes of phosphatidylethanolamine fraction of synaptosomal plasma membrane vesicles (SPMVPE) isolated from bovine cerebral cortex. In the SPMVPE, barbiturates decreased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and increased the fluorescence polarization (P), anisotropy (r), limiting anisotropy $(r_{8})$, order parameter (S) and rotational relaxation time $({\bar{P}})$ of DPH in a dose-dependent manner. The relative potencies of barbiturates to order the SPMVPE were in the order: pentobarbital > hexobarbital > amobarbital > phenobarbital. Hence, it is concluded that barbiturates have ordering effects on the SPMVPE. And the membrane-ordering potencies of barbiturates appear to be correlated with the potencies for enhancement of GABA-stimulated chloride influx and with the anesthetic effects of barbiturates.

  • PDF

Relationship between the DNA content of human chromosome and their contribution to radiation-induced chromosome aberration analysed by fluorescence in situ hybridization(FISH) (방사선에 의한 염색체이상과 DNA 함량과의 관계)

  • Chung, Hai-Won;Kim, Su-Young;Ha, Sung-Whan;Kim, Tae-Hwan;Cho, Chul-Koo
    • Journal of Radiation Protection and Research
    • /
    • v.26 no.2
    • /
    • pp.101-111
    • /
    • 2001
  • To study the relationship between the DNA content of human chromosomes and their involvement in radiation-induced structural chromosome aberrations, the frequencies of translocations and dicentrics induced in human lymphocytes after in vitro exposure to 2Gy of x-tay were analysed by fluorescence in situ hybridization(FISH). Single whole chromosome probes(WCP), specific for chromosomes 1, 2, 4, 7, 8, 9, and 21 and triple combination of probes tot chromosome 1, 2 & 4 were used separately. A significant deviation of the frequency of symmetrical translocation and dicentrics from a DNA-proportional distribution was observed. Chromosomes 2, 7, 8, 9 and 21 were less frequently involved in the formation of symmetrical translocations and dicentrics than expected, whereas chromosomes 1 and 4 were more frequently involved. Chromosome 2 and 4 showed a higher frequency of acentric fragments. When triple combination probe for chromosome 1, 2 & 4 was used, no differences were found between the observed and expected frequency of exchange type aberrations. The results showed that the frequency of radiation-induced chromosome aberrations was not proportional to DNA contents, suggesting the difference in the susceptibility to specific aberrations among individual chromosomes. The results also indicated that the FISH technique with combination of probes for chromosome 1, 2 & 4 was useful for radiation biodosimetry.

  • PDF

Effect of Lidocaine-HCl on Microviscosity of Phosphatidylcholine Model Membrane

  • Chung, In-Kyo;Kim, Inn-Se;Choi, Chang-Hwa;Cho, Goon-Jae;Kim, Jin-Bom;Son, Woo-Sung;Jang, Hye-Ock;Yun, Il
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.4 no.3
    • /
    • pp.243-251
    • /
    • 2000
  • In order to provide a basis for studying the molecular mechanism of pharmacological action of local anesthetics and to develop a fluorescence spectroscopic method which can detect the microviscosity of native and model membranes using intramolecular excimerization of 1,3-di(l-pyrenyl)propane (Py-3-Py), we examined the effect of lidocaine HCl on the microviscosity of model membranes of phosphatidylcholine fraction extracted from synaptosomal plasma membrane vesicles (SPMVPC). The excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in liquid paraffin was a simple linear function of $T/{\eta}.$ Based on this calibration curve, the microviscosity values of the direct probe environment in SPMVPC model membranes ranged from $234.97{\pm}48.85$ cP at $4^{\circ}C$ to %19.21{\pm}1.11$ cP at $45^{\circ}C.$ At $37^{\circ}C,$ a value of $27.25{\pm}0.44$ cP was obtained. The lidocaine HCl decreased the microviscosity of SPMVPC model membranes in a concentration-dependent manner, with a significant decrease in microviscosity value by injecting the local anesthetic even at the concentration of 0.5 mM. These results indicate that the direct environment of Py-3-Py in the SPMVPC model membranes is significantly fluidized by the lidocaine HCl. Also, the present study explicitly shows that an interaction between local anesthetics and membrane lipids is of importance in the molecular mechanism of pharmacological action of lidocaine HCl.

  • PDF

The Production of Sex Determined Cattle by Embryonic Sexing Using Fluorescence In Situ Hybridization Technique (FISH 기법을 이용한 소 수정란의 성감별과 산자 생산)

  • Sohn, S.H.;Park, H.
    • Proceedings of the Korean Society of Embryo Transfer Conference
    • /
    • 2007.05a
    • /
    • pp.39-50
    • /
    • 2007
  • Sexing from bovine embryos fertilized in vitro implicates a possibility of the sex controlled cattle production. This study was carried out to produce the sex determined cattle through the embryonic sexing by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe constructed from the btDYZ-1 sequence. Using this probe, a male-specific signal was detected on 100% of Y-chromosome bearing metaphase specimens. The analyzable rate of embryonic sexing by FISH technique was about 93% (365/393) regardless of embryonic stages. As tested single blastomere by FISH and then karyotype with their biopsied embryos, the accuracy of sex determination with FISH was 97.6%. We tried the embryo transfer with sex determined embryos on 15 cattle. Among them, the 5 cattle delivered calf with expected sex last year.

  • PDF

Rapid Detection of Cochlodinium polykrikoides by Sandwich Hybridization Integrated with Invertase Assay (Invertase 분석이 통합된 sandwich hybridization에 의한 Cochlodinium polykrikoides의 신속 검출)

  • Kang, Mingyeong;Park, Mirye;Lee, Taek-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
    • /
    • v.20 no.8
    • /
    • pp.627-633
    • /
    • 2019
  • Rapid and accurate identification of marine microalgae causing harmful algal blooms (HABs) is a crucial tool for predicting and managing HABs. We previously developed a nuclease protection assay sandwich hybridization (NPA-SH) method for the in situ detection of blooming microalgae Cochlodinium polykrikoides. In this study, we improved the applicability of the NPA-SH method for the detection of C. polykrikoides by simplifying the reaction step. For this purpose, invertase (INV) was conjugated to the signal probe instead of using fluorescence, and sucrose was used as a reactant to induce a color reaction. The INV-signal probe conjugation was confirmed by SDS-PAGE and epifluoromicroscopy. The treatment time and appropriate amounts of the probe and sucrose that optimized the reaction were determined. As a result, the developed INV-SH reduced the treatment time in the field compared with NPA-SH, and also enabled the use of a relatively small volume and low-priced personal glucose meter, as well as an absorbance meter. INV-SH is the first C. polykrikoides species identification technology to which INV has been applied and could be an improved field technique.

Effects of Chlorhexidine Digluconate on Rotational Rate of n-(9-Anthroyloxy)stearic Acid in Porphyromonas ginginvalis Outer Membranes

  • Jang, Hye-Ock;Cha, Seong-Kweon;Lee, Chang;Choi, Min-Gak;Huh, Sung-Ryul;Shin, Sang-Hun;Chung, In-Kyo;Yun, Il
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.7 no.3
    • /
    • pp.125-130
    • /
    • 2003
  • The aim of this study was to provide a basis for studying the molecular mechanism of pharmacological action of chlorhexidine digluconate. Fluorescence polarization of n-(9-anthroyloxy)stearic acid was used to examine the effect of chlorhexidine digluconate on differential rotational mobility of different positions of the number of membrane bilayer phospholipid carbon atoms. The six membrane components differed with respect to 2, 3, 6, 9, 12, and 16-(9-anthroyloxy)stearic acid (2-AS, 3-AS, 6-AS, 9-AS, 12-AS and 16-AP) probes, indicating different membrane fluidity. Chlorhexidine digluconate increased the rate of rotational mobility of hydrocarbon interior of the cultured Porphyromonas gingivalis outer membranes (OPG) in a dose-dependent manner, but decreased the mobility of surface region (membrane interface) of the OPG. Disordering or ordering effects of chlorhexidine digluconate on membrane lipids may be responsible for some, but not all of its bacteriostatic and bactericidal actions.

Differential Effects of Local Anesthetics on Rate of Rotational Mobility between Hydrocarbon Interior and Surface Region of Model Membrane Outer Monolayer

  • Chung, In-Kyo;Cha, Seong-Kweon;Chung, Yong-Za;Kim, Bong-Sun;Choi, Chang-Hwa;Cho, Goon-Jae;Jang, Hye-Ock;Yun, Il
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.4 no.1
    • /
    • pp.41-46
    • /
    • 2000
  • Using fluorescence polarization of 12-(9-anthroyloxy)stearic acid (12-AS) and 2-(9-anthroyloxy)stearic acid (2-AS), we evaluated the differential effects of local anesthetics on differential rotational rate between the surface (in carbon number 2 and its surroundings including the head group) and the hydrocarbon interior (in carbon number 12 and its surroundings) of the outer monolayer of the total lipid fraction liposome extracted from synaptosomal plasma membrane vesicles. The anisotropy (r) values for the hydrocarbon interior and the surface region of the liposome outer monolayer were $0.078{\pm}0.001$ and $0.114{\pm}0.001,$ respectively. This means that the rate of rotational mobility in the hydrocarbon interior is faster than that of the surface region. In a dose-dependent manner, the local anesthetics decreased the anisotropy of 12-AS in the hydrocarbon interior of the liposome outer monolayer but increased the anisotropy of 2-AS in the surface region of the monolayer. These results indicate that local anesthetics have significant disordering effects on the hydrocarbon interior but have significant ordering effects on the surface region of the liposome outer monolayer.

  • PDF

Amphiphilic graft copolymers: Effect of graft chain length and content on colloid gel

  • Nitta, Kyohei;Kimoto, Atsushi;Watanabe, Junji;Ikeda, Yoshiyuki
    • Biomaterials and Biomechanics in Bioengineering
    • /
    • v.2 no.2
    • /
    • pp.97-109
    • /
    • 2015
  • A series of amphiphilic graft copolymers were synthesized by varying the number of graft chains and graft chain lengths. The polarity of the hydrophobic graft chain on the copolymers was varied their solution properties. The glass transition temperature of the copolymers was in the low-temperature region, because of the amorphous nature of poly (trimethylene carbonate) (PTMC). The surface morphology of the lyophilized colloid gel had a bundle structure, which was derived from the combination of poly(N-hydroxyethylacrylamide)( poly(HEAA)) and PTMC. The solution properties were evaluated using dynamic light scattering and fluorescence measurements. The particle size of the graft copolymers was about 30-300 nm. The graft copolymers with a higher number of repeating units attributed to the TMC (trimethylene carbonate) component and with a lower macromonomer ratio showed high thermal stability. The critical association concentration was estimated to be between $2.2{\times}10^{-3}$ and $8.9{\times}10^{-2}mg/mL$, using the pyrene-based fluorescence probe technique. These results showed that the hydrophobic chain of the graft copolymer having a long PTMC segment had a low polarity, dependent on the number of repeating units of TMC and the macromonomer composition ratio. These results demonstrated that a higher number of repeating units of TMC, with a lower macromonomer composition, was preferable for molecular encapsulation.

Different Effects of Dopamine on Differential Rotational Mobility between Inner and Outer Monolayer of Synaptosomal Plasma Membrane Vesicles Isolated from Bovine Brain

  • Kim, Hyun-Gang;Choi, Chang-Hwa;Kim, Inn-Se;Chung, In-Kyo;Cho, Goon-Jae;Jang, Hye-Ock;Yun, Il
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.4 no.5
    • /
    • pp.409-415
    • /
    • 2000
  • Fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) was used to evaluate the effects of dopamine HCl on the range of the rotatioanl mobility of bulk bilayer structure of the synaptosomal plasma membrane vesicles (SPMV) isolated from whole bovine brain. In a dose-dependent manner, dopamine decreased the anisotropy $({\gamma}),$ limiting anisotropy $({\gamma}{infty})$ and order parameter (S) of DPH in the membranes. These indicate that dopamine increased the rotational mobility of the probe in the neuronal membranes. Cationic 1-[4-(trimethylammonio)-phenyl]-6-phenylhexa-1,3,5-hexatriene (TMA-DPH) and anionic 3-[p-(6-phenyl)-1,3,5-hexatrienyl]-phenylpropionic acid (PRO-DPH) were utilized to examine the range of transbilayer asymmetric rotational mobility of the neuronal membranes. Dopamine had a greater increasing effect on the mobility of the inner monolayer as compared to the outer monolayer of the neuronal membranes. It has been proven that dopamine exhibits a selective rather than nonselective fluidizing effect within the transbilayer domains of the SPMV.

  • PDF