Metastatic hidradenocarcinoma with demonstration of Her-2/neu gene amplification by fluorescence in situ hybridization: potential treatment implications.

Science.gov (United States)

Nash, Jason W; Barrett, Terry L; Kies, Merrill; Ross, Merrick I; Sneige, Nour; Diwan, A Hafeez; Lazar, Alexander J F

2007-01-01

A 44-year-old man was referred for a right chest nodule of 3 months duration. A 'benign' nodule had been excised from this location 8 years prior. On examination, palpable nodes were noted in the right axilla. Radiographic studies were significant only for right axillary lymphadenopathy. Histologically, a nodular dermal proliferation composed of poorly differentiated epithelioid cells in nests and focally forming ducts with pseudopapillary architecture comprised the primary tumor. Features of a clear cell hidradenoma were noted focally. Immunohistochemical (IHC) analysis revealed reactivity for HMW cytokeratins, CK5 and CK7, p53, p63, CEA (focal), androgen receptor, EGFR, estrogen receptor (ER), MUC5AC, and strong/diffuse membranous staining for Her-2/neu. Negative stains included villin, TTF-1, CDX2, S-100 protein, vimentin, gross cystic disease fluid protein 15 (GCDFP-15), mammoglobulin, and MUC2. A wide local excision and axillary node dissection was performed. Metastatic tumor involved nine of 28 nodes. Interphase fluorescence in situ hybridization (FISH) demonstrated chromosomal amplification of the Her-2/neu locus within the tumor and a nodal metastasis. The patient has completed adjuvant and radiotherapy, including trastuzumab, and is asymptomatic. We believe this to be the first demonstration of Her-2/neu amplification in a malignant skin adnexal tumor. In analogy to breast carcinoma, these findings suggest the applicability of trastuzumab for patients with metastatic adnexal carcinomas demonstrating Her-2/neu amplification.

Comparison of Chromogenic In Situ Hybridization and Fluorescence In Situ Hybridization for the Evaluation of MDM2 Amplification in Adipocytic Tumors.

Science.gov (United States)

Mardekian, Stacey K; Solomides, Charalambos C; Gong, Jerald Z; Peiper, Stephen C; Wang, Zi-Xuan; Bajaj, Renu

2015-11-01

Atypical lipomatous tumor/well-differentiated liposarcoma (ALT-WDLPS) and dedifferentiated liposarcoma (DDLPS) are characterized cytogenetically by a 12q13-15 amplification involving the mouse double minute 2 (MDM2) oncogene. Fluorescence in situ hybridization (FISH) is used frequently to detect this amplification and aid with the diagnosis of these entities, which is difficult by morphology alone. Recently, bright-field in situ hybridization techniques such as chromogenic in situ hybridization (CISH) have been introduced for the determination of MDM2 amplification status. The present study compared the results of FISH and CISH for detecting MDM2 amplification in 41 cases of adipocytic tumors. Amplification was defined in both techniques as a MDM2/CEN12 ratio of 2 or greater. Eleven cases showed amplification with both FISH and CISH, and 26 cases showed no amplification with both methods. Two cases had discordant results between CISH and FISH, and two cases were not interpretable by CISH. CISH is advantageous for allowing pathologists to evaluate the histologic and molecular alterations occurring simultaneously in a specimen. Moreover, CISH is found to be more cost- and time-efficient when used with automation, and the signals do not quench over time. CISH technique is a reliable alternative to FISH in the evaluation of adipocytic tumors for MDM2 amplification. © 2014 Wiley Periodicals, Inc.

In situ biosynthesis of bacterial nanocellulose-CaCO3 hybrid bionanocomposite: One-step process

International Nuclear Information System (INIS)

Mohammadkazemi, Faranak; Faria, Marisa; Cordeiro, Nereida

2016-01-01

In this work, a simple and green route to the synthesis of the bacterial nanocellulose-calcium carbonate (BNC/CaCO 3 ) hybrid bionanocomposites using one-step in situ biosynthesis was studied. The CaCO 3 was incorporated in the bacterial nanocellulose structure during the cellulose biosynthesis by Gluconacetobacter xylinus PTCC 1734 bacteria. Hestrin-Schramm (HS) and Zhou (Z) culture media were used to the hybrid bionanocomposites production and the effect of ethanol addition was investigated. Attenuated total reflection Fourier transform infrared spectroscopy, field emission scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray spectroscopy, inverse gas chromatography and thermogravimetric analysis were used to characterize the samples. The experimental results demonstrated that the ethanol and culture medium play an important role in the BNC/CaCO 3 hybrid bionanocomposites production, structure and properties. The BNC/CaCO 3 biosynthesized in Z culture medium revealed higher O/C ratio and amphoteric surface character, which justify the highest CaCO 3 content incorporation. The CaCO 3 was incorporated into the cellulosic matrix decreasing the bacterial nanocellulose crystallinity. This work reveals the high potential of in situ biosynthesis of BNC/CaCO 3 hybrid bionanocomposites and opens a new way to the high value-added applications of bacterial nanocellulose. - Graphical Abstract: Display Omitted - Highlights: • BNC/CaCO 3 hybrid bionanocomposites were produced using in situ biosynthesis process. • Ethanol and culture medium play an important role in the production and properties. • Z-BNC/CaCO 3 bionanocomposites revealed higher O/C ratio and amphoteric surface character. • CaCO 3 incorporated into the BNC decreased crystallinity.

The Use of Whole-Mount "in Situ" Hybridization to Illustrate Gene Expression Regulation

Science.gov (United States)

Llamusí, Beatriz; Muñoz-Soriano, Verónica; Paricio, Nuria; Artero, Rubén

2014-01-01

"In situ" hybridization is a widely used technique for studying gene expression. Here, we describe two experiments addressed to postgraduate genetics students in which the effect of transcription factors on gene expression is analyzed in "Drosophila embryos of different genotypes by whole-mount in situ hybridization. In one of the…

Resolution-improved in situ DNA hybridization detection based on microwave photonic interrogation.

Science.gov (United States)

Cao, Yuan; Guo, Tuan; Wang, Xudong; Sun, Dandan; Ran, Yang; Feng, Xinhuan; Guan, Bai-ou

2015-10-19

In situ bio-sensing system based on microwave photonics filter (MPF) interrogation method with improved resolution is proposed and experimentally demonstrated. A microfiber Bragg grating (mFBG) is used as sensing probe for DNA hybridization detection. Different from the traditional wavelength monitoring technique, we use the frequency interrogation scheme for resolution-improved bio-sensing detection. Experimental results show that the frequency shift of MPF notch presents a linear response to the surrounding refractive index (SRI) change over the range of 1.33 to 1.38, with a SRI resolution up to 2.6 × 10(-5) RIU, which has been increased for almost two orders of magnitude compared with the traditional fundamental mode monitoring technique (~3.6 × 10(-3) RIU). Due to the high Q value (about 27), the whole process of DNA hybridization can be in situ monitored. The proposed MPF-based bio-sensing system provides a new interrogation method over the frequency domain with improved sensing resolution and rapid interrogation rate for biochemical and environmental measurement.

Localization of mRNA in vertebrate axonal compartments by in situ hybridization.

Science.gov (United States)

Sotelo-Silveira, José Roberto; Calliari, Aldo; Kun, Alejandra; Elizondo, Victoria; Canclini, Lucía; Sotelo, José Roberto

2011-01-01

The conclusive demonstration of RNA in vertebrate axons by in situ hybridization (ISH) has been elusive. We review the most important reasons for difficulties, including low concentration of axonal RNAs, localization in specific cortical domains, and the need to isolate axons. We demonstrate the importance of axon micro-dissection to obtain a whole mount perspective of mRNA distribution in the axonal territory. We describe a protocol to perform fluorescent ISH in isolated axons and guidelines for the preservation of structural and molecular integrity of cortical RNA-containing domains (e.g., Periaxoplasmic Ribosomal Plaques, or PARPs) in isolated axoplasm.

In situ hybridization of superparamagnetic iron-biomolecule nanoparticles.

Science.gov (United States)

Moghimi, Nafiseh; Donkor, Apraku David; Mohapatra, Mamata; Thomas, Joseph Palathinkal; Su, Zhengding; Tang, Xiaowu Shirley; Leung, Kam Tong

2014-07-23

The increase in interest in the integration of organic-inorganic nanostructures in recent years has promoted the use of hybrid nanoparticles (HNPs) in medicine, energy conversion, and other applications. Conventional hybridization methods are, however, often long, complicated, and multistepped, and they involve biomolecules and discrete nanostructures as separate entities, all of which hinder the practical use of the resulting HNPs. Here, we present a novel, in situ approach to synthesizing size-specific HNPs using Fe-biomolecule complexes as the building blocks. We choose an anticancer peptide (p53p, MW 1.8 kDa) and an enzyme (GOx, MW 160 kDa) as model molecules to demonstrate the versatility of the method toward different types of molecules over a large size range. We show that electrostatic interaction for complex formation of metal hydroxide ion with the partially charged side of biomolecule in the solution is the key to hybridization of metal-biomolecule materials. Electrochemical deposition is then used to produce hybrid NPs from these complexes. These HNPs with controllable sizes ranging from 30 nm to 3.5 μm are found to exhibit superparamagnetic behavior, which is a big challenge for particles in this size regime. As an example of greatly improved properties and functionality of the new hybrid material, in vitro toxicity assessment of Fe-GOx HNPs shows no adverse effect, and the Fe-p53p HNPs are found to selectively bind to cancer cells. The superparamagnetic nature of these HNPs (superparamagnetic even above the size regime of 15-20 nm!), their biocompatibility, and the direct integration approach are fundamentally important to biomineralization and general synthesis strategy for bioinspired functional materials.

Design of Hybrid Steam-In Situ Combustion Bitumen Recovery Processes

International Nuclear Information System (INIS)

Yang Xiaomeng; Gates, Ian D.

2009-01-01

Given enormous capital costs, operating expenses, flue gas emissions, water treatment and handling costs of thermal in situ bitumen recovery processes, improving the overall efficiency by lowering energy requirements, environmental impact, and costs of these production techniques is a priority. Steam-assisted gravity drainage (SAGD) is the most widely used in situ recovery technique in Athabasca reservoirs. Steam generation is done on surface and consequently, because of heat losses, the energy efficiency of SAGD can never be ideal with respect to the energy delivered to the sandface. An alternative to surface steam generation is in situ combustion (ISC) where heat is generated within the formation through injection of oxygen at a sufficiently high pressure to initiate combustion of bitumen. In this manner, the heat from the combustion reactions can be used directly to mobilize the bitumen. As an alternative, the heat can be used to generate steam within the formation which then is the agent to move heat in the reservoir. In this research, alternative hybrid techniques with simultaneous and sequential steam-oxygen injection processes are examined to maximize the thermal efficiency of the recovery process. These hybrid processes have the advantage that during ISC, steam is generated within the reservoir from injected and formation water and as a product of oxidation. This implies that ex situ steam generation requirements are reduced and if there is in situ storage of combustion gases, that overall gas emissions are reduced. In this research, detailed reservoir simulations are done to examine the dynamics of hybrid processes to enable design of these processes. The results reveal that hybrid processes can lower emitted carbon dioxide-to-oil ratio by about 46%, decrease the consumed natural gas-to-oil ratio by about 73%, reduce the cumulative energy-to-oil ratio by between 40% and 70% compared to conventional SAGD, and drop water consumption per unit oil produced

Detection of denitrification genes by in situ rolling circle amplification - fluorescence in situ hybridization (in situ RCA-FISH) to link metabolic potential with identity inside bacterial cells

DEFF Research Database (Denmark)

Hoshino, Tatsuhiko; Schramm, Andreas

2010-01-01

target site. Finally, the RCA product inside the cells was detected by standard fluorescence in situ hybridization (FISH). The optimized protocol showed high specificity and signal-to-noise ratio but low detection frequency (up to 15% for single-copy genes and up to 43% for the multi-copy 16S rRNA gene...... as Candidatus Accumulibacter phosphatis by combining in situ RCA-FISH with 16S rRNA-targeted FISH. While not suitable for quantification because of its low detection frequency, in situ RCA-FISH will allow to link metabolic potential with 16S rRNA (gene)-based identification of single microbial cells.......). Nevertheless, multiple genes (nirS and nosZ; nirS and the 16S rRNA gene) could be detected simultaneously in P. stutzeri. Environmental application of in situ RCA-FISH was demonstrated on activated sludge by the differential detection of two types of nirS-defined denitrifiers; one of them was identified...

In situ biosynthesis of bacterial nanocellulose-CaCO{sub 3} hybrid bionanocomposite: One-step process

Energy Technology Data Exchange (ETDEWEB)

Mohammadkazemi, Faranak, E-mail: f_mkazemi@sbu.ac.ir [Department of Cellulose and Paper Technology, Faculty of New Technologies Engineering, Shahid Beheshti University, Science and Research Campus, Zirab, Savadkooh, Mazandaran (Iran, Islamic Republic of); Faria, Marisa; Cordeiro, Nereida [Faculty of Exact Science and Engineering, University of Madeira, Funchal (Portugal)

2016-08-01

In this work, a simple and green route to the synthesis of the bacterial nanocellulose-calcium carbonate (BNC/CaCO{sub 3}) hybrid bionanocomposites using one-step in situ biosynthesis was studied. The CaCO{sub 3} was incorporated in the bacterial nanocellulose structure during the cellulose biosynthesis by Gluconacetobacter xylinus PTCC 1734 bacteria. Hestrin-Schramm (HS) and Zhou (Z) culture media were used to the hybrid bionanocomposites production and the effect of ethanol addition was investigated. Attenuated total reflection Fourier transform infrared spectroscopy, field emission scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray spectroscopy, inverse gas chromatography and thermogravimetric analysis were used to characterize the samples. The experimental results demonstrated that the ethanol and culture medium play an important role in the BNC/CaCO{sub 3} hybrid bionanocomposites production, structure and properties. The BNC/CaCO{sub 3} biosynthesized in Z culture medium revealed higher O/C ratio and amphoteric surface character, which justify the highest CaCO{sub 3} content incorporation. The CaCO{sub 3} was incorporated into the cellulosic matrix decreasing the bacterial nanocellulose crystallinity. This work reveals the high potential of in situ biosynthesis of BNC/CaCO{sub 3} hybrid bionanocomposites and opens a new way to the high value-added applications of bacterial nanocellulose. - Graphical Abstract: Display Omitted - Highlights: • BNC/CaCO{sub 3} hybrid bionanocomposites were produced using in situ biosynthesis process. • Ethanol and culture medium play an important role in the production and properties. • Z-BNC/CaCO{sub 3} bionanocomposites revealed higher O/C ratio and amphoteric surface character. • CaCO{sub 3} incorporated into the BNC decreased crystallinity.

The correlation between dual-color chromogenic in situ hybridization and fluorescence in situ hybridization in assessing HER2 gene amplification in breast cancer.

Science.gov (United States)

Pedersen, Marianne; Rasmussen, Birgitte Bruun

2009-06-01

Fluorescence in situ hybridization (FISH) is regarded as the gold standard method for detecting HER2 gene amplification. Chromogenic in situ hybridization (CISH) is a promising alternative to FISH because CISH has the advantages of being a method evaluated by bright-field microscopy and the generated chromogenic signals are also stable. This study presents a dual color CISH for simultaneous detection of the HER2 gene and chromosome 17. The CISH method performs a chromogenic detection "on top" of the Food and Drug Administration (FDA)-approved HER2 FISH pharmDx method, where the fluorochrome-labeled probes are detected using enzyme-labeled antibodies and visualized by chromogenic enzymatic reactions. The HER2 status (amplified/not amplified and HER2 ratios) was evaluated by the CISH method and compared with results obtained by the FDA-approved FISH method. Of the 72 successfully investigated invasive breast carcinomas, both FISH and CISH detected HER2 amplification in 24 cases and nonamplification was detected in 47 cases. One case showed a discrepancy between FISH and CISH. The concordance between CISH and FISH was found to be almost perfect (98.6%). The correlation between the HER2 ratios obtained by the 2 methods showed excellent correlation (correlation coefficient 0.95). In conclusion, it is possible by dual-color CISH method to demonstrate HER2 genes and chromosome 17 genes, in the same tissue section and reliably assess HER2 status. The CISH method is a very promising alternative to the FISH method.

Common Fluorescence In Situ Hybridization Applications in Cytology.

Science.gov (United States)

Savic, Spasenija; Bubendorf, Lukas

2016-12-01

- Fluorescence in situ hybridization (FISH) is a well-established method for detection of genomic aberrations in diagnostic, prognostic, and predictive marker testing. - To review common applications of FISH in cytology. - The published literature was reviewed. - Cytology is particularly well suited for all kinds of FISH applications, which is highlighted in respiratory tract cytology with an increasing demand for predictive FISH testing in lung cancer. Fluorescence in situ hybridization is the gold standard for detection of predictive anaplastic lymphoma kinase gene (ALK) rearrangements, and the same evaluation criteria as in histology apply to cytology. Several other gene rearrangements, including ROS proto-oncogene 1 receptor tyrosine kinase (ROS1), are becoming clinically important and share the same underlining cytogenetic mechanisms with ALK. MET amplification is one of the most common mechanisms of acquired resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors and can be targeted by crizotinib. As genomic aberrations are a hallmark of malignant cells, FISH is a valuable objective ancillary diagnostic tool. In urinary tract cytology, atypical urothelial cells equivocal for malignancy are a common diagnostic dilemma and multitarget FISH can help clarify such cells. Diagnosis of malignant mesothelioma remains one of the most challenging fields in effusion cytology, and ancillary FISH is useful in establishing the diagnosis. Fluorescence in situ hybridization is a morphology-based technique, and the prerequisite for reliable FISH results is a targeted evaluation of the cells in question (eg, cancer or atypical cells). Cytopathologists and cytotechnicians should therefore be involved in molecular testing in order to select the best material and to provide their morphologic expertise.

Localization of insulin receptor mRNA in rat brain by in situ hybridization

International Nuclear Information System (INIS)

Marks, J.L.; Porte, D. Jr.; Stahl, W.L.; Baskin, D.G.

1990-01-01

Insulin receptor mRNA was demonstrated in rat brain slices by in situ hybridization with three 35 S-oligonucleotide probes and contact film autoradiography. Specificity was confirmed by showing that (a) excess unlabeled probe abolished the signal, (b) an oligonucleotide probe for rat neuropeptide Y mRNA showed a different distribution of hybridization signal, and (c) the distribution of insulin receptor binding was consistent with the distribution of insulin receptor mRNA. Insulin receptor mRNA was most abundant in the granule cell layers of the olfactory bulb, cerebellum and dentate gyrus, in the pyramidal cell body layers of the pyriform cortex and hippocampus, in the choroid plexus and in the arcuate nucleus of the hypothalamus

In situ biosynthesis of bacterial nanocellulose-CaCO3 hybrid bionanocomposite: One-step process.

Science.gov (United States)

Mohammadkazemi, Faranak; Faria, Marisa; Cordeiro, Nereida

2016-08-01

In this work, a simple and green route to the synthesis of the bacterial nanocellulose-calcium carbonate (BNC/CaCO3) hybrid bionanocomposites using one-step in situ biosynthesis was studied. The CaCO3 was incorporated in the bacterial nanocellulose structure during the cellulose biosynthesis by Gluconacetobacter xylinus PTCC 1734 bacteria. Hestrin-Schramm (HS) and Zhou (Z) culture media were used to the hybrid bionanocomposites production and the effect of ethanol addition was investigated. Attenuated total reflection Fourier transform infrared spectroscopy, field emission scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray spectroscopy, inverse gas chromatography and thermogravimetric analysis were used to characterize the samples. The experimental results demonstrated that the ethanol and culture medium play an important role in the BNC/CaCO3 hybrid bionanocomposites production, structure and properties. The BNC/CaCO3 biosynthesized in Z culture medium revealed higher O/C ratio and amphoteric surface character, which justify the highest CaCO3 content incorporation. The CaCO3 was incorporated into the cellulosic matrix decreasing the bacterial nanocellulose crystallinity. This work reveals the high potential of in situ biosynthesis of BNC/CaCO3 hybrid bionanocomposites and opens a new way to the high value-added applications of bacterial nanocellulose. Copyright © 2016 Elsevier B.V. All rights reserved.

A novel fluorescent in situ hybridization technique for detection of Rickettsia spp. in archival samples

DEFF Research Database (Denmark)

Svendsen, Claus Bo; Boye, Mette; Struve, Carsten

2009-01-01

A novel, sensitive and specific method for detecting Rickettsia spp. in archival samples is described. The method involves the use of fluorescently marked oligonucleotide probes for in situ hybridization. Specific hybridization of Ricekttsia was found without problems of cross-reactions with bact......A novel, sensitive and specific method for detecting Rickettsia spp. in archival samples is described. The method involves the use of fluorescently marked oligonucleotide probes for in situ hybridization. Specific hybridization of Ricekttsia was found without problems of cross...

Image files of planarians analyzed by in situ hybridication and immunohistochemical staining - Plabrain DB | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available List Contact us Plabrain DB Image files of planarians analyzed by in situ hybridication and immunohistochemical... staining Data detail Data name Image files of planarians analyzed by in situ hybridication and immunohistochemical...sion patterns by whole-mount in situ hybridication and also protein distribution by immunohistochemical...Images are displayed in A list of image files of planarians analyzed by in situ hybridication and immunohistochemical...le search URL - Data acquisition method Whole-mount in situ hybridication, immunohistochemical staining Data

Detection and quantitative analysis of actin mRNA by in situ hybridization with an oligodeoxynucleotide probe

International Nuclear Information System (INIS)

Taneja, K.; Singer, R.

1987-01-01

In situ hybridization is a useful method for localizing specific nucleic acid sequences intracellularly and for studying regulation of gene expression. Recently synthetic oligonucleotides have been successfully used as probes in this technique. Since they can be made easily to specific nucleic acid regions, they may be the best approach for analysis of a gene family of highly conserved sequences. They have analyzed these probes for the development of an in situ hybridization method. Oligonucleotides were made to different regions of chick beta-actin mRNA and used for detection of these sequences in a culture of chicken fibroblasts and myoblasts. They found that synthetic DNAs have different efficiencies of hybridization, indicating that not all target sequences are equivalent. They have investigated in detail a particular probe to the actin mRNA coding region and have optimized hybridization parameters. When hybridization was quantitated it was found that an oligonucleotide end labelled with 35 S or 32 P was capable of detecting several thousand messages per cell with a signal-to-noise ratio of 10:1. In situ hybridization confirmed the specificity of the hybridization as well as the background level. Increase in the number of oligonucleotides used should increase the signal-to-noise ratio-proportionately. Under particular circumstances the specificity of oligonucleotides make them an important reagent for in situ hybridization

Fluorescence in situ hybridization on formalin-fixed and paraffin-embedded tissue

DEFF Research Database (Denmark)

Laub Petersen, Bodil; Zeuthen, Mette Christa; Pedersen, Sanni

2004-01-01

Fluorescence in situ hybridization (FISH) is widely used to study numerical and structural genetic abnormalities in both metaphase and interphase cells. The technique is based on the hybridization of labeled probes to complementary sequences in the DNA or RNA of the cells. Interphase FISH is most...... in time lapse between removal of tissue and fixation, duration of fixation, enzymatic pretreatment, hybridization conditions, and posthybridization washing conditions are important factors in the hybridization. In this study, we have listed the results of a systematic approach to improve FISH on isolated...

In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold.

Science.gov (United States)

Hutchison, N J; Langer-Safer, P R; Ward, D C; Hamkalo, B A

1982-11-01

In situ hybridization has become a standard method for localizing DNA or RNA sequences in cytological preparations. We developed two methods to extend this technique to the transmission electron microscope level using mouse satellite DNA hybridization to whole mount metaphase chromosomes as the test system. The first method devised is a direct extension of standard light microscope level using mouse satellite DNA hybridization to whole mount metaphase chromosomes as the test system. The first method devised is a direct extension of standard light microscope in situ hybridization. Radioactively labeled complementary RNA (cRNA) is hybridized to metaphase chromosomes deposited on electron microscope grids and fixed in 70 percent ethanol vapor; hybridixation site are detected by autoradiography. Specific and intense labeling of chromosomal centromeric regions is observed even after relatively short exposure times. Inerphase nuclei present in some of the metaphase chromosome preparations also show defined paatterms of satellite DNA labeling which suggests that satellite-containing regions are associate with each other during interphase. The sensitivity of this method is estimated to at least as good as that at the light microscope level while the resolution is improved at least threefold. The second method, which circumvents the use of autoradiogrphic detection, uses biotin-labeled polynucleotide probes. After hybridization of these probes, either DNA or RNA, to fixed chromosomes on grids, hybrids are detected via reaction is improved at least threefold. The second method, which circumvents the use of autoradiographic detection, uses biotin-labeled polynucleotide probes. After hybridization of these probes, either DNA or RNA, to fixed chromosomes on grids, hybrids are detected via reaction with an antibody against biotin and secondary antibody adsorbed to the surface of over centromeric heterochromatin and along the associated peripheral fibers. Labeling is on average

Hard and transparent hybrid polyurethane coatings using in situ incorporation of calcium carbonate nanoparticles

International Nuclear Information System (INIS)

Yao Lu; Yang Jie; Sun Jing; Cai Lifang; He Linghao; Huang Hui; Song Rui; Hao Yongmei

2011-01-01

Highlights: → In situ mineralization via gas diffusion was adopted for a good dispersion of calcium carbonate nanoparticles in the polymeric PU matrix. → Hybrid films with high dispersion, transparency, robust and thermal stability can be obtained by controlling the CaCO 3 loading. → The hybrid films display a significant improvement in its water resistance, surface hardness, scratch resistance and flexibility, with the introduction of CaCO 3 , and all coatings exhibited excellent chemical resistance and adhesion. - Abstract: The combination of hardness, scratch resistance, and flexibility is a highly desired feature in many coating applications. The aim of this study is to achieve this goal through the in situ introduction of an unmodified calcium carbonate (CaCO 3 ) into a water-soluble polyurethane (PU) matrix. Smooth and (semi-) transparent films were prepared from both the neat PU and the CaCO 3 -filled composites. As evidenced by the measurements from scanning electron microscopy (SEM), optical microscopy, dynamic mechanical analysis (DMA) and thermogravimetric analysis (TGA), hybrid films with high dispersion, transparency, robustness and thermal stability could be obtained by controlling the CaCO 3 loading. The storage modulus could increase from 441 MPa of neat PU matrix to 1034 MPa of hybrid film containing 2% (w/w) CaCO 3 . In addition, the same hybrid films displayed a significant improvement in its water resistance. In this case, the water-uptake ratio decreased from 41.54% of PU to 2.21% of hybrid film containing 2% (w/w) CaCO 3 . Moreover, with the introduction of CaCO 3 , conventional coating characterization methods demonstrated an increase in the surface hardness, scratch resistance and flexibility, and all coatings exhibited excellent chemical resistance and adhesion.

Manitoba plug-in hybrid electric vehicle (PHEV) demonstration

Energy Technology Data Exchange (ETDEWEB)

Hoemsen, R. [Red River College, Winnipeg, MB (Canada); Parsons, R. [Government of Manitoba, Winnipeg, MB (Canada). Centre for Emerging Renewable Energy

2010-07-01

Manitoba has low electricity rates, the highest proportion of renewables, and a legislated commitment to reduce greenhouse gases. However, the province still relies heavily on oil as everyone else. The mix of energy opportunities in Manitoba were highlighted in this presentation, with particular reference to the commercialization of electric vehicles. Several photographs were presented of the Toyota plug-in hybrid vehicle and a plug-in hybrid electric demonstration vehicle. A demonstration project overview was offered that used technology from A123 Systems Inc. The conversion module and vehicle users were profiled. Topics that were presented related to the demonstration project included monitoring; gasoline fuel economy results; fuel economy variability; cold weather operation; cold weather issues; battery upgrade solutions; and highly qualified personnel. It was concluded that in terms of follow-up, there is a need to combine findings of current plug-in hybrid electric vehicle demonstration with those for the new Toyota production plug-in hybrid vehicles. Key next steps for the demonstration are to address cabin heating requirements; better characterizing winter performance; and implementation of IPLC units on all plug-in hybrid electric vehicles for electricity consumption. figs.

Chromosome identification by new molecular markers and genomic in situ hybridization in the Triticum-Secale-Thinopyrum trigeneric hybrids.

Science.gov (United States)

Dai, Yi; Duan, Yamei; Chi, Dawn; Liu, Huiping; Huang, Shuai; Cao, Wenguang; Gao, Yong; Fedak, George; Chen, Jianmin

2017-08-01

It is very important to use chromosome-specific markers for identifying alien chromosomes in advanced generations of distant hybridization. The chromosome-specific markers of rye and Thinopyrum elongatum, as well as genomic in situ hybridization, were used to identify the alien chromosomes in eight lines that were derived from the crossing between Triticum trititrigia (AABBEE) and triticale (AABBRR). The results showed that four lines contained all rye chromosomes but no Th. elongatum chromosomes. The line RE36-1 contained all of the rye chromosomes except for chromosome 2R. The lines RE33-2 and RE62-1 contained all rye chromosomes and 1E and 5E translocated chromosome, respectively. The line RE24-4 contained 12 rye chromosomes plus a 7E chromosome or 12 rye chromosomes plus one R-E translocated chromosome. Chromosome identification in the above lines was consistent using chromosome-specific markers and genomic in situ hybridization. These chromosome-specific markers provide useful tools for detecting alien chromosomes in trigeneric hybrids, and these lines could be utilized as valuable germplasm in wheat improvement.

Biomarkers for ALK and ROS1 in Lung Cancer: Immunohistochemistry and Fluorescent In Situ Hybridization.

Science.gov (United States)

Luk, Peter P; Selinger, Christina I; Mahar, Annabelle; Cooper, Wendy A

2018-06-14

- A small proportion of non-small cell lung cancers harbor rearrangements of ALK or ROS1 genes, and these tumors are sensitive to targeted tyrosine kinase inhibitors. It is crucial for pathologists to accurately identify tumors with these genetic alterations to enable patients to access optimal treatments and avoid unnecessary side effects of less effective agents. Although a number of different techniques can be used to identify ALK- and ROS1-rearranged lung cancers, immunohistochemistry and fluorescence in situ hybridization are the mainstays. - To review the role of immunohistochemistry in assessment of ALK and ROS1 rearrangements in lung cancer, focusing on practical issues in comparison with other modalities such as fluorescence in situ hybridization. - This manuscript reviews the current literature on ALK and ROS1 detection using immunohistochemistry and fluorescence in situ hybridization as well as current recommendations. - Although fluorescence in situ hybridization remains the gold standard for detecting ALK and ROS1 rearrangement in non-small cell lung cancer, immunohistochemistry plays an important role and can be an effective screening method for detection of these genetic alterations, or a diagnostic test in the setting of ALK.

Tracking alien chromosome in sativa background by genomic in situ hybridization

International Nuclear Information System (INIS)

Abbasi, F.M.; Iqbal, M.; Salim, M.

2004-01-01

Genomic in situ hybridization (GISH) was used to look into the genomic constitution of monosomic alien -addition line derived from O. sativa x O. brachyantha. Biotin label genomic DNA from O. brachyantha was used as probe. The probe hybridized to the brachyantha chromosome. No detectable hybridization signal was observed on sativa chromosomes. This differential painting of chromosome enables us to unequivocally discriminate brachyantha chromosome from those of sativa. Results showed the usefulness of GISH in the identification of a single alien chromosome in the sativa background. (author)

In situ hybridization of nucleus basalis neurons shows increased β-amyloid mRNA in Alzheimer disease

International Nuclear Information System (INIS)

Cohen, M.L.; Golde, T.E.; Usiak, M.F.; Younkin, L.H.; Younkin, S.G.

1988-01-01

To determine which cells within the brain produce β-amyloid mRNA and to assess expression of the β-amyloid gene in Alzheimer disease, the authors analyzed brain tissue from Alzheimer and control patients by in situ hybridization. The results demonstrate that β-amyloid mRNA is produced by neurons in the nucleus basalis of Meynert and cerebral cortex and that nuclues basalis perikarya from Alzheimer patients consistently hybridize more β-amyloid probe than those from controls. These observations support the hypothesis that increased expression of the β-amyloid gene plays an important role in the deposition of amyloid in the brains of patients with Alzheimer disease

Oligonucleotide PIK3CA/Chromosome 3 Dual in Situ Hybridization Automated Assay with Improved Signals, One-Hour Hybridization, and No Use of Blocking DNA.

Science.gov (United States)

Zhang, Wenjun; Hubbard, Antony; Baca-Parkinson, Leslie; Stanislaw, Stacey; Vladich, Frank; Robida, Mark D; Grille, James G; Maxwell, Daniel; Tsao, Tsu-Shuen; Carroll, William; Gardner, Tracie; Clements, June; Singh, Shalini; Tang, Lei

2015-09-01

The PIK3CA gene at chromosome 3q26.32 was found to be amplified in up to 45% of patients with squamous cell carcinoma of the lung. The strong correlation between PIK3CA amplification and increased phosphatidylinositol 3-kinase (PI3K) pathway activities suggested that PIK3CA gene copy number is a potential predictive biomarker for PI3K inhibitors. Currently, all microscopic assessments of PIK3CA and chromosome 3 (CHR3) copy numbers use fluorescence in situ hybridization. PIK3CA probes are derived from bacterial artificial chromosomes whereas CHR3 probes are derived mainly from the plasmid pHS05. These manual fluorescence in situ hybridization assays mandate 12- to 18-hour hybridization and use of blocking DNA from human sources. Moreover, fluorescence in situ hybridization studies provide limited morphologic assessment and suffer from signal decay. We developed an oligonucleotide-based bright-field in situ hybridization assay that overcomes these shortcomings. This assay requires only a 1-hour hybridization with no need for blocking DNA followed by indirect chromogenic detection. Oligonucleotide probes produced discrete and uniform CHR3 stains superior to those from the pHS05 plasmid. This assay achieved successful staining in 100% of the 195 lung squamous cell carcinoma resections and in 94% of the 33 fine-needle aspirates. This robust automated bright-field dual in situ hybridization assay for the simultaneous detection of PIK3CA and CHR3 centromere provides a potential clinical diagnostic method to assess PIK3CA gene abnormality in lung tumors. Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

Hierarchically structured transparent hybrid membranes by in situ growth of mesostructured organosilica in host polymer

Science.gov (United States)

Vallé, Karine; Belleville, Philippe; Pereira, Franck; Sanchez, Clément

2006-02-01

The elaborate performances characterizing natural materials result from functional hierarchical constructions at scales ranging from nanometres to millimetres, each construction allowing the material to fit the physical or chemical demands occurring at these different levels. Hierarchically structured materials start to demonstrate a high input in numerous promising applied domains such as sensors, catalysis, optics, fuel cells, smart biologic and cosmetic vectors. In particular, hierarchical hybrid materials permit the accommodation of a maximum of elementary functions in a small volume, thereby optimizing complementary possibilities and properties between inorganic and organic components. The reported strategies combine sol-gel chemistry, self-assembly routes using templates that tune the material's architecture and texture with the use of larger inorganic, organic or biological templates such as latex, organogelator-derived fibres, nanolithographic techniques or controlled phase separation. We propose an approach to forming transparent hierarchical hybrid functionalized membranes using in situ generation of mesostructured hybrid phases inside a non-porogenic hydrophobic polymeric host matrix. We demonstrate that the control of the multiple affinities existing between organic and inorganic components allows us to design the length-scale partitioning of hybrid nanomaterials with tuned functionalities and desirable size organization from ångström to centimetre. After functionalization of the mesoporous hybrid silica component, the resulting membranes have good ionic conductivity offering interesting perspectives for the design of solid electrolytes, fuel cells and other ion-transport microdevices.

Detection of human papillomavirus in oral warts using in situ hybridization

Directory of Open Access Journals (Sweden)

Suzana Orsini Machado de Sousa

2008-01-01

Full Text Available Objective: The human papillomavirus is a group of DNA epitheliotrophic viruses associated with the etiology of benign and malignant oral warts. More than 100 types have been identified and among them, 24 have been found into the oral cavity. The aim of this study was to analyze human papillomavirus prevalence and its subtypes in 50 oral warts, of which 20 were squamous papillomas, 17 condylomaacuminatum and 13 verruca vulgaris. Method: In situ hybridization was used with biotinylated DNA probes for wide-spectrum HPV and with specific probes for human papillomavirus 6/11, human papillomavirus 16/18 and human papillomavirus 31/33. Results: Human papillomavirus was present in ten (20% of the 50 oral wart cases, 03 (3/20 squamous papillomas, 05 (5/17 condyloma acuminatum and 02 (2/13 verruca vulgaris. Of these, 8 (16% were positive to the HPV probe 6/11 being 5 condyloma acuminatum, 1 squamous papilloma and 2 verruca vulgaris. Three cases (6% demonstrated positivity to the human papillomavirus probe 16/18, with 2 being cases of condyloma and the other a case of squamous papilloma. Of the six positive cases to the human papillomavirus probe 31/33, (12% 4 were condyloma acuminatum and 2 squamous papillomas. Conclusion: The human papillomavirus expression (20% found in this study was low, but within the average found in the literature. Nonetheless, in addition to in situ hybridization, other methods may be necessary for confirming the presence of human papillomavirus.

Demonstration of constant upregulation of the telomerase RNA component in human gastric carcinomas using in situ hybridization.

Science.gov (United States)

Heine, B; Hummel, M; Demel, G; Stein, H

1998-06-01

Upregulation of the ribonucleoprotein telomerase seems to be a prerequisite for immortality, a feature of malignant cells. Using a polymerase chain reaction (PCR)-based assay, it is possible to demonstrate telomerase activity (TA) in specimens of most human malignancies, whereas it is absent from most normal tissues. It remains unclear, however, why between 5 and 50 per cent of various malignant tumour samples give negative results when TA is measured by the telomeric repeat amplification protocol (TRAP). The expectation that reverse transcription (RT)-PCR for detection of the telomerase RNA component (hTR) would be able to complement or to replace the TRAP assay failed, since malignant as well as non-malignant tissue samples gave positive results in most instances. In the present study, in situ hybridization (ISH) was developed to demonstrate the RNA component of human telomerase at the single cell level. With this method, 13 specimens of fresh frozen gastric carcinoma and four of normal, dysplastic, or inflamed gastric mucosa were investigated and the results were compared with those obtained by RT-PCR and the TRAP assay. In addition, ISH was performed on formalin-fixed sections of the same cases. The TRAP assay revealed positive results in 8 out of 13 gastric carcinomas and was negative in all non-malignant tissues. RT-PCR led to amplification of the telomerase RNA component in all specimens tested, irrespective of the presence or absence of malignant cells. By ISH, all gastric carcinomas showed strong telomerase RNA component-specific signals over malignant cells, whereas only a few grains were detectable over some types of normal somatic cells, including activated lymphocytes. In conclusion, high expression of the telomerase RNA component was restricted to the malignant cells of all the gastric carcinomas investigated, as shown by ISH. This indicates that the absence of TA in a proportion of carcinomas is due to methodological problems of the TRAP assay and is

Comparison of Fluorescence In Situ Hybridization and Chromogenic In Situ Hybridization for Low and High Throughput HER2 Genetic Testing

Science.gov (United States)

Poulsen, Tim S.; Espersen, Maiken L. M.; Kofoed, Vibeke; Dabetic, Tanja; Høgdall, Estrid; Balslev, Eva

2013-01-01

The purpose was to evaluate and compare 5 different HER2 genetic assays with different characteristics that could affect the performance to analyze the human epidermal growth factor 2 (HER2) gene copy number under low and high throughput conditions. The study included 108 tissue samples from breast cancer patients with HER2 immunohistochemistry (IHC) results scored as 0/1+, 2+, and 3+. HER2 genetic status was analysed using chromogenic in situ hybridization (CISH) and fluorescence in situ hybridization (FISH). Scoring results were documented through digital image analysis. The cancer region of interest was identified from a serial H&E stained slide following tissue cores were transferred to a tissue microarrays (TMA). When using TMA in a routine flow, all patients will be tested for HER2 status with IHC followed by CISH or FISH, thereby providing individual HER2 results. In conclusion, our results show that the differences between the HER2 genetic assays do not have an effect on the analytic performance and the CISH technology is superior to high throughput HER2 genetic testing due to scanning speed, while the IQ-FISH may still be a choice for fast low throughput HER2 genetic testing. PMID:24383005

Fluorescent In Situ Hybridization to Detect Transgene Integration into Plant Genomes

Science.gov (United States)

Schwarzacher, Trude

Fluorescent chromosome analysis technologies have advanced our understanding of genome organization during the last 30 years and have enabled the investigation of DNA organization and structure as well as the evolution of chromosomes. Fluorescent chromosome staining allows even small chromosomes to be visualized, characterized by their composition and morphology, and counted. Aneuploidies and polyploidies can be established for species, breeding lines, and individuals, including changes occurring during hybridization or tissue culture and transformation protocols. Fluorescent in situ hybridization correlates molecular information of a DNA sequence with its physical location on chromosomes and genomes. It thus allows determination of the physical position of sequences and often is the only means to determine the abundance and distribution of DNA sequences that are difficult to map with any other molecular method or would require segregation analysis, in particular multicopy or repetitive DNA. Equally, it is often the best way to establish the incorporation of transgenes, their numbers, and physical organization along chromosomes. This chapter presents protocols for probe and chromosome preparation, fluorescent in situ hybridization, chromosome staining, and the analysis of results.

Determination of HER2 amplification in primary breast cancer using dual-colour chromogenic in situ hybridization is comparable to fluorescence in situ hybridization: a European multicentre study involving 168 specimens

Science.gov (United States)

García-Caballero, Tomás; Grabau, Dorthe; Green, Andrew R; Gregory, John; Schad, Arno; Kohlwes, Elke; Ellis, Ian O; Watts, Sarah; Mollerup, Jens

2010-01-01

García-Caballero T, Grabau D, Green A R, Gregory J, Schad A, Kohlwes E, Ellis I O, Watts S & Mollerup J (2010) Histopathology56, 472–480 Determination of HER2 amplification in primary breast cancer using dual-colour chromogenic in situ hybridization is comparable to fluorescence in situ hybridization: a European multicentre study involving 168 specimens Aims: Fluorescence in situ hybridization (FISH) can be used to reveal several genomic imbalances relevant to proper cancer diagnosis and to the correct treatment regime. However, FISH requires expensive and advanced fluorescence microscopes in addition to expertise in fluorescence microscopy. To determine whether a newly developed dual-colour chromogenic in situ hybridization (CISH) method is a suitable alternative to FISH, we analysed the human epidermal growth factor receptor 2 gene (HER2) amplification level of 168 breast cancer specimens using dual-colour CISH and FISH and compared the results. Methods and results: We found 100% agreement between HER2 status determined by FISH and dual-colour CISH. Furthermore, we observed that the time used to score slides was significantly reduced by 28% in dual-colour CISH compared with the FISH protocol. Concordance between HER2 protein status and dual-colour CISH or FISH was equally good with an overall agreement of 96.8%. Correlation between the HER2/centromere 17 gene ratios obtained with dual-colour CISH and FISH was highly significant with an overall correlation coefficient (ρ) of 0.96. Conclusions: We conclude that dual-colour CISH and bright field microscopy are excellent alternatives to FISH when analysing the HER2 status of primary breast cancer. PMID:20459554

A list of image files of planarians analyzed by in situ hybridication and immunohistochemical staining - Plabrain DB | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available List Contact us Plabrain DB A list of image files of planarians analyzed by in situ hybridication and immunohistochemical...tu hybridication and also protein distribution by immunohistochemical staining in intact planarians or plana...planarians analyzed by In situ hybridication and immunohistochemical staining . D..._image#en Data acquisition method Whole-mount in situ hybridication, immunohistochemical...te Policy | Contact Us A list of image files of planarians analyzed by in situ hybridication and immunohistochemical staining - Plabrain DB | LSDB Archive ...

RNA Imaging with Multiplexed Error Robust Fluorescence in situ Hybridization

Science.gov (United States)

Moffitt, Jeffrey R.; Zhuang, Xiaowei

2016-01-01

Quantitative measurements of both the copy number and spatial distribution of large fractions of the transcriptome in single-cells could revolutionize our understanding of a variety of cellular and tissue behaviors in both healthy and diseased states. Single-molecule Fluorescence In Situ Hybridization (smFISH)—an approach where individual RNAs are labeled with fluorescent probes and imaged in their native cellular and tissue context—provides both the copy number and spatial context of RNAs but has been limited in the number of RNA species that can be measured simultaneously. Here we describe Multiplexed Error Robust Fluorescence In Situ Hybridization (MERFISH), a massively parallelized form of smFISH that can image and identify hundreds to thousands of different RNA species simultaneously with high accuracy in individual cells in their native spatial context. We provide detailed protocols on all aspects of MERFISH, including probe design, data collection, and data analysis to allow interested laboratories to perform MERFISH measurements themselves. PMID:27241748

Detection of human papillomavirus type 6/11 DNA in conjunctival papillomas by in situ hybridization with radioactive probes

International Nuclear Information System (INIS)

McDonnell, P.J.; McDonnell, J.M.; Kessis, T.; Green, W.R.; Shah, K.V.

1987-01-01

Twenty-three conjunctival papillomas and 28 conjunctival dysplasias were examined for human papillomavirus (HPV)-DNA sequences by in situ hybridization with nick-translated 35 S-labeled HPV probes. Adjacent paraffin sections were hybridized with HPV type 2, 6, 16, and 18 probes at Tm - 17 degrees C. Fifteen tissues, all papillomas, displayed positive hybridization with the HPV-6 probe. Infection with HPV-6 (or the closely related HPV-11) appeared to be responsible for most of the conjunctival papillomas of children and young adults. The presence of genital tract HPV-6 in these lesions suggests that some of the infections were acquired during passage through an infected birth canal. The lack of hybridization in adult conjunctival dysplasias indicates either that HPVs are not associated with this condition or that the probes and the technique utilized were not adequate for demonstration of this association

Chromogenic in situ hybridization for Her-2/neu-oncogene in breast cancer: comparison of a new dual-colour chromogenic in situ hybridization with immunohistochemistry and fluorescence in situ hybridization.

Science.gov (United States)

Mayr, Doris; Heim, Sibylle; Weyrauch, Kerstin; Zeindl-Eberhart, Evelyn; Kunz, Anne; Engel, Jutta; Kirchner, Thomas

2009-12-01

Her-2/neu testing is used as a marker for Herceptin therapy. The aim was to investigate new dual-colour chromogenic in situ hybridization (CISH), in a large number of breast carcinomas (n = 205) with DNA-specific dual-colour probes (ZytoVision, Bremerhaven, Germany) and to compare the results with immunohistochemistry (n = 205) and fluorescence in situ hybridization (FISH) (n = 129). Paraffin-embedded tissue of 205 patients was used. After immunohistochemistry with a focus on immunohistochemically uncertain cases, Her-2/neu amplification using dual-colour CISH (ZytoVision) was analysed. Validation by FISH was performed. The results were: immunohistochemistry, 27.8% with strong expression, 53.7% with uncertain overexpression and 18.5% with no expression; FISH, 25.6% amplified and 74.4% negative; CISH, 35.6% amplified, 62.9% negative and 1.5% not evaluable. Comparison of immunohistochemistry with CISH: CISH negative in 100% with immunohistochemistry 0/1+, amplified in 82.5% with immunohistochemistry 3+; 5.9% contradictory results: 4.4% immunohistochemistry 3+ and negative by CISH, 1.5% negative in immunohistochemistry but amplified by CISH; FISH (129 cases), 8.5% contradictory results to immunohistochemistry, 6.2% immunohistochemistry 3+ and negative by FISH, 2.3% negative by immunohistochemistry and amplified by FISH; comparison of CISH and FISH, 94.6% same results, 3.9% different ones, 1.6% CISH not analysable. CISH, using dual-colour probes (ZytoVision) is as good as FISH for Her-2/neu analysis. The few discrepant results are likely to be caused by polysomy or tumour heterogeneity.

Design and optimization of hybrid ex situ/in situ steam generation recovery processes for heavy oil and bitumen

Energy Technology Data Exchange (ETDEWEB)

Yang, X.; Gates, I.D. [Calgary Univ., AB (Canada). Dept. of Chemical and Petroleum Engineering; Larter, S.R. [Calgary Univ., AB (Canada). Dept. of Geoscience]|[Alberta Ingenuity Centre for In Situ Energy, Edmonton, AB (Canada)

2008-10-15

Hybrid steam-air based oil recovery techniques were investigated using advanced 3-D reactive thermal reservoir simulations. The hybrid techniques combined ex situ steam and in situ steam generation processes in order to raise efficiency, lower natural gas consumption, and reduce gas emissions. The steam-air based processes used 70 per cent of the energy of conventional steam assisted gravity drainage (SAGD) techniques to recover the same amount of oil. The process used an SAGD wellpair arrangement, where steam and air were injected through the top injection well. The kinetic parameters used in the study were developed by history matching a combustion tube experiments with Athabasca bitumen conducted to predict cumulative bitumen and gas production volumes and compositions. A total of 6 SAGD and 6 in situ combustion simulations were conducted with steam oxygen volume ratios set at 50 per cent steam and 50 per cent oxygen. Various case studies were considered over a 5 year period. Carbon dioxide (CO{sub 2}) emissions were also measured as well as cumulative water and methane consumption rates. Results of the study were used to develop an optimized hybrid operation that consisted of a SAGD well pair arrangement operating with cyclic steam-oxygen injection at high pressures. It was concluded that the high pressure operation increased the steam partial pressure within the reservoir and enhanced combustion performance. A 29 per cent improvement in the cumulative energy to oil ratio was obtained. 23 refs., 2 tabs., 9 figs.

Comparative genomic and in situ hybridization of germ cell tumors of the infantile testis

NARCIS (Netherlands)

Mostert, M; Rosenberg, C; Stoop, H; Schuyer, M; Timmer, A; Oosterhuis, W; Looijenga, L

Chromosomal information on germ cell tumors of the infantile testis, ie, teratomas and yolk sac tumors, is limited and controversial. We studied two teratomas and four yolk sac tumors using comparative genomic hybridization (CGH) and in situ hybridization. No chromosomal anomalies were found in the

Exploring the origin of the D genome of oat by fluorescence in situ hybridization.

Science.gov (United States)

Luo, Xiaomei; Zhang, Haiqin; Kang, Houyang; Fan, Xing; Wang, Yi; Sha, Lina; Zhou, Yonghong

2014-09-01

Further understanding of the origin of cultivated oat would accelerate its genetic improvement. In particular, it would be useful to clarify which diploid progenitor contributed the D genome of this allohexaploid species. In this study, we demonstrate that the landmarks produced by fluorescence in situ hybridization (FISH) of species of Avena using probes derived from Avena sativa can be used to explore the origin of the D genome. Selected sets of probes were hybridized in several sequential experiments performed on exactly the same chromosome spreads, with multiple probes of cytological preparations. Probes pITS and A3-19 showed there might be a similar distribution of pITS between the Ac and D genomes. These results indicated that the Ac genome is closely related to the D genome, and that Avena canariensis (AcAc) could be the D-genome donor of cultivated oat.

In Situ Synthesis of Metal Nanoparticle Embedded Hybrid Soft Nanomaterials.

Science.gov (United States)

Divya, Kizhmuri P; Miroshnikov, Mikhail; Dutta, Debjit; Vemula, Praveen Kumar; Ajayan, Pulickel M; John, George

2016-09-20

The allure of integrating the tunable properties of soft nanomaterials with the unique optical and electronic properties of metal nanoparticles has led to the development of organic-inorganic hybrid nanomaterials. A promising method for the synthesis of such organic-inorganic hybrid nanomaterials is afforded by the in situ generation of metal nanoparticles within a host organic template. Due to their tunable surface morphology and porosity, soft organic materials such as gels, liquid crystals, and polymers that are derived from various synthetic or natural compounds can act as templates for the synthesis of metal nanoparticles of different shapes and sizes. This method provides stabilization to the metal nanoparticles by the organic soft material and advantageously precludes the use of external reducing or capping agents in many instances. In this Account, we exemplify the green chemistry approach for synthesizing these materials, both in the choice of gelators as soft material frameworks and in the reduction mechanisms that generate the metal nanoparticles. Established herein is the core design principle centered on conceiving multifaceted amphiphilic soft materials that possess the ability to self-assemble and reduce metal ions into nanoparticles. Furthermore, these soft materials stabilize the in situ generated metal nanoparticles and retain their self-assembly ability to generate metal nanoparticle embedded homogeneous organic-inorganic hybrid materials. We discuss a remarkable example of vegetable-based drying oils as host templates for metal ions, resulting in the synthesis of novel hybrid nanomaterials. The synthesis of metal nanoparticles via polymers and self-assembled materials fabricated via cardanol (a bioorganic monomer derived from cashew nut shell liquid) are also explored in this Account. The organic-inorganic hybrid structures were characterized by several techniques such as UV-visible spectroscopy, scanning electron microscopy (SEM), and

The effect of tissue decalcification on mRNA retention within bone for in-situ hybridization studies.

Science.gov (United States)

Walsh, L; Freemont, A J; Hoyland, J A

1993-06-01

Tissue decalcification is a routine part of the preparation of bone tissue for histological studies. Although in-situ hybridization has been employed to localize mRNA of collagenous and non-collagenous bone related proteins in skeletal tissue, little is known regarding the effects of decalcifying agents on mRNA retention within tissue. In this study in-situ hybridization using an oligonucleotide probe (i.e. a poly d(T) probe) to detect total messenger RNA has been employed to investigate the effects of the decalcifying agents nitric acid, formic acid and EDTA on mRNA retention compared to undeacalcified tissue. The results show that formalin fixation and EDTA decalcification preserve substantial amounts of mRNA within the tissue. In particular, this study illustrates that it is possible to perform in-situ hybridization on formalin fixed decalcified paraffin embedded tissue.

Hybrid Life Support System Technology Demonstrations

Science.gov (United States)

Morrow, R. C.; Wetzel, J. P.; Richter, R. C.

2018-02-01

Demonstration of plant-based hybrid life support technologies in deep space will validate the function of these technologies for long duration missions, such as Mars transit, while providing dietary variety to improve habitability.

Demonstration of variation in chondrocyte activity in different zones of articular cartilage: an assessment of the value of in-situ hybridization.

Science.gov (United States)

Marles, P J; Hoyland, J A; Parkinson, R; Freemont, A J

1991-04-01

Several methods have been described for investigating chondrocyte metabolism in vitro. In this study, in-situ hybridization (ISH) using an oligonucleotide probe (i.e. a poly-d(T) probe) to detect total messenger RNA (mRNA) in cartilage explants has been compared with radiosulphate and radioleucine uptake studies in an attempt to assess the value of ISH in investigating chondrocyte metabolism. The relative results of the three parameters indicate qualitative similarities in cells in the intermediate, deep and calcified zones but differences in the superficial zone. The relative levels of mRNA and leucine and sulphate uptake in the midzone areas could be construed as indicating that the bulk of cellular activity was directed towards the synthesis of proteoglycans. A similar relation between the three parameters, but at a lower level, was seen in chondrocytes in the calcified zone demonstrating that these cells are viable and biosynthetic. Both quantitative and qualitative differences between the three methods were observed in the superficial chondrocytes regarding the amount of mRNA compared to sulphate and leucine uptake. The results suggest that ISH can detect differences in the amount of mRNA present in chondrocytes in differing zones of cartilage and, like the radioleucine and radiosulphate studies, particularly emphasizes their functional heterogeneity.

Implementation of the Fluorescent in Situ Hybridization technique in the Faculty of Medicine, UdelaR

Directory of Open Access Journals (Sweden)

Andrea Cairus

2017-11-01

Full Text Available The Cytogenetic Laboratory of the Faculty of Medicine processes, on average, 300 annual samples of public and private healthcare centers by conventional cytogenetics. It is essential to implement new techniques to improve the quality of the service offered. The purpose of this work was to implement the Fluorescent in situ Hybridization technique (FISH. An observational, cross-sectional, analytical study was performed. Peripheral blood samples from patients with sex chromosomopathies diagnosed by conventional cytogenetics were analyzed. Fluorescent in situ hybridization technique was applied, comparing results with FISH and with conventional cytogenetics. The percentage of mosaicism detected by conventional cytogenetics and Fluorescent in situ Hybridization was studied: 24 samples were analyzed; 19 presented numerical alterations, 3 structural and 2 both. Numerical alterations were Turner syndrome, Klinefelter syndrome, XXX syndrome and XYY syndrome. Concordance in diagnoses was found for both techniques. For Turner syndrome, 8 of 12 samples corresponded to mosaicism, and there were no significant differences between conventional cytogenetics and the technique studied (p0.05. Klinefelter syndrome and XYY were both presented in a non-mosaic karyotype. For XXX syndrome, a normal line (46, XX was observed in three of the samples, in a percentage close to the cut off. From this research, it will be possible to implement Fluorescent in situ Hybridization in this service, to extend it to other pathologies and to enable the training of human resources; consolidating this laboratory as a national academic reference center.

Detection of c-myc amplification in formalin-fixed paraffin-embedded tumor tissue by chromogenic in situ hybridization (CISH).

Science.gov (United States)

Todorović-Raković, Nataša

2013-01-01

In situ hybridization (ISH) allows evaluation of genetic abnormalities, such as changes in chromosome number, chromosome translocations or gene amplifications, by hybridization of tagged DNA (or RNA) probes with complementary DNA (or RNA) sequences in interphase nuclei of target tissue. However, chromogenic in situ hybridization (CISH) is also applicable to formalin-fixed, paraffin-embedded (FFPE) tissues, besides metaphase chromosome spreads. CISH is similar to fluorescent in situ hybridization (FISH) regarding pretreatments and hybridization protocols but differs in the way of visualization. Indeed, CISH signal detection is similar to that used in immunohistochemistry, making use of a peroxidase-based chromogenic reaction instead of fluorescent dyes. In particular, tagged DNA probes are indirectly detected using an enzyme-conjugated antibody targeting the tags. The enzymatic reaction of the chromogenic substrate leads to the formation of strong permanent brown signals that can be visualized by bright-field microscopy at 40 × magnification. The advantage of CISH is that it allows the simultaneous observation of gene amplification and tissue morphology and the slides can be stored for a long time.

Dramatically improved RNA in situ hybridization signals using LNA-modified probes

DEFF Research Database (Denmark)

Thomsen, Rune; Nielsen, Peter Stein; Jensen, Torben Heick

2005-01-01

. This increases the thermal stability of hybrids formed with RNA. The LNA-based probes detect specific RNAs in fixed yeast cells with an efficiency far better than conventional DNA oligonucleotide probes of the same sequence. Using this probe design, we were also able to detect poly(A)+ RNA accumulation within......In situ detection of RNA by hybridization with complementary probes is a powerful technique. Probe design is a critical parameter in successful target detection. We have evaluated the efficiency of fluorescent DNA oligonucleotides modified to contain locked nucleic acid (LNA) residues...

In situ hybridization for the detection of infectious laryngotracheitis virus in sections of trachea from experimentally infected chickens

DEFF Research Database (Denmark)

Nielsen, O.L.; Handberg, Kurt; Jørgensen, Poul Henrik

1998-01-01

An in situ hybridization procedure for the detection of infectious laryngotracheitis virus (ILTV) in experimentally infected chickens is described. Formalin-fixed, paraffin-embedded sections of trachea, taken from chickens on days 3-10 post-inoculation (p.i.) with ILTV were hybridized with a mixt......An in situ hybridization procedure for the detection of infectious laryngotracheitis virus (ILTV) in experimentally infected chickens is described. Formalin-fixed, paraffin-embedded sections of trachea, taken from chickens on days 3-10 post-inoculation (p.i.) with ILTV were hybridized...... on day 5 p.i. No hybridization was observed in 3 out of 3 chickens examined on day 10 p.i. ILTV nucleic acid was detected in nuclei of degenerated tracheal epithelial cells and in intranuclear inclusion bodies of syncytia....

Analysis of experimental mink enteritis virus infection in mink: in situ hybridization, serology, and histopathology

DEFF Research Database (Denmark)

Uttenthal, Åse; Larsen, S; Lund, E

1990-01-01

Strand-specific hybridization probes were used in in situ hybridization studies to localize cells containing mink enteritis virus (MEV) virion DNA or MEV replicative-form DNA and mRNA. Following the experimental MEV infection of 3-month-old unvaccinated mink, a significant increase in serum antib...

Preparation of epoxy/zirconia hybrid materials via in situ polymerization using zirconium alkoxide coordinated with acid anhydride

International Nuclear Information System (INIS)

Ochi, Mitsukazu; Nii, Daisuke; Harada, Miyuki

2011-01-01

Highlights: → Novel epoxy/zirconia hybrid materials were synthesized via in situ polymerization using zirconium alkoxide coordinated with acid anhydride. → The half-ester compound of acid anhydride desorbed from zirconium played as curing agent of epoxy resin. → The zirconia was uniformly dispersed in the epoxy matrix on the nanometer or sub-nanometer scale by synchronizing the epoxy curing and sol-gel reactions. → The refractive indices of the hybrid materials significantly improved with an increase in the zirconia content. - Abstract: Novel epoxy/zirconia hybrid materials were synthesized using a bisphenol A epoxy resin (diglycidyl ether of bisphenol A; DGEBA), zirconium(IV)-n-propoxide (ZTNP), and hexahydrophthalic anhydride (HHPA) via in situ polymerization. HHPA played two roles in this system: it acted as a modifier to control the hydrolysis and condensation reactions of zirconium alkoxide and also as a curing agent - the half-ester compound of HHPA desorbed from zirconium reacted with the epoxy resin to form the epoxy network. As a result, both the sol-gel reaction and epoxy curing occurred simultaneously in a homogeneous solution, and organic-inorganic hybrid materials were readily obtained. Further, the zirconia produced by the in situ polymerization was uniformly dispersed in the epoxy matrix on the nanometer or sub-nanometer scale; thus, hybrid materials that exhibited excellent optical transparency were obtained. Furthermore, the heat resistance of the hybrid materials could be improved by hybridization with zirconia. And, the refractive indices of the hybrid materials significantly improved with an increase in the zirconia content.

Novel RNA hybridization method for the in situ detection of ETV1, ETV4, and ETV5 gene fusions in prostate cancer.

Science.gov (United States)

Kunju, Lakshmi P; Carskadon, Shannon; Siddiqui, Javed; Tomlins, Scott A; Chinnaiyan, Arul M; Palanisamy, Nallasivam

2014-09-01

The genetic basis of 50% to 60% of prostate cancer (PCa) is attributable to rearrangements in E26 transformation-specific (ETS) (ERG, ETV1, ETV4, and ETV5), BRAF, and RAF1 genes and overexpression of SPINK1. The development and validation of reliable detection methods are warranted to classify various molecular subtypes of PCa for diagnostic and prognostic purposes. ETS gene rearrangements are typically detected by fluorescence in situ hybridization and reverse-transcription polymerase chain reaction methods. Recently, monoclonal antibodies against ERG have been developed that detect the truncated ERG protein in immunohistochemical assays where staining levels are strongly correlated with ERG rearrangement status by fluorescence in situ hybridization. However, specific antibodies for ETV1, ETV4, and ETV5 are unavailable, challenging their clinical use. We developed a novel RNA in situ hybridization-based assay for the in situ detection of ETV1, ETV4, and ETV5 in formalin-fixed paraffin-embedded tissues from prostate needle biopsies, prostatectomy, and metastatic PCa specimens using RNA probes. Further, with combined RNA in situ hybridization and immunohistochemistry we identified a rare subset of PCa with dual ETS gene rearrangements in collisions of independent tumor foci. The high specificity and sensitivity of RNA in situ hybridization provides an alternate method enabling bright-field in situ detection of ETS gene aberrations in routine clinically available PCa specimens.

An Optimized Small Tissue Handling System for Immunohistochemistry and In Situ Hybridization.

Directory of Open Access Journals (Sweden)

Giovanni Anthony

Full Text Available Recent development in 3D printing technology has opened an exciting possibility for manufacturing 3D devices on one's desktop. We used 3D modeling programs to design 3D models of a tissue-handling system and these models were "printed" in a stereolithography (SLA 3D printer to create precision histology devices that are particularly useful to handle multiple samples with small dimensions in parallel. Our system has been successfully tested for in situ hybridization of zebrafish embryos. Some of the notable features include: (1 A conveniently transferrable chamber with 6 mesh-bottomed wells, each of which can hold dozens of zebrafish embryos. This design allows up to 6 different samples to be treated per chamber. (2 Each chamber sits in a well of a standard 6-well tissue culture plate. Thus, up to 36 different samples can be processed in tandem using a single 6 well plate. (3 Precisely fitting lids prevent solution evaporation and condensation, even at high temperatures for an extended period of time: i.e., overnight riboprobe hybridization. (4 Flat bottom mesh maximizes the consistent treatment of individual tissue samples. (5 A magnet-based lifter was created to handle up to 6 chambers (= 36 samples in unison. (6 The largely transparent resin aids in convenient visual inspection both with eyes and using a stereomicroscope. (7 Surface engraved labeling enables an accurate tracking of different samples. (8 The dimension of wells and chambers minimizes the required amount of precious reagents. (9 Flexible parametric modeling enables an easy redesign of the 3D models to handle larger or more numerous samples. Precise dimensions of 3D models and demonstration of how we use our devices in whole mount in situ hybridization are presented. We also provide detailed information on the modeling software, 3D printing tips, as well as 3D files that can be used with any 3D printer.

Modulation of tyrosine hydroxylase gene expression in the central nervous system visualized by in situ hybridization

International Nuclear Information System (INIS)

Berod, A.; Biguet, N.F.; Dumas, S.; Bloch, B.; Mallet, J.

1987-01-01

cDNA probe was used for in situ hybridization studies on histological sections through the locus coeruleus, substantia nigra, and the ventral tegmental area of the rat brain. Experimental conditions were established that yielded no background and no signal when pBR322 was used as control probe. Using the tyrosine hydroxylase probe, the authors ascertained the specificity of the labeling over catecholaminergic cells by denervation experiments and comparison of the hybridization pattern with that of immunoreactivity. The use of 35 S-labeled probe enabled the hybridization signal to be resolved at the cellular level. A single injection of reserpine into the rat led to an increase of the intensity of the autoradiographic signal over the locus coeruleus area, confirming an RNA gel blot analysis. The potential of in situ hybridization to analyze patterns of modulation of gene activity as a result of nervous activity is discussed

Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications.

Science.gov (United States)

Cui, Chenghua; Shu, Wei; Li, Peining

2016-01-01

Fluorescence in situ hybridization (FISH) is a macromolecule recognition technology based on the complementary nature of DNA or DNA/RNA double strands. Selected DNA strands incorporated with fluorophore-coupled nucleotides can be used as probes to hybridize onto the complementary sequences in tested cells and tissues and then visualized through a fluorescence microscope or an imaging system. This technology was initially developed as a physical mapping tool to delineate genes within chromosomes. Its high analytical resolution to a single gene level and high sensitivity and specificity enabled an immediate application for genetic diagnosis of constitutional common aneuploidies, microdeletion/microduplication syndromes, and subtelomeric rearrangements. FISH tests using panels of gene-specific probes for somatic recurrent losses, gains, and translocations have been routinely applied for hematologic and solid tumors and are one of the fastest-growing areas in cancer diagnosis. FISH has also been used to detect infectious microbias and parasites like malaria in human blood cells. Recent advances in FISH technology involve various methods for improving probe labeling efficiency and the use of super resolution imaging systems for direct visualization of intra-nuclear chromosomal organization and profiling of RNA transcription in single cells. Cas9-mediated FISH (CASFISH) allowed in situ labeling of repetitive sequences and single-copy sequences without the disruption of nuclear genomic organization in fixed or living cells. Using oligopaint-FISH and super-resolution imaging enabled in situ visualization of chromosome haplotypes from differentially specified single-nucleotide polymorphism loci. Single molecule RNA FISH (smRNA-FISH) using combinatorial labeling or sequential barcoding by multiple round of hybridization were applied to measure mRNA expression of multiple genes within single cells. Research applications of these single molecule single cells DNA and RNA FISH

Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications

Directory of Open Access Journals (Sweden)

Chenghua Cui

2016-09-01

Full Text Available Fluorescence in situ hybridization (FISH is a macromolecule recognition technology based on the complementary nature of DNA or DNA/RNA double strands. Selected DNA strands incorporated with fluorophore-coupled nucleotides can be used as probes to hybridize onto the complementary sequences in tested cells and tissues and then visualized through a fluorescence microscope or an imaging system. This technology was initially developed as a physical mapping tool to delineate genes within chromosomes. Its high analytical resolution to a single gene level and high sensitivity and specificity enabled an immediate application for genetic diagnosis of constitutional common aneuploidies, microdeletion/microduplication syndromes and subtelomeric rearrangements. FISH tests using panels of gene-specific probes for somatic recurrent losses, gains and translocations have been routinely applied for hematologic and solid tumors and are one of the fastest-growing areas in cancer diagnosis. FISH has also been used to detect infectious microbials and parasites like malaria in human blood cells. Recent advances in FISH technology involve various methods for improving probe labeling efficiency and the use of super resolution imaging systems for direct visualization of intra-nuclear chromosomal organization and profiling of RNA transcription in single cells. Cas9-mediated FISH (CASFISH allowed in situ labeling of repetitive sequences and single-copy sequences without the disruption of nuclear genomic organization in fixed or living cells. Using oligopaint-FISH and super-resolution imaging enabled in situ visualization of chromosome haplotypes from differentially specified single-nucleotide polymorphism loci. Single molecule RNA FISH (smRNA-FISH using combinatorial labeling or sequential barcoding by multiple round of hybridization were applied to measure mRNA expression of multiple genes within single cells. Research applications of these single molecule single cells

Demonstration testing and evaluation of in situ heating of soil

International Nuclear Information System (INIS)

1995-01-01

This document describes the Quality Assurance Project Plan (QAPP) for IITRI Project C06787 entitled open-quotes Demonstration Testing and Evaluation of In Situ Heating of Soilclose quotes. A work plan for the above mentioned work was previously submitted. This QAPP describes the sampling and analysis of soil core-samples obtained from the K-25 Site (Oak Ridge Gaseous Diffusion Plant) where an in-situ heating and soil decontamination demonstration experiment will be performed. Soil samples taken before and after the experiment will be analyzed for selected volatile organic compounds. The Work Plan mentioned above provides a complete description of the demonstration site, the soil sampling plan, test plan, etc

Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Energy Technology Data Exchange (ETDEWEB)

Rosa, F.E.; Santos, R.M. [Departamento de Patologia, Hospital das Clínicas, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil); Rogatto, S.R. [Departamento de Urologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil); Hospital A.C. Camargo, CIPE, São Paulo, SP (Brazil); Domingues, M.A.C. [Departamento de Patologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil)

2013-03-19

Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Science.gov (United States)

Rosa, F.E.; Santos, R.M.; Rogatto, S.R.; Domingues, M.A.C.

2013-01-01

Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer. PMID:23558859

Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

International Nuclear Information System (INIS)

Rosa, F.E.; Santos, R.M.; Rogatto, S.R.; Domingues, M.A.C.

2013-01-01

Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer

In situ synthesis of silver benzene-dithiolate hybrid films

Energy Technology Data Exchange (ETDEWEB)

Brenier, Roger, E-mail: roger.brenier@univ-lyon1.fr [Institut Lumière Matière, UMR 5306, Université Lyon 1-CNRS, Université de Lyon, Domaine Scientifique de La Doua, Batiment Kastler, 10 rue Ada Byron, 69622 Villeurbanne, Cedex (France); Piednoir, Agnès, E-mail: agnes.piednoir@univ-lyon1.fr [Institut Lumière Matière, UMR 5306, Université Lyon 1-CNRS, Université de Lyon, Domaine Scientifique de La Doua, Batiment Kastler, 10 rue Ada Byron, 69622 Villeurbanne, Cedex (France); Bertorelle, Franck, E-mail: franck.bertorelle@univ-lyon1.fr [Institut Lumière Matière, UMR 5306, Université Lyon 1-CNRS, Université de Lyon, Domaine Scientifique de La Doua, Batiment Kastler, 10 rue Ada Byron, 69622 Villeurbanne, Cedex (France); Penuelas, José, E-mail: jose.penuelas@ec-lyon.fr [Université de Lyon, Institut des Nanotechnologies de Lyon, Ecole Centrale de Lyon, CNRS, UMR 5270, 36 rue Guy de Collongues, F69134 Ecully (France); Grenet, Geneviève, E-mail: genevieve.grenet@ec-lyon.fr [Université de Lyon, Institut des Nanotechnologies de Lyon, Ecole Centrale de Lyon, CNRS, UMR 5270, 36 rue Guy de Collongues, F69134 Ecully (France)

2016-02-01

In this article, a method for in situ synthesis of silver benzene-dithiolate hybrid films is presented. Silver nanoparticles, generated on ZrO{sub 2} films, are transformed into silver benzene 1,4-dithiolate or, partially, into silver benzene 1,2-dithiolate after sample immersion in the corresponding thiol solutions. These transformations occur at room temperature owing to the catalytic action of ZrO{sub 2}. It is also shown that TiO{sub 2} in place of ZrO{sub 2} is very efficient, both for the catalytic generation of silver nanoparticles and for their further transformation in benzene 1,4-dithiolate compound. This latter semiconductor has an optical bandgap of about 3 eV and the film is made of touching nanoparticles in an amorphous state. Our work has potential applications in the electronic and photovoltaic fields. - Highlights: • A method for in situ synthesis of silver benzene-dithiolate hybrid semiconductor films is presented. • Silver nanoparticles are, first, generated on ZrO{sub 2} or on TiO{sub 2} coated silica substrates. • The samples are immersed in benzene dithiol solution for two days at room temperature. • During the immersion, the silver nanoparticles are transformed into silver benzene dithiolate. • The silver benzene dithiolate film is made of amorphous nanoparticles with a banbgap of 3 eV.

Evaluation of Myc Gene Amplification in Prostate Cancer Using a Dual Color Chromogenic in-Situ Hybridization (Dual CISH Assay

Directory of Open Access Journals (Sweden)

Daniel Lerda

2013-04-01

Full Text Available Objetive: The overall purpose of the study was to demonstrate applicability of the Dako dual-color chromogenic in situ hybridization (CISH assay (DAKO Denmark, Glostrup with respect to fluorescence in situ hybridization (FISH probes MYC-C. Methods: MYC gene amplification by FISH and Dako dual-color CISH Results: The study showed that the dual-color CISH assay can convert Texas red and fluorescein isothiocyanate (FITC signals into chromogenic signals. The dual –color CISH assay was performed on 40 cases of prostate cancer. Amplification was identified in 12 of 40 (30% tumors. No amplification was seen in 28 of 40 (70% tumors. FISH data were available in total of 40 tumors. All tumors showed concordant results between dual-color CISH and FISH for classifying a tumor as MYC amplified or not amplified. Conclusions: We conclude that dual-color Dako CISH assay is an accurate method for determining MYC gene amplification with added advantages that make it a more practically useful method. [J Interdiscipl Histopathol 2013; 1(2.000: 81-84

Chromosome translocations measured by fluorescence in-situ hybridization: A promising biomarker

International Nuclear Information System (INIS)

Lucas, J.N.; Straume, T.

1995-10-01

A biomarker for exposure and risk assessment would be most useful if it employs an endpoint that is highly quantitative, is stable with time, and is relevant to human risk. Recent advances in chromosome staining using fluorescence in situ hybridization (FISH) facilitate fast and reliable measurement of reciprocal translocations, a kind of DNA damage linked to both prior exposure and risk. In contrast to other biomarkers available, the frequency of reciprocal translocations in individuals exposed to whole-body radiation is stable with time post exposure, has a rather small inter-individual variability, and can be measured accurately at the low levels. Here, the authors discuss results from their studies demonstrating that chromosome painting can be used to reconstruct radiation dose for workers exposed within the dose limits, for individuals exposed a long time ago, and even for those who have been diagnosed with leukemia but not yet undergone therapy

Fluorescence in situ hybridization evaluation of chromosome deletion patterns in prostate cancer.

Science.gov (United States)

Huang, S F; Xiao, S; Renshaw, A A; Loughlin, K R; Hudson, T J; Fletcher, J A

1996-11-01

Various nonrandom chromosomal aberrations have been identified in prostate carcinoma. These aberrations include deletions of several chromosome regions, particularly the chromosome 8 short arm. Large-scale numerical aberrations, reflected in aberrant DNA ploidy, are also found in a minority of cases. However, it is unclear whether prostate carcinomas contain aberrations of certain chromosome regions that are deleted frequently in other common types of cancer. In this study, we performed dual-color fluorescence in situ hybridization on intact nuclei from touch preparations of 16 prostate cancers. Chromosome copy number was determined using pericentromeric probes, whereas potential chromosome arm deletions were evaluated using yeast artificial chromosome (YAC) and P1 probes. Two YAC probes targeted chromosome 8 short arm regions known to be deleted frequently in prostate cancer. Other YACs and P1s were for chromosome regions, including 1p22, 3p14, 6q21, 9p21, and 22q12, that are deletion targets in a variety of cancers although not extensively studied in prostate cancer. Hybridization efficiencies and signal intensities were excellent for both repeat sequence (alpha-satellite) and single, copy (YAC and P1) fluorescence in situ hybridization probes. Of 16 prostate cancers, 11 had clonal aberrations of 1 or more of the 13 chromosome regions evaluated, and 10 cases (62.5%) had 8p deletions, including 4 cases with 8p deletion in virtually all cells and aneuploidy in only a subset of those deleted cells. Deletions at 3p14, 6q21, and 22q12 were identified in 2, 1, and 1 case, respectively, and each of those cases had a similarly sized cell population with 8p deletion. These studies confirm 8p deletion in the majority of prostate carcinomas. 8p deletions appear to be early events in prostate tumorigenesis, often antedating aneuploidy. Fluorescence in situ hybridization strategies incorporating pericentromeric and single-copy regional chromosome probes offer a powerful and

Utility of chromogenic in situ hybridization (CISH) for detection of EGFR amplification in glioblastoma: comparison with fluorescence in situ hybridization (FISH).

Science.gov (United States)

Fischer, Ingeborg; de la Cruz, Clarissa; Rivera, Andreana L; Aldape, Kenneth

2008-12-01

In this study, we test the reliability of chromogenic in situ hybridization (CISH) for the detection of epidermal growth factor receptor (EGFR) gene amplification in glioblastoma. Earlier reports have described EGFR CISH in glioblastoma multiforme, but a comparison of CISH with a "gold standard" testing method, such as fluorescence in situ hybridization (FISH), has not been described. Therapies targeting the EGFR-signaling pathway might increase the importance of assessment of EGFR-amplification status. CISH is a potential alternative to FISH as a testing method. To test its reliability, EGFR-amplification status by CISH was assessed in 89 cases of glioblastoma and compared with FISH results, and correlated with the protein expression using immunohistochemistry (IHC) for EGFR. FISH was scored as being EGFR-amplified in 47/89 tumors, CISH as being amplified in 43/89 tumors. The CISH and FISH results were in agreement in 83/89 cases (93%). Four glioblastomas were scored as being amplified by FISH, but not by CISH; whereas amplification was detected in 2 tumors by CISH that were not amplified using FISH. Forty-eight of the 89 cases were positive for EGFR expression by IHC. EGFR amplification was highly correlated with protein expression by IHC, as 40/48 (83%) EGFR IHC-positive cases were found to be EGFR-amplified. The high concordance of CISH and FISH for the assessment of EGFR gene-amplification status indicates that CISH is a viable alternative to FISH for the detection of EGFR gene amplification in glioblastoma. Detectable EGFR expression by IHC can occur in the absence of gene amplification, but is uncommon.

Application of Pseudomurein Endoisopeptidase to Fluorescence In Situ Hybridization of Methanogens within the Family Methanobacteriaceae▿

Science.gov (United States)

Nakamura, Kohei ; Terada, Takeshi; Sekiguchi, Yuji; Shinzato, Naoya; Meng, Xian-Ying; Enoki, Miho; Kamagata, Yoichi

2006-01-01

In situ detection of methanogens within the family Methanobacteriaceae is sometimes known to be unsuccessful due to the difficulty in permeability of oligonucleotide probes. Pseudomurein endoisopeptidase (Pei), a lytic enzyme that specifically acts on their cell walls, was applied prior to 16S rRNA-targeting fluorescence in situ hybridization (FISH). For this purpose, pure cultured methanogens within this family, Methanobacterium bryantii, Methanobrevibacter ruminantium, Methanosphaera stadtmanae, and Methanothermobacter thermautotrophicus together with a Methanothermobacter thermautotrophicus-containing syntrophic acetate-oxidizing coculture, endosymbiotic Methanobrevibacter methanogens within an anaerobic ciliate, and an upflow anaerobic sludge blanket (UASB) granule were examined. Even without the Pei treatment, Methanobacterium bryantii and Methanothermobacter thermautotrophicus cells are relatively well hybridized with oligonucleotide probes. However, almost none of the cells of Methanobrevibacter ruminantium, Methanosphaera stadtmanae, cocultured Methanothermobacter thermautotrophicus, and the endosymbiotic methanogens and the cells within UASB granule were hybridized. Pei treatment was able to increase the probe hybridization ratio in every specimen, particularly in the specimen that had shown little hybridization. Interestingly, the hybridizing signal intensity of Methanothermobacter thermautotrophicus cells in coculture with an acetate-oxidizing H2-producing syntroph was significantly improved by Pei pretreatment, whereas the probe was well hybridized with the cells of pure culture of the same strain. We found that the difference is attributed to the differences in cell wall thicknesses between the two culture conditions. These results indicate that Pei treatment is effective for FISH analysis of methanogens that show impermeability to the probe. PMID:16950902

In situ DNA-RNA hybridization using in vitro 125I-labeled ribosomal RNA of higher plant

International Nuclear Information System (INIS)

Sato, Seiichi; Kikuchi, Tadatoshi; Ishida, M.R.; Tanaka, Ryuso.

1975-01-01

In situ hybridization using 125 I-labeled ribosomal RNA was applied to plant cells. Cytoplasmic 25 s rRNA, which was eluted from acrylamide gels after electrophoretic separation, was labeled in vitro with carrier-free 125 I and hybridized with the interphase nuclei in root tips of Vicia faba. In most of the preparations, the nucleoli were more heavily labeled than the other regions within nuclei, and several types of grain distribution were observed on the nucleoli. From these results, it was confirmed that in situ hybridization using 125 I-labeled rRNA can be used very effectively to detect the annealing sites of different molecular species of rRNA within the nuclei of plant cells, for which it is not as easy to obtain high specific radioactive rRNA in vivo as it is in the case of cultured animal cells. (auth.)

Determination of HER2 gene amplification by chromogenic in situ hybridization (CISH) in archival breast carcinoma.

Science.gov (United States)

Zhao, Jianxin; Wu, Rina; Au, Alfred; Marquez, Abbey; Yu, Yibing; Shi, Zuorong

2002-06-01

To compare the efficacy of chromogenic in situ hybridization (CISH(TM)) with fluorescence in situ (FISH) hybridization and immunohistochemistry (IHC) in determination of the HER2 status in human breast cancer. HER2 gene amplification was determined on formalin-fixed paraffin-embedded (FFPE) sections of 62 invasive breast cancers by FISH and followed by CISH using a digoxigenin (DIG)-labeled HER2 DNA probe generated by Subtraction Probe Technology (SPT(TM)), and a biotin-labeled chromosome 17 centromeric (chr.17cen) probe. The sections were heat treated and enzyme digested. After in situ hybridization, the HER2 probe was detected with fluorescein (FITC)-anti-DIG for FISH, followed by peroxidase-anti-FITC and diaminobenzidine (DAB) for CISH. The chr.17cen probe was detected with peroxidase-streptavidin and DAB. For CISH application, HER2 gene copies or chromosome 17 centromeres and morphology of cells were easily visualized simultaneously with a 40x objective under bright-field microscope in hematoxylin-counterstained sections. IHC study of HER2 overexpression was performed on adjacent sections using a panel of three HER2 antibodies (TAB 250, CB11, A0485), and staining was scored according to the criteria specified in the HercepTest. HER2 gene amplification detected by CISH was visualized typically as large DAB-stained clusters or by many dots in the nucleus. FISH and CISH identified HER2 gene amplification in 19% of the tumors. Chromosome 17 polysomy was detected in 31% of the tumors. HER2 overexpression was demonstrated in 19% (TAB 250), 23% (CB11), and 36% (A0485) of the tumors. Complete concordance between the results of CISH with FISH, TAB 250, CB11, and A0485 was seen in 100%, 97%, 94%, and 84% of the cases, respectively. By permitting observation of morphology using a bright-field microscope, CISH is an accurate, practical, and economical approach to screen HER2 status in breast cancers. It is a useful methodology for confirming ambiguous IHC results.

Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Directory of Open Access Journals (Sweden)

F.E. Rosa

Full Text Available Human epidermal growth factor receptor 2 (HER2 has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC. HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH in moderate immunoexpression (IHC 2+ cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH, which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

Directory of Open Access Journals (Sweden)

F.E. Rosa

2013-03-01

Full Text Available Human epidermal growth factor receptor 2 (HER2 has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC. HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH in moderate immunoexpression (IHC 2+ cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH, which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

Assignment of electron transfer flavoprotein-ubiquinone oxidoreductase (ETF-QO) to human chromosome 4q33 by fluorescence in situ hybridization and somatic cell hybridization.

Science.gov (United States)

Spector, E B; Seltzer, W K; Goodman, S I

1999-08-01

Electron transfer flavoprotein-ubiquinone oxidoreductase (ETF-QO) is a nuclear-encoded protein located in the inner mitochondrial membrane. Inherited defects of ETF-QO cause glutaric acidemia type II. We here describe the localization of the ETF-QO gene to human chromosome 4q33 by somatic cell hybridization and fluorescence in situ hybridization. Copyright 1999 Academic Press.

A safer, urea-based in situ hybridization method improves detection of gene expression in diverse animal species.

Science.gov (United States)

Sinigaglia, Chiara; Thiel, Daniel; Hejnol, Andreas; Houliston, Evelyn; Leclère, Lucas

2018-02-01

In situ hybridization is a widely employed technique allowing spatial visualization of gene expression in fixed specimens. It has greatly advanced our understanding of biological processes, including developmental regulation. In situ protocols are today routinely followed in numerous laboratories, and although details might change, they all include a hybridization step, where specific antisense RNA or DNA probes anneal to the target nucleic acid sequence. This step is generally carried out at high temperatures and in a denaturing solution, called hybridization buffer, commonly containing 50% (v/v) formamide - a hazardous chemical. When applied to the soft-bodied hydrozoan medusa Clytia hemisphaerica, we found that this traditional hybridization approach was not fully satisfactory, causing extensive deterioration of morphology and tissue texture which compromised our observation and interpretation of results. We thus tested alternative solutions for in situ detection of gene expression and, inspired by optimized protocols for Northern and Southern blot analysis, we substituted the 50% formamide with an equal volume of 8M urea solution in the hybridization buffer. Our new protocol not only yielded better morphologies and tissue consistency, but also notably improved the resolution of the signal, allowing more precise localization of gene expression and reducing aspecific staining associated with problematic areas. Given the improved results and reduced manipulation risks, we tested the urea protocol on other metazoans, two brachiopod species (Novocrania anomala and Terebratalia transversa) and the priapulid worm Priapulus caudatus, obtaining a similar reduction of aspecific probe binding. Overall, substitution of formamide by urea during in situ hybridization offers a safer alternative, potentially of widespread use in research, medical and teaching contexts. We encourage other workers to test this approach on their study organisms, and hope that they will also

Fluorescence in situ hybridization of old G-banded and mounted chromosome preparations

DEFF Research Database (Denmark)

Gerdes, A M; Pandis, N; Bomme, L

1997-01-01

the coverslips detach spontaneously; any mechanical manipulation will jeopardize the results. The success of chromosome painting is improved by excluding the regular RNase treatment step prior to hybridization. Additional changes compared with standard FISH protocols are that the 2 x SSC step is omitted......An improved method for fluorescence in situ hybridization (FISH) investigation of old, previously G-banded, mounted chromosome preparations with chromosome specific painting probes and centromere-specific probes is described. Before hybridization, the slides are incubated in xylene until......, that the amount of added probe is increased approximately 2.5 times, and that the amplification of signals is performed twice. The applicability of the method, which allows double painting with two differently labeled probes using two differently fluorescing colors, was tested on 11 cases involving different...

Demonstration testing and evaluation of in situ soil heating

International Nuclear Information System (INIS)

Sresty, G.C.

1994-01-01

A Treatability Study planned for the demonstration of the in situ electromagnetic (EM) heating process to remove organic solvents is described in this Work Plan. The treatability study will be conducted by heating subsurface vadose-zone soils in an organic plume adjacent to the Classified Burial Ground K-1070-D located at K-25 Site, Oak Ridge. The test is scheduled to start during the fourth quarter of FY94 and will be completed during the first quarter of FY95. The EM heating process for soil decontamination is based on volumetric heating technologies developed during the '70s for the recovery of fuels from shale and tar sands by IIT Research Institute (IITRI) under a co-operative program with the US Department of Energy (DOE). Additional modifications of the technology developed during the mid '80s are currently used for the production of heavy oil and waste treatment. Over the last nine years, a number of Government agencies (EPA, Army, AF, and DOE) and industries sponsored further development and testing of the in situ heating and soil decontamination process for the remediation of soils containing hazardous organic contaminants. In this process the soil is heated in situ using electrical energy. The contaminants are removed from the soil due to enhanced vaporization, steam distillation and stripping. IITRI will demonstrate the EM Process for in situ soil decontamination at K-25 Site under the proposed treatability study. Most of the contaminants of concern are volatile organics which can be removed by heating the soil to a temperature range of 85 to 95 C. The efficiency of the treatment will be determined by comparing the concentration of contaminants in soil samples. Samples will be obtained before and after the demonstration for a measurement of the concentration of contaminants of concern

Spatial Exploration and Characterization of Endozoicomonas spp. Bacteria in Stylophora pistillata Using Fluorescence In Situ Hybridization

KAUST Repository

Alsheikh-­Hussain, Areej

2011-12-12

Studies of coral-­associated bacterial communities have repeatedly demonstrated that the microbial assemblages of the coral host are highly specific and complex. In particular, bacterial community surveys of scleractinian and soft corals from geographically diverse reefs continually uncover a high abundance of sequences affiliated with the Gammaproteobacteria genus Endozoicomonas. The role of these bacteria within the complex coral holobiont is currently unknown. In order to localize these cells and gain an understanding of their potential interactions within the coral, we developed a fluorescence in situ hybridization(FISH) approach for reef-­building coral tissues. Using a custom small-­subunit ribosomal RNA gene database, we developed two Endozoicomonas-­specific probes that cover almost all known coral-­associated Endozoicomonas sequences. Probe hybridization conditions were quantitatively evaluated against target and non-­target bacterial cultures using fluorescence microscopy. Using these experimentally tested conditions, probes were then hybridized to the branching coral Stylophora pistillata, obtained from the Red Sea, using whole mount and paraffin embedding techniques. This study allowed preliminary spatial exploration and characterization of Endozoicomonas in coral, which has provided insight into their functional role and interactions within the coral holobiont.

Chromogenic in situ hybridization (CISH) to detect HER2 gene amplification in breast and gastric cancer: comparison with immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH).

Science.gov (United States)

Kiyose, Shinichiro; Igarashi, Hisaki; Nagura, Kiyoko; Kamo, Takaharu; Kawane, Kazunori; Mori, Hiroki; Ozawa, Takachika; Maeda, Matsuyoshi; Konno, Keisuke; Hoshino, Hideaki; Konno, Hiroyuki; Ogura, Hiroyuki; Shinmura, Kazuya; Hattori, Naohiko; Sugimura, Haruhiko

2012-11-01

The chromogenic in situ hybridization (CISH) assay, designed to detect the amplification of the HER2 gene in formalin-fixed, paraffin-embedded (FFPE) breast cancer (BC) and gastric cancer (GC) tissue specimens, was evaluated in 125 FFPE BC cases and 198 FFPE GC cases for which the HER2 status had been predetermined using immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). In the 125 BC cases and the 198 gastric cases, we found a very good concordance (98.4% and 99.0%, respectively) between CISH and FISH. In particular, we evaluated the polysomy cases, as these cases often have ambiguous treatment options in clinical practice. The polysomy of chromosome 17 was defined as the presence of three or more CEP17 signals in at least 10% of the tumor cells. In the 50 BC cases and 54 GC cases displaying chromosome 17 polysomy, the concordance between FISH and CISH was 98.0% and 98.1%, respectively. These results indicate that CISH could provide an accurate and practical alternative to FISH for the clinical diagnosis of HER2 gene amplification in FFPE BC and FFPE GC samples. © 2012 The Authors. Pathology International © 2012 Japanese Society of Pathology and Wiley Publishing Asia Pty Ltd.

Combination of microautoradiography and fluorescence in situ hybridization for identification of microorganisms degrading xenobiotic contaminants.

Science.gov (United States)

Yang, Yanru; Zarda, Annatina; Zeyer, Josef

2003-12-01

One of the central topics in environmental bioremediation research is to identify microorganisms that are capable of degrading the contaminants of interest. Here we report application of combined microautoradiography (MAR) and fluorescence in situ hybridization (FISH). The method has previously been used in a number of systems; however, here we demonstrate its feasibility in studying the degradation of xenobiotic compounds. With a model system (coculture of Pseudomonas putida B2 and Sphingomonas stygia incubated with [14C] o-nitrophenol), combination of MAR and FISH was shown to be able to successfully identify the microorganisms degrading o-nitrophenol. Compared with the conventional techniques, MAR-FISH allows fast and accurate identification of the microorganisms involved in environmental contaminant degradation.

Demonstration of Hybrid DSMC-CFD Capability for Nonequilibrium Reacting Flow

Science.gov (United States)

2018-02-09

AFRL-RV-PS- TR-2018-0056 AFRL-RV-PS- TR-2018-0056 DEMONSTRATION OF HYBRID DSMC-CFD CAPABILITY FOR NONEQUILIBRIUM REACTING FLOW Thomas E...4. TITLE AND SUBTITLE Demonstration of Hybrid DSMC-CFD Capability for Nonequilibrium Reacting Flow 5a. CONTRACT NUMBER 5b. GRANT NUMBER FA9453-17-1...simulation codes. The models are based on new ab-intio rate data obtained using state -of-the-art potential energy surfaces for air species. A probability

Chromogenic in situ hybridization (CISH): a novel alternative in screening archival breast cancer tissue samples for HER-2/neu status

OpenAIRE

Madrid, Manuelito A; Lo, Raymundo W

2004-01-01

Background Chromogenic in situ hybridization (CISH) is emerging as a practical, cost-effective, and valid alternative to fluorescent in situ hybridization in testing for gene alteration, especially in centers primarily working with immunohistochemistry (IHC). Methods We assessed Her-2/neu alteration using CISH on formalin-fixed paraffin-embedded primary invasive ductal carcinoma tumors in which IHC (CB11 antibody) had previously been performed, and we compared the results with IHC. The 160 se...

Karyotypes and Distribution of Tandem Repeat Sequences in Brassica nigra Determined by Fluorescence in situ Hybridization

Czech Academy of Sciences Publication Activity Database

Wang, G.; He, Q.; Macas, Jiří; Novák, Petr; Neumann, Pavel; Meng, D.; Zhao, H.; Guo, N.; Han, S.; Zong, M.; Jin, W.; Liu, F.

2017-01-01

Roč. 152, č. 3 (2017), s. 158-165 ISSN 1424-8581 R&D Projects: GA ČR GBP501/12/G090 Institutional support: RVO:60077344 Keywords : asymmetric somatic hybridization * Fluorescence in situ hybridization * Karyotype * (Peri) centromere Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Genetics and heredity (medical genetics to be 3) Impact factor: 1.354, year: 2016

Regulatory pathway analysis by high-throughput in situ hybridization.

Directory of Open Access Journals (Sweden)

Axel Visel

2007-10-01

Full Text Available Automated in situ hybridization enables the construction of comprehensive atlases of gene expression patterns in mammals. Such atlases can become Web-searchable digital expression maps of individual genes and thus offer an entryway to elucidate genetic interactions and signaling pathways. Towards this end, an atlas housing approximately 1,000 spatial gene expression patterns of the midgestation mouse embryo was generated. Patterns were textually annotated using a controlled vocabulary comprising >90 anatomical features. Hierarchical clustering of annotations was carried out using distance scores calculated from the similarity between pairs of patterns across all anatomical structures. This process ordered hundreds of complex expression patterns into a matrix that reflects the embryonic architecture and the relatedness of patterns of expression. Clustering yielded 12 distinct groups of expression patterns. Because of the similarity of expression patterns within a group, members of each group may be components of regulatory cascades. We focused on the group containing Pax6, an evolutionary conserved transcriptional master mediator of development. Seventeen of the 82 genes in this group showed a change of expression in the developing neocortex of Pax6-deficient embryos. Electromobility shift assays were used to test for the presence of Pax6-paired domain binding sites. This led to the identification of 12 genes not previously known as potential targets of Pax6 regulation. These findings suggest that cluster analysis of annotated gene expression patterns obtained by automated in situ hybridization is a novel approach for identifying components of signaling cascades.

In situ hybridization studies of hepatitis A viral RNA in patients with acute hepatitis A

Energy Technology Data Exchange (ETDEWEB)

Taylor, M; Goldin, R D; Ladva, S [Department of Histopathology, St. Mary' s Hospital Medical School, Imperial College of Science, Technology and Medicine, London (United Kingdom); Scheuer, P J [Department of Histopathology, Royal Free Hospital and School of Medicine, London (United Kingdom); Thomas, H C [Department of Medicine, St. Mary' s Hospital Medical School, Imperial College of Science, Technology and Medicine, London (United Kingdom)

1994-01-01

In situ hybridization with oligonucleotide probes has been used to localise hepatitis A virus RNA genomic sequences in formalin-fixed and routinely processed human liver biopsies from three patients. Using radiolabelled Sulphur-35 antisense probes, viral genomic sequences were found in all three cases, but signal intensity was greatest in cases 1 and 2 with fulminant hepatitis, and was minimal in the third case of resolving hepatitis biopsied 2 months after acute illness. Localisation showed the viral RNA to be present in hepatocytes, sinusoidal cells and inflammatory cells in and around the portal tracts. Both cases showed signal in similar cell types, but the distribution of staining was predominantly periportal in case 1, whereas lobular staining was more apparent in case 2. Hybridization with sense polarity probes failed to detect any evidence of replicative intermediates of antigenomic viral RNA. The presence of hepatitis A RNA in phagocytic cells was confirmed using immunohistochemistryfor a macrophage marker, CD68, combined with in situ hybridization. In all cases the signal was predominantly cytoplasmic, and this was confirmed with the use of tritiated probes. (au).

In situ hybridization studies of hepatitis A viral RNA in patients with acute hepatitis A

International Nuclear Information System (INIS)

Taylor, M.; Goldin, R.D.; Ladva, S.; Scheuer, P.J.; Thomas, H.C.

1994-01-01

In situ hybridization with oligonucleotide probes has been used to localise hepatitis A virus RNA genomic sequences in formalin-fixed and routinely processed human liver biopsies from three patients. Using radiolabelled Sulphur-35 antisense probes, viral genomic sequences were found in all three cases, but signal intensity was greatest in cases 1 and 2 with fulminant hepatitis, and was minimal in the third case of resolving hepatitis biopsied 2 months after acute illness. Localisation showed the viral RNA to be present in hepatocytes, sinusoidal cells and inflammatory cells in and around the portal tracts. Both cases showed signal in similar cell types, but the distribution of staining was predominantly periportal in case 1, whereas lobular staining was more apparent in case 2. Hybridization with sense polarity probes failed to detect any evidence of replicative intermediates of antigenomic viral RNA. The presence of hepatitis A RNA in phagocytic cells was confirmed using immunohistochemistryfor a macrophage marker, CD68, combined with in situ hybridization. In all cases the signal was predominantly cytoplasmic, and this was confirmed with the use of tritiated probes. (au)

Human Papilloma Virus 16 and 18 Association in Cervical Intraepithelial Lesions and Cervical Cancers by In Situ Hybridization

Directory of Open Access Journals (Sweden)

Mohanty Manisa

2017-03-01

Full Text Available Objective: To correlate the association of high risk Human Papilloma Virus (HPV 16, 18 in cervical intraepithelial lesions and cervical cancers by in-situ hybridization (ISH technique. Study Group: Cervical biopsy and hysterectomy specimen of 78 young and adult women, attending Hi-Tech Medical College and Hospital, Bhubaneswar, who were clinically or cytologically suspected of cervical intraepithelial lesion or cervical cancer were taken as source of target viral DNA. Material: Formalin 10% as fixative H & E stain as routine staining agent In-situ hybridization kit for HPV 16 and 18 DNA. Method: After following standard protocol for surgical grossing, HPV 16, 18 In-situ hybridization kit was used on paraffin embedded tissue sections. Results: The percentage of positive cases was highest in cervical cancer patients followed by cervical intraepithelial lesions, high grade, and low grade. Conclusion: This study has been carried out for the first in our state and our results show high degree of positivity of HPV 16/18 in females with cervical intraepithelial lesions and cervical cancers attending our tertiary care hospital.

Differential in situ hybridization for herpes simplex virus typing in routine skin biopsies

NARCIS (Netherlands)

Botma, H. J.; Dekker, H.; van Amstel, P.; Cairo, I.; van den Berg, F. M.

1995-01-01

A herpes simplex virus (HSV) type 2 specific recombinant plasmid probe designated pH2S3 was constructed from non-HSV-1 crossreactive regions of the HSV-2 genome. DNA in situ hybridization on in vitro reconstructed tissue samples of sheep collagen matrix impregnated with herpes virus-infected human

Whole-slide imaging is a robust alternative to traditional fluorescent microscopy for fluorescence in situ hybridization imaging using break-apart DNA probes.

Science.gov (United States)

Laurent, Camille; Guérin, Maxime; Frenois, François-Xavier; Thuries, Valérie; Jalabert, Laurence; Brousset, Pierre; Valmary-Degano, Séverine

2013-08-01

Fluorescence in situ hybridization is an indispensable technique used in routine pathology and for theranostic purposes. Because fluorescence in situ hybridization techniques require sophisticated microscopic workstations and long procedures of image acquisition with sometimes subjective and poorly reproducible results, we decided to test a whole-slide imaging system as an alternative approach. In this study, we used the latest generation of Pannoramic 250 Flash digital microscopes (P250 Flash digital microscopes; 3DHISTECH, Budapest, Hungary) to digitize fluorescence in situ hybridization slides of diffuse large B cells lymphoma cases for detecting MYC rearrangement. The P250 Flash digital microscope was found to be precise with better definition of split signals in cells containing MYC rearrangement with fewer truncated signals as compared to traditional fluorescence microscopy. This digital technique is easier thanks to the preview function, which allows almost immediate identification of the tumor area, and the panning and zooming functionalities as well as a shorter acquisition time. Moreover, fluorescence in situ hybridization analyses using the digital technique appeared to be more reproducible between pathologists. Finally, the digital technique also allowed prolonged conservation of photos. In conclusion, whole-slide imaging technologies represent rapid, robust, and highly sensitive methods for interpreting fluorescence in situ hybridization slides with break-apart probes. In addition, these techniques offer an easier way to interpret the signals and allow definitive storage of the images for pathology expert networks or e-learning databases. Copyright © 2013 Elsevier Inc. All rights reserved.

In situ gas treatment technology demonstration test plan

International Nuclear Information System (INIS)

Thornton, E.C.; Miller, R.D.

1996-01-01

This document defines the objectives and requirements associated with undertaking a field demonstration of an in situ gas treatment appoach to remediation chromate-contaminated soil. The major tasks presented in this plan include the design and development of the surface gas treatment system, performance of permitting activities, and completion of site preparation and field testing activities

10p Duplication characterized by fluorescence in situ hybridization

Energy Technology Data Exchange (ETDEWEB)

Wiktor, A.; Feldman, G.L.; Van Dyke, D.L.; Kratkoczki, P.; Ditmars, D.M. Jr. [Henry Ford Hospital, Detroit, MI (United States)

1994-09-01

We describe a patient with severe failure to thrive, mild-moderate developmental delay, cleft lip and palate, and other anomalies. Routine cytogenetic analysis documented a de novo chromosome rearrangement involving chromosome 4, but the origin of the derived material was unknown. Using chromosome specific painting probes, the karyotype was defined as 46,XY,der(4)t(4;10)(q35;p11.23). Characterization of the dup(10p) by fluorescence in situ hybridization (FISH) analysis provides another example of the usefulness of this technology in identifying small deletions, duplications, or supernumerary marker chromosomes. 19 refs., 4 figs.

Detection of a variable number of ribosomal DNA loci by fluorescent in situ hybridization in Populus species.

Science.gov (United States)

Prado, E A; Faivre-Rampant, P; Schneider, C; Darmency, M A

1996-10-01

Fluorescent in situ hybridization (FISH) was applied to related Populus species (2n = 19) in order to detect rDNA loci. An interspecific variability in the number of hybridization sites was revealed using as probe an homologous 25S clone from Populus deltoides. The application of image analysis methods to measure fluorescence intensity of the hybridization signals has enabled us to characterize major and minor loci in the 18S-5.8S-25S rDNA. We identified one pair of such rDNA clusters in Populus alba; two pairs, one major and one minor, in both Populus nigra and P. deltoides; and three pairs in Populus balsamifera, (two major and one minor) and Populus euroamericana (one major and two minor). FISH results are in agreement with those based on RFLP analysis. The pBG13 probe containing 5S sequence from flax detected two separate clusters corresponding to the two size classes of units that coexist within 5S rDNA of most Populus species. Key words : Populus spp., fluorescent in situ hybridization, FISH, rDNA variability, image analysis.

Fluorescence in situ hybridization for phytoplasma and endophytic bacteria localization in plant tissues.

Science.gov (United States)

Bulgari, Daniela; Casati, Paola; Faoro, Franco

2011-11-01

In the present study, we developed a rapid and efficient fluorescence in situ hybridization assay (FISH) in non-embedded tissues of the model plant Catharanthus roseus for co-localizing phytoplasmas and endophytic bacteria, opening new perspectives for studying the interaction between these microorganisms. Copyright © 2011 Elsevier B.V. All rights reserved.

Hydraulic Hybrid Parcel Delivery Truck Deployment, Testing & Demonstration

Energy Technology Data Exchange (ETDEWEB)

Gallo, Jean-Baptiste [Calstart Incorporated, Pasadena, CA (United States)

2014-03-07

Although hydraulic hybrid systems have shown promise over the last few years, commercial deployment of these systems has primarily been limited to Class 8 refuse trucks. In 2005, the Hybrid Truck Users Forum initiated the Parcel Delivery Working Group including the largest parcel delivery fleets in North America. The goal of the working group was to evaluate and accelerate commercialization of hydraulic hybrid technology for parcel delivery vehicles. FedEx Ground, Purolator and United Parcel Service (UPS) took delivery of the world’s first commercially available hydraulic hybrid parcel delivery trucks in early 2012. The vehicle chassis includes a Parker Hannifin hydraulic hybrid drive system, integrated and assembled by Freightliner Custom Chassis Corp., with a body installed by Morgan Olson. With funding from the U.S. Department of Energy, CALSTART and its project partners assessed the performance, reliability, maintainability and fleet acceptance of three pre-production Class 6 hydraulic hybrid parcel delivery vehicles using information and data from in-use data collection and on-road testing. This document reports on the deployment of these vehicles operated by FedEx Ground, Purolator and UPS. The results presented provide a comprehensive overview of the performance of commercial hydraulic hybrid vehicles in parcel delivery applications. This project also informs fleets and manufacturers on the overall performance of hydraulic hybrid vehicles, provides insights on how the technology can be both improved and more effectively used. The key findings and recommendations of this project fall into four major categories: -Performance, -Fleet deployment, -Maintenance, -Business case. Hydraulic hybrid technology is relatively new to the market, as commercial vehicles have been introduced only in the past few years in refuse and parcel delivery applications. Successful demonstration could pave the way for additional purchases of hydraulic hybrid vehicles throughout the

In situ green synthesis of antimicrobial carboxymethyl chitosan-nanosilver hybrids with controlled silver release.

Science.gov (United States)

Huang, Siqi; Yu, Zhiming; Zhang, Yang; Qi, Chusheng; Zhang, Shifeng

2017-01-01

In order to fabricate antimicrobial carboxymethyl chitosan-nanosilver (CMC-Ag) hybrids with controlled silver release, this study demonstrated comparable formation via three synthetic protocols: 1) carboxymethyl chitosan (CMC) and glucose (adding glucose after AgNO 3 ), 2) CMC and glucose (adding glucose before AgNO 3 ), and 3) CMC only. Under principles of green chemistry, the synthesis was conducted in an aqueous medium exposed to microwave irradiation for 10 minutes with nontoxic chemicals. The structure and formation mechanisms of the three CMC-Ag hybrids were explored using X-ray diffraction, ultraviolet-visible spectroscopy, transmission electron microscopy, and Fourier-transform infrared analyses. Additionally, antimicrobial activity and in vitro silver release of the three synthesized hybrids were investigated in detail. The results revealed that a large number of stable, uniform, and small silver nanoparticles (AgNPs) were synthesized in situ on CMC chains via protocol 1. AgNPs were well dispersed with narrow size distribution in the range of 6-20 nm, with mean diameter only 12.22±2.57 nm. The addition of glucose resulted in greater AgNP synthesis. The order of addition of glucose and AgNO 3 significantly affected particle size and size distribution of AgNPs. Compared to CMC alone and commercially available AgNPs, the antimicrobial activities of three hybrids were significantly improved. Of the three hybrids, CMC-Ag1 synthesized via protocol 1 exhibited better antimicrobial activity than CMC-Ag2 and CMC-Ag3, and showed more effective inhibition of Staphylococcus aureus than Escherichia coli . Due to strong coordination and electrostatic interactions between CMC and silver and good steric protection provided by CMC, CMC-Ag1 displayed stable and continuous silver release and better performance in retaining silver for prolonged periods than CMC-Ag2 and CMC-Ag3.

In situ green synthesis of antimicrobial carboxymethyl chitosan–nanosilver hybrids with controlled silver release

Science.gov (United States)

Huang, Siqi; Yu, Zhiming; Zhang, Yang; Qi, Chusheng; Zhang, Shifeng

2017-01-01

In order to fabricate antimicrobial carboxymethyl chitosan–nanosilver (CMC-Ag) hybrids with controlled silver release, this study demonstrated comparable formation via three synthetic protocols: 1) carboxymethyl chitosan (CMC) and glucose (adding glucose after AgNO3), 2) CMC and glucose (adding glucose before AgNO3), and 3) CMC only. Under principles of green chemistry, the synthesis was conducted in an aqueous medium exposed to microwave irradiation for 10 minutes with nontoxic chemicals. The structure and formation mechanisms of the three CMC-Ag hybrids were explored using X-ray diffraction, ultraviolet-visible spectroscopy, transmission electron microscopy, and Fourier-transform infrared analyses. Additionally, antimicrobial activity and in vitro silver release of the three synthesized hybrids were investigated in detail. The results revealed that a large number of stable, uniform, and small silver nanoparticles (AgNPs) were synthesized in situ on CMC chains via protocol 1. AgNPs were well dispersed with narrow size distribution in the range of 6–20 nm, with mean diameter only 12.22±2.57 nm. The addition of glucose resulted in greater AgNP synthesis. The order of addition of glucose and AgNO3 significantly affected particle size and size distribution of AgNPs. Compared to CMC alone and commercially available AgNPs, the antimicrobial activities of three hybrids were significantly improved. Of the three hybrids, CMC-Ag1 synthesized via protocol 1 exhibited better antimicrobial activity than CMC-Ag2 and CMC-Ag3, and showed more effective inhibition of Staphylococcus aureus than Escherichia coli. Due to strong coordination and electrostatic interactions between CMC and silver and good steric protection provided by CMC, CMC-Ag1 displayed stable and continuous silver release and better performance in retaining silver for prolonged periods than CMC-Ag2 and CMC-Ag3. PMID:28458539

Characterization of a de novo duplication of 11p14----p13, using fluorescent in situ hybridization and southern hybridization

NARCIS (Netherlands)

Speleman, F.; Mannens, M.; Redeker, B.; Vercruyssen, M.; van Oostveldt, P.; Leroy, J.; Slater, R.

1991-01-01

A de novo 11p+ chromosome was found in a child with mild mental retardation but no other remarkable dysmorphic characteristics. Banding studies suggested a duplication of regions 11p13 and 11p14 or regions 11p14 and 11p15. Using fluorescent in situ hybridization and digital imaging microscopy, we

Presence and localization of bacteria in the bovine endometrium postpartum using fluorescence in situ hybridization

DEFF Research Database (Denmark)

Karstrup, C. C.; Agerholm, J. S.; Jensen, Tim Kåre

2017-01-01

The aim of this study was to investigate bacterial invasiveness of the bovine endometrium during the postpartum period. Fluorescence in situ hybridization was applied to endometrial biopsies using probes for Fusobacterium necrophorum, Porphyromonas levii, Trueperella pyogenes, Escherichia coli...

  In situ identification of streptococci and other bacteria in initial dental biofilm by confocal laser scanning microscopy and fluorescence in situ hybridization

DEFF Research Database (Denmark)

Dige, Irene; Kilian, Mogens; Nilsson, Holger

2007-01-01

Confocal laser scanning microscopy (CLSM) has been employed as a method for studying intact natural biofilm. When combined with fluorescence in situ hybridization (FISH) it is possible to analyze spatial relationships and changes of specific members of microbial populations over time. The aim...

Reliability of chromogenic in situ hybridization for epidermal growth factor receptor gene copy number detection in non-small-cell lung carcinomas: a comparison with fluorescence in situ hybridization study.

Science.gov (United States)

Yoo, Seol Bong; Lee, Hyun Ju; Park, Jung Ok; Choe, Gheeyoung; Chung, Doo Hyun; Seo, Jeong-Wook; Chung, Jin-Haeng

2010-03-01

Fluorescence in situ hybridization (FISH) has been known to be the most representative and standardized test for assessing gene amplification. However, FISH requires a fluorescence microscope, the signals are labile and rapidly fade over time. Recently, chromogenic in situ hybridization (CISH) has emerged as a potential alternative to FISH. The aim of this study is to test the reliability of CISH technique for the detection of epidermal growth factor receptor (EGFR) gene amplification in non-small-cell lung carcinomas (NSCLC), to compare CISH results with FISH. A total of 277 formalin-fixed and paraffin embedded NSCLC tissue samples were retrieved from the surgical pathology archives at Seoul National University Bundang Hospital. CISH and FISH examinations were performed to test EGFR gene amplification status. There was high concordance in the assessment of EGFR gene copy number between CISH and FISH tests (Kappa coefficient=0.83). Excellent concordance was shown between two observers on the interpretation of the CISH results (Kappa coefficient=0.90). In conclusion, CISH result is highly reproducible, accurate and practical method to determine EGFR gene amplification in NSCLC. In addition, CISH allows a concurrent analysis of histological features of the tumors and gene copy numbers.

Numerical simulations in support of the in situ bioremediation demonstration at Savannah River

International Nuclear Information System (INIS)

Travis, B.J.; Rosenberg, N.D.

1994-06-01

This report assesses the performance of the in situ bioremediation technology demonstrated at the Savannah River Integrated Demonstration (SRID) site in 1992--1993. The goal of the technology demonstration was to stimulate naturally occurring methanotrophic bacteria at the SRID site with injection of methane, air and air-phase nutrients (nitrogen and phosphate) such that significant amounts of the chlorinated solvent present in the subsurface would be degraded. Our approach is based on site-specific numerical simulations using the TRAMP computer code. In this report, we discuss the interactions among the physical and biochemical processes involved in in situ bioremediation. We also investigate improvements to technology performance, make predictions regarding the performance of this technology over long periods of time and at different sites, and compare in situ bioremediation with other remediation technologies

A double labeling technique for performing immunocytochemistry and in situ hybridization in virus infected cell cultures and tissues

International Nuclear Information System (INIS)

Gendelman, H.E.; Moench, T.R.; Narayan, O.; Griffin, D.E.; Clements, J.E.

1985-01-01

This report describes a combined immunocytochemical and in situ hybridization procedure which allows visualization of cellular or viral antigens and viral RNA in the same cell. Cultures infected with visna or measles virus were fixed in periodate-lysine-paraformaldehyde-glutaraldehyde, stained by the avidin-biotin-peroxidase technique using antibodies to viral or cellular proteins and then incubated with radiolabeled specific DNA probes (in situ hybridization). This technique provides a new approach to the study of viral pathogenesis by: (1) identifying the types of cells which are infected in the host and (2) identifying points of blockade in the virus life cycle during persistent infections. (Auth.)

Development and use of fluorescent 16S rRNA-targeted probes for the specific detection of Methylophaga species by in situ hybridization in marine sediments.

Science.gov (United States)

Janvier, Monique; Regnault, Béatrice; Grimont, Patrick

2003-09-01

Methylotrophic bacteria are widespread in nature. They may play an important role in the cycling of carbon and in the metabolism of dimethylsulfide in a marine environment. Bacteria belonging to the genus Methylophaga are a unique group of aerobic, halophilic, non-methane-utilizing methylotrophs. Two 16S rRNA-targeted oligonucleotide probes were developed for the specific detection of Methylophaga species, marine methylobacteria, by fluorescence in situ hybridization. Probe MPH-730 was highly specific for all members of the genus Methylophaga while probe MPHm-994 targeted exclusively M. marina. The application of these probes were demonstrated by the detection of Methylophaga species in enrichment cultures from various marine sediments. All isolates recovered were visualized by using the genus specific probe MPH-730. The results were confirmed by 16S rDNA sequencing which demonstrated that all selected isolates belong to Methylophaga. Five isolates could be detected by the M. marina-specific probe MPHm-994 and were confirmed by rRNA gene restriction pattern (ribotyping). With the development of these specific probes, fluorescence in situ hybridization shows that the genus Methylophaga is widespread in marine samples.

Motivational study for an hybrid demonstrator

International Nuclear Information System (INIS)

Boidron, M.; Fiorini, G.L.; Thomas, J.B.

2001-02-01

This document recalls first the role of hybrid accelerator driven systems (ADS) in the domain of transmutation of long-lived fission products and minor actinides. It presents the specific contribution of these systems in the management of radioactive wastes and their technical feasibility and safety aspects. Then, follows a motivational analysis for the construction of a demonstration facility with its specifications and R and D needs: feasibility, schedule, links with other ADS-related programs, cost, international cooperation, recommendations. (J.S.)

Study of the frequency of translocations and dicentrics in human spermatozoid using fluorescent in situ hybridization

International Nuclear Information System (INIS)

Alvarez, R.; Ponsa, I.; Tusell, L.; Genesca, A.; Miro, R.; Egozcue, J.

1998-01-01

Present study has intended to analyze the induction translocations and dicentrics in human sperms irradiated in vitro to the dose 4Gy by means of the use technical in situ hybridization with probes marked fluorescently

Fluorescence in situ hybridization on human metaphase chromosomes detected by near-field scanning optical microscopy

NARCIS (Netherlands)

Moers, M.H.P.; Moers, M.H.P.; Kalle, W.H.J.; Kalle, W.H.J.; Ruiter, A.G.T.; Wiegant, J.C.A.G.; Raap, A.K.; Greve, Jan; de Grooth, B.G.; van Hulst, N.F.

1996-01-01

Fluorescence in situ hybridization o­n human metaphase chromosomes is detected by near-field scanning optical microscopy. This combination of cytochemical and scanning probe techniques enables the localization and identification of several fluorescently labelled genomic DNA fragments o­n a single

City of Las Vegas Plug-in Hybrid Electric Vehicle Demonstration Program

Energy Technology Data Exchange (ETDEWEB)

None

2013-12-31

The City of Las Vegas was awarded Department of Energy (DOE) project funding in 2009, for the City of Las Vegas Plug-in Hybrid Electric Vehicle Demonstration Program. This project allowed the City of Las Vegas to purchase electric and plug-in hybrid electric vehicles and associated electric vehicle charging infrastructure. The City anticipated the electric vehicles having lower overall operating costs and emissions similar to traditional and hybrid vehicles.

Cytogenetic analysis using fluorescence in situ hybridization (FISH) to evaluate occupational exposure to carcinogens

Czech Academy of Sciences Publication Activity Database

Šrám, Radim; Beskid, Olena; Binková, Blanka; Rössner, P.; Šmerhovský, Zdeněk

2004-01-01

Roč. 149, - (2004), s. 335-344 ISSN 0378-4274 R&D Projects: GA MŽP SI/340/2/00 Institutional research plan: CEZ:AV0Z5039906 Keywords : Chromosomal aberrations * Fluorescence in situ hybridization Subject RIV: DN - Health Impact of the Environment Quality Impact factor: 2.571, year: 2004

Summary performance assessment of in situ remediation technologies demonstrated at Savannah River

International Nuclear Information System (INIS)

Rosenberg, N.D.; Robinson, B.A.; Birdsell, K.H.; Travis, B.J.

1994-06-01

The Office of Technology Development (OTD) in the Department of Energy's (DOE) Office of Environmental Restoration and Waste Management is investigating new technologies for ''better, faster, cheaper, safer'' environmental remediation. A program at DOE's Savannah River site was designed to demonstrate innovative technologies for the remediation of volatile organic compounds (VOCs) at nonarid sites. Two remediation technologies, in situ air stripping and in situ bioremediation--both using horizontal wells, were demonstrated at the site between 1990--1993. This brief report summarizes the conclusions from three separate modeling studies on the performance of these technologies

Field demonstration and transition of SCAPS direct push VOC in-situ sensing technologies

International Nuclear Information System (INIS)

Davis, William M.

1999-01-01

This project demonstrated two in-situ volatile organic compound (VOC) samplers in combination with the direct sampling ion trap mass spectrometer (DSITMS). The technologies chosen were the Vadose Sparge and the Membrane Interface Probe (MIP) sensing systems. Tests at two demonstration sites showed the newer VOC technologies capable of providing in situ contaminant measurements at two to four times the rate of the previously demonstrated Hydrosparge sensor. The results of this project provide initial results supporting the utility of these new technologies to provide rapid site characterization of VOC contaminants in the subsurface

Demonstration of Hepatitis C Virus RNA with In Situ Hybridization Employing a Locked Nucleic Acid Probe in Humanized Liver of Infected Chimeric Mice and in Needle-Biopsied Human Liver

Directory of Open Access Journals (Sweden)

Kazuya Shiogama

2013-01-01

Full Text Available Background. In situ hybridization (ISH with high sensitivity has been requested to demonstrate hepatitis C virus (HCV RNA in formalin-fixed, paraffin-embedded (FFPE sections of the liver. Methods. ISH employing a locked-nucleic-acid- (LNA-modified oligonucleotide probe and biotin-free catalyzed signal amplification system (CSAII was applied to HCV-RNA detection in the liver tissue. Nested reverse-transcription polymerase chain reaction (RT-PCR was performed for HCV genotyping using total RNA extracted from FFPE sections. The target tissues included FFPE tissue sections of humanized livers in HCV-infected chimeric mice (HCV genotypes 1a, 1b, and 2a and noninfected and of needle-biopsied livers from HCV-infected patients. Results. HCV-RNA was demonstrated with the ISH technique in HCV-infected liver tissues from both chimeric mice and 9 (82% of 11 patients with HCV infection. The HCV signals were sensitive to RNase. Nested RT-PCR confirmed the genotype in 8 (73% of 11 livers (type 1b: 6 lesions and type 2a: 2 lesions. HCV-RNA was not identified in chronic hepatitis B lesions, fatty liver, autoimmune hepatitis, and hepatocellular carcinoma. Conclusion. ISH using the LNA-modified oligonucleotide probe and CSAII was applicable to detecting HCV-RNA in routinely prepared FFPE liver specimens.

Detection of human papillomavirus in laryngeal lesions by in situ hybridization

DEFF Research Database (Denmark)

Multhaupt, H A; Fessler, J N; Warhol, M J

1994-01-01

Human papillomavirus (HPV) is associated with human neoplasms of squamous epithelium. Squamous papillomas and verrucous carcinomas are two types of squamous neoplasms of the larynx that present difficult problems in differential diagnosis. Using in situ hybridization with biotinylated DNA probes......, we examined benign squamous papillomas and verrucous squamous carcinomas of the larynx for the presence of HPV. Forty-two biopsy specimens from 18 patients with laryngeal papillomas and 11 biopsy specimens from seven patients with verrucous carcinomas were obtained from the files of Pennsylvania...... Hospital, Philadelphia, PA. Tissue sections were hybridized with an HPV DNA cocktail. The HPV-positive cases then were subtyped further with DNA probes specific for HPV subtypes 6/11, 16/18, and 31/33/35. All benign squamous papillomas (42 of 42) were positive for HPV subtype 6/11. None of the verrucous...

Efficient in situ growth of enzyme-inorganic hybrids on paper strips for the visual detection of glucose.

Science.gov (United States)

Li, WanYun; Lu, ShiYu; Bao, ShuJuan; Shi, ZhuanZhuan; Lu, Zhisong; Li, ChangMing; Yu, Ling

2018-01-15

A visual colorimetric microfluidic paper-based analytical device (μPAD) was constructed following the direct synthesis of enzyme-inorganic hybrid nanomaterials on the paper matrix. An inorganic solution of MnSO 4 and KH 2 PO 4 containing a diluted enzyme (glucose oxidase, GOx) was subsequently pipetted onto cellulose paper for the in situ growth of GOx@Mn 3 (PO 4 ) 2 hybrid functional materials. The characterization of the morphology and chemical composition validated the presence of hybrid materials roots in the paper fiber, while the Mn 3 (PO 4 ) 2 of the hybrid provided both a surface for enzyme anchoring and a higher peroxidase-like catalytic activity as compared to the Mn 3 (PO 4 ) 2 crystal that was synthesized without enzyme modulation. This new approach for the in situ growth of an enzyme-inorganic hybrid on a paper matrix eliminates centrifugation and the dry process by casting the solution on paper. The sensing material loading was highly reproducible because of the accuracy and stability of pipetting, which eventually contributed to the reliability of the μPAD. The self-assembled natural and artificial enzyme hybrid on the μPADs specifically detected glucose from a group of interferences, which shows great specificity using this method. Moreover, the colorimetric signal exhibited detection limitation for glucose is 0.01mM, which lies in the physiological range of glucose in biological samples. Copyright © 2017 Elsevier B.V. All rights reserved.

[HER-2 oncogene amplification assessment in invasive breast cancer by dual-color in situ hybridization (dc-CISH): a comparative study with fluorescent in situ hybridization (FISH)].

Science.gov (United States)

Akhdar, Abbas; Bronsard, Marc; Lemieux, Renald; Geha, Sameh

2011-12-01

The amplification of the gene encoding for the human epidermal growth factor receptor 2 (HER-2 oncogene), located on chromosome 17 (17q21-q22), or the overexpression of this receptor have prognostic and therapeutic implications in invasive breast cancer. An evaluation of the HER-2 status by immunohistochemistry (IHC) is performed on all invasive breast cancer cases. Fluorescent in situ hybridization (FISH) is considered as the gold standard for the detection of HER-2 gene amplification for IHC equivocal cases (score 2+). A more recent in situ hybridization technique, the dual-color chromogenic in situ hybridization (dc-CISH), has been proposed as an alternative to FISH. The aim of this study was to measure the correlation between dc-CISH and FISH for HER-2 oncogene amplification assessment in invasive breast cancer. We built four tissue micro-array (TMA) blocs with 100 breast invasive cancer cases that had been previously tested by IHC for HER-2 detection: 10 score 0 cases, 10 score 3+cases, 39 score 1+and 41 score 2+cases. Both FISH and dc-CISH techniques were applied on all TMA cases as well as on two additional slides serving as controls. Interpretation of dc-CISH was carried out by a pathologist using an optical microscope. For FISH, the interpretation was done by a professional from the medical genetics department using a fluorescent microscope linked to a computer system for image capturing and analysis. The interpretation of the HER-2/CEN-17 ratio for both tests was in accordance with the values of the updated recommendations from the Canadian National Consensus Meeting on HER-2/neu testing in breast cancer and from the ASCO/CAP. Among the 100 cases initially included in the study, eight were excluded from the analysis due to sampling or technical flaws. From the 92 remaining cases, we obtained a concordance of 97.8% (90/92 cases) between the two techniques (Kappa coefficient 0.97, 95% confidence interval). The correlation coefficient (rho) between ratios

Using Whole Mount in situ Hybridization to Link Molecular and Organismal Biology

OpenAIRE

Jacobs, Nicole L.; Albertson, R. Craig; Wiles, Jason R.

2011-01-01

Whole mount in situ hybridization (WISH) is a common technique in molecular biology laboratories used to study gene expression through the localization of specific mRNA transcripts within whole mount specimen. This technique (adapted from Albertson and Yelick, 2005) was used in an upper level undergraduate Comparative Vertebrate Biology laboratory classroom at Syracuse University. The first two thirds of the Comparative Vertebrate Biology lab course gave students the opportunity to study the ...

Demonstration testing and evaluation of in situ soil heating. Revision 1, Demonstration system design

International Nuclear Information System (INIS)

Dev, H.

1994-01-01

Over the last nine years IIT Research Institute (IITRI) has been developing and testing the in situ heating and soil decontamination process for the remediation of soils containing hazardous organic contaminants. In this process the soil is heated in situ using electrical energy. The contaminants are removed from the soil due to enhanced vaporization, steam distillation and stripping. The vaporized contaminants, water vapor and air are recovered from the heated zone by means of a vacuum manifold system which collects gases from below surface as well as from the soil surface. A vapor barrier is used to prevent fugitive emissions of the contaminants and to control air infiltration to minimize dilution of the contaminant gases and vapors. The recovered gases and vapors are conveyed to an on site vapor treatment system for the clean up of the vent gases. Electrical energy is applied to the soil by forming an array of electrodes in the soil which are electrically interconnected and supplied with power. The electrodes are placed in drilled bore holes which are made through the contaminated zone. There are two versions of the in situ heating and soil treatment process: the f irst version is called the In Situ Radio Frequency (RF) Soil Decontamination Process and the second version is called the In Situ Electromagnetic (EM) Soil Decontamination Process. The first version, the RF Process is capable of heating the soil in a temperature range of 100 degrees to 400 degrees C. The soil temperature in the second version, the EM Process, is limited to the boiling point of water under native conditions. Thus the soil will be heated to a temperature of about 85 degrees to 95 degrees C. In this project IITRI will demonstrate the EM Process for in situ soil decontamination at K-25 Site due to the fact that most of the contaminants of concern are volatile organics which can be removed by heating the soil to a temperature range of 85 degrees to 95 degrees C

Demonstration testing and evaluation of in situ soil heating. Revision 1, Demonstration system design

Energy Technology Data Exchange (ETDEWEB)

Dev, H.

1994-08-16

Over the last nine years IIT Research Institute (IITRI) has been developing and testing the in situ heating and soil decontamination process for the remediation of soils containing hazardous organic contaminants. In this process the soil is heated in situ using electrical energy. The contaminants are removed from the soil due to enhanced vaporization, steam distillation and stripping. The vaporized contaminants, water vapor and air are recovered from the heated zone by means of a vacuum manifold system which collects gases from below surface as well as from the soil surface. A vapor barrier is used to prevent fugitive emissions of the contaminants and to control air infiltration to minimize dilution of the contaminant gases and vapors. The recovered gases and vapors are conveyed to an on site vapor treatment system for the clean up of the vent gases. Electrical energy is applied to the soil by forming an array of electrodes in the soil which are electrically interconnected and supplied with power. The electrodes are placed in drilled bore holes which are made through the contaminated zone. There are two versions of the in situ heating and soil treatment process: the f irst version is called the In Situ Radio Frequency (RF) Soil Decontamination Process and the second version is called the In Situ Electromagnetic (EM) Soil Decontamination Process. The first version, the RF Process is capable of heating the soil in a temperature range of 100{degrees} to 400{degrees}C. The soil temperature in the second version, the EM Process, is limited to the boiling point of water under native conditions. Thus the soil will be heated to a temperature of about 85{degrees} to 95{degrees}C. In this project IITRI will demonstrate the EM Process for in situ soil decontamination at K-25 Site due to the fact that most of the contaminants of concern are volatile organics which can be removed by heating the soil to a temperature range of 85{degrees} to 95{degrees}C.

In-situ fabrication of hybrid polyoxometalate nanoparticles composite films

International Nuclear Information System (INIS)

Lan Yang; Mao Baodong; Wang Enbo; Song Yonghai; Kang Zhenhui; Wang Chunlei; Tian Chungui; Zhang Chao; Xu Lin; Li Zhuang

2007-01-01

Inorganic-organic hybrid nanoparticles multilayer films were fabricated by extending the method of nucleation and growth of particles in polymer assemblies. The polyelectrolyte matrix was constructed by layer-by-layer self-assembly method. Synthesis of polyoxometalate nanoparticles was achieved by alternately dipping the precursor polyelectrolyte matrix into AgNO 3 and H 4 SiW 12 O 40 aqueous solutions. Repeating the above synthesis process, Ag 4 SiW 12 O 40 nanoparticles with controllable diameters of 20 to 77 nm were synthesized in the multilayer films in-situ. UV-vis absorption spectra indicate that the nanoparticles grew gradually in the synthesis process. Transmission electron microscopy was used to observe the size and morphology of the nanoparticles

Study of lower hybrid current drive for the demonstration reactor

Energy Technology Data Exchange (ETDEWEB)

Molavi-Choobini, Ali Asghar [Dept. of Physics, Faculty of Engineering, Islamic Azad University, Shahr-e-kord Branch, Shahr-e-kord (Iran, Islamic Republic of); Naghidokht, Ahmed [Dept. of Physics, Urmia University, Urmia (Iran, Islamic Republic of); Karami, Zahra [Dept. of Engineering, Islamic Azad University, Zanjan Branch, Zanjan (Iran, Islamic Republic of)

2016-06-15

Steady-state operation of a fusion power plant requires external current drive to minimize the power requirements, and a high fraction of bootstrap current is required. One of the external sources for current drive is lower hybrid current drive, which has been widely applied in many tokamaks. Here, using lower hybrid simulation code, we calculate electron distribution function, electron currents and phase velocity changes for two options of demonstration reactor at the launched lower hybrid wave frequency 5 GHz. Two plasma scenarios pertaining to two different demonstration reactor options, known as pulsed (Option 1) and steady-state (Option 2) models, have been analyzed. We perceive that electron currents have major peaks near the edge of plasma for both options but with higher efficiency for Option 1, although we have access to wider, more peripheral regions for Option 2. Regarding the electron distribution function, major perturbations are at positive velocities for both options for flux surface 16 and at negative velocities for both options for flux surface 64.

Subcellular localization of low-abundance human immunodeficiency virus nucleic acid sequences visualized by fluorescence in situ hybridization

International Nuclear Information System (INIS)

Lawrence, J.B.; Marselle, L.M.; Byron, K.S.; Johnson, C.V.; Sullivan, J.L.; Singer, R.H.

1990-01-01

Detection and subcellular localization of human immunodeficiency virus (HIV) were investigated using sensitive high-resolution in situ hybridization methodology. Lymphocytes infected with HIV in vitro or in vivo were detected by fluorescence after hybridization with either biotin or digoxigenin-labeled probes. At 12 hr after infection in vitro, a single intense signal appeared in the nuclei of individual cells. Later in infection, when cytoplasmic fluorescence became intense, multiple nuclear foci frequently appeared. The nuclear focus consisted of newly synthesized HIV RNA as shown by hybridization in the absence of denaturation and by susceptibility to RNase and actinomycin D. Virus was detected in patient lymphocytes and it was shown that a singular nuclear focus also characterizes cells infected in vivo. The cell line 8E5/LAV containing one defective integrated provirus revealed a similar focus of nuclear RNA, and the single integrated HIV genome was unequivocally visualized on a D-group chromosome. This demonstrates an extremely sensitive single-cell assay for the presence of a single site of HIV transcription in vitro and in vivo and suggests that it derives from one (or very few) viral genomes per cell. In contrast, productive Epstein-Barr virus infection exhibited many foci of nuclear RNA per cell

Submicroscopic deletions at the WAGR locus, revealed by nonradioactive in situ hybridization.

OpenAIRE

Fantes, J A; Bickmore, W A; Fletcher, J M; Ballesta, F; Hanson, I M; van Heyningen, V

1992-01-01

Fluorescence in situ hybridization (FISH) with biotin-labeled probes mapping to 11p13 has been used for the molecular analysis of deletions of the WAGR (Wilms tumor, aniridia, genitourinary abnormalities, and mental retardation) locus. We have detected a submicroscopic 11p13 deletion in a child with inherited aniridia who subsequently presented with Wilms tumor in a horseshoe kidney, only revealed at surgery. The mother, who has aniridia, was also found to carry a deletion including both the ...

Project summary, 116-B-6-1 crib ISV [in situ vitrification] demonstration project

International Nuclear Information System (INIS)

Koegler, S.S.

1989-01-01

The 116-B Crib Demonstration Project is intended to demonstrate the emerging in situ vitrification (ISV) technology to immobilize or destroy hazardous and radioactive chemicals at an actual site. In situ vitrification is the conversion of contaminated soil into a durable glass and crystalline product through joule heating. The 116-B crib site was chosen for the demonstration because it contains both radioactive and hazardous chemicals (e.g., chromium) and presents a potential threat to environment. The project will involve sampling and analysis of the soil beneath the crib, a small-scale ISV test to verify operating parameters, vitrification of the crib, and analysis of the vitrified soil. 5 figs

Fluorescence in situ hybridization karyotyping reveals the presence of two distinct genomes in the taxon Aegilops tauschii

OpenAIRE

Zhao, Laibin; Ning, Shunzong; Yi, Yingjin; Zhang, Lianquan; Yuan, Zhongwei; Wang, Jirui; Zheng, Youliang; Hao, Ming; Liu, Dengcai

2018-01-01

Background Aegilops tauschii is the donor of the bread wheat D genome. Based on spike morphology, the taxon has conventionally been subdivided into ssp. tauschii and ssp. strangulata. The present study was intended to address the poor match between this whole plant morphology-based subdivision and genetic relationships inferred from genotyping by fluorescence in situ hybridization karyotyping a set of 31 Ae. tauschii accessions. Results The distribution of sites hybridizing to the two probes ...

Automated Image Analysis for Quantitative Fluorescence In Situ Hybridization with Environmental Samples▿ †

OpenAIRE

Zhou, Zhi; Pons, Marie Noëlle; Raskin, Lutgarde; Zilles, Julie L.

2007-01-01

When fluorescence in situ hybridization (FISH) analyses are performed with complex environmental samples, difficulties related to the presence of microbial cell aggregates and nonuniform background fluorescence are often encountered. The objective of this study was to develop a robust and automated quantitative FISH method for complex environmental samples, such as manure and soil. The method and duration of sample dispersion were optimized to reduce the interference of cell aggregates. An au...

Fluorescence in situ hybridization: an improved method of quantitating chromosome damage and repair

International Nuclear Information System (INIS)

Brown, J.M.; Evans, J.W.

1993-01-01

The authors combined fluorescence in situ hybridization (FISH) with specific full-length chromosome probes using the premature chromosome condensation (PCC) technique chromosome condensation (PCC) technique to simplify scoring chromosome damage and its repair. They have shown the technique works well and enables breaks and exchanges to be readily detected and scored in individual chromosomes. A chromosome 4 full-length specific library has been used in initial studies. (UK)

The Diagnosis of Gastric Mucosa-associated Lymphoid Tissue Lymphoma by Flow Cytometry and Fluorescence in situ Hybridization of Biopsy Specimens.

Science.gov (United States)

Matsueda, Katsunori; Omote, Sizuma; Sakata, Masahiro; Fujita, Isao; Horii, Jouichiro; Toyokawa, Tatsuya

2018-04-15

Mucosa-associated lymphoid tissue (MALT) lymphoma and reactive inflammatory lymphoid changes are frequently difficult to distinguish based on a routine histological differential diagnosis. We were unable to diagnose gastric MALT lymphoma histologically using specimens obtained by endoscopy, although a flow cytometry (FCM) analysis demonstrated clonality of neoplastic cells by separating cells by CD45 gating. Furthermore, a fluorescence in situ hybridization (FISH) analysis showed trisomy 18. We therefore diagnosed gastric MALT lymphoma with trisomy 18. We recommend that FCM and FISH analyses of biopsy specimens be considered for diagnosing gastric MALT lymphoma if this diagnosis is suspected based on endoscopic findings.

Reviews of the In-situ Demonstration Test of the Engineered Barrier System in Many Countries

Energy Technology Data Exchange (ETDEWEB)

Lee, Minsoo; Choi, Heui Joo [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

2013-10-15

Many nations considering the deep geologic disposal of HLW are now planning or executing in-situ demonstration experiments on their regional EBS (Engineering barrier system) at their deep underground research facilities. The main purpose of the in-situ EBS test is the experimental confirmation of its performance, and the prediction of its long-term evolution through the modeling of EBS based on the experimental data. Additionally, the engineering feasibility for the construction of an engineering barrier system can also be checked through full scale construction of an in-situ test. KAERI is currently preparing an in-situ test at a large 1/3 scale, called IN-DEBS (In-situ Demonstration of EBS) at KURT (KAERI Underground Research Tunnel) for the generic EBS suggested in A-KRS (Advanced KAERI Reference System), which was developed to treat the HLW from pyroprocessing. As the first step for the design of IN-DEBS, the foreign in-situ demonstrations of EBS were reviewed in this paper. The demonstration projects, which were completed or are still being executed in some countries such as Sweden, France, Finland, Canada, Belgium, Switzerland, Spain, and Japan, were surveyed and summarized. In particular, hardware constitutions such as the heating element or compact bentonite, and the experimental procedures, have focused more on reviews than on experimental results in this survey, since their hardware information is very important for the design of the IN-DEBS.

Reviews of the In-situ Demonstration Test of the Engineered Barrier System in Many Countries

International Nuclear Information System (INIS)

Lee, Minsoo; Choi, Heui Joo

2013-01-01

Many nations considering the deep geologic disposal of HLW are now planning or executing in-situ demonstration experiments on their regional EBS (Engineering barrier system) at their deep underground research facilities. The main purpose of the in-situ EBS test is the experimental confirmation of its performance, and the prediction of its long-term evolution through the modeling of EBS based on the experimental data. Additionally, the engineering feasibility for the construction of an engineering barrier system can also be checked through full scale construction of an in-situ test. KAERI is currently preparing an in-situ test at a large 1/3 scale, called IN-DEBS (In-situ Demonstration of EBS) at KURT (KAERI Underground Research Tunnel) for the generic EBS suggested in A-KRS (Advanced KAERI Reference System), which was developed to treat the HLW from pyroprocessing. As the first step for the design of IN-DEBS, the foreign in-situ demonstrations of EBS were reviewed in this paper. The demonstration projects, which were completed or are still being executed in some countries such as Sweden, France, Finland, Canada, Belgium, Switzerland, Spain, and Japan, were surveyed and summarized. In particular, hardware constitutions such as the heating element or compact bentonite, and the experimental procedures, have focused more on reviews than on experimental results in this survey, since their hardware information is very important for the design of the IN-DEBS

Development of an in situ hybridization assay for the detection of ostreid herpesvirus type 1 mRNAs in the Pacific oyster, Crassostrea gigas.

Science.gov (United States)

Corbeil, Serge; Faury, Nicole; Segarra, Amélie; Renault, Tristan

2015-01-01

An in situ hybridization protocol for detecting mRNAs of ostreid herpesvirus type 1 (OsHV-1) which infects Pacific oysters, Crassostrea gigas, was developed. Three RNA probes were synthesized by cloning three partial OsHV-1 genes into plasmids using three specific primer pairs, and performing a transcription in the presence of digoxigenin dUTP. The RNA probes were able to detect the virus mRNAs in paraffin sections of experimentally infected oysters 26 h post-injection. The in situ hybridization showed that the OsHV-1 mRNAs were mainly present in connective tissues in gills, mantle, adductor muscle, digestive gland and gonads. DNA detection by in situ hybridization using a DNA probe and viral DNA quantitation by real-time PCR were also performed and results were compared with those obtained using RNA probes. Copyright © 2014 Elsevier B.V. All rights reserved.

In-Space Hybrid Energy Storage System Demonstration on CSUNSat1

Data.gov (United States)

National Aeronautics and Space Administration — Objective: Flight demonstrate the JPL Hybrid Energy Storage System (low temperature, high power) using the SSTP-funded CSUNSat1 CubeSat: Environmental testing...

Management Plan: Demonstration testing and evaluation of in situ soil heating

International Nuclear Information System (INIS)

Dev, H.

1993-01-01

In this project IITRI will demonstrate an in situ soil heating technology for the removal of hazardous organic contaminants present in the soil. In Situ heating will be accomplished by the application of 60 Hz AC power to the soil. The soil will be heated to a temperature of about 90 degree C. This technology is suited for the removal of those organic compounds which have a normal boiling point in the range of 100 degree to 210 degree C, or else for those which exhibit a pure component vapor pressure of at least 10 mm Hg in the 90 degree to 100 degree C temperature range. For example, perchloroethylene, dichlorobenzene, trichlorobenzene, etc. may be removed by in situ AC heating. It is planned to demonstrate the technology by heating approximately 400 tons of soil in the K-1070 Classified Burial Ground located at DOE's K-25 Site located in Oak Ridge, TN. It is estimated that the heating portion of the demonstration will take approximately 3 weeks at an average power input rate of 150 to 175 kill. IITRI expects to spend considerable time in the front end reviewing site characteristics, preparing detail design, developing Health and Safety Plans and other documents needed to obtain regulatory approval for the demonstration, arranging for site sampling, infrastructure development and document preparation. It is anticipated that site activities will begin in approximately 5 to 6 months. This contract was signed on September 30, 1993. IITRI started work on it in October 1993. It is planned to complete the demonstration and submit approved final reports by September 30, 1994. This project has 12 tasks and four major milestones. The major milestones and their planned completion dates are presented

Detection of genetic changes in Barrett's adenocarcinoma and Barrett's esophagus by DNA in situ hybridization and immunohistochemistry

NARCIS (Netherlands)

Krishnadath, K. K.; Tilanus, H. W.; Alers, J. C.; Mulder, A. H.; van Dekken, H.

1994-01-01

We have investigated the occurrence of chromosomal DNA and cell cycle-related protein changes in Barrett's epithelium and adenocarcinoma. The presence of numerical chromosomal aberrations was studied by applying nonisotopic in situ hybridization (ISH) with (peri-)centromeric DNA probes, specific for

Synthesis and luminescence properties of hybrid organic-inorganic transparent titania thin film activated by in-situ formed lanthanide complexes

International Nuclear Information System (INIS)

Wang Yige; Wang Li; Li Huanrong; Liu Peng; Qin Dashan; Liu Binyuan; Zhang Wenjun; Deng Ruiping; Zhang Hongjie

2008-01-01

Stable transparent titania thin films were fabricated at room temperature by combining thenoyltrifluoroacetone (TTFA)-modified titanium precursors with amphiphilic triblock poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO, P123) copolymers. The obtained transparent titania thin films were systematically investigated by IR spectroscopy, PL emission and excitation spectroscopy and transmission electron microscopy. IR spectroscopy indicates that TTFA coordinates the titanium center during the process of hydrolysis and condensation. Luminescence spectroscopy confirms the in-situ formation of lanthanide complexes in the transparent titania thin film. TEM image shows that the in-situ formed lanthanide complexes were homogeneously distributed throughout the whole thin film. The quantum yield and the number of water coordinated to lanthanide metal center have been theoretically determined based on the luminescence data. - Graphical abstract: Novel stable luminescent organic-inorganic hybrid titania thin film with high transparency activated by in-situ formed lanthanide complexes have been obtained at room temperature via a simple one-pot synthesis approach by using TTFA-modified titanium precursor with amphiphilic triblock copolymer P123. The obtained hybrid thin film displays bright red (or green), near-monochromatic luminescence due to the in-situ formed lanthanide complex

Applications of ribosomal in situ hybridization for the study of bacterial cells in the mouse intestine

DEFF Research Database (Denmark)

Licht, Tine Rask; Poulsen, Lars Kongsbak; Molin, Søren

1997-01-01

Localization of E. coli and S. typhimurium in the large and small intestine of streptomycin-treated mice was visualized by in situ hybridization with specific rRNA target probes and epi-fluorescence microscopy. Growth rates of E. coli BJ4 colonizing the large intestine of streptomycin-treated mic...

Parker Hybrid Hydraulic Drivetrain Demonstration

Energy Technology Data Exchange (ETDEWEB)

Collett, Raymond [Parker-Hannifin Corporation, Cleveland, OH (United States); Howland, James [Parker-Hannifin Corporation, Cleveland, OH (United States); Venkiteswaran, Prasad [National Energy Technology Lab. (NETL), Morgantown, WV (United States)

2014-03-31

This report examines the benefits of Parker Hannifin hydraulic hybrid brake energy recovery systems used in commercial applications for vocational purposes. A detailed background on the problem statement being addressed as well as the solution set specific for parcel delivery will be provided. Objectives of the demonstration performed in high start & stop applications included opportunities in fuel usage reduction, emissions reduction, vehicle productivity, and vehicle maintenance. Completed findings during the demonstration period and parallel investigations with NREL, CALSTART, along with a literature review will be provided herein on this research area. Lastly, results identified in the study by third parties validated the savings potential in fuel reduction of on average of 19% to 52% over the baseline in terms of mpg (Lammert, 2014, p11), Parker data for parcel delivery vehicles in the field parallels this at a range of 35% - 50%, emissions reduction of 17.4% lower CO2 per mile and 30.4% lower NOx per mile (Gallo, 2014, p15), with maintenance improvement in the areas of brake and starter replacement, while leaving room for further study in the area of productivity in terms of specific metrics that can be applied and studied.

In situ 3D nanoprinting of free-form coupling elements for hybrid photonic integration

Science.gov (United States)

Dietrich, P.-I.; Blaicher, M.; Reuter, I.; Billah, M.; Hoose, T.; Hofmann, A.; Caer, C.; Dangel, R.; Offrein, B.; Troppenz, U.; Moehrle, M.; Freude, W.; Koos, C.

2018-04-01

Hybrid photonic integration combines complementary advantages of different material platforms, offering superior performance and flexibility compared with monolithic approaches. This applies in particular to multi-chip concepts, where components can be individually optimized and tested. The assembly of such systems, however, requires expensive high-precision alignment and adaptation of optical mode profiles. We show that these challenges can be overcome by in situ printing of facet-attached beam-shaping elements. Our approach allows precise adaptation of vastly dissimilar mode profiles and permits alignment tolerances compatible with cost-efficient passive assembly techniques. We demonstrate a selection of beam-shaping elements at chip and fibre facets, achieving coupling efficiencies of up to 88% between edge-emitting lasers and single-mode fibres. We also realize printed free-form mirrors that simultaneously adapt beam shape and propagation direction, and we explore multi-lens systems for beam expansion. The concept paves the way to automated assembly of photonic multi-chip systems with unprecedented performance and versatility.

Two-dimensional Semiconductor-Superconductor Hybrids

DEFF Research Database (Denmark)

Suominen, Henri Juhani

This thesis investigates hybrid two-dimensional semiconductor-superconductor (Sm-S) devices and presents a new material platform exhibiting intimate Sm-S coupling straight out of the box. Starting with the conventional approach, we investigate coupling superconductors to buried quantum well....... To overcome these issues we integrate the superconductor directly into the semiconducting material growth stack, depositing it in-situ in a molecular beam epitaxy system under high vacuum. We present a number of experiments on these hybrid heterostructures, demonstrating near unity interface transparency...

Conceptual design of a Demonstration Tokamak Hybrid Reactor (DTHR), September 1978

International Nuclear Information System (INIS)

Kelley, J.L.

1978-12-01

The flexibility of the fusion hybrid reactor to function as a fuel production facility, power plant, waste disposal burner or combinations of all of these, as well as the reactor's ability to use proliferation resistant fuel cycles, has provided the incentive to assess the feasibility of a near-term demonstration plant. The goals for a Demonstration Tokamak Hybrid Reactor (DTHR) were established and an initial conceptual design was selected. Reactor performance and economics were evaluated and key developmental issues were assessed. The study has shown that a DTHR is feasible in the late 1980's, a significant quantity of fissile fuel could be produced from fertile thorium using present day fission reactor blanket technology, and a large number of commercially prototypical components and systems could be developed and operationally verified. The DTHR concept would not only serve as proof-of-principle for hybrid technology, but could be operated in the ignited mode and provide major advancements for pure fusion technology

Immunoglobulin heavy-chain fluorescence in situ hybridization-chromogenic in situ hybridization DNA probe split signal in the clonality assessment of lymphoproliferative processes on cytological samples.

Science.gov (United States)

Zeppa, Pio; Sosa Fernandez, Laura Virginia; Cozzolino, Immacolata; Ronga, Valentina; Genesio, Rita; Salatiello, Maria; Picardi, Marco; Malapelle, Umberto; Troncone, Giancarlo; Vigliar, Elena

2012-12-25

The human immunoglobulin heavy-chain (IGH) locus at chromosome 14q32 is frequently involved in different translocations of non-Hodgkin lymphoma (NHL), and the detection of any breakage involving the IGH locus should identify a B-cell NHL. The split-signal IGH fluorescence in situ hybridization-chromogenic in situ hybridization (FISH-CISH) DNA probe is a mixture of 2 fluorochrome-labeled DNAs: a green one that binds the telomeric segment and a red one that binds the centromeric segment, both on the IGH breakpoint. In the current study, the authors tested the capability of the IGH FISH-CISH DNA probe to detect IGH translocations and diagnose B-cell lymphoproliferative processes on cytological samples. Fifty cytological specimens from cases of lymphoproliferative processes were tested using the split-signal IGH FISH-CISH DNA probe and the results were compared with light-chain assessment by flow cytometry (FC), IGH status was tested by polymerase chain reaction (PCR), and clinicohistological data. The signal score produced comparable results on FISH and CISH analysis and detected 29 positive, 15 negative, and 6 inadequate cases; there were 29 true-positive cases (66%), 9 true-negative cases (20%), 6 false-negative cases (14%), and no false-positive cases (0%). Comparing the sensitivity of the IGH FISH-CISH DNA split probe with FC and PCR, the highest sensitivity was obtained by FC, followed by FISH-CISH and PCR. The split-signal IGH FISH-CISH DNA probe is effective in detecting any translocation involving the IGH locus. This probe can be used on different samples from different B-cell lymphoproliferative processes, although it is not useful for classifying specific entities. Cancer (Cancer Cytopathol) 2012;. © 2012 American Cancer Society. Copyright © 2012 American Cancer Society.

Sensor Network Demonstration for In Situ Decommissioning - 13332

Energy Technology Data Exchange (ETDEWEB)

Lagos, L.; Varona, J.; Awwad, A. [Applied Research Center, Florida International University, 10555 West Flagler Street, Suite 2100, Miami, FL 33174 (United States); Rivera, J.; McGill, J. [Department of Energy - DOE, Environmental Management Office (United States)

2013-07-01

Florida International University's (FIU's) Applied Research Center is currently supporting the Department of Energy's (DOE) Environmental Management Office of D and D and Facility Engineering program. FIU is supporting DOE's initiative to improve safety, reduce technical risks, and limit uncertainty within D and D operations by identifying technologies suitable to meet specific facility D and D requirements, assessing the readiness of those technologies for field deployment, and conducting feasibility studies and large scale demonstrations of promising technologies. During FY11, FIU collaborated with Savannah River National Laboratory in the development of an experimental test site for the demonstration of multiple sensor systems for potential use in the in situ decommissioning process. In situ decommissioning is a process in which the above ground portion of a facility is dismantled and removed, and the underground portion is filled with a cementious material such as grout. In such a scenario, the question remains on how to effectively monitor the structural health of the grout (cracking, flexing, and sinking), as well as track possible migration of contaminants within and out of the grouted monolith. The right types of sensors can aid personnel in better understanding the conditions within the entombed structure. Without sensors embedded in and around the monolith, it will be very difficult to estimate structural integrity and contaminant transport. Yet, to fully utilize the appropriate sensors and the provided data, their performance and reliability must be evaluated outside a laboratory setting. To this end, a large scale experimental setup and demonstration was conducted at FIU. In order to evaluate a large suite of sensor systems, FIU personnel designed and purchased a pre-cast concrete open-top cube, which served as a mock-up of an in situ DOE decommissioned facility. The inside of the cube measures 10 ft x 10 ft x 8 ft. In order to

RNA detection in situ with FISH-STICs.

Science.gov (United States)

Sinnamon, John R; Czaplinski, Kevin

2014-02-01

The ability to detect RNA molecules in situ has long had important applications for molecular biological studies. Enzyme or dye-labeled antisense in vitro runoff transcripts and synthetic oligodeoxynucleotides (ODN) both have a proven track record of success, but each of these also has scientific and practical drawbacks and limitations to its use. We devised a means to use commercially synthesized oligonucleotides as RNA-FISH probes without further modification and show that such probes work well for detection of RNA in cultured cells. This approach can bind a high concentration of fluorescent ODN to a short stretch of an RNA using commercial DNA synthesis outlets available to any laboratory. We call this approach for creating in situ hybridization probes Fluorescence In Situ Hybridization with Sequential Tethered and Intertwined ODN Complexes (FISH-STICs). We demonstrate that one FISH-STIC probe can detect mRNA molecules in culture, and that probe detection can be improved by the addition of multiple probes that can be easily adapted for robust mRNA quantification. Using FISH-STICs, we demonstrate a nonoverlapping distribution for β-actin and γ-actin mRNA in cultured fibroblasts, and the detection of neuron-specific transcripts within cultured primary hippocampal neurons.

Detection of Methylobacterium radiotolerans IMBG290 in potato plants by in situ hybridization

OpenAIRE

Pirttila A. M.; Kozyrovska N. O.; Ovcharenko L. P.; Podolich O. V.

2009-01-01

A new bacterial strain of pink-pigmented facultative methylotroph (M. radiotolerans IMBG290) which was previously isolated from in vitro grown potato plantlets after their inoculation with Pseudomonas fluorescens IMBG163 was detected in tissues by in situ hybridization method (ISH/FISH). The presence of Methylobacterium rRNA was observed in leaves and stems of potato plantlets, whereas no signal was detected in potato roots. The signal was less abundant in the untreated plants than in the pla...

Dealing with the problem of non-specific in situ mRNA hybridization signals associated with plant tissues undergoing programmed cell death

Directory of Open Access Journals (Sweden)

Jokela Anne

2010-02-01

Full Text Available Abstract Background In situ hybridization is a general molecular method typically used for the localization of mRNA transcripts in plants. The method provides a valuable tool to unravel the connection between gene expression and anatomy, especially in species such as pines which show large genome size and shortage of sequence information. Results In the present study, expression of the catalase gene (CAT related to the scavenging of reactive oxygen species (ROS and the polyamine metabolism related genes, diamine oxidase (DAO and arginine decarboxylase (ADC, were localized in developing Scots pine (Pinus sylvestris L. seeds. In addition to specific signals from target mRNAs, the probes continually hybridized non-specifically in the embryo surrounding region (ESR of the megagametophyte tissue, in the remnants of the degenerated suspensors as well as in the cells of the nucellar layers, i.e. tissues exposed to cell death processes and extensive nucleic acid fragmentation during Scots pine seed development. Conclusions In plants, cell death is an integral part of both development and defence, and hence it is a common phenomenon in all stages of the life cycle. Our results suggest that extensive nucleic acid fragmentation during cell death processes can be a considerable source of non-specific signals in traditional in situ mRNA hybridization. Thus, the visualization of potential nucleic acid fragmentation simultaneously with the in situ mRNA hybridization assay may be necessary to ensure the correct interpretation of the signals in the case of non-specific hybridization of probes in plant tissues.

A Storable, Hybrid Mars Ascent Vehicle Technology Demonstrator for the 2020 Launch Opportunity

Science.gov (United States)

Chandler, A. A.; Karabeyoglu, M. A.; Cantwell, B. J.; Reeve, R.; Goldstein, B. G.; Hubbard, G. S.

2012-06-01

A Phoenix sized mission including a reduced payload, two-stage, hybrid Mars Ascent Vehicle technology demonstrator is proposed for the 2020 opportunity. The hybrid MAV is storable on Mars and would retire risk for a Mars Sample Return campaign.

Metaphase FISH on a Chip: Miniaturized Microfluidic Device for Fluorescence in situ Hybridization

Directory of Open Access Journals (Sweden)

Niels Tommerup

2010-11-01

Full Text Available Fluorescence in situ Hybridization (FISH is a major cytogenetic technique for clinical genetic diagnosis of both inherited and acquired chromosomal abnormalities. Although FISH techniques have evolved and are often used together with other cytogenetic methods like CGH, PRINS and PNA-FISH, the process continues to be a manual, labour intensive, expensive and time consuming technique, often taking over 3–5 days, even in dedicated labs. We have developed a novel microFISH device to perform metaphase FISH on a chip which overcomes many shortcomings of the current laboratory protocols. This work also introduces a novel splashing device for preparing metaphase spreads on a microscope glass slide, followed by a rapid adhesive tape-based bonding protocol leading to rapid fabrication of the microFISH device. The microFISH device allows for an optimized metaphase FISH protocol on a chip with over a 20-fold reduction in the reagent volume. This is the first demonstration of metaphase FISH on a microfluidic device and offers a possibility of automation and significant cost reduction of many routine diagnostic tests of genetic anomalies.

Identification of MYCN gene amplification in neuroblastoma using chromogenic in situ hybridization (CISH): an alternative and practical method.

Science.gov (United States)

Bhargava, Rohit; Oppenheimer, Orit; Gerald, William; Jhanwar, Suresh C; Chen, Beiyun

2005-06-01

Chromogenic in situ hybridization (CISH) is a recently developed technique, which utilizes the general principles of in situ hybridization and a detection system similar to immunohistochemistry. To assess the utility of CISH for analysis of MYCN gene amplification, we compared this assay with established diagnostic assays such as Southern blot analysis (SB) and fluorescent in situ hybridization (FISH). CISH was performed on 67 cases of neuroblastoma using tissue microarray (65 cases) and whole tissue sections (2 cases). Unequivocal, high-level amplification (> or =10 gene copies per tumor nucleus) was identified in 19 of 67 (28.4%) tumors. Two (3%) tumors showed low-level amplification (6-9 gene copies per tumor nucleus). No amplification was seen in 46 of 67 (68.6%) tumors. SB data were available in 44 tumors. Forty-one of the 44 tumors (93%) showed concordant results between CISH and SB. Three tumors showed MYCN amplification by CISH but no amplification by SB, most likely due to dilution effect of nonneoplastic tissue in the test samples. Two of these three tumors also showed MYCN amplification by FISH, and the third tumor was not analyzed by FISH. FISH data were available in total of 30 tumors. All 30 tumors showed concordant results between CISH and FISH for classifying a tumor as MYCN amplified or not amplified. We conclude that CISH is an accurate method for determining MYCN gene amplification, with added advantages that make it a more practically useful method.

Localization of tRNAsup(asp)2 genes from Drosophila melanogaster by 'in situ' hybridization

International Nuclear Information System (INIS)

Schmidt, T.; Egg, A.H.; Kubli, E.

1978-01-01

Transfer RNAsup(asp) 2 delta was isolated from Drosophila melanogaster by affinity chromatography on concanavalin A-Sepharose. The tRNA was iodinated 'in vitro' with Na[ 125 I] and hybridized 'in situ' to salivary gland chromosomes from Drosophila. Subsequent autoradiography allowed the localization of the genes for tRNAsup(asp) 2 delta to the left arm of the second chromosome in the regions 29 D and E. (orig.) [de

High sensitive diagnostic technique for virus gene using radioisotope. Development of PCR in situ hybridization and its application

Energy Technology Data Exchange (ETDEWEB)

Iwasaki, Takuya; Sada, Tetsutaro; Terai, Masanori; Sato, Yuko; Kurata, Takeshi [National Inst. of Infectious Diseases, Tokyo (Japan); Yamaguchi, Kazuyoshi; Yanagisawa, Akio; Sakai, Yuzo

1998-02-01

An animal model, experimentally induced squamous cell carcinoma was produced in hamster mucosa to establish a carcinogenic system inducible by interaction of a virus and chemical(s). Human hydatid mole and cholioepithelioma were chosen as the target subjects. Several tumor cells and host cells were isolated under microscopy and DNA was extracted from these cells to indentify the respective origins (maternal, paternal or host origin). The base sequence of HLADRB region was analyzed by PCR using synthetic primer and the tissue localization was examined by PCR in situ hybridization. A PCR product of 82 bp was found in 15 of 17 samples and there were 2 samples in which the product was not detectable with the constructed primer and 6 samples were negative. While significant differences were not observed by in situ hybridization compared with the negative control. (M.N.)

High sensitive diagnostic technique for virus gene using radioisotope. Development of PCR in situ hybridization and its application

International Nuclear Information System (INIS)

Iwasaki, Takuya; Sada, Tetsutaro; Terai, Masanori; Sato, Yuko; Kurata, Takeshi; Yamaguchi, Kazuyoshi; Yanagisawa, Akio; Sakai, Yuzo

1998-01-01

An animal model, experimentally induced squamous cell carcinoma was produced in hamster mucosa to establish a carcinogenic system inducible by interaction of a virus and chemical(s). Human hydatid mole and cholioepithelioma were chosen as the target subjects. Several tumor cells and host cells were isolated under microscopy and DNA was extracted from these cells to indentify the respective origins (maternal, paternal or host origin). The base sequence of HLADRB region was analyzed by PCR using synthetic primer and the tissue localization was examined by PCR in situ hybridization. A PCR product of 82 bp was found in 15 of 17 samples and there were 2 samples in which the product was not detectable with the constructed primer and 6 samples were negative. While significant differences were not observed by in situ hybridization compared with the negative control. (M.N.)

Diagnosis, prognosis and disease management using in situ hybridization

International Nuclear Information System (INIS)

Kucheria, K.; Talwar, R.

2002-01-01

The year 2001 saw unveiling of anatomy of the human genome with sequencing of 90% of the euchromatic region. But the ultimate goal of the Human Genome Project to delineate the positions of all genes is yet to be achieved. In Situ Hybridization (ISH) is one of the methods that help in localizing genes on chromosomes. The present study aimed to use radioactive- and fluorescent-labeled probes for screening various congenital anomalies (sex chromosomal and autosomal), for prenatal diagnosis and cancer genetics. Standard techniques were used for hybridization with radioactively and fluorescent labeled probes. Sex chromosome aneuploidies (XXY, XO, XXX, XYY etc.) were analyzed using centromeric probes for chromosomes X and Y. The cases with ambiguous genitalia were further analyzed using probe specific for the sex-determining region (SRY) on the Y chromosome. Suspected cases of Down syndrome were analyzed using probe specific for centromeric region of chromosome 21 to confirm trisomy 21. Prenatal diagnosis included screening aneuploidies of chromosomes 13, 18, 21, X and Y on uncultured cells and metaphases obtained from amniotic fluid and chorionic villi samplings. Gene alterations were also studied in Retinoblastoma patients, Chronic Myeloid Leukemia (CML) and Acute Promyelocytic Leukemia (APML) using probes specific for Rb1, bcr/abl and PML/RARα genes respectively. Response to therapy was assessed by evaluating minimal residual disease (MRD) in leukemia patients. Attempts were also made to analyze cells obtained from buccal mucosa and bladder epithelium that could facilitate rapid screening of sex chromosome anomalies and bladder cancer without painful invasive techniques. Prenatal diagnosis using ISH on uncultured cells could provide an accurate and rapid result. These results of prenatal diagnosis were in conformation with results of conventional cytogenetics obtained after long-term cultures. Molecular rearrangements that could not be detected with conventional

Distribution of 45S rDNA in Modern Rose Cultivars (Rosa hybrida), Rosa rugosa, and Their Interspecific Hybrids Revealed by Fluorescence in situ Hybridization.

Science.gov (United States)

Ding, Xiao-Liu; Xu, Ting-Liang; Wang, Jing; Luo, Le; Yu, Chao; Dong, Gui-Min; Pan, Hui-Tang; Zhang, Qi-Xiang

2016-01-01

To elucidate the evolutionary dynamics of the location and number of rDNA loci in the process of polyploidization in the genus Rosa, we examined 45S rDNA sites in the chromosomes of 6 modern rose cultivars (R. hybrida), 5 R. rugosa cultivars, and 20 hybrid progenies by fluorescence in situ hybridization. Variation in the number of rDNA sites in parents and their interspecific hybrids was detected. As expected, 4 rDNA sites were observed in the genomes of 4 modern rose cultivars, while 3 hybridization sites were observed in the 2 others. Two expected rDNA sites were found in 2 R. rugosa cultivars, while in the other 3 R. rugosa cultivars 4 sites were present. Among the 20 R. hybrida × R. rugosa offspring, 13 carried the expected number of rDNA sites, and 1 had 6 hybridization sites, which exceeded the expected number by far. The other 6 offspring had either 2 or 3 hybridization sites, which was less than expected. Differences in the number of rDNA loci were observed in interspecific offspring, indicating that rDNA loci exhibit instability after distant hybridization events. Abnormal chromosome pairing may be the main factor explaining the variation in rDNA sites during polyploidization. © 2016 S. Karger AG, Basel.

Combined Confocal and Wide-Field High-Resolution Cytometry of Fluorescent In Situ Hybridization-Stained Cells

Czech Academy of Sciences Publication Activity Database

Kozubek, Michal; Kozubek, Stanislav; Lukášová, Emilie; Bártová, Eva; Skalníková, M.; Matula, Pa.; Matula, Pe.; Jirsová, Pavla; Cafourková, Alena; Koutná, Irena

2001-01-01

Roč. 45, č. 1 (2001), s. 1-12 ISSN 0196-4763 R&D Projects: GA MŠk VS97031; GA ČR GA202/99/P008; GA AV ČR IBS5004010 Institutional research plan: CEZ:AV0Z5004920 Keywords : high-resolution cytometry * fluorescence in situ hybridization * interphase nuclei Subject RIV: BO - Biophysics Impact factor: 2.220, year: 2001

Fluorescence In Situ Hybridization for MicroRNA Detection in Archived Oral Cancer Tissues

Directory of Open Access Journals (Sweden)

Zonggao Shi

2012-01-01

Full Text Available The noncoding RNA designated as microRNA (miRNA is a large group of small single-stranded regulatory RNA and has generated wide-spread interest in human disease studies. To facilitate delineating the role of microRNAs in cancer pathology, we sought to explore the feasibility of detecting microRNA expression in formalin-fixed paraffin-embedded (FFPE tissues. Using FFPE materials, we have compared fluorescent in situ hybridization (FISH procedures to detect miR-146a with (a different synthetic probes: regular custom DNA oligonucleotides versus locked nucleic acid (LNA incorporated DNA oligonucleotides; (b different reporters for the probes: biotin versus digoxigenin (DIG; (c different visualization: traditional versus tyramide signal amplification (TSA system; (d different blocking reagents for endogenous peroxidase. Finally, we performed miR-146a FISH on a commercially available oral cancer tissue microarray, which contains 40 cases of oral squamous cell carcinoma (OSCC and 10 cases of normal epithelia from the human oral cavity. A sample FISH protocol for detecting miR-146a is provided. In summary, we have established reliable in situ hybridization procedures for detecting the expression of microRNA in FFPE oral cancer tissues. This method is an important tool for studies on the involvement of microRNA in oral cancer pathology and may have potential prognostic or diagnostic value.

Ultrastructural localization of human papilloma virus by nonradioactive in situ hybridization on tissue of human cervical intraepithelial neoplasia

DEFF Research Database (Denmark)

Multhaupt, H A; Rafferty, P A; Warhol, M J

1992-01-01

BACKGROUND: A nonradioactive in situ hybridization was developed to localize human papilloma virus (HPV) at the ultrastructural level. EXPERIMENTAL DESIGN: Cervical biopsies from human uterine cervices clinically suspicious of condyloma were embedded in Lowicryl K4M at low temperature...

Heavy ion-induced chromosomal aberrations analyzed by fluorescence in situ hybridization

International Nuclear Information System (INIS)

Durante, M.; Gialanella, G.; Grossi, G.; Pugliese, M.; Cella, L.; Greco, O.; George, K.; Yang, T.C.

1997-01-01

We have investigated the effectiveness of heavy ions in the induction of chromosomal aberrations in mammalian cells by the recent technique of fluorescence in situ hybridization (FISH) with whole-chromosome probes. FISH-painting was used both in metaphase and interphase (prematurely condensed) chromosomes. The purpose of our experiments was to address the following problems: (a) the ratio of different types of aberrations as a function of radiation quality (search for biomarkers); (b) the ratio between aberrations scored in interphase and metaphase as a function of radiation quality (role of apoptosis); (c) differences between cytogenetic effects produced by different ions at the same LET (role of track structure). (orig./MG)

Heavy ion-induced chromosomal aberrations analyzed by fluorescence in situ hybridization

Energy Technology Data Exchange (ETDEWEB)

Durante, M; Gialanella, G; Grossi, G; Pugliese, M [Univ. ` ` Federico II` ` , Naples (Italy). Dept. of Physics; [INFN, Naples (Italy); Cella, L; Greco, O [Univ. ` ` Federico II` ` , Naples (Italy). Dept. of Physics; Furusawa, Y [NIRS, Chiba (Japan); George, K; Yang, T C [NASA Lyndon B. Johnson Space Center, Houston, TX (United States)

1997-09-01

We have investigated the effectiveness of heavy ions in the induction of chromosomal aberrations in mammalian cells by the recent technique of fluorescence in situ hybridization (FISH) with whole-chromosome probes. FISH-painting was used both in metaphase and interphase (prematurely condensed) chromosomes. The purpose of our experiments was to address the following problems: (a) the ratio of different types of aberrations as a function of radiation quality (search for biomarkers); (b) the ratio between aberrations scored in interphase and metaphase as a function of radiation quality (role of apoptosis); (c) differences between cytogenetic effects produced by different ions at the same LET (role of track structure). (orig./MG)

In-situ preparation of NaA zeolite/chitosan porous hybrid beads for removal of ammonium from aqueous solution.

Science.gov (United States)

Yang, Kai; Zhang, Xiang; Chao, Cong; Zhang, Bing; Liu, Jindun

2014-07-17

Inorganic/organic hybrid materials play important roles in removal of contaminants from wastewater. Herein, we used the natural materials of halloysite and chitosan to prepare a new adsorbent of NaA zeolite/chitosan porous hybrid beads by in-situ hydrothermal synthesis method. SEM indicated that the porous hybrid beads were composed of 6-8 μm sized cubic NaA zeolite particles congregated together with chitosan. The adsorption behavior of NH4(+) from aqueous solution onto hybrid beads was investigated at different conditions. The Langmuir and Freundlich adsorption models were applied to describe the equilibrium isotherms. A maximum adsorption capacity of 47.62 mg/g at 298 K was achieved according to Langmuir model. The regenerated or reused experiments indicated that the adsorption capacity of the hybrid beads could maintain in 90% above after 10 successive adsorption-desorption cycles. The high adsorption and reusable ability implied potential application of the hybrid beads for removing NH4(+) pollutants from wastewater. Copyright © 2014 Elsevier Ltd. All rights reserved.

Detection of Alien Oryza punctata Kotschy Chromosomes in Rice, Oryza sativa L., by Genomic in situ Hybridization

OpenAIRE

Yasui, Hideshi; Nonomura, Ken-ichi; Iwata, Nobuo; 安井, 秀; 野々村, 賢一; 岩田, 伸夫

1997-01-01

Genomic in situ hybridization (GIS H) using total Oryza punctata Kotschy genomic DNA as a probe was applied to detect alien chromosomes transferred from O. punctata (W1514: 2n=2x=24: BB) to O. sativa Japonica cultivar, Nipponbare (2n=2x=24: AA). Only 12 chromosomes in the interspecific hybrids (2n=3x=36: AAB) between autotetraploid of O. sativa cultivar Nipponbare and a diploid strain of O. punctata (W1514) showed intense staining by FITC in mitotic metaphase spreads. Only one homologous pair...

Nerve growth factor mRNA in brain: localization by in situ hybridization

International Nuclear Information System (INIS)

Rennert, P.D.; Heinrich, G.

1986-01-01

Nerve Growth Factor is a 118 amino acid polypeptide that plays an important role in the differentiation and survival of neurons. The recent discovery that a mRNA that encodes beta Nerve Growth Factor is present in brain suggests that the Nerve Growth Factor gene may not only regulate gene expression of peripheral but also of central neurons. To identify the site(s) of Nerve Growth Factor mRNA production in the brain and to determine which cells express the Nerve Growth Factor gene, the technique of in situ hybridization was employed. A 32P-labeled RNA probe complementary to Nerve Growth Factor mRNA hybridized to cells in the stratum granulosum of the dentate gyrus and the stratum pyramidale of the hippocampus. These observations identify for the first time cellular sites of Nerve Growth Factor gene expression in the central nervous system, and suggest that Nerve Growth Factor mRNA is produced by neurons

Putting the colours into chromogenic in situ hybridization (CISH).

Science.gov (United States)

Shipley, J

2006-09-01

Recurrent genomic alterations are the hallmarks of particular cancers. Application of molecular cytogenetic technologies to tumour material in order to detect these alterations has become important for molecular diagnostics and research. A dual-colour chromogenic in situ hybridization (dc-CISH) method described recently in the Journal of Pathology has the advantage of visualizing two probes simultaneously with the ability to discern morphological features. In addition, the bright field microscopy required is readily accessible to many laboratories. The approach has been validated by comparison of results with standard analyses for HER-2 amplification status in formalin-fixed, paraffin-embedded breast cancers and is applicable to the analysis of other clinically relevant genomic aberrations as well as of use in research investigations. Copyright (c) 2006 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Identification of Cannabis sativa L. using the 1-kbTHCA synthase-fluorescence in situ hybridization probe.

Science.gov (United States)

Jeangkhwoa, Pattraporn; Bandhaya, Achirapa; Umpunjun, Puangpaka; Chuenboonngarm, Ngarmnij; Panvisavas, Nathinee

2017-03-01

This study reports a successful application of fluorescence in situ hybridization (FISH) technique in the identification of Cannabis sativa L. cells recovered from fresh and dried powdered plant materials. Two biotin-16-dUTP-labeled FISH probes were designed from the Cannabis-specific tetrahydrocannabinolic acid synthase (THCAS) gene and the ITS region of the 45S rRNA gene. Specificity of probe-target hybridization was tested against the target and 4 non-target plant species, i.e., Humulus lupulus, Mitragyna speciosa, Papaver sp., and Nicotiana tabacum. The 1-kb THCA synthase hybridization probe gave Cannabis-specific hybridization signals, unlike the 700-bp Cannabis-ITS hybridization probe. Probe-target hybridization was also confirmed against 20 individual Cannabis plant samples. The 1-kb THCA synthase and 700-bp Cannabis-ITS hybridization probes clearly showed 2 hybridization signals per cell with reproducibility. The 1-kb THCA synthase probe did not give any FISH signal when tested against H. lupulus, its closely related member of the Canabaceae family. It was also showed that 1-kb THCA synthase FISH probe can be applied to identify small amount of dried powdered Cannabis material with an addition of rehydration step prior to the experimental process. This study provided an alternative identification method for Cannabis trace. Copyright © 2016. Published by Elsevier B.V.

Hybrid Perovskite Thin-Film Photovoltaics: In Situ Diagnostics and Importance of the Precursor Solvate Phases

KAUST Repository

Munir, Rahim

2016-11-07

Solution-processed hybrid perovskite semiconductors attract a great deal of attention, but little is known about their formation process. The one-step spin-coating process of perovskites is investigated in situ, revealing that thin-film formation is mediated by solid-state precursor solvates and their nature. The stability of these intermediate phases directly impacts the quality and reproducibility of thermally converted perovskite films and their photovoltaic performance.

Hybrid Perovskite Thin-Film Photovoltaics: In Situ Diagnostics and Importance of the Precursor Solvate Phases

KAUST Repository

Munir, Rahim; Sheikh, Arif D.; Abdelsamie, Maged; Hu, Hanlin; Yu, Liyang; Zhao, Kui; Kim, Taesoo; El Tall, Omar; Li, Ruipeng; Smilgies, Detlef M.; Amassian, Aram

2016-01-01

Solution-processed hybrid perovskite semiconductors attract a great deal of attention, but little is known about their formation process. The one-step spin-coating process of perovskites is investigated in situ, revealing that thin-film formation is mediated by solid-state precursor solvates and their nature. The stability of these intermediate phases directly impacts the quality and reproducibility of thermally converted perovskite films and their photovoltaic performance.

Evaluation of potential blanket concepts for a Demonstration Tokamak Hybrid Reactor

International Nuclear Information System (INIS)

Chapin, D.L.; Chi, J.W.H.; Kelly, J.L.

1978-01-01

An evaluation has been made of several different blanket concepts for use in a near-term Demonstration Tokamak Hybrid Reactor (DTHR), whose main objective would be to produce a significant amount of fissile fuel while demonstrating the feasibility of the tokamak hybrid reactor concept. The desirability of a simple design using proven technology plus a proliferation resistant fuel cycle led to the selection of a low temperature and pressure water-cooled, zircaloy clad ThO 2 blanket concept to breed 233 U. The nuclear performance and thermal-hydraulics characteristics of the blanket were evaluated to arrive at a consistent design. The blanket was found to be feasible for producing a significant amount of fissile fuel even with the limited operating conditions and blanket coverage in the DTHR

EnVision+, a new dextran polymer-based signal enhancement technique for in situ hybridization (ISH).

Science.gov (United States)

Wiedorn, K H; Goldmann, T; Henne, C; Kühl, H; Vollmer, E

2001-09-01

Seventy paraffin-embedded cervical biopsy specimens and condylomata were tested for the presence of human papillomavirus (HPV) by conventional in situ hybridization (ISH) and ISH with subsequent signal amplification. Signal amplification was performed either by a commercial biotinyl-tyramide-based detection system [GenPoint (GP)] or by the novel two-layer dextran polymer visualization system EnVision+ (EV), in which both EV-horseradish peroxidase (EV-HRP) and EV-alkaline phosphatase (EV-AP) were applied. We could demonstrate for the first time, that EV in combination with preceding ISH results in a considerable increase in signal intensity and sensitivity without loss of specificity compared to conventional ISH. Compared to GP, EV revealed a somewhat lower sensitivity, as measured by determination of the integrated optical density (IOD) of the positively stained cells. However, EV is easier to perform, requires a shorter assay time, and does not raise the background problems that may be encountered with biotinyl-tyramide-based amplification systems. (J Histochem Cytochem 49:1067-1071, 2001)

Human chromosome-specific changes in a human-hamster hybrid cell line (AL) assessed by fluorescent in situ hybridization (fish)

International Nuclear Information System (INIS)

Geard, Charles R.; Jenkins, Gloria

1995-01-01

Purpose: To quantitatively assess all gamma-ray induced chromosomal changes confined to one human chromosome using fluorescence microscopy and in situ hybridization with a fluorescently labeled human chromosome specific nucleic acid probe. Methods and Materials: Synchronized human-hamster hybrid cells containing human chromosome 11 were obtained by a modified mitotic shake-off procedure. G1 phase cells (> 95%) were irradiated with 137 Cs gamma rays (0, 0.5, 1.0, 1.5, 2.0, 4.0, 6.0, 8.0, and 10.0 Gy) at a dose rate of 1.1 Gy/min and mitotic cells collected 16-20 h later; chromosomal spreads were prepared, denatured, and hybridized with a fluorescein-tagged nucleic acid probe against total human DNA. Chromosomes were examined by fluorescence microscopy and all categories of change involving the human chromosome 11 as target, recorded. Results: Overall, of the 3104 human-hamster hybrid cells examined, 82.1% were euploid, of which 88.6% contained one copy of human chromosome 11, 6.2% contained two copies, and 5.2% contained 0 copies. This is compatible with mitotic nondisjunction in a small fraction of cells. Of the remaining 17.9% of cells, 85.2% were tetraploid cells with two copies of human chromosome 11. For all aberrations involving human chromosome 11 there was a linear relationship between yield and absorbed dose of 0.1 aberrations per chromosome per Gy. The yield of dicentrics, translocations, and terminal deletions that involve one lesion on the human chromosome was linear, while the yield of interstitial deletions that arise from two interacting lesions on the human chromosome was curvilinear. The frequencies of dicentrics and translocations were about equal, while there was a high (40-60%) incidence of incomplete exchanges between human and hamster chromosomes. Conclusions: Fluorescent in situ hybridization (FISH) procedures allow for the efficient detection of a broad range of induced changes in target chromosomes. Symmetrical exchanges induced in G1

Detection of Methylobacterium radiotolerans IMBG290 in potato plants by in situ hybridization

Directory of Open Access Journals (Sweden)

Pirttila A. M.

2009-04-01

Full Text Available A new bacterial strain of pink-pigmented facultative methylotroph (M. radiotolerans IMBG290 which was previously isolated from in vitro grown potato plantlets after their inoculation with Pseudomonas fluorescens IMBG163 was detected in tissues by in situ hybridization method (ISH/FISH. The presence of Methylobacterium rRNA was observed in leaves and stems of potato plantlets, whereas no signal was detected in potato roots. The signal was less abundant in the untreated plants than in the plantlets infected with M. radiotolerans IMBG290.

Application of locked nucleic acid-based probes in fluorescence in situ hybridization

DEFF Research Database (Denmark)

Fontenete, Sílvia; Carvalho, Daniel R; Guimarães, Nuno

2016-01-01

of nucleic acid mimics used as mixmers in LNA-based probes strongly influence the efficiency of detection. LNA probes with 10 to 15 mers showed the highest efficiency. Additionally, the combination of 2′-OMe RNA with LNA allowed an increase on the fluorescence intensities of the probes. Overall......Fluorescence in situ hybridization (FISH) employing nucleic acid mimics as probes is becoming an emerging molecular tool in the microbiology area for the detection and visualization of microorganisms. However, the impact that locked nucleic acid (LNA) and 2′-O-methyl (2′-OMe) RNA modifications have...

Comparison of in situ hybridization methods for the assessment of HER-2/neu gene amplification status in breast cancer using a tissue microarray.

Science.gov (United States)

Malicka-Durczak, Anna; Korski, Konstanty; Ibbs, Matthew

2012-01-01

This project compared HER-2/neu gene status in breast cancers, as demonstrated by FISH (fluorescent in situ hybridization) and CISH (chromogenic in situ hybridization) and using a tissue microarray (TMA). The study also aimed to show whether the TMA technique could be used in clinical diagnostics, rather than remain a scientific tool. A TMA was constructed using 121 breast cancer specimens, 6 cores from each specimen. Demonstration and assessment of HER-2/neu gene status was by FISH (Vysis Path) and CISH (DAKO Duo CISH). The 121 breast cancer specimens were divided into 3 groups by HER-2 status, as determined by immunohistochemistry. In the HER-2 negative group no amplification was observed in 36 out of 40 cases. 3 cases showed amplification by both methods and one by CISH alone. The equivocal HER-2 group showed no amplification in 30 out of 41 cases and amplification in 9 cases. One case was FISH negative CISH positive and one was discarded. In the HER-2 positive group, amplification was confirmed in 37 of the 40 cases by both methods. 3 cases were unsuitable for assessment. This study indicated that CISH is a sensitive alternative to FISH in detecting HER2 gene amplification and may replace FISH in HER2 testing. Good agreement was observed between methods (98.5% - 119 out of 121 cases). Furthermore, as only 4 out of 121 cases were unsuitable for assessment (no signal or missing TMA cores) - it may be feasible to use TMA in diagnostics.

Structure and Optical Properties of Titania-PDMS Hybrid Nanocomposites Prepared by In Situ Non-Aqueous Synthesis

Directory of Open Access Journals (Sweden)

Antoine R. M. Dalod

2017-12-01

Full Text Available Organic-inorganic hybrid materials are attractive due to the combination of properties from the two distinct types of materials. In this work, transparent titania-polydimethylsiloxane hybrid materials with up to 15.5 vol. % TiO2 content were prepared by an in situ non-aqueous method using titanium (IV isopropoxide and hydroxy-terminated polydimethylsiloxane as precursors. Spectroscopy (Fourier transform infrared, Raman, Ultraviolet-visible, ellipsometry and small-angle X-ray scattering analysis allowed to describe in detail the structure and the optical properties of the nanocomposites. Titanium alkoxide was successfully used as a cross-linker and titania-like nanodomains with an average size of approximately 4 nm were shown to form during the process. The resulting hybrid nanocomposites exhibit high transparency and tunable refractive index from 1.42 up to 1.56, depending on the titania content.

Facile In Situ Fabrication of Nanostructured Graphene–CuO Hybrid with Hydrogen Sulfide Removal Capacity

Institute of Scientific and Technical Information of China (English)

Sunil P.Lonkar; Vishnu V.Pillai; Samuel Stephen; Ahmed Abdala; Vikas Mittal

2016-01-01

A simple and scalable synthetic approach for one-step synthesis of graphene–Cu O(TRGC) nanocomposite by an in situ thermo-annealing method has been developed.Using graphene oxide(GO) and copper hydroxide as a precursors reagent,the reduction of GO and the uniform deposition of in situ formed Cu O nanoparticles on graphene was simultaneously achieved.The method employed no solvents,toxic-reducing agents,or organic modifiers.The resulting nanostructured hybrid exhibited improved H2 S sorption capacity of 1.5 mmol H2S/g-sorbent(3 g S/100 g-sorbent).Due to its highly dispersed sub-20 nm Cu O nanoparticles and large specific surface area,TRGC nanocomposite exhibits tremendous potential for energy and environment applications.

Plug-In Hybrid Medium-Duty Truck Demonstration and Evaluation

Energy Technology Data Exchange (ETDEWEB)

Miyasato, Matt [Electric Power Research Institute (EPRI), Palo Alto, CA (United States); Kosowski, Mark [Electric Power Research Institute (EPRI), Palo Alto, CA (United States)

2015-10-01

The Plug-In Hybrid Medium-Duty Truck Demonstration and Evaluation Program was sponsored by the United States Department of Energy (DOE) using American Recovery and Reinvestment Act of 2009 (ARRA) funding. The purpose of the program is to develop a path to migrate plug-in hybrid electric vehicle (PHEV) technology to medium-duty vehicles by demonstrating and evaluating vehicles in diverse applications. The program also provided three production-ready PHEV systems—Odyne Systems, Inc. (Odyne) Class 6 to 8 trucks, VIA Motors, Inc. (VIA) half-ton pickup trucks, and VIA three-quarter-ton vans. The vehicles were designed, developed, validated, produced, and deployed. Data were gathered and tests were run to understand the performance improvements, allow cost reductions, and provide future design changes. A smart charging system was developed and produced during the program. The partnerships for funding included the DOE; the California Energy Commission (CEC); the South Coast Air Quality Management District (SCAQMD); the Electric Power Research Institute (EPRI); Odyne; VIA; Southern California Edison; and utility and municipal industry participants. The reference project numbers are DOE FOA-28 award number EE0002549 and SCAQMD contract number 10659.

Performance of in situ chemical oxidation field demonstrations at DOE sites

International Nuclear Information System (INIS)

Cline, S.R.; West, O.R.; Siegrist, R.L.; Holden, W.L.

1997-01-01

Researchers at the Oak Ridge National Laboratory (ORNL) have been investigating the use of in situ chemical oxidation to remediate organic contaminants (VOCs, SVOCs, and PCBs) in soils and groundwater at the laboratory and field scales. Field scale design parameters (e.g., oxidant loading rates and oxidant delivery techniques) are often dictated by site conditions (e.g., soil properties and initial contaminant concentrations). Chemical destruction of organic compounds can be accomplished using a variety of oxidants. Recent research has involved field scale in situ chemical oxidation demonstrations using H 2 O 2 and KMnO 4 in conjunction with soil mixing as the oxidant delivery mechanism. A description of some of these fields activities and future field-scale work is presented here

MMP activity in the hybrid layer detected with in situ zymography.

Science.gov (United States)

Mazzoni, A; Nascimento, F D; Carrilho, M; Tersariol, I; Papa, V; Tjäderhane, L; Di Lenarda, R; Tay, F R; Pashley, D H; Breschi, L

2012-05-01

Dentinal proteases are believed to play an important role in the degradation of hybrid layers (HL). This study investigated the HL gelatinolytic activity by in situ zymography and functional enzyme activity assay. The hypotheses were that HLs created by an etch-and-rinse adhesive exhibit active gelatinolytic activity, and MMP-2 and -9 activities in dentin increase during adhesive procedures. Etched-dentin specimens were bonded with Adper Scotchbond 1XT and restored with composite. Adhesive/dentin interface slices were placed on microscope slides, covered with fluorescein-conjugated gelatin, and observed with a multi-photon confocal microscope after 24 hrs. Human dentin powder aliquots were prepared and assigned to the following treatments: A, untreated; B, etched with 10% phosphoric acid; or C, etched with 10% phosphoric acid and mixed with Scotchbond 1XT. The MMP-2 and -9 activities of extracts of dentin powder were measured with functional enzyme assays. Intense and continuous enzyme activity was detected at the bottom of the HL, while that activity was more irregular in the upper HL. Both acid-etching and subsequent adhesive application significantly increased MMP-2 and -9 activities (p < 0.05). The results demonstrate, for the first time, intrinsic MMP activity in the HL, and intense activation of matrix-bound MMP activity with both etching and adhesive application.

Test plan for in situ bioremediation demonstration of the Savannah River Integrated Demonstration Project DOE/OTD TTP No.: SR 0566-01. Revision 3

Energy Technology Data Exchange (ETDEWEB)

Hazen, T.C.

1991-09-18

This project is designed to demonstrate in situ bioremediation of groundwater and sediment contaminated with chlorinated solvents. Indigenous microorganisms will be simulated to degrade trichloroethylene (TCE), tetrachloroethylene (PCE) and their daughter products in situ by addition of nutrients to the contaminated zone. in situ biodegradation is a highly attractive technology for remediation because contaminants are destroyed, not simply moved to another location or immobilized, thus decreasing costs, risks, and time, while increasing efficiency and public and regulatory acceptability. Bioremediation has been found to be among the least costly technologies in applications where it will work.

Erythrovirus B19 infection in acquired immunodeficiency syndrome: screening by histopathology, immunohistochemistry, and in situ hybridization

Directory of Open Access Journals (Sweden)

Sérgio Setúbal

2006-06-01

Full Text Available Erythrovirus B19 infects erythrocytic progenitors, transiently interrupting erythropoiesis. In AIDS patients it causes chronic anemia amenable to treatment. We looked for evidences of B19 infection in stored bone marrow material from patients with acquired immunodeficiency syndrome. Histological sections were made from stored paraffin blocks from 33 autopsies (39 blocks and 35 biopsies (45 blocks, 30 patients performed from 1988 to 2002. They were examined after hematoxylin-eosin (HE staining, immunohistochemical (IHC, and in situ hybridization. HE revealed intra-nuclear inclusion bodies ("lantern cells" suggesting B19 infection in 19 sections corresponding to 19 of 63 patients examined with this test. Seven of 78 sections subjected to immunohistochemistry were positive, corresponding to 7 of 58 patients examined with this test. Fourteen sections corresponding to 13 of the 20 HE and/or IHC positive patients were subjected to in situ hybridization, with six positives results. Among the 13 patients subjected to the three techniques, only one gave unequivocal positive results in all and was considered a true positive. The frequency of B19 infection (1/63 patients in the material examined can be deemed low.

One-step synthesis of graphene/polyaniline hybrids by in situ intercalation polymerization and their electromagnetic properties

Science.gov (United States)

Chen, Xiangnan; Meng, Fanchen; Zhou, Zuowan; Tian, Xin; Shan, Liming; Zhu, Shibu; Xu, Xiaoling; Jiang, Man; Wang, Li; Hui, David; Wang, Yong; Lu, Jun; Gou, Jihua

2014-06-01

A new method is introduced for the preparation of graphene/polyaniline hybrids using a one-step intercalation polymerization of aniline inside the expanded graphite. The structural and morphological characterizations were performed by X-ray diffraction analysis, transmission electron microscopy and field emission scanning electron microscopy. Both the experimental and first-principles simulated results show that the aniline cation formed by aniline and H+ tends to be drawn towards the electron-enriched zone and to intercalate into the interlayer of graphite. Subsequently, an in situ polymerization leads to the separation of graphite into graphene sheet, resulting from the exothermic effect and more vigorous movements of the chain molecules of polyaniline. The interactions between polyaniline and graphene were confirmed by Fourier transform infrared spectroscopy and Raman spectra. In addition, the graphene/polyaniline hybrid exhibited a breakthrough in the improvement of microwave absorption.A new method is introduced for the preparation of graphene/polyaniline hybrids using a one-step intercalation polymerization of aniline inside the expanded graphite. The structural and morphological characterizations were performed by X-ray diffraction analysis, transmission electron microscopy and field emission scanning electron microscopy. Both the experimental and first-principles simulated results show that the aniline cation formed by aniline and H+ tends to be drawn towards the electron-enriched zone and to intercalate into the interlayer of graphite. Subsequently, an in situ polymerization leads to the separation of graphite into graphene sheet, resulting from the exothermic effect and more vigorous movements of the chain molecules of polyaniline. The interactions between polyaniline and graphene were confirmed by Fourier transform infrared spectroscopy and Raman spectra. In addition, the graphene/polyaniline hybrid exhibited a breakthrough in the improvement of

Motivational study for an hybrid demonstrator; Dossier de motivation pour un demonstrateur hybride

Energy Technology Data Exchange (ETDEWEB)

Boidron, M.; Fiorini, G.L.; Thomas, J.B. [CEA Cadarache, 13 - Saint Paul lez Durance (France). Direction des Reacteurs Nucleaires; Flocard, H. [Institut National de Physique Nucleaire et de Physique des Particules (CNRS/IN2P3), 75 - Paris (France)

2001-02-15

This document recalls first the role of hybrid accelerator driven systems (ADS) in the domain of transmutation of long-lived fission products and minor actinides. It presents the specific contribution of these systems in the management of radioactive wastes and their technical feasibility and safety aspects. Then, follows a motivational analysis for the construction of a demonstration facility with its specifications and R and D needs: feasibility, schedule, links with other ADS-related programs, cost, international cooperation, recommendations. (J.S.)

Use of Hybridization Chain Reaction-Fluorescent In Situ Hybridization To Track Gene Expression by Both Partners during Initiation of Symbiosis.

Science.gov (United States)

Nikolakakis, K; Lehnert, E; McFall-Ngai, M J; Ruby, E G

2015-07-01

The establishment of a productive symbiosis between Euprymna scolopes, the Hawaiian bobtail squid, and its luminous bacterial symbiont, Vibrio fischeri, is mediated by transcriptional changes in both partners. A key challenge to unraveling the steps required to successfully initiate this and many other symbiotic associations is characterization of the timing and location of these changes. We report on the adaptation of hybridization chain reaction-fluorescent in situ hybridization (HCR-FISH) to simultaneously probe the spatiotemporal regulation of targeted genes in both E. scolopes and V. fischeri. This method revealed localized, transcriptionally coregulated epithelial cells within the light organ that responded directly to the presence of bacterial cells while, at the same time, provided a sensitive means to directly show regulated gene expression within the symbiont population. Thus, HCR-FISH provides a new approach for characterizing habitat transition in bacteria and for discovering host tissue responses to colonization. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Demonstration testing and evaluation of in situ soil heating. Treatability study work plan (Revision 2)

International Nuclear Information System (INIS)

Sresty, G.C.

1994-01-01

A Treatability Study planned for the demonstration of the in situ electromagnetic (EM) heating process to remove organic solvents is described in this Work Plan. The treatability study will be conducted by heating subsurface vadose-zone soils in an organic plume adjacent to the Classified Burial Ground K-1070-D located at K-25 Site, Oak Ridge. The test is scheduled to start during the fourth quarter of FY94 and will be completed during the first quarter of FY95. Over the last nine years, a number of Government agencies (EPA, Army, AF, and DOE) and industries sponsored further development and testing of the in situ heating and soil decontamination process for the remediation of soils containing hazardous organic contaminants. In this process the soil is heated in situ using electrical energy. The contaminants are removed from the soil due to enhanced vaporization, steam distillation and stripping. IITRI will demonstrate the EM Process for in situ soil decontamination at K-25 Site under the proposed treatability study. Most of the contaminants of concern are volatile organics which can be removed by heating the soil to a temperature range of 85 degrees to 95 degrees C. The efficiency of the treatment will be determined by comparing the concentration of contaminants in soil samples. Samples will be obtained before and after the demonstration for a measurement of the concentration of contaminants of concern. This document is a Treatability Study Work Plan for the demonstration program. The document contains a description of the proposed treatability study, background of the EM heating process, description of the field equipment, and demonstration test design

Human cDNA mapping using fluorescence in situ hybridization

Energy Technology Data Exchange (ETDEWEB)

Korenberg, J.R.

1993-03-04

Genetic mapping is approached using the techniques of high resolution fluorescence in situ hybridization (FISH). This technology and the results of its application are designed to rapidly generate whole genome as tool box of expressed sequence to speed the identification of human disease genes. The results of this study are intended to dovetail with and to link the results of existing technologies for creating backbone YAC and genetic maps. In the first eight months, this approach generated 60--80% of the expressed sequence map, the remainder expected to be derived through more long-term, labor-intensive, regional chromosomal gene searches or sequencing. The laboratory has made significant progress in the set-up phase, in mapping fetal and adult brain and other cDNAs, in testing a model system for directly linking genetic and physical maps using FISH with small fragments, in setting up a database, and in establishing the validity and throughput of the system.

Hybrid aerogels and bioactive aerogels under uniaxial compression: an in situ SAXS study

Directory of Open Access Journals (Sweden)

Esquivias, L.

2010-12-01

Full Text Available The complex structure of hybrid organic/inorganic aerogels is composed by an inorganic phase covalently bonded to an organic chain forming a copolymer. Conventional hybrid aerogels were studied as well as bioactive hybrid aerogels, that is, aerogels with a calcium active phase added. In this work, the relationship between mechanical response and nanostructure was studied, using a specifically designed sample-holder for in situ uniaxial compression obtaining at the same time the small-angle X-ray pattern from synchrotron radiation (SAXS. Structural elements can be described as a particulated silica core surrounded by the organic chains. These chains are compressed on the direction parallel to the load, and a relationship between macroscopic uniaxial compression and particle and pore deformations can be established.

La compleja estructura de los aerogeles híbridos orgánico/inorgánicos está compuesta por una fase inorgánica de sílice, unida mediante enlaces covalentes a una red de cadenas orgánicas. Se han estudiado composites híbridos convencionales y bioactivos, esto es, con una fase activa de calcio añadida. En este trabajo se ha investigado la relación entre la respuesta mecánica y la nanoestructura, con ayuda de un portamuestras específicamente diseñado para el estudio in situ de muestras bajo compresión uniaxial, a la vez que se obtiene el espectro de rayos-X a bajo-ángulo de radiación sincrotrón (SAXS. Los elementos estructurales se pueden describir como núcleos particulados de sílice rodeados de las cadenas orgánicas. Estas, se comprimen en la dirección paralela a la carga pudiéndose establecer una relación entre la compresión uniaxial macroscópica y la deformación de las partículas y poros que forman la estructura.

HER-2 protein concentrations in breast cancer cells increase before immunohistochemical and fluorescence in situ hybridization analysis turn positive

DEFF Research Database (Denmark)

Olsen, Dorte A; Østergaard, Birthe; Bokmand, Susanne

2007-01-01

BACKGROUND: The level of HER-2/neu in breast cancer cells is normally measured by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH). It determines whether patients should be treated with trastuzumab (Herceptin). In this study, HER-2 protein in breast cancer tissue...

Environmental assessment for the electric and hybrid vehicle demonstration project, performance standards and financial incentives

Energy Technology Data Exchange (ETDEWEB)

LaBelle, S. J.

1978-10-01

The assessment is concerned with the impacts of the demonstration of electric and hybrid vehicles acquired to fulfill certain requirements of the Electric and Hybrid Vehicle Research, Development, and Demonstration Act, PL 94-413 as amended. The financial incentives programs and vehicle performance standards associated with the demonstration are also covered. Not included is an assessment of the long term effects of EHV commercialization and of the research and development program being carried out simultaneously with the demonstration, also in response to PL 94-413. These federal actions will be included in a programmatic environmental assessment scheduled for completion in FY 79.

Transmission electron microscopic method for gene mapping on polytene chromosomes by in situ hybridization

OpenAIRE

Wu, Madeline; Davidson, Norman

1981-01-01

A transmission electron microscope method for gene mapping by in situ hybridization to Drosophila polytene chromosomes has been developed. As electron-opaque labels, we use colloidal gold spheres having a diameter of 25 nm. The spheres are coated with a layer of protein to which Escherichia coli single-stranded DNA is photochemically crosslinked. Poly(dT) tails are added to the 3' OH ends of these DNA strands, and poly(dA) tails are added to the 3' OH ends of a fragmented cloned Drosophila DN...

In situ hybridization of somatolactin transcripts in the pituitary glands from acclimatized carp (Cyprinus carpio

Directory of Open Access Journals (Sweden)

MAURICIO LÓPEZ

2001-01-01

Full Text Available We isolated and cloned a carp somatolactin SL DNA fragment, of which 78% of the nucleotides were identical to the corresponding salmon SL sequence. The results obtained upon Northern blot hybridization of carp pituitary RNA allowed the identification of two transcripts as described for other fish. When the content of SL transcripts in pituitary sections from summer- and winter- acclimatized carp was quantified by in situ hybridization assays, we found no significant differences between the two seasons. In salmonids, plasma SL reaches higher levels in summer than in winter in synchrony with the water temperature cycle; in the eurythermal carp, however, the complex adaptive responses imposed by seasonal environmental changes do not seem to include the regulation of the somatolactin detected with the probe used at the transcriptional level in pituitary glands

In-situ fabrication of halloysite nanotubes/silica nano hybrid and its application in unsaturated polyester resin

Science.gov (United States)

Lin, Jing; Zhong, Bangchao; Jia, Zhixin; Hu, Dechao; Ding, Yong; Luo, Yuanfang; Jia, Demin

2017-06-01

Silica nanoparticles was in-situ grown on the surface of halloysite nanotubes (HNTs) by a facile one-step approach to prepare a unique nano-structured hybrid (HNTs-g-Silica). The structure, morphology and composition of HNTs-g-Silica were investigated. It was confirmed that silica nanoparticles with the diameter of 10-20 nm were chemically grafted through Sisbnd O bonds and uniformly dispersed onto the surface of HNTs, leading to the formation of nano-protrusions on the nanotube surface. Due to the significantly improved interface strength between HNTs-g-Silica and polymer matrix, HNTs-g-Silica effectively toughened unsaturated polyester resin (UPE) and endowed UPE with superior thermal stability compared to HNTs. Based on the unique hybrid architecture and the improved properties of UPE nanocomposites, it is envisioned that HNTs-g-Silica may be a promising filler for more high performance and functional polymers composites and the fabrication method may have implications in the synthesis of nano hybrid materials.

In situ vitrification: Demonstrated capabilities and potential applications

International Nuclear Information System (INIS)

Luey, J.K.

1993-01-01

A large-scale demonstration of the in situ vitrification (ISV) process was performed in April 1990 on the 116-B-6A Crib in the 100 Area of the Hanford Site in southeastern Washington. The 116-B-6A Crib is a radioactive mixed waste site and was selected to demonstrate the applicability of ISV to soils contaminated with mixed wastes common to many US Department of Energy (DOE) sites. Results from the demonstration show that the ISV process is a viable remediation technology for contaminated soils. The demonstration of the ISV process on an actual contaminated soil site followed research and development efforts by the Pacific Northwest Laboratory (PNL) over the last 10 years. PNL's research has led to the development of the ISV process as a viable remediation technology for contaminated soils and the creation of a commercial supplier of ISV services, Geosafe Corporation. Development efforts for ISV applications other than treatment of contaminated soils, by PNL and in collaboration with Oak Ridge National Laboratory (ORNL) and Idaho National Engineering Laboratory (INEL), show the ISV process has potential applicability for remediating buried waste sites, remediating underground storage tanks, and enabling the placement of subsurface vitrified barriers and engineered structures. This paper discusses the results from the April 1990 large-scale demonstration and provides a general overview of the current capabilities of the ISV process for contaminated soils. In addition, this paper outlines some of the technical issues associated with other ISV applications and provides a qualitative discussion of the level of effort needed to resolve these technical issues

Hybrid Electrodes by In-Situ Integration of Graphene and Carbon-Nanotubes in Polypyrrole for Supercapacitors

Science.gov (United States)

Aphale, Ashish; Maisuria, Krushangi; Mahapatra, Manoj K.; Santiago, Angela; Singh, Prabhakar; Patra, Prabir

2015-09-01

Supercapacitors also known as electrochemical capacitors, that store energy via either Faradaic or non-Faradaic processes, have recently grown popularity mainly because they complement, and can even replace, conventional energy storage systems in variety of applications. Supercapacitor performance can be improved significantly by developing new nanocomposite electrodes which utilizes both the energy storage processes simultaneously. Here we report, fabrication of the freestanding hybrid electrodes, by incorporating graphene and carbon nanotubes (CNT) in pyrrole monomer via its in-situ polymerization. At the scan rate of 5 mV s-1, the specific capacitance of the polypyrrole-CNT-graphene (PCG) electrode film was 453 F g-1 with ultrahigh energy and power density of 62.96 W h kg-1 and 566.66 W kg-1 respectively, as shown in the Ragone plot. A nanofibrous membrane was electrospun and effectively used as a separator in the supercapacitor. Four supercapacitors were assembled in series to demonstrate the device performance by lighting a 2.2 V LED.

Hybrid Electrodes by In-Situ Integration of Graphene and Carbon-Nanotubes in Polypyrrole for Supercapacitors.

Science.gov (United States)

Aphale, Ashish; Maisuria, Krushangi; Mahapatra, Manoj K; Santiago, Angela; Singh, Prabhakar; Patra, Prabir

2015-09-23

Supercapacitors also known as electrochemical capacitors, that store energy via either Faradaic or non-Faradaic processes, have recently grown popularity mainly because they complement, and can even replace, conventional energy storage systems in variety of applications. Supercapacitor performance can be improved significantly by developing new nanocomposite electrodes which utilizes both the energy storage processes simultaneously. Here we report, fabrication of the freestanding hybrid electrodes, by incorporating graphene and carbon nanotubes (CNT) in pyrrole monomer via its in-situ polymerization. At the scan rate of 5 mV s(-1), the specific capacitance of the polypyrrole-CNT-graphene (PCG) electrode film was 453 F g(-1) with ultrahigh energy and power density of 62.96 W h kg(-1) and 566.66 W kg(-1) respectively, as shown in the Ragone plot. A nanofibrous membrane was electrospun and effectively used as a separator in the supercapacitor. Four supercapacitors were assembled in series to demonstrate the device performance by lighting a 2.2 V LED.

mathFISH, a web tool that uses thermodynamics-based mathematical models for in silico evaluation of oligonucleotide probes for fluorescence in situ hybridization.

Science.gov (United States)

Yilmaz, L Safak; Parnerkar, Shreyas; Noguera, Daniel R

2011-02-01

Mathematical models of RNA-targeted fluorescence in situ hybridization (FISH) for perfectly matched and mismatched probe/target pairs are organized and automated in web-based mathFISH (http://mathfish.cee.wisc.edu). Offering the users up-to-date knowledge of hybridization thermodynamics within a theoretical framework, mathFISH is expected to maximize the probability of success during oligonucleotide probe design.

MicroRNA Detection by Whole-Mount In Situ Hybridization in C. elegans.

Science.gov (United States)

Andachi, Yoshiki; Kohara, Yuji

2018-01-01

MicroRNAs (miRNAs) loaded on argonaute proteins guide RNA-induced silencing complexes to target mRNAs. An excellent method to decipher the spatiotemporal expression patterns of miRNAs is whole-mount in situ hybridization (WISH), which has been successfully used in vertebrate embryos but still remains unavailable for many animal species. Here, we describe a WISH method for miRNA detection in Caenorhabditis elegans at both embryonic and post-embryonic stages. Strategies devised for detection include fixation of animals with carbodiimide at a high temperature and subsequent partial digestion of the fixed animals with an extremely high concentration of proteinase. WISH signals are visualized by staining with a chromogenic substrate or a fluorescent dye.

Fluorescent whole-mount RNA in situ hybridization (F-WISH) in plant germ cells and the fertilized ovule.

Science.gov (United States)

Bleckmann, Andrea; Dresselhaus, Thomas

2016-04-01

First evidence on gene function and regulation is provided by the cellular expression pattern in complex tissues. However, to understand the activity of a specific gene, it is essential to analyze the regulatory network, which controls the spatio-temporal translation pattern during the entire life span of the transcribed mRNA. To explore mechanisms which control mRNA abundance and localization in space and time, it is necessary to visualize mRNAs quantitatively with a subcellular resolution, without sectioning the tissues. We have adapted and optimized a protocol for colorimetric whole-mount RNA in situ hybridization (WISH) using egg cell-specific digoxigenin (DIG) labeled probes (Hejátko et al., 2006) [1] on ovules and early seeds of Arabidopsis. Furthermore, we established a fluorescent whole-mount RNA in situ hybridization (F-WISH) protocol, which allows mRNA visualization on a subcellular level. The polar localized mRNA of SBT4.13, encoding a subtilase, was identified using this protocol. Both methods are described and discussed in detail. Additionally a (F)-WISH flow-chart is provided along with a troubleshooting table. Copyright © 2015 Elsevier Inc. All rights reserved.

Detection of ALK gene rearrangement in non-small cell lung cancer: a comparison of fluorescence in situ hybridization and chromogenic in situ hybridization with correlation of ALK protein expression.

Science.gov (United States)

Kim, Hyojin; Yoo, Seol-Bong; Choe, Ji-Young; Paik, Jin Ho; Xu, Xianhua; Nitta, Hiroaki; Zhang, Wenjun; Grogan, Thomas M; Lee, Choon-Taek; Jheon, Sanghoon; Chung, Jin-Haeng

2011-08-01

Accurate determination of ALK rearrangement is important in lung cancer patients, especially in determining their eligibility for crizotinib therapy. Fluorescence in situ hybridization (FISH) has been regarded as the gold standard method for detecting ALK rearrangement. However, FISH requires a fluorescence microscope, and the signals are labile and rapidly fade over time. This study evaluates the concordance between ALK gene rearrangement in non-small cell lung cancer assessed by ALK FISH and a newly developed ALK chromogenic in situ hybridization (CISH) and correlates the results with ALK protein expression assessed by immunohistochemistry. A total of 465 formalin-fixed, paraffin-embedded non-small cell lung cancer samples were analyzed by ALK FISH (PathVysion, Vysis, Abbott) and ALK CISH. For comparison, all specimens were stained by immunohistochemistry (clone 5A4, Novocastra) and interobserver reproducibility was assessed. We found that agreement between the pathologists on the CISH-determined ALK status was achieved in 449 patients (96.6%), and ALK rearrangement was identified in 18 patients (4.0%) in CISH method. Among these cases, 443 cases (95.3%) had results matching the corresponding FISH results: 17 rearranged, 425 wild types, and 1 discordant case. There was high concordance in the assessment of ALK gene rearrangement between FISH and CISH techniques (κ = 0.92) and between observers (κ = 0.97). In addition, there was high concordance in the ALK gene status and ALK protein expression between CISH and IHC tests (κ = 0.82). CISH is a highly reproducible and practical method to detect ALK gene rearrangement and correlated well with ALK protein expression. Here, we present a diagnostic algorithm (Chung's SNUBH ALK protocol) to detect lung cancer with ALK rearrangements using IHC, FISH and CISH. Because CISH allows a concurrent analysis of histological features of the tumors and gene rearrangement, it appears to be a useful method in determining ALK gene

Bright-field in situ hybridization for HER2 gene amplification in breast cancer using tissue microarrays: correlation between chromogenic (CISH) and automated silver-enhanced (SISH) methods with patient outcome.

Science.gov (United States)

Francis, Glenn D; Jones, Mark A; Beadle, Geoffrey F; Stein, Sandra R

2009-06-01

HER2 gene amplification or overexpression occurs in 15% to 25% of breast cancers and has implications for treatment and prognosis. The most commonly used methods for HER2 testing are fluorescence in situ hybridization (FISH) and immunohistochemistry. FISH is considered to be the reference standard and more accurately predicts response to trastuzumab, but is technically demanding, expensive, and requires specialized equipment. In situ hybridization is required to be eligible for adjuvant treatment with trastuzumab in Australia. Bright-field in situ hybridization is an alternative to FISH and uses a combination of in situ methodology and a peroxidase-mediated chromogenic substrate such as diaminobenzidine [chromogenic in situ hybridization (CISH)] or multimer technology coupled with enzyme metallography [silver-enhanced in situ hybridization (SISH)] to create a marker visible under bright-field microscopy. CISH was introduced into diagnostic testing in Australia in October 2006. SISH methodology is a more recent introduction into the testing repertoire. An evaluation of CISH and SISH performance to assess patient outcome were performed using tissue microarrays. Tissue microarrays were constructed in duplicate using material from 593 patients with invasive breast carcinoma and assessed using CISH and SISH. Gene amplification was assessed using the American Society of Clinical Oncology/College of American Pathologists guideline and Australian HER2 Advisory Board criteria (single probe: diploid, 1 to 2.5 copies/nucleus; polysomy >2.5 to 4 copies/nucleus; equivocal, >4 to 6 copies/nucleus; low-level amplification, >6 to 10 copies/nucleus and high-level amplification >10 copies/nucleus; dual probe HER2/CHR17 ratio: nonamplified 2.2). Results were informative for 337 tissue cores comprising 230 patient samples. Concordance rates were 96% for HER2 single probe CISH and SISH and 95.5% for single probe CISH and dual probe HER2/CHR17 SISH. Both bright-field methods correlated

Demonstration of in situ-constructed horizontal soil containment barrier at Fernald

International Nuclear Information System (INIS)

Pettit, P.J.; Ridenour, D.; Walker, J.; Saugier, K.

1994-01-01

A new design of jet grouting tool that can be guided by horizontal well casings and that operates in the horizontal plane has been used for the in situ placement of grout and construction of a prototype horizontal barrier that is free of windows. Jet grouting techniques have been advanced to permit construction of horizontal barriers underneath contaminated soil without having to excavate or disturb the waste. The paper describes progress on the Fernald Environmental Restoration Management Corporation (FERMCO) In Situ Land Containment Project which is sponsored by the US Department of Energy's (DOE) Office of Technology Development (OTD) for DOE's Fernald Environmental Management Project (FEMP). The Fernald project is to demonstrate a novel, enabling technology for the controlled underground placement of horizontal panels of grout, and the joining of adjacent panels to construct practical, extensive barriers. Construction strategy, equipment mechanics and operating details of this new method are described

Partial trisomy 13 in an infant with a mild phenotype: application of fluorescence in situ hybridization in cytogenetic syndromes.

Science.gov (United States)

Begovic, D; Hitrec, V; Lasan, R; Letica, L; Baric, I; Sarnavka, V; Galic, S

1998-06-01

We report on a month-old infant with dysmorphic face and several anomalies known to be associated with trisomy 13. Fluorescence in situ hybridization (FISH) studies performed on metaphase cells allowed us to identify an extra material on the short arm of the chromosome 13 as a duplication of 13q22-qter.

Comparative Demonstration of Active and Semi-Passive In Situ Bioremediation Approaches for Perchlorate Impacted Groundwater: Active In Situ Bioremediation Demonstration

Science.gov (United States)

2013-04-01

http://www.itrcweb.org/Documents/PERC-1.pdf • ITRC Perchlorate Team. 2008. Remediation Technologies for Perchlorate Contamination in Water and Soil ...pdf • Solutions EIS. 2006. Protocol for Enhanced In Situ Bioremediation Using Emulsified Vegetable Oil . Prepared for ESTCP. May 2006. • http...Air Force. 2007. Protocol for In Situ Bioremediation of Chlorinated Solvents Using Edible Oil . Prepared for AFCEC - Environmental Science Division

Plug-In Hybrid Urban Delivery Truck Technology Demonstration

Energy Technology Data Exchange (ETDEWEB)

Miyasato, Matt [South Coast Air Quality Management District Building Corporation, Diamond Bar, CA (United States); Impllitti, Joseph [South Coast Air Quality Management District Building Corporation, Diamond Bar, CA (United States); Pascal, Amar [South Coast Air Quality Management District Building Corporation, Diamond Bar, CA (United States)

2015-07-31

The I-710 and CA-60 highways are key transportation corridors in the Southern California region that are heavily used on a daily basis by heavy duty drayage trucks that transport the cargo from the ports to the inland transportation terminals. These terminals, which include store/warehouses, inland-railways, are anywhere from 5 to 50 miles in distance from the ports. The concentrated operation of these drayage vehicles in these corridors has had and will continue to have a significant impact on the air quality in this region whereby significantly impacting the quality of life in the communities surrounding these corridors. To reduce these negative impacts it is critical that zero and near-zero emission technologies be developed and deployed in the region. A potential local market size of up to 46,000 trucks exists in the South Coast Air Basin, based on near- dock drayage trucks and trucks operating on the I-710 freeway. The South Coast Air Quality Management District (SCAQMD), California Air Resources Board (CARB) and Southern California Association of Governments (SCAG) — the agencies responsible for preparing the State Implementation Plan required under the federal Clean Air Act — have stated that to attain federal air quality standards the region will need to transition to broad use of zero and near zero emission energy sources in cars, trucks and other equipment (Southern California Association of Governments et al, 2011). SCAQMD partnered with Volvo Trucks to develop, build and demonstrate a prototype Class 8 heavy-duty plug-in hybrid drayage truck with significantly reduced emissions and fuel use. Volvo’s approach leveraged the group’s global knowledge and experience in designing and deploying electromobility products. The proprietary hybrid driveline selected for this proof of concept was integrated with multiple enhancements to the complete vehicle in order to maximize the emission and energy impact of electrification. A detailed review of all

Application of Fluorescence In Situ Hybridization (FISH) Technique for the Detection of Genetic Aberration in Medical Science

OpenAIRE

Ratan, Zubair Ahmed; Zaman, Sojib Bin; Mehta, Varshil; Haidere, Mohammad Faisal; Runa, Nusrat Jahan; Akter, Nasrin

2017-01-01

Fluorescence in situ hybridization (FISH) is a macromolecule recognition technique, which is considered as a new advent in the field of cytology.?Initially, it was developed as a physical mapping tool to delineate genes within chromosomes. The accuracy and versatility of FISH were subsequently capitalized upon in biological and medical research. This visually appealing technique provides an intermediate degree of resolution between DNA analysis and chromosomal investigations. FISH consists of...

Expression and localization of ionotropic glutamate receptor subunits in the goldfish retina--an in situ hybridization and immunocytochemical study

NARCIS (Netherlands)

Vandenbranden, C. A.; Kamphuis, W.; Nunes Cardozo, B.; Kamermans, M.

2000-01-01

The expression and distribution of AMPA, kainate and NMDA glutamate receptor subunits was studied in the goldfish retina. For the immunocytochemical localization of the AMPA receptor antisera against GluR2, GluR2/3 and GluR4 were used, and for in situ hybridization rat specific probes for GluR1 and

In Situ Hybridization Method Reveals (Pro)renin Receptor Expressing Cells in the Pituitary Gland of Rats: Correlation with Anterior Pituitary Hormones.

Science.gov (United States)

Takahashi, Kazuhiro; Yatabe, Megumi; Fujiwara, Ken; Hirose, Takuo; Totsune, Kazuhito; Yashiro, Takashi

2013-02-28

Expression of (pro)renin receptor ((P)RR), a specific receptor for renin and prorenin, was studied in rat pituitary gland. In situ hybridization showed that cells expressing (P)RR mRNA were widely distributed in the anterior lobe and intermediate lobe of the pituitary gland. Double-staining using in situ hybridization for (P)RR mRNA and immunohistochemistry for the pituitary hormones showed that (P)RR mRNA was expressed in most of the GH cells and ACTH cells in the anterior lobe. (P)RR mRNA was also expressed in a few prolactin cells and TSH cells, but not in LH cells. The present study has shown for the first time the distribution of (P)RR mRNA expressing cells in the rat pituitary gland. These findings suggest that (P)RR plays physiological roles in the pituitary gland, such as the modulation of the pituitary hormone secretion.

In Situ Hybridization Method Reveals (Pro)renin Receptor Expressing Cells in the Pituitary Gland of Rats: Correlation with Anterior Pituitary Hormones

International Nuclear Information System (INIS)

Takahashi, Kazuhiro; Yatabe, Megumi; Fujiwara, Ken; Hirose, Takuo; Totsune, Kazuhito; Yashiro, Takashi

2013-01-01

Expression of (pro)renin receptor ((P)RR), a specific receptor for renin and prorenin, was studied in rat pituitary gland. In situ hybridization showed that cells expressing (P)RR mRNA were widely distributed in the anterior lobe and intermediate lobe of the pituitary gland. Double-staining using in situ hybridization for (P)RR mRNA and immunohistochemistry for the pituitary hormones showed that (P)RR mRNA was expressed in most of the GH cells and ACTH cells in the anterior lobe. (P)RR mRNA was also expressed in a few prolactin cells and TSH cells, but not in LH cells. The present study has shown for the first time the distribution of (P)RR mRNA expressing cells in the rat pituitary gland. These findings suggest that (P)RR plays physiological roles in the pituitary gland, such as the modulation of the pituitary hormone secretion

Identification of bacteria used for microbial enhanced oil recovery process by fluorescence in situ hybridization technique

Energy Technology Data Exchange (ETDEWEB)

Fujiwara, K.; Tanaka, S.; Otsuka, M. [Kansai Research Institute, Kyoto (Japan). Lifescience Lab.; Yonebayashi, H. [Japan National Oil Corp., Chiba (Japan). Tech. Research Center; Enomoto, H. [Tohoku University, Sendai (Japan). Dept. of Geoscience and Tech.

2000-01-01

A fluorescence in situ hybridization (FISH) technique using 16S rRNA-targeted oligonucleotide probes was developed for rapid detection of microorganisms for use in the microbial enhancement of oil recovery (MEOR) process. Two microorganisms, Enterobacter cloacae TRC-322 and Bacillus licheniformis TRC-18-2-a, were selected from a collection of Enterobacter sp. and Bacillus sp. which were screened in previous studies as candidate microorganisms for injection, and were used for this experiment. Oligonucleotide probes, design based on specific sequences in the 16S rRNA gene were labeled with either fluorescein isothiocyanate (FITC), or 6-car-boxy-X-rhodamine (ROX), and were allowed to hybridize with fixed cells of the two microorganisms noted above. The fluorescence signal emitted from each microorganism cells could clearly be detected by an epifluorescence microscope. Moreover, E. cloacae TRC-322 and B, licheniformis TRC-18-2-a, suspended in actual reservoir brine, including inorganic salts, oil and aboriginal cells of the reservoir brine, could be detected directly by this hybridization method, without the need for cultivation and isolation. (author)

Fluorescence in situ hybridization with reference to biodosimetry: a review

International Nuclear Information System (INIS)

Venkatachalam, P.; Paul, S.F.D.; Jeevanram, R.K.

1996-01-01

Many advances have taken place in the field of radiation biodosimetry in the recent past. Measurement of dicentric chromosome aberrations, was first developed and followed by micronuclei scoring. These, however, are unstable type aberrations and the cells carrying such aberrations are eliminated from the body in few years. They are therefore of use primarily in case of accidental exposures. The challenge is to measure the cumulative radiation exposure resulting from normal operations by measuring stable chromosome aberrations. Banding technique can measure stable chromosome aberration but require long time to analyse the banding pattern to study translocations. On the other hand fluorescence in situ hybridization (FISH) technique is sensitive, fast and easy to identify the translocations as the chromosomes involved in translocation are painted with different colours. This review brings out the requirements of various materials, their preparations, method of detection of fluorescence etc. for carrying out FISH. The experience of various laboratories using FISH in the monitoring of radiation absorbed dose is discussed. (author)

Microstructure and wear of in-situ Ti/(TiN + TiB) hybrid composite layers produced using liquid phase process

Energy Technology Data Exchange (ETDEWEB)

Yazdi, R., E-mail: ryazdi@ut.ac.ir; Kashani-Bozorg, S.F.

2015-02-15

Tungsten inert gas (TIG) technique was conducted on commercially pure (CP)-Ti substrate, which was coated with h-BN-based powder mixture prior to the treatment. The treated surfaces were evaluated and characterized by means of scanning electron microscope (SEM), X-ray diffraction analysis, and electron dispersive spectrometry (EDS). The microhardness and wear experiment were also performed by using a microhardness machine and pin-on-disk tribometer. As h-BN reacted with titanium, an in-situ hybrid composite layer was formed showing near stoichiometric dendrites of TiN, platelets of TiB and interdendritic regions of α′-Ti martensite crystal structures. The population level of TiN and TiB regions were found to increase using a pre-placed powder mixture with greater h-BN content. However, the fabricated layers exhibited cracking and porosity; these were minimized by adjusting arc energy density and h-BN content of powder mixture. The microhardness value of the fabricated hybrid composite layers was found to be in the range of ∼650 HV{sub 0.2}–1000 HV{sub 0.2}; this is three to five times higher than that of the untreated CP-Ti substrate. In addition, the in-situ hybrid composite layers exhibited superior wear behavior over CP-Ti substrate; this is attributed to the formation of newly formed ceramic phases in the solidified surface layers and good coherent interface between the composite layer and CP-substrate. Meanwhile, severe adhesive wear mechanism of CP-titanium surface changed to mild abrasive one as a result of surface treatment. - Highlights: • In-situ Ti/(TiN + TiB) hybrid composite layers were synthesized by TIG processing on commercially pure titanium. • The microstructure features were characterized by several methods. • Microhardness enhanced three to five times higher than that of the CP-Ti substrate after surface modification. • The fabricated composite layers improved wear resistance of CP-titanium. • Severe adhesive wear mechanism of

Demonstration testing and evaluation of in situ soil heating. Health and safety plan (Revision 2)

Energy Technology Data Exchange (ETDEWEB)

Dev, H.

1994-12-28

This document is the Health and Safety Plan (HASP) for the demonstration of IITRI`s EM Treatment Technology. In this process, soil is heated in situ by means of electrical energy for the removal of hazardous organic contaminants. This process will be demonstrated on a small plot of contaminated soil located in the Pit Area of Classified Burial Ground K-1070-D, K-25 Site, Oak Ridge, TN. The purpose of the demonstration is to remove organic contaminants present in the soil by heating to a temperature range of 85{degrees} to 95{degrees}C. The soil will be heated in situ by applying 60-Hz AC power to an array of electrodes placed in boreholes drilled through the soil. In this section a brief description of the process is given along with a description of the site and a listing of the contaminants found in the area.

Demonstration testing and evaluation of in situ soil heating. Health and safety plan (Revision 2)

International Nuclear Information System (INIS)

Dev, H.

1994-01-01

This document is the Health and Safety Plan (HASP) for the demonstration of IITRI's EM Treatment Technology. In this process, soil is heated in situ by means of electrical energy for the removal of hazardous organic contaminants. This process will be demonstrated on a small plot of contaminated soil located in the Pit Area of Classified Burial Ground K-1070-D, K-25 Site, Oak Ridge, TN. The purpose of the demonstration is to remove organic contaminants present in the soil by heating to a temperature range of 85 degrees to 95 degrees C. The soil will be heated in situ by applying 60-Hz AC power to an array of electrodes placed in boreholes drilled through the soil. In this section a brief description of the process is given along with a description of the site and a listing of the contaminants found in the area

Fluorescence in situ hybridization and molecular studies in infertile men with dysplasia of the fibrous sheath.

Science.gov (United States)

Baccetti, Baccio; Collodel, Giulia; Gambera, Laura; Moretti, Elena; Serafini, Francesca; Piomboni, Paola

2005-07-01

To perform fluorescence in situ hybridization (FISH) and molecular analysis in patients with the genetic sperm defect "dysplasia of the fibrous sheath" (DFS). Retrospective study. Regional Referral Center for Male Infertility, Siena, Italy. Twelve infertile patients with DFS sperm defects. Family history, lymphocytic karyotype, physical and hormonal assays, semen analysis. The DFS sperm phenotype was defined by light, fluorescent, and electron microscopy. Sperm chromosomal constitution was examined by FISH. Gene deletions were tested by polymerase chain reaction. The genetic sperm defect DFS was determined by transmission and scanning electron microscopy. Immunofluorescence staining of A-kinase anchoring protein 4 (AKAP4) showed a moderate and diffuse signal, revealing a disorganized and incompletely assembled fibrous sheath. In 11 of 12 DFS patients, polymerase chain reaction for detecting the presence of partial sequence of AKAP4/AKAP3 binding regions gave positive results. Fluorescence in situ hybridization was performed in decondensed sperm nuclei with probes for chromosomes 18, X, and Y. The mean disomy frequency of chromosome 18 was in the normal range, whereas the mean disomy frequencies of sex chromosomes and diploidies were twice those of controls. These results should be considered when DFS sperm are used in assisted reproductive technology, owing to the high risk of transmission of chromosomal unbalance and of DFS sperm defects to male offspring.

Preimplantation genetic diagnosis by fluorescence in situ hybridization of reciprocal and Robertsonian translocations.

Science.gov (United States)

Chen, Chun-Kai; Wu, Dennis; Yu, Hsing-Tse; Lin, Chieh-Yu; Wang, Mei-Li; Yeh, Hsin-Yi; Huang, Hong-Yuan; Wang, Hsin-Shin; Soong, Yung-Kuei; Lee, Chyi-Long

2014-03-01

The presence of reciprocal and Robertsonian chromosomal rearrangement is often related to recurrent miscarriage. Using preimplantation genetic diagnosis, the abortion rate can be decreased. Cases treated at our center were reviewed. A retrospective analysis for either Robertsonian or reciprocal translocations was performed on all completed cycles of preimplantation genetic diagnosis at our center since the first reported case in 2004 until the end of 2010. Day 3 embryo biopsies were carried out, and the biopsied cell was checked by fluorescent in situ hybridization using relevant informative probes. Embryos with a normal or balanced translocation karyotype were transferred on Day 4. Thirty-eight preimplantation genetic diagnosis cycles involving 17 couples were completed. A total of 450 (82.6%) of the total oocytes were MII oocytes, and 158 (60.0%) of the two-pronuclei embryos were biopsied. In 41.4% of the fluorescent in situ hybridization analyses, the results were either normal or balanced. Embryos were transferred back after 21 cycles. Three babies were born from Robertsonian translocation carriers and another two from reciprocal translocation carriers. The miscarriage rate was 0%. Among the reciprocal translocation group, the live delivery rate was 8.3% per ovum pick-up cycle and 18.2% per embryo transfer cycle. Among the Robertsonian translocation group, the live delivery rate was 14.3% per ovum pick-up cycle and 20.0% per embryo transfer cycle. There is a trend whereby the outcome for Robertsonian translocation group carriers is better than that for reciprocal translocation group carriers. Aneuploidy screening may possibly be added in order to improve the outcome, especially for individuals with an advanced maternal age. The emergence of an array-based technology should help improve this type of analysis. Copyright © 2014. Published by Elsevier B.V.

Experimental Investigation on Mechanical and Turning Behavior of Al 7075/x% wt. TiB2-1% Gr In Situ Hybrid Composite

Directory of Open Access Journals (Sweden)

K. R. Ramkumar

2015-01-01

Full Text Available The present research work involves the study of AA 7075-TiB2-Gr in situ composite through stir casting route. This in situ method involves formation of reinforcements within the matrix by the chemical reaction of two or more compounds which also produces some changes in the matrix material within the vicinity. Titanium Diboride (TiB2 and graphite were the reinforcement in a matrix of AA 7075 alloy. The composite was prepared with the formation of the reinforcement inside the molten matrix by adding salts of Potassium Tetrafluoroborate (KBF4 and Potassium Hexafluorotitanate (K2TiF6. The samples were taken under casted condition and the properties of the composite were tested by conducting characterization using X-ray diffraction (XRD, hardness test, flexural strength by using three-point bend test, scanning electron microscope (SEM, optical microstructure, grain size analysis, and surface roughness. It was found that good/excellent mechanical properties were obtained in AA 7075-TiB2-Gr reinforced in situ hybrid composite compared to alloy due to particulate strengthening of ceramic particles of TiB2 in the matrix. Further, Al 7075-3% TiB2-1% Gr hybrid in situ composite exhibited improved machinability over the alloy and composites due to self-lubricating property given by the Gr particles in the materials.

One-pot in situ redox synthesis of hexacyanoferrate/conductive polymer hybrids as lithium-ion battery cathodes.

Science.gov (United States)

Wong, Min Hao; Zhang, Zixuan; Yang, Xianfeng; Chen, Xiaojun; Ying, Jackie Y

2015-09-14

An efficient and adaptable method is demonstrated for the synthesis of lithium hexacyanoferrate/conductive polymer hybrids for Li-ion battery cathodes. The hybrids were synthesized via a one-pot method, involving a redox-coupled reaction between pyrrole monomers and the Li3Fe(CN)6 precursor. The hybrids showed much better cyclability relative to reported Prussian Blue (PB) analogs.

Clinical application of fluorescence in situ hybridization for prenatal diagnosis

Directory of Open Access Journals (Sweden)

Shu-fang JIANG

2012-07-01

Full Text Available Objective To establish and optimize the procedures of fluorescence in situ hybridization（FISH）, and evaluate its clinical value in rapid prenatal diagnosis of fetal numerical abnormality of chromosomes 21, 18, 13, X, Y. Methods Amniotic fluid or fetal blood was sampled by routine invasive procedures. After the amniotic fluid cells or fetal blood cells were separated and sequentially processed with hypotonic solution, fixation solution, smear and high temperature, they were hybridized in situ with two panels of specific fluorescence probes to detect numerical abnormality of chromosomes 21, 18, 13, X, Y. All the samples were also cultured and analyzed for their karyotype by conventional methods. Results When it was used as a diagnostic criterion of chromosomal number that the fluorescence signals were observed in ≥90% cells, GLP 13/GLP 21 probe panel showed 2 green/2 red fluorescence signals and CSP18/CSP X/CSP Y probe panel showed 2 blue/2 yellow (female or 2 blue/1 yellow/1 red fluorescence signals (male under normal condition. The test reports of all 196 cases were sent out in 72-96 hours, and 7 cases of Down syndrome, 2 cases of trisomy 18 and 1 case of sex chromosomal numerical abnormality were detected, which were accordant with karyotype analysis results reported one month later. Conclusions FISH has potential for clinical application, and is applicable to rapid prenatal diagnosis of fetal numerical abnormality of chromosomes 21, 18, 13, X, Y. The rapid FISH, together with conventional karyotyping, offer a valuable means for prenatal diagnosis of fetal aneuploidies.

Specific identification of human papillomavirus type in cervical smears and paraffin sections by in situ hybridization with radioactive probes: a preliminary communication

International Nuclear Information System (INIS)

Gupta, J.; Gendelman, H.E.; Naghashfar, Z.; Gupta, P.; Rosenshein, N.; Sawada, E.; Woodruff, J.D.; Shah, K.

1985-01-01

Cervical Papanicolaou smears and paraffin sections of biopsy specimens obtained from women attending dysplasia clinics were examined for viral DNA sequences by in situ hybridization technique using 35 S-labeled cloned recombinant DNA probes of human papillomavirus (HPV) types 6, 11, and 16. These and one unrelated DNA probe complementary to measles virus RNA were labeled by nick translation using either one or two 35 S-labeled nucleotides. Paraffin sections and cervical smears were collected on pretreated slides, hybridized with the probes under stringent or nonstringent conditions for 50 h, and autoradiographed. Additional cervical specimens from the same women were examined for the presence of genus-specific papillomavirus capsid antigen by the immunoperoxidase technique. Preliminary results may be summarized as follows. The infecting virus could be identified in smears as well as in sections. Viral DNA sequences were detected only when there were condylomatous cells in the specimen and in only a proportion of the condylomatous cells. Even under stringent conditions, some specimens reacted with both HPV-6 and HPV-11. In some instances, the cells did not hybridize with any of the three probes even when duplicate specimens contained frankly condylomatous, capsid antigen-positive cells. In situ hybridization of Papanicolaou smears or of tissue sections is a practical method for diagnosis and follow-up of specific papillomavirus infection using routinely collected material

Localization of the Norrie disease gene mRNA by in situ hybridization.

Science.gov (United States)

Hartzer, M K; Cheng, M; Liu, X; Shastry, B S

1999-07-15

Norrie disease is a rare X-linked recessive neurodevelopmental disorder. The affected males manifest congenital blindness, which is often associated with hearing loss, mental retardation and psychiatric problems. Genetic linkage studies have localized the gene to the short arm of the X-chromosome and the gene has been isolated recently. The encoded protein is a member of the superfamily of growth factors containing a cystine knot motif and may be involved in cell adhesion and neurodevelopment. Molecular genetic analysis revealed a large number of missense, nonsense, deletion, and splice-site mutations among Norrie patients. In order to further determine the role of the Norrie disease gene, we studied the distribution pattern of its mRNA in the retina and in brain by in situ hybridization. The results show abundant hybridization signals in outer nuclear, inner nuclear, and ganglion cell layers of the retina in all three species (mice, rabbit, and human) examined. There was no significant expression in the vitreous body, lens, and rod outer segment. High expression levels were also observed in the cerebellar granular layer, hippocampus, olfactory bulb, cortex, and epithelium of the rabbit brain. These data suggest that the Norrie disease gene could play a critical role in the differentiation or maintenance of the differentiated state of the retina.

Experimental demonstration of optical data links using a hybrid CAP/QAM modulation scheme.

Science.gov (United States)

Wei, J L; Ingham, J D; Cheng, Q; Cunningham, D G; Penty, R V; White, I H

2014-03-15

The first known experimental demonstrations of a 10  Gb/s hybrid CAP-2/QAM-2 and a 20  Gb/s hybrid CAP-4/QAM-4 transmitter/receiver-based optical data link are performed. Successful transmission over 4.3 km of standard single-mode fiber (SMF) is achieved, with a link power penalty ∼0.4  dBo for CAP-2/QAM-2 and ∼1.5  dBo for CAP-4/QAM-4 at BER=10(-9).

Electric and hybrid vehicle self-certification and verification procedures: Market Demonstration Program

Energy Technology Data Exchange (ETDEWEB)

None

1980-03-01

The process by which a manufacturer of an electric or hybrid vehicle certifies that his vehicle meets the DOE Performance Standards for Demonstration is described. Such certification is required for any vehicles to be purchased under the Market Demonstration Program. It also explains the verification testing process followed by DOE for testing to verify compliance. Finally, the document outlines manufacturer responsibilities and presents procedures for recertification of vehicles that have failed verification testing.

DEMONSTRATION OF THE GENUINE ISO-12P CHARACTER OF THE STANDARD MARKER CHROMOSOME OF TESTICULAR GERM-CELL TUMORS AND IDENTIFICATION OF FURTHER CHROMOSOME-12 ABERRATIONS BY COMPETITIVE INSITU HYBRIDIZATION

NARCIS (Netherlands)

SUIJKERBUIJK, RF; VANDEVEEN, AY; VANECHTEN, J; BUYS, CHCM; DEJONG, B; OOSTERHUIS, JW; WARBURTON, DA; CASSIMAN, JJ; SCHONK, D; VANKESSEL, AG

The recently developed competitive in situ hybridization (CISH) strategy was applied to the analysis of chromosome 12 aberrations in testicular germ cell tumors (TGCTs). DNAs from two rodent-human somatic cell hybrids, containing either a normal chromosome 12 or the p arm of chromosome 12 as their

A whole-mount in situ hybridization method for microRNA detection in Caenorhabditis elegans.

Science.gov (United States)

Andachi, Yoshiki; Kohara, Yuji

2016-07-01

Whole-mount in situ hybridization (WISH) is an outstanding method to decipher the spatiotemporal expression patterns of microRNAs (miRNAs) and provides important clues for elucidating their functions. The first WISH method for miRNA detection was developed in zebrafish. Although this method was quickly adapted for other vertebrates and fruit flies, WISH analysis has not been successfully used to detect miRNAs in Caenorhabditis elegans Here, we show a novel WISH method for miRNA detection in C. elegans Using this method, mir-1 miRNA was detected in the body-wall muscle where the expression and roles of mir-1 miRNA have been previously elucidated. Application of the method to let-7 family miRNAs, let-7, mir-48, mir-84, and mir-241, revealed their distinct but partially overlapping expression patterns, indicating that miRNAs sharing a short common sequence were distinguishably detected. In pash-1 mutants that were depleted of mature miRNAs, signals of mir-48 miRNA were greatly reduced, suggesting that mature miRNAs were detected by the method. These results demonstrate the validity of WISH to detect mature miRNAs in C. elegans. © 2016 Andachi and Kohara; Published by Cold Spring Harbor Laboratory Press for the RNA Society.

Evidence for Integrity of Parental Genomes in the Diploid Hybridogenetic Water Frog Pelophylax esculentus by Genomic in situ Hybridization

Czech Academy of Sciences Publication Activity Database

Zalésna, A.; Choleva, Lukáš; Ogielska, M.; Rábová, Marie; Marec, František; Ráb, Petr

2011-01-01

Roč. 134, č. 3 (2011), s. 206-212 ISSN 1424-8581 R&D Projects: GA MŠk LC06073; GA ČR GA523/09/2106 Institutional research plan: CEZ:AV0Z50450515; CEZ:AV0Z50070508 Keywords : Amphibia * Chromosomes * Genomic in situ hybridization (GISH) Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.533, year: 2011

Filter-Adapted Fluorescent In Situ Hybridization (FA-FISH) for Filtration-Enriched Circulating Tumor Cells.

Science.gov (United States)

Oulhen, Marianne; Pailler, Emma; Faugeroux, Vincent; Farace, Françoise

2017-01-01

Circulating tumor cells (CTCs) may represent an easily accessible source of tumor material to assess genetic aberrations such as gene-rearrangements or gene-amplifications and screen cancer patients eligible for targeted therapies. As the number of CTCs is a critical parameter to identify such biomarkers, we developed fluorescent in situ hybridization (FISH) for CTCs enriched on filters (filter-adapted-FISH, FA-FISH). Here, we describe the FA-FISH protocol, the combination of immunofluorescent staining (DAPI/CD45) and FA-FISH techniques, as well as the semi-automated microscopy method that we developed to improve the feasibility and reliability of FISH analyses in filtration-enriched CTC.

Analysis of the Ceratitis capitata y chromosome using in situ hybridization to mitotic chromosomes

International Nuclear Information System (INIS)

Willhoeft, U.; Franz, G.

1998-01-01

In Ceratitis capitata the Y chromosome is responsible for sex-determination. We used fluorescence in situ hybridization (FISH) for cytogenetic analysis of mitotic chromosomes. FISH with the wild-type strain EgyptII and two repetitive DNA probes enabled us to differentiate between the short and the long arm of the Y chromosome and gives a much better resolution than C-banding of mitotic chromosomes. We identified the Y-chromosomal breakpoints in Y-autosome translocations using FISH. Even more complex rearrangements i.e. deletions and insertions in some translocation strains were detected by this method. A strategy for mapping the primary sex determination factor in Ceratitis capitata by FISH is presented. (author)

Demonstration testing and evaluation of in situ soil heating. Treatability study work plan, Revision 1

Energy Technology Data Exchange (ETDEWEB)

Sresty, G.C.

1994-07-07

A Treatability Study planned for the demonstration of the in situ electromagnetic (EM) heating process to remove organic solvents is described in this Work Plan. The treatability study will be conducted by heating subsurface vadose-zone soils in an organic plume adjacent to the Classified Burial Ground K-1070-D located at K-25 Site, Oak Ridge. The test is scheduled to start during the fourth quarter of FY94 and will be completed during the first quarter of FY95. The EM heating process for soil decontamination is based on volumetric heating technologies developed during the `70s for the recovery of fuels from shale and tar sands by IIT Research Institute (IITRI) under a co-operative program with the US Department of Energy (DOE). Additional modifications of the technology developed during the mid `80s are currently used for the production of heavy oil and waste treatment. Over the last nine years, a number of Government agencies (EPA, Army, AF, and DOE) and industries sponsored further development and testing of the in situ heating and soil decontamination process for the remediation of soils containing hazardous organic contaminants. In this process the soil is heated in situ using electrical energy. The contaminants are removed from the soil due to enhanced vaporization, steam distillation and stripping. IITRI will demonstrate the EM Process for in situ soil decontamination at K-25 Site under the proposed treatability study. Most of the contaminants of concern are volatile organics which can be removed by heating the soil to a temperature range of 85 to 95 C. The efficiency of the treatment will be determined by comparing the concentration of contaminants in soil samples. Samples will be obtained before and after the demonstration for a measurement of the concentration of contaminants of concern.

A simple protocol for attenuating the auto-fluorescence of cyanobacteria for optimized fluorescence in situ hybridization (FISH) imaging.

Science.gov (United States)

Zeller, Perrine; Ploux, Olivier; Méjean, Annick

2016-03-01

Cyanobacteria contain pigments, which generate auto-fluorescence that interferes with fluorescence in situ hybridization (FISH) imaging of cyanobacteria. We describe simple chemical treatments using CuSO4 or H2O2 that significantly reduce the auto-fluorescence of Microcystis strains. These protocols were successfully applied in FISH experiments using 16S rRNA specific probes and filamentous cyanobacteria. Copyright © 2016 Elsevier B.V. All rights reserved.

Detection of airborne Legionella while showering using liquid impingement and fluorescent in situ hybridization (FISH).

Science.gov (United States)

Deloge-Abarkan, Magali; Ha, Thi-Lan; Robine, Enric; Zmirou-Navier, Denis; Mathieu, Laurence

2007-01-01

Aerosols of water contaminated with Legionella bacteria constitute the only mode of exposure for humans. However, the prevention strategy against this pathogenic bacteria risk is managed through the survey of water contamination. No relationship linked the Legionella bacteria water concentration and their airborne abundance. Therefore, new approaches in the field of the metrological aspects of Legionella bioaerosols are required. This study was aimed at testing the main principles for bioaerosol collection (solid impaction, liquid impingement and filtration) and the in situ hybridization (FISH) method, both in laboratory and field assays, with the intention of applying such methodologies for airborne Legionella bacteria detection while showering. An aerosolization chamber was developed to generate controlled and reproducible L. pneumophila aerosols. This tool allowed the identification of the liquid impingement method as the most appropriate one for collecting airborne Legionella bacteria. The culturable fraction of airborne L. pneumophila recovered with the liquid impingement principle was 4 and 700 times higher compared to the impaction and filtration techniques, respectively. Moreover, the concentrations of airborne L. pneumophila in the impinger fluid were on average 7.0 x 10(5) FISH-cells m(-3) air with the fluorescent in situ hybridization (FISH) method versus 9.0 x 10(4) CFU m(-3) air with the culture method. These results, recorded under well-controlled conditions, were confirmed during the field experiments performed on aerosols generated by hot water showers in health institutions. This new approach may provide a more accurate characterization of aerobiocontamination by Legionella bacteria.

Utility and impact of early t(15;17) identification by Fluorescence In Situ Hybridization (FISH) in clinical decision making for patients in Acute Promyelocytic Leukemia (APL).

Science.gov (United States)

Kolhe, R; Mangaonkar, A; Mansour, J; Clemmons, A; Shaw, J; Dupont, B; Walczak, L; Mondal, A; Rojiani, A; Jillella, A; Kota, V

2015-08-01

Acute Promyelocytic Leukemia (APL) is a curable malignancy with studies showing above 90% survival. However, population-based studies looking at survival suggest that approximately 30% of patients with APL die during induction. Early demonstration of t(15;17) will lead to accurate decision making regarding treatment. The aim of this project was to validate earlier time frames for the Abbott Molecular Vysis LSI promyelocytic leukemia (PML)/ retinoic acid receptor alpha (RARA) fluorescence in situ hybridization (FISH) probe (ASR 6-16 h). Twenty patients (15 APL cases and five non-APL cases) were selected for validating various hybridization times for the FISH probe. Expected normal signal pattern was two red and two green signals (2R2G), and the most common expected abnormal signal pattern was two fusion (yellow) signals, one red and one green (2F1R1G) and/or one fusion, one red and one green (1F1R1G). The specificity of the probe ranged from 84% at 2 h, 86% at 4 h, 84% at 6 h, and 87% for overnight hybridization. The sensitivity increased from 79% at 2 h, 80% at 4 h, 81% at 6 h to 87% for overnight hybridization. Based on the validation studies, we recommend reading of FISH results at the 4-h incubation mark for a preliminary diagnosis and confirmation with overnight hybridization. © 2015 John Wiley & Sons Ltd.

Establishment of 60Co dose calibration curve using fluorescent in situ hybridization assay technique: Result of preliminary study

International Nuclear Information System (INIS)

Rahimah Abdul Rahim; Noriah Jamal; Noraisyah Mohd Yusof; Juliana Mahamad Napiah; Nelly Bo Nai Lee

2010-01-01

This study aims at establishing an in-vitro 60 Co dose calibration curve using Fluorescent In-Situ Hybridization assay technique for the Malaysian National Bio dosimetry Laboratory. Blood samples collected from a female healthy donor were irradiated with several doses of 60 Co radiation. Following culturing of lymphocytes, microscopic slides are prepared, denatured and hybridized. The frequencies of translocation are estimated in the metaphases. A calibration curve was then generated using a regression technique. It shows a good fit to a linear-quadratic model. The results of this study might be useful in estimating absorbed dose for the individual exposed to ionizing radiation retrospectively. This information may be useful as a guide for medical treatment for the assessment of possible health consequences. (author)

In situ hybridization analysis of the temporospatial expression of the midkine/pleiotrophin family in rat embryonic pituitary gland.

Science.gov (United States)

Fujiwara, Ken; Maliza, Rita; Tofrizal, Alimuddin; Batchuluun, Khongorzul; Ramadhani, Dini; Tsukada, Takehiro; Azuma, Morio; Horiguchi, Kotaro; Kikuchi, Motoshi; Yashiro, Takashi

2014-07-01

Pituitary gland development is controlled by numerous signaling molecules, which are produced in the oral ectoderm and diencephalon. A newly described family of heparin-binding growth factors, namely midkine (MK)/pleiotrophin (PTN), is involved in regulating the growth and differentiation of many tissues and organs. Using in situ hybridization with digoxigenin-labeled cRNA probes, we detected cells expressing MK and PTN in the developing rat pituitary gland. At embryonic day 12.5 (E12.5), MK expression was localized in Rathke's pouch (derived from the oral ectoderm) and in the neurohypophyseal bud (derived from the diencephalon). From E12.5 to E19.5, MK mRNA was expressed in the developing neurohypophysis, and expression gradually decreased in the developing adenohypophysis. To characterize MK-expressing cells, we performed double-staining of MK mRNA and anterior pituitary hormones. At E19.5, no MK-expressing cells were stained with any hormone. In contrast, PTN was expressed only in the neurohypophysis primordium during all embryonic stages. In situ hybridization clearly showed that MK was expressed in primitive (immature/undifferentiated) adenohypophyseal cells and neurohypophyseal cells, whereas PTN was expressed only in neurohypophyseal cells. Thus, MK and PTN might play roles as signaling molecules during pituitary development.

In Situ Hybridization of Pulp Fibers Using Mg-Al Layered Double Hydroxides

Directory of Open Access Journals (Sweden)

Carl-Erik Lange

2015-04-01

Full Text Available Inorganic Mg2+ and Al3+ containing layered double hydroxide (LDH particles were synthesised in situ from aqueous solution onto chemical pulp fibers of pine (Pinus sylvestris. High super saturated (hss solution with sodium carbonate produced LDH particles with an average diameter of 100–200 nm. Nano-size (70 nm LDH particles were found from fibers external surface and, to a lesser degree, from the S2 cell wall after synthesis via low super saturated (lss route. The synthesis via slow urea hydrolysis (Uhyd yielded micron and clay sized LDH (2–5 μm and enabled efficient fiber densification via mineralization of S2 fiber wall layer as indicated by TEM and compliance analysis. The Uhyd method decreased fiber compliance up to 50%. Reduction in the polymerisation degree of cellulose was observed with capillary viscometry. Thermogravimetric analysis showed that the hybridization with LDH reduced the exothermic heat, indicating, that this material can be incorporated in flame retardant applications. Fiber charge was assessed by Fibers 2015, 3 104 adsorption expermients with methylene blue (MB and metanil yellow (MY. Synthesis via lss route retained most of the fibres original charge and provided the highest capacity (10 μmol/g for anionic MY, indicating cationic character of hybrid fibers. Our results suggested that mineralized fibers can be potentially used in advanced applications such as biocomposites and adsorbent materials.

Visualization of mcr mRNA in a methanogen by fluorescence in situ hybridization with an oligonucleotide probe and two-pass tyramide signal amplification (two-pass TSA-FISH).

Science.gov (United States)

Kubota, Kengo; Ohashi, Akiyoshi; Imachi, Hiroyuki; Harada, Hideki

2006-09-01

Two-pass tyramide signal amplification-fluorescence in situ hybridization (two-pass TSA-FISH) with a horseradish peroxidase (HRP)-labeled oligonucleotide probe was applied to detect prokaryotic mRNA. In this study, mRNA of a key enzyme for methanogenesis, methyl coenzyme M reductase (mcr), in Methanococcus vannielii was targeted. Applicability of mRNA-targeted probes to in situ hybridization was verified by Clone-FISH. It was observed that sensitivity of two-pass TSA-FISH was significantly higher than that of TSA-FISH, which was further increased by the addition of dextran sulphate in TSA working solution. Signals from two-pass TSA-FISH were more reliable compared to the weak, spotty signals yielded by TSA-FISH.

Demonstration, testing, & evaluation of in situ heating of soil. Draft final report, Volume I

Energy Technology Data Exchange (ETDEWEB)

Dev, H.; Enk, J.; Jones, D.; Saboto, W.

1996-02-12

This document is a draft final report (Volume 1) for US DOE contract entitled, {open_quotes}Demonstration Testing and Evaluation of In Situ Soil Heating,{close_quotes} Contract No. DE-AC05-93OR22160, IITRI Project No. C06787. This report is presented in two volumes. Volume I contains the technical report and Volume II contains appendices with background information and data. In this project approximately 300 cu. yd. of clayey soil containing a low concentration plume of volatile organic chemicals was heated in situ by the application of electrical energy. It was shown that as a result of heating the effective permeability of soil to air flow was increased such that in situ soil vapor extraction could be performed. The initial permeability of soil was so low that the soil gas flow rate was immeasurably small even at high vacuum levels. When scaled up, this process can be used for the environmental clean up and restoration of DOE sites contaminated with VOCs and other organic chemicals boiling up to 120{degrees} to 130{degrees}C in the vadose zone. Although it may applied to many types of soil formations, it is particularly attractive for low permeability clayey soil where conventional in situ venting techniques are limited by low air flow.

Rapid molecular cytogenetic analysis of X-chromosomal microdeletions: Fluorescence in situ hybridization (FISH) for complex glycerol kinase deficiency

Energy Technology Data Exchange (ETDEWEB)

Worley, K.C.; Lindsay, E.A.; McCabe, E.R.B. [Baylor College of Medicine, Houston, TX (United States)] [and others

1995-07-17

Diagnosis of X-chromosomal microdeletions has relied upon the traditional methods of Southern blotting and DNA amplification, with carrier identification requiring time-consuming and unreliable dosage calculations. In this report, we describe rapid molecular cytogenetic identification of deleted DNA in affected males with the Xp21 contiguous gene syndrome (complex glycerol kinase deficiency, CGKD) and female carriers for this disorder. CGKD deletions involve the genes for glycerol kinase, Duchenne muscular dystrophy, and/or adrenal hypoplasia congenita. We report an improved method for diagnosis of deletions in individuals with CGKD and for identification of female carriers within their families using fluorescence in situ hybridization (FISH) with a cosmid marker (cosmid 35) within the glycerol kinase gene. When used in combination with an Xq control probe, affected males demonstrate a single signal from the control probe, while female carriers demonstrate a normal chromosome with two signals, as well as a deleted chromosome with a single signal from the control probe. FISH analysis for CGKD provides the advantages of speed and accuracy for evaluation of submicroscopic X-chromosome deletions, particularly in identification of female carriers. In addition to improving carrier evaluation, FISH will make prenatal diagnosis of CGKD more readily available. 17 refs., 2 figs.

Detection of TTV-DNA in PBMC using digoxigenin labelled probe by in situ hybridization

International Nuclear Information System (INIS)

Liu Yang; Qi Qige

2002-01-01

To determine TTV-DNA in PBMC in patients with viral hepatitis, a study of in situ hybridization using digoxigenin labelled probe by PCR method to the TTV ORF1 region was performed on PBMC. Results showed that the detection rate of TTV-DNA using double-stranded probe in TTV-DNA positive group in sera was 58.06 (18/31), and the detection rate of TTV-DNA using double-stranded probe in TTV-DNA negative group in sera was 27.59 (8/29). For TTV-DNA positive group detected by double- stranded probe, then we use negative- stranded probe to detect their replication. The detection rate was 22.2%(4/18). Conclusions: TTV can infect PBMC and replicate in PBMC

Fabrication of nanostructured graphene/polyaniline hybrid material for supercapacitors

Energy Technology Data Exchange (ETDEWEB)

Wang, H.L.; Hao, Q.L.; Wang, X.; Lu, L.D.; Yang, X.J. [Nanjing Univ. of Science and Technology (China). Key Laboratory for Soft Chemistry and Functional Materials, Ministry of Education

2010-07-01

In this study, a flexible graphene/polyaniline hybrid material was prepared using an in situ polymerization-reduction/dedoping-redoping process for use as a supercapacitor electrode. Graphene oxide and a single layer of graphite oxide were used as a substrate material for the graphene oxide-polyaniline composite using an in situ polymerization method. The composite was then treated with a hot sodium hydroxide solution in order to produce a reduced graphene oxide/polyaniline hybrid material. The sodium hydroxide was also used as a dedoping reagent for the polyaniline in the composite. A thin, uniform and flexible conducting graphene/polyaniline product with an unchanged morphology was obtained using the process. Analyses of the material demonstrated that the composite showed an improved electrochemical performance than the pure individual components, with a specific capacitance of 1126 F per g and a retention life of 84 per cent after 1000 cycles. 4 refs., 1 fig.

Development of a 16S rRNA-targeted fluorescence in situ hybridization probe for quantification of the ammonia-oxidizer Nitrosotalea devanaterra and its relatives.

Science.gov (United States)

Restrepo-Ortiz, C X; Merbt, S N; Barrero-Canossa, J; Fuchs, B M; Casamayor, E O

2018-04-28

The Thaumarchaeota SAGMCG-1 group and, in particular, members of the genus Nitrosotalea have high occurrence in acidic soils, the rhizosphere, groundwater and oligotrophic lakes, and play a potential role in nitrogen cycling. In this study, the specific oligonucleotide fluorescence in situ hybridization probe SAG357 was designed for this Thaumarchaeota group based on the available 16S rRNA gene sequences in databases, and included the ammonia-oxidizing species Nitrosotalea devanaterra. Cell permeabilization for catalyzed reporter deposition fluorescence in situ detection and the hybridization conditions were optimized on enrichment cultures of the target species N. devanaterra, as well as the non-target ammonia-oxidizing archaeon Nitrosopumilus maritimus. Probe specificity was improved with a competitor oligonucleotide, and fluorescence intensity and cell visualization were enhanced by the design and application of two adjacent helpers. Probe performance was tested in soil samples along a pH gradient, and counting results matched the expected in situ distributions. Probe SAG357 and the CARD-FISH protocol developed in the present study will help to improve the current understanding of the ecology and physiology of N. devanaterra and its relatives in natural environments. Copyright © 2018 Elsevier GmbH. All rights reserved.

The effect of aging of formalin-fixed paraffin-embedded tissues on the in situ hybridization and immunohistochemistry signals in cervical lesions.

Science.gov (United States)

Nuovo, Allison J; Garofalo, Michela; Mikhail, Alexandria; Nicol, Alcina F; Vianna-Andrade, Cecilia; Nuovo, Gerard J

2013-09-01

Formalin-fixed, paraffin-embedded tissues are widely used in biomedical research but little is known about the effect of the age of the block or unstained slides on the in situ hybridization or immunohistochemistry signal. We compared the in situ-based and immunohistochemistry-based signals for cervical intraepithelial neoplasia samples that ranged from 0 to 15 years of age. There was a progressive and statistically significant decrease in the strength of the p16 signal when comparing tissues prepared from recent unstained slides (0 to 1 y old, mean score of 92%) to those of intermediate age (5 to 7 y old, mean score of 49%) to old unstained slides (cut 13 to 15 y ago, mean score of 10%). Equivalent, progressive, and significant decreases in the intensity of the signals for microRNAs, CD45, and human papillomavirus DNA were seen in tissues stored on slides from 5 to 7 years and 13 to 15 years, respectively. However, the diminution of signal was much less, although still statistically significant, if the sections from the 13- to 15-year-old paraffin blocks were prepared in 2012. The data likely does not represent degradation of the targets as extraction of several microRNA from the old blocks showed no detectable degradation, despite the markedly weakened in situ hybridization signal. It is concluded that in situ-based signal for DNA, microRNAs, and proteins in paraffin-embedded tissues are significantly reduced over time, especially when stored long term on glass slides which, in turn, can lead to a significant underestimation of the amount and presence of the nucleic acid or protein target.

Quantitative fluorescence in situ hybridization measurement of telomere length in skin with/without sun exposure or actinic keratosis.

Science.gov (United States)

Ikeda, Hiroyuki; Aida, Junko; Hatamochi, Atsushi; Hamasaki, Yoichiro; Izumiyama-Shimomura, Naotaka; Nakamura, Ken-Ichi; Ishikawa, Naoshi; Poon, Steven S; Fujiwara, Mutsunori; Tomita, Ken-Ichiro; Hiraishi, Naoki; Kuroiwa, Mie; Matsuura, Masaaki; Sanada, Yukihiro; Kawano, Youichi; Arai, Tomio; Takubo, Kaiyo

2014-03-01

Chromosomal and genomic instability due to telomere dysfunction is known to play an important role in carcinogenesis. To study telomere shortening in the epidermis surrounding actinic keratosis, we measured telomere lengths of basal, parabasal, and suprabasal cells in epidermis with actinic keratosis (actinic keratosis group, n = 18) and without actinic keratosis (sun-protected, n = 15, and sun-exposed, n = 13 groups) and in actinic keratosis itself as well as in dermal fibroblasts in the 3 groups, using quantitative fluorescence in situ hybridization. Among the 3 cell types, telomeres of basal cells were not always the longest, suggesting that tissue stem cells are not necessarily located among basal cells. Telomeres of basal cells in the sun-exposed group were shorter than those in the sun-protected group. Telomeres in the background of actinic keratosis and in actinic keratosis itself and those of fibroblasts in actinic keratosis were significantly shorter than those in the controls. Our findings demonstrate that sun exposure induces telomere shortening and that actinic keratosis arises from epidermis with shorter telomeres despite the absence of any histologic atypia. © 2014.

Identification of supernumerary ring chromosome 1 mosaicism using fluorescence in situ hybridization.

Science.gov (United States)

Chen, H; Tuck-Muller, C M; Batista, D A; Wertelecki, W

1995-03-27

We report on a 15-year-old black boy with severe mental retardation, multiple congenital anomalies, and a supernumerary ring chromosome mosaicism. Fluorescence in situ hybridization with a chromosome 1 painting probe (pBS1) identified the ring as derived from chromosome 1. The karyotype was 46,XY/47,XY,+r(1)(p13q23). A review showed 8 reports of ring chromosome 1. In 5 cases, the patients had a non-supernumerary ring chromosome 1 resulting in partial monosomies of the short and/or long arm of chromosome 1. In 3 cases, the presence of a supernumerary ring resulted in partial trisomy of different segments of chromosome 1. In one of these cases the supernumerary ring was composed primarily of the centromere and the heterochromatic region of chromosome 1, resulting in normal phenotype. Our patient represents the third report of a supernumerary ring chromosome 1 resulting in abnormal phenotype.

Assignment of Alzheimer's presenilin-2 (PS-2) gene to 1q42.1 by fluorescence in situ hybridization.

Science.gov (United States)

Takano, T; Sahara, N; Yamanouchi, Y; Mori, H

1997-01-17

Presenilin-2 (PS-2) was suggested to be localized on 1q31-42 based on linkage analysis and cDNA cloning. The final identification of PS-2 as the causal gene for early-onset familial Alzheimer's disease in Voga-German pedigrees was concluded based on the point mutation found in the candidate cDNA isolated from this familial AD. We present evidence of its physical genome mapping of PS-2 on chromosome 1q42.1 by fluorescence in situ hybridization method.

DEMONSTRATION BULLETIN: IN SITU ELECTROKINETIC EXTRACTION SYSTEM - SANDIA NATIONAL LABORATORIES

Science.gov (United States)

Sandia National Laboratories (SNL) has developed an in situ soil remediation system that uses electrokinetic principles to remediate hexavalent chromium-contaminated unsaturated or partially saturated soils. The technology involves the in situ application of direct current to the...

Synthetic Strategies in the Preparation of Polymer/Inorganic Hybrid Nanoparticles

Science.gov (United States)

Hood, Matthew A.; Mari, Margherita; Muñoz-Espí, Rafael

2014-01-01

This article reviews the recent advances and challenges in the preparation of polymer/inorganic hybrid nanoparticles. We mainly focus on synthetic strategies, basing our classification on whether the inorganic and the polymer components have been formed in situ or ex situ, of the hybrid material. Accordingly, four types of strategies are identified and described, referring to recent examples: (i) ex situ formation of the components and subsequent attachment or integration, either by covalent or noncovalent bonding; (ii) in situ polymerization in the presence of ex situ formed inorganic nanoparticles; (iii) in situ precipitation of the inorganic components on or in polymer structures; and (iv) strategies in which both polymer and inorganic component are simultaneously formed in situ. PMID:28788665

In situ intercalation strategies for device-quality hybrid inorganic-organic self-assembled quantum wells

Science.gov (United States)

Pradeesh, K.; Baumberg, J. J.; Prakash, G. Vijaya

2009-07-01

Thin films of self-organized quantum wells of inorganic-organic hybrid perovskites of (C6H9C2H4NH3)2PbI4 are formed from a simple intercalation strategy to yield well-ordered uniform films over centimeter-size scales. These films compare favorably with traditional solution-chemistry-synthesized thin films. The hybrid films show strong room-temperature exciton-related absorption and photoluminescence, which shift with fabrication protocol. We demonstrate the potential of this method for electronic and photonic device applications.

Demonstration, testing, and evaluation of in situ heating of soil. Volume 1, Final report

International Nuclear Information System (INIS)

Dev, H.; Enk, J.; Jones, D.; Sabato, W.

1996-01-01

This document is a final reports in two volumes. Volume I contains the technical report and Volume II contains appendices with background information and data. In this project approximately 300 cubic yards of clayey soil containing a low concentration plume of volatile organic chemicals was heated in situ by the application of electrical energy. It was shown that as a result of heating the effective permeability of soil to air flow was increased such that in situ soil vapor extraction could be performed. The initial permeability of soil was so low that the soil gas flow rate was immeasurably small even at high vacuum levels. It was demonstrated that the mass flow rate of the volatile organic chemicals was enhanced in the recovered soil gas as a result of heating

Demonstration, testing, and evaluation of in situ heating of soil. Volume 1, Final report

Energy Technology Data Exchange (ETDEWEB)

Dev, H.; Enk, J.; Jones, D.; Sabato, W.

1996-04-05

This document is a final reports in two volumes. Volume I contains the technical report and Volume II contains appendices with background information and data. In this project approximately 300 cubic yards of clayey soil containing a low concentration plume of volatile organic chemicals was heated in situ by the application of electrical energy. It was shown that as a result of heating the effective permeability of soil to air flow was increased such that in situ soil vapor extraction could be performed. The initial permeability of soil was so low that the soil gas flow rate was immeasurably small even at high vacuum levels. It was demonstrated that the mass flow rate of the volatile organic chemicals was enhanced in the recovered soil gas as a result of heating.

Analysis of Single-cell Gene Transcription by RNA Fluorescent In Situ Hybridization (FISH)

DEFF Research Database (Denmark)

Ronander, Elena; Bengtsson, Dominique C; Joergensen, Louise

2012-01-01

Adhesion of Plasmodium falciparum infected erythrocytes (IE) to human endothelial receptors during malaria infections is mediated by expression of PfEMP1 protein variants encoded by the var genes. The haploid P. falciparum genome harbors approximately 60 different var genes of which only one has...... been believed to be transcribed per cell at a time during the blood stage of the infection. How such mutually exclusive regulation of var gene transcription is achieved is unclear, as is the identification of individual var genes or sub-groups of var genes associated with different receptors...... fluorescent in situ hybridization (FISH) analysis of var gene transcription by the parasite in individual nuclei of P. falciparum IE(1). Here, we present a detailed protocol for carrying out the RNA-FISH methodology for analysis of var gene transcription in single-nuclei of P. falciparum infected human...

Fluorescence in situ hybridization detection of cytogenetic abnormalities and prognosis in multiple myeloma

Directory of Open Access Journals (Sweden)

Zhou Xu

2015-01-01

Full Text Available We evaluated the prognosis of patients with newly diagnosed multiple myeloma (MM and attempted to find a suitable treatment strategy for them. Interphase fluorescence in situ hybridization (FISH detection was performed on 57 patients with MM. The following probes: IgH, p53, 1q21, RB1, and D13S319 specific for the rearrangements of 14q32, 17p13, 1q21 and 13q14 were used. Fluorescent hybridization signals were observed using an Olympus BX60 epifluorescence microscope equipped with filters for detecting fluoroisothiocyanate (FITC, Texas red, and 4'-6-Diamidino-2-phenylindole (DAPI. Triple color clone-specific images were captured using a Quips XL genetic workstation. The mortalities in patients with moderate prognosis (66.7% and poor prognosis (50% were significantly higher compared with that in patients with good prognosis (15%, P<0.05. All the patients in good and moderate prognosis groups achieved complete remission (CR/very good partial remission (VGPR/partial remission (PR, whereas only half of the cases in the poor prognosis group reached this level. Patients 2 supported by autologous hematopoietic stem-cell transplantation presented CR/PR and long survival. For those with poor prognosis, a proper therapeutic regimen and timely transplantation, especially tandem transplantation, was necessary due to the rapid progression and complications.

Fluorescence In Situ Hybridization (FISH Signal Analysis Using Automated Generated Projection Images

Directory of Open Access Journals (Sweden)

Xingwei Wang

2012-01-01

Full Text Available Fluorescence in situ hybridization (FISH tests provide promising molecular imaging biomarkers to more accurately and reliably detect and diagnose cancers and genetic disorders. Since current manual FISH signal analysis is low-efficient and inconsistent, which limits its clinical utility, developing automated FISH image scanning systems and computer-aided detection (CAD schemes has been attracting research interests. To acquire high-resolution FISH images in a multi-spectral scanning mode, a huge amount of image data with the stack of the multiple three-dimensional (3-D image slices is generated from a single specimen. Automated preprocessing these scanned images to eliminate the non-useful and redundant data is important to make the automated FISH tests acceptable in clinical applications. In this study, a dual-detector fluorescence image scanning system was applied to scan four specimen slides with FISH-probed chromosome X. A CAD scheme was developed to detect analyzable interphase cells and map the multiple imaging slices recorded FISH-probed signals into the 2-D projection images. CAD scheme was then applied to each projection image to detect analyzable interphase cells using an adaptive multiple-threshold algorithm, identify FISH-probed signals using a top-hat transform, and compute the ratios between the normal and abnormal cells. To assess CAD performance, the FISH-probed signals were also independently visually detected by an observer. The Kappa coefficients for agreement between CAD and observer ranged from 0.69 to 1.0 in detecting/counting FISH signal spots in four testing samples. The study demonstrated the feasibility of automated FISH signal analysis that applying a CAD scheme to the automated generated 2-D projection images.

Detection of Bacteria by Fluorescence in Situ Hybridization in Culture-Negative Soft Tissue Filler Lesions

DEFF Research Database (Denmark)

Bjarnsholt, Thomas; Tolker-Nielsen, Tim; Givskov, Michael

2009-01-01

BACKGROUND Adverse reactions to polyacrylamide gel occur as swellings or nodules, and controversy exists whether these are due to bacterial infection or an autoimmune reaction to the filler. OBJECTIVES Biopsies from culture-negative long-lasting nodules after injection with different types...... of polyacrylamide gel were examined with a combination of Gram stain and fluorescence in situ hybridization. RESULTS Bacteria were detected in biopsies from seven of eight patients. They inhabited gel and intervening tissue and tended to lie in aggregates. CONCLUSION This study supports the assumption...... that infection with bacteria in aggregates causes culture-negative late adverse reactions to polyacrylamide gel, suggesting a biofilm environment. The authors have indicated no significant interest with commercial supporters....

Demonstration, testing, ampersand evaluation of in situ heating of soil. Draft final report, Volume I

International Nuclear Information System (INIS)

Dev, H.; Enk, J.; Jones, D.; Saboto, W.

1996-01-01

This document is a draft final report (Volume 1) for US DOE contract entitled, open-quotes Demonstration Testing and Evaluation of In Situ Soil Heating,close quotes Contract No. DE-AC05-93OR22160, IITRI Project No. C06787. This report is presented in two volumes. Volume I contains the technical report and Volume II contains appendices with background information and data. In this project approximately 300 cu. yd. of clayey soil containing a low concentration plume of volatile organic chemicals was heated in situ by the application of electrical energy. It was shown that as a result of heating the effective permeability of soil to air flow was increased such that in situ soil vapor extraction could be performed. The initial permeability of soil was so low that the soil gas flow rate was immeasurably small even at high vacuum levels. When scaled up, this process can be used for the environmental clean up and restoration of DOE sites contaminated with VOCs and other organic chemicals boiling up to 120 degrees to 130 degrees C in the vadose zone. Although it may applied to many types of soil formations, it is particularly attractive for low permeability clayey soil where conventional in situ venting techniques are limited by low air flow

PRINS and in situ PCR protocols

National Research Council Canada - National Science Library

Gosden, John R

1997-01-01

... mapping of DNA sequences on chromosomes and location of gene expression followed the invention and refinement of in situ hybridization. Among the most recent technical developments has been the use of oligonucleotide primers to detect and amplify or extend complementary sequences in situ, and it is to this novel field that PRINS and In S...

A populational survey of 45S rDNA polymorphism in the Jefferson salamander Ambystoma jeffersonianum revealed by fluorescence in situ hybridization (FISH

Directory of Open Access Journals (Sweden)

Jinzhong FU

2009-04-01

Full Text Available The chromosomal localization of 45S ribosomal RNA genes in Ambystoma jeffersonianum was determined by fluorescence in situ hybridization with 18S rDNA fragment as a probe (FISH-rDNA. Our results revealed the presence of rDNA polymorphism among A.jeffersonianum populations in terms of number, location and FISH signal intensity on the chromosomes. Nine rDNA cytotypes were found in ten geographically isolated populations and most of them contained derivative rDNA sites. Our preliminary study provides strong indication of karyotypic diversification of A.jeffersonianum that is demonstrated by intraspecific variation of 45S rDNA cytotypes. rDNA cytotype polymorphism has been described in many other caudate amphibians. We predict that habitat isolation, low dispersal ability and decline of effective population size could facilitate the fixation and accumulation of variable rDNA cytotypes during their chromosome evolution.

Evaluation of HER2 gene amplification in invasive breast cancer using a dual-color chromogenic in situ hybridization (dual CISH).

Science.gov (United States)

Kato, Nobuaki; Itoh, Hitoshi; Serizawa, Akihiko; Hatanaka, Yutaka; Umemura, Shinobu; Osamura, R Yoshiyuki

2010-07-01

Fluorescence in situ hybridization (FISH) assay is considered the 'gold standard' for evaluation of HER2/neu (HER2) gene status, however, it is difficult to recognize morphologic features of tumors using fluorescence microscopy. Thus, chromogenic in situ hybridization (CISH) has been proposed as an alternative method to evaluate HER2 gene amplification. Here, we examined the dual color CISH (dual CISH) method which provides information regarding the copy number of the HER2 gene and chromosome 17 centromere from a single slide. We examined 40 cases of invasive ductal carcinomas of the breast that were resected surgically. HER2 gene status was assessed with FISH (Abbott) and dual CISH (Dako). HER2 gene amplification status was classified according to the guidelines of the American Society of Clinical Oncology and College of American Pathologists (ASCO/CAP). Comparison of the cut-off values for HER2/chromosome 17 centromere copy number ratio obtained by dual CISH and FISH showed that there was almost perfect agreement between two methods (Kappa coefficient 0.96). The results of the two commercial products were almost consistent for evaluation of HER2 gene counts on the sections. The current study proved that dual CISH is comparable with FISH for evaluating HER2 gene status.

Magnetic Criticality Enhanced Hybrid Nanodiamond Thermometer under Ambient Conditions

Science.gov (United States)

Wang, Ning; Liu, Gang-Qin; Leong, Weng-Hang; Zeng, Hualing; Feng, Xi; Li, Si-Hong; Dolde, Florian; Fedder, Helmut; Wrachtrup, Jörg; Cui, Xiao-Dong; Yang, Sen; Li, Quan; Liu, Ren-Bao

2018-01-01

Nitrogen-vacancy (NV) centers in diamond are attractive as quantum sensors owing to their superb coherence under ambient conditions. However, the NV center spin resonances are relatively insensitive to some important parameters such as temperature and pressure. Here we design and experimentally demonstrate a hybrid nanothermometer composed of NV centers and a magnetic nanoparticle (MNP), in which the temperature sensitivity is enhanced by the critical magnetization of the MNP near the ferromagnetic-paramagnetic transition temperature. The temperature susceptibility of the NV center spin resonance reaches 14 MHz /K , nearly 200 times larger than that of bare NV centers. The sensitivity of a hybrid nanothermometer composed of a Cu1 -xNix MNP and a nanodiamond is measured to be 11 mK /√{Hz } under ambient conditions. The working range of the hybrid thermometer can be designed from cryogenic temperature to about 600 K by tuning the chemical composition of the Cu1 -xNix MNP. We demonstrate in situ detection of the magnetic phase transition of a single magnetic nanoparticle using the hybrid nanothermometer. This hybrid nanothermometer provides a novel approach to studying a broad range of thermal processes at nanoscales such as nanoplasmonics, heat-stimulated subcellular processes, and thermodynamics of nanosystems.

Magnetic Criticality Enhanced Hybrid Nanodiamond Thermometer under Ambient Conditions

Directory of Open Access Journals (Sweden)

Ning Wang

2018-03-01

Full Text Available Nitrogen-vacancy (NV centers in diamond are attractive as quantum sensors owing to their superb coherence under ambient conditions. However, the NV center spin resonances are relatively insensitive to some important parameters such as temperature and pressure. Here we design and experimentally demonstrate a hybrid nanothermometer composed of NV centers and a magnetic nanoparticle (MNP, in which the temperature sensitivity is enhanced by the critical magnetization of the MNP near the ferromagnetic-paramagnetic transition temperature. The temperature susceptibility of the NV center spin resonance reaches 14  MHz/K, nearly 200 times larger than that of bare NV centers. The sensitivity of a hybrid nanothermometer composed of a Cu_{1-x}Ni_{x} MNP and a nanodiamond is measured to be 11  mK/sqrt[Hz] under ambient conditions. The working range of the hybrid thermometer can be designed from cryogenic temperature to about 600 K by tuning the chemical composition of the Cu_{1-x}Ni_{x} MNP. We demonstrate in situ detection of the magnetic phase transition of a single magnetic nanoparticle using the hybrid nanothermometer. This hybrid nanothermometer provides a novel approach to studying a broad range of thermal processes at nanoscales such as nanoplasmonics, heat-stimulated subcellular processes, and thermodynamics of nanosystems.

Spatial gene expression quantification: a tool for analysis of in situ hybridizations in sea anemone Nematostella vectensis

Directory of Open Access Journals (Sweden)

Botman Daniel

2012-10-01

Full Text Available Abstract Background Spatial gene expression quantification is required for modeling gene regulation in developing organisms. The fruit fly Drosophila melanogaster is the model system most widely applied for spatial gene expression analysis due to its unique embryonic properties: the shape does not change significantly during its early cleavage cycles and most genes are differentially expressed along a straight axis. This system of development is quite exceptional in the animal kingdom. In the sea anemone Nematostella vectensis the embryo changes its shape during early development; there are cell divisions and cell movement, like in most other metazoans. Nematostella is an attractive case study for spatial gene expression since its transparent body wall makes it accessible to various imaging techniques. Findings Our new quantification method produces standardized gene expression profiles from raw or annotated Nematostella in situ hybridizations by measuring the expression intensity along its cell layer. The procedure is based on digital morphologies derived from high-resolution fluorescence pictures. Additionally, complete descriptions of nonsymmetric expression patterns have been constructed by transforming the gene expression images into a three-dimensional representation. Conclusions We created a standard format for gene expression data, which enables quantitative analysis of in situ hybridizations from embryos with various shapes in different developmental stages. The obtained expression profiles are suitable as input for optimization of gene regulatory network models, and for correlation analysis of genes from dissimilar Nematostella morphologies. This approach is potentially applicable to many other metazoan model organisms and may also be suitable for processing data from three-dimensional imaging techniques.

Geology, hydrology, chemistry, and microbiology of the in situ bioremediation demonstration site

International Nuclear Information System (INIS)

Newcomer, D.R.; Doremus, L.A.; Hall, S.H.; Truex, M.J.; Vermeul, V.R.; Engelman, R.E.

1995-03-01

This report summarizes characterization information on the geology, hydrology, microbiology, contaminant distribution, and ground-water chemistry to support demonstration of in situ bioremediation at the Hanford Site. The purpose of this information is to provide baseline conditions, including a conceptual model of the aquifer being utilized for in situ bioremediation. Data were collected from sampling and other characterization activities associated with three wells drilled in the upper part of the suprabasalt aquifer. Results of point-dilution tracer tests, conducted in the upper 9 m (30 ft) of the aquifer, showed that most ground-water flow occurs in the upper part of this zone, which is consistent with hydraulic test results and geologic and geophysical data. Other tracer test results indicated that natural ground-water flow velocity is equal to or less than about 0.03 m/d (0.1 ft/d). Laboratory hydraulic conductivity measurements, which represent the local distribution of vertical hydraulic conductivity, varied up to three orders of magnitude. Based on concentration data from both the vadose and saturated zone, it is suggested that most, if not all, of the carbon tetrachloride detected is representative of the aqueous phase. Concentrations of carbon tetrachloride, associated with a contaminant plume in the 200-West Area, ranged from approximately 500 to 3,800 μg/L in the aqueous phase and from approximately 10 to 290 μg/L in the solid phase at the demonstration site. Carbon tetrachloride gas was detected in the vadose zone, suggesting volatilization and subsequent upward migration from the saturated zone

Distribution and evolution of repeated sequences in genomes of Triatominae (Hemiptera-Reduviidae inferred from genomic in situ hybridization.

Directory of Open Access Journals (Sweden)

Sebastian Pita

Full Text Available The subfamily Triatominae, vectors of Chagas disease, comprises 140 species characterized by a highly homogeneous chromosome number. We analyzed the chromosomal distribution and evolution of repeated sequences in Triatominae genomes by Genomic in situ Hybridization using Triatoma delpontei and Triatoma infestans genomic DNAs as probes. Hybridizations were performed on their own chromosomes and on nine species included in six genera from the two main tribes: Triatomini and Rhodniini. Genomic probes clearly generate two different hybridization patterns, dispersed or accumulated in specific regions or chromosomes. The three used probes generate the same hybridization pattern in each species. However, these patterns are species-specific. In closely related species, the probes strongly hybridized in the autosomal heterochromatic regions, resembling C-banding and DAPI patterns. However, in more distant species these co-localizations are not observed. The heterochromatic Y chromosome is constituted by highly repeated sequences, which is conserved among 10 species of Triatomini tribe suggesting be an ancestral character for this group. However, the Y chromosome in Rhodniini tribe is markedly different, supporting the early evolutionary dichotomy between both tribes. In some species, sex chromosomes and autosomes shared repeated sequences, suggesting meiotic chromatin exchanges among these heterologous chromosomes. Our GISH analyses enabled us to acquire not only reliable information about autosomal repeated sequences distribution but also an insight into sex chromosome evolution in Triatominae. Furthermore, the differentiation obtained by GISH might be a valuable marker to establish phylogenetic relationships and to test the controversial origin of the Triatominae subfamily.

Determination of the ruminant origin of bone particles using fluorescence in situ hybridization (FISH).

Science.gov (United States)

Lecrenier, M C; Ledoux, Q; Berben, G; Fumière, O; Saegerman, C; Baeten, V; Veys, P

2014-07-17

Molecular biology techniques such as PCR constitute powerful tools for the determination of the taxonomic origin of bones. DNA degradation and contamination by exogenous DNA, however, jeopardise bone identification. Despite the vast array of techniques used to decontaminate bone fragments, the isolation and determination of bone DNA content are still problematic. Within the framework of the eradication of transmissible spongiform encephalopathies (including BSE, commonly known as "mad cow disease"), a fluorescence in situ hybridization (FISH) protocol was developed. Results from the described study showed that this method can be applied directly to bones without a demineralisation step and that it allows the identification of bovine and ruminant bones even after severe processing. The results also showed that the method is independent of exogenous contamination and that it is therefore entirely appropriate for this application.

Control Demonstration of Multiple Doubly-Fed Induction Motors for Hybrid Electric Propulsion

Science.gov (United States)

Sadey, David J.; Bodson, Marc; Csank, Jeffrey T.; Hunker, Keith R.; Theman, Casey J.; Taylor, Linda M.

2017-01-01

The Convergent Aeronautics Solutions (CAS) High Voltage-Hybrid Electric Propulsion (HVHEP) task was formulated to support the move into future hybrid-electric aircraft. The goal of this project is to develop a new AC power architecture to support the needs of higher efficiency and lower emissions. This proposed architecture will adopt the use of the doubly-fed induction machine (DFIM) for propulsor drive motor application.The Convergent Aeronautics Solutions (CAS) High Voltage-Hybrid Electric Propulsion (HVHEP) task was formulated to support the move into future hybrid-electric aircraft. The goal of this project is to develop a new AC power architecture to support the needs of higher efficiency and lower emissions. This proposed architecture will adopt the use of the doubly-fed induction machine (DFIM) for propulsor drive motor application. DFIMs are attractive for several reasons, including but not limited to the ability to self-start, ability to operate sub- and super-synchronously, and requiring only fractionally rated power converters on a per-unit basis depending on the required range of operation. The focus of this paper is based specifically on the presentation and analysis of a novel strategy which allows for independent operation of each of the aforementioned doubly-fed induction motors. This strategy includes synchronization, soft-start, and closed loop speed control of each motor as a means of controlling output thrust; be it concurrently or differentially. The demonstration of this strategy has recently been proven out on a low power test bed using fractional horsepower machines. Simulation and hardware test results are presented in the paper.

Electron microscopic in situ hybridization and autoradiography: Localization and transcription of rDNA in human lymphocyte nucleoli

International Nuclear Information System (INIS)

Wachtler, F.; Mosgoeller, W.S.; Schwarzacher, H.G.

1990-01-01

The distribution of ribosomal DNA (rDNA) in the nucleoli of human lymphocytes was revealed by in situ hybridization with a nonautoradiographic procedure at the electron microscopic level. rDNA is located in the dense fibrillar component of the nucleolus but not in the fibrillar centers. In the same cells the incorporation of tritiated uridine takes place in the dense fibrillar component of the nucleolus as seen by autoradiography followed by gold latensification. From these findings it can be concluded that the transcription of ribosomal DNA takes place in the dense fibrillar component of the nucleolus

In situ hybridization of iodinated 5S and 18/25S RNA to Vicia faba metaphase chromosomes

International Nuclear Information System (INIS)

Schubert, I.; Baeumlein, H.; Wobus, U.

1978-01-01

In vitro labelled 125 I ribosomal RNA fractions (18/25S and 5S) were in situ hybridized to metaphase chromosomes of a reconstructed karyotype of Vicia faba (characterized by two translocations and one pericentric inversion, each being present homozygously). The sites of 18S and 25S RNA were found to be confined to the nucleolus organizing secondary constriction. Two loci of 5S RNA were recognized on the satellite of nucleolus bearing chromosome. Possible correlations between the location of ribosomal genes, heterochromatic G-bands and clusters of mutagen induced chromatid aberrations are discussed. (author)

Small supernumerary marker chromosome derived from proximal p-arm of chromosome 2: identification by fluorescent in situ hybridization.

Science.gov (United States)

Lasan Trcić, Ruzica; Hitrec, Vlasta; Letica, Ljiljana; Cuk, Mario; Begović, Davor

2003-08-01

Conventional cytogenetics detected an interstitial deletion of proximal region of p-arm of chromosome 2 in a 6-month-old boy with a phenotype slightly resembling Down's syndrome. The deletion was inherited from the father, whose karyotype revealed a small ring-shaped marker chromosome, in addition to interstitial deletion. Fluorescence in situ hybridization identified the marker, which consisted of the proximal region of the p-arm of chromosome 2, including a part of its centromere. This case shows that molecular cytogenetic analysis can reveal the mechanism of the formation of the marker chromosome.

Association of Serpulina hyodysenteriae with the colonic mucosa in experimental swine dysentery studied by fluorescent in situ hybridization

DEFF Research Database (Denmark)

Jensen, Tim Kåre; Boye, Mette; Møller, Kristian

1998-01-01

The localization of Serpulina hyodysenteriae in experimental swine dysentery was studied by fluorescent in situ hybridization (FISH) using an oligonucleotide probe targeting the 23S rRNA of S. hyodysenteriae. Nine 8-week-old pigs were challenged. Seven of the pigs were intragastrically dosed with 1......x10(9) cfu S. hyodysenteriae for 3 consecutive days, whereas two pigs were infected by contact. Six non-challenged pigs served as negative controls. The challenged pigs developed clinical swine dysentery from 8 to 14 days postinfection with typical gross lesions. By FISH S. hyodysenteriae cells...

Luminescent hybrid lanthanide sulfates and lanthanide sulfonate-carboxylates with 1,10-phenanthroline involving in-situ oxidation of 2-mercaptonbenzoic acid

International Nuclear Information System (INIS)

Zhong, Jie-Cen; Wan, Fang; Sun, Yan-Qiong; Chen, Yi-Ping

2015-01-01

A series of lanthanide sulfates and lanthanide sulfonate-carboxylates, [Ln 2 (phen) 2 (SO 4 ) 3 (H 2 O) 2 ] n (I:Ln=Nd(1a), Sm(1b), Eu(1c), phen=1,10-phenanthroline) and [Ln(phen)(2-SBA)(BZA)] n (II: Ln=Sm(2a), Eu(2b), Dy(2c), 2-SBA=2-sulfobenzoate, BZA=benzoate) have been hydrothermally synthesized from lanthanide oxide, 2-mercaptonbenzoic acid with phen as auxiliary ligand and characterized by single-crystal X-ray diffraction, elemental analyses, IR spectra, TG analyses and luminescence spectroscopy. Interestingly, SO 4 2− anions in I came from the in situ deep oxidation of thiol groups of 2-mercaptonbenzoic acid while 2-sulfobenzoate and benzoate ligands in II from the middle oxidation and desulfuration reactions of 2-mercaptonbenzoic acid. Compounds I are organic–inorganic hybrid lanthanide sulfates, which have rare one-dimensional column-like structures. Complexes II are binuclear lanthanide sulfonate-carboxylates with 2-sulfobenzoate and benzoate as bridges and 1,10-phenanthroline as terminal. Photoluminescence studies reveal that complexes I and II exhibit strong lanthanide characteristic emission bands in the solid state at room temperature. - Graphical abstract: Lanthanide sulfates and lanthanide sulfonate-carboxylates have been hydrothermally synthesized. Interestingly, sulfate anions, 2-sulfobenzoate and benzoate ligands came from the in situ oxidation and desulfuration reactions of 2-mercaptonbenzoic acid. - Highlights: • In situ oxidation and desulfuration reactions of 2-mercaptonbenzoic acid. • The organic–inorganic hybrid lanthanide sulfates with one-dimensional column-like structure. • The dinuclear lanthanide sulfonate-carboxylates. • The emission spectra exhibit the characteristic transition of 5 D 0 → 7 F J (J=0–4) of the Eu(III)

Standardization and optimization of fluorescence in situ hybridization (FISH) for HER-2 assessment in breast cancer: A single center experience.

Science.gov (United States)

Bogdanovska-Todorovska, Magdalena; Petrushevska, Gordana; Janevska, Vesna; Spasevska, Liljana; Kostadinova-Kunovska, Slavica

2018-05-20

Accurate assessment of human epidermal growth factor receptor 2 (HER-2) is crucial in selecting patients for targeted therapy. Commonly used methods for HER-2 testing are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Here we presented the implementation, optimization and standardization of two FISH protocols using breast cancer samples and assessed the impact of pre-analytical and analytical factors on HER-2 testing. Formalin fixed paraffin embedded (FFPE) tissue samples from 70 breast cancer patients were tested for HER-2 using PathVysion™ HER-2 DNA Probe Kit and two different paraffin pretreatment kits, Vysis/Abbott Paraffin Pretreatment Reagent Kit (40 samples) and DAKO Histology FISH Accessory Kit (30 samples). The concordance between FISH and IHC results was determined. Pre-analytical and analytical factors (i.e., fixation, baking, digestion, and post-hybridization washing) affected the efficiency and quality of hybridization. The overall hybridization success in our study was 98.6% (69/70); the failure rate was 1.4%. The DAKO pretreatment kit was more time-efficient and resulted in more uniform signals that were easier to interpret, compared to the Vysis/Abbott kit. The overall concordance between IHC and FISH was 84.06%, kappa coefficient 0.5976 (p characteristics. Differences in the pre-analytical and analytical steps can affect the hybridization quality and efficiency. The use of DAKO pretreatment kit is time-saving and cost-effective.

Fluorescence in situ hybridization techniques (FISH) to detect changes in CYP19a gene expression of Japanese medaka (Oryzias latipes)

International Nuclear Information System (INIS)

Park, June-Woo; Tompsett, Amber; Zhang, Xiaowei; Newsted, John L.; Jones, Paul D.; Au, Doris; Kong, Richard; Wu, Rudolf S.S.; Giesy, John P.; Hecker, Markus

2008-01-01

The aim of this study was to develop a sensitive in situ hybridization methodology using fluorescence-labeled riboprobes (FISH) that allows for the evaluation of gene expression profiles simultaneously in multiple target tissues of whole fish sections of Japanese medaka (Oryzias latipes). To date FISH methods have been limited in their application due to autofluorescence of tissues, fixatives or other components of the hybridization procedure. An optimized FISH method, based on confocal fluorescence microscopy was developed to reduce the autofluorescence signal. Because of its tissue- and gender-specific expression and relevance in studies of endocrine disruption, gonadal aromatase (CYP19a) was used as a model gene. The in situ hybridization (ISH) system was validated in a test exposure with the aromatase inhibitor fadrozole. The optimized FISH method revealed tissue-specific expression of the CYP19a gene. Furthermore, the assay could differentiate the abundance of CYP19a mRNA among cell types. Expression of CYP19a was primarily associated with early stage oocytes, and expression gradually decreased with increasing maturation. No expression of CYP19a mRNA was observed in other tissues such as brain, liver, or testes. Fadrozole (100 μg/L) caused up-regulation of CYP19a expression, a trend that was confirmed by RT-PCR analysis on excised tissues. In a combination approach with gonad histology, it could be shown that the increase in CYP19a expression as measured by RT-PCR on a whole tissue basis was due to a combination of both increases in numbers of CYP19a-containing cells and an increase in the amount of CYP19a mRNA present in the cells

Detection and quantification of Epstein-Barr virus EBER1 in EBV-infected cells by fluorescent in situ hybridization and flow cytometry

Science.gov (United States)

Stowe, R. P.; Cubbage, M. L.; Sams, C. F.; Pierson, D. L.; Barrett, A. D.

1998-01-01

A rapid and highly sensitive fluorescent in situ hybridization (FISH) assay was developed to detect Epstein Barr virus (EBV)-infected cells in peripheral blood. Multiple fluorescein-labeled antisense oligonucleotide probes were designed to hybridize to the EBER1 transcript, which is highly expressed in latently infected cells. After a rapid (30 min) hybridization, the cells were analyzed by flow cytometry. EBER1 was detected in several positive control cell lines that have variable numbers of EBV genome copies. No EBER1 was detected in two known EBV-negative cell lines. Northern blot analyses confirmed the presence and quantity of EBER1 transcripts in each cell line. This method was used to quantify the number of EBV-infected cells in peripheral blood from a patient with chronic mononucleosis. These results indicate that EBV-infected cells can be detected at the single cell level, and that this assay can be used to quantify the number of EBV-infected cells in clinical samples.

DEMONSTRATION OF A FULL-SCALE RETROFIT OF THE ADVANCED HYBRID PARTICULATE COLLECTOR TECHNOLOGY

Energy Technology Data Exchange (ETDEWEB)

Tom Hrdlicka; William Swanson

2005-12-01

The Advanced Hybrid Particulate Collector (AHPC), developed in cooperation between W.L. Gore & Associates and the Energy & Environmental Research Center (EERC), is an innovative approach to removing particulates from power plant flue gas. The AHPC combines the elements of a traditional baghouse and electrostatic precipitator (ESP) into one device to achieve increased particulate collection efficiency. As part of the Power Plant Improvement Initiative (PPII), this project was demonstrated under joint sponsorship from the U.S. Department of Energy and Otter Tail Power Company. The EERC is the patent holder for the technology, and W.L. Gore & Associates was the exclusive licensee for this project. The project objective was to demonstrate the improved particulate collection efficiency obtained by a full-scale retrofit of the AHPC to an existing electrostatic precipitator. The full-scale retrofit was installed on an electric power plant burning Powder River Basin (PRB) coal, Otter Tail Power Company's Big Stone Plant, in Big Stone City, South Dakota. The $13.4 million project was installed in October 2002. Project related testing concluded in December 2005. The following Final Technical Report has been prepared for the project entitled ''Demonstration of a Full-Scale Retrofit of the Advanced Hybrid Particulate Collector Technology'' as described in DOE Award No. DE-FC26-02NT41420. The report presents the operation and performance results of the system.

Expression and distribution of octopus gonadotropin-releasing hormone in the central nervous system and peripheral organs of the octopus (Octopus vulgaris) by in situ hybridization and immunohistochemistry.

Science.gov (United States)

Iwakoshi-Ukena, Eiko; Ukena, Kazuyoshi; Takuwa-Kuroda, Kyoko; Kanda, Atshuhiro; Tsutsui, Kazuyoshi; Minakata, Hiroyuki

2004-09-20

We recently purified a peptide with structural features similar to vertebrate gonadotropin-releasing hormone (GnRH) from the brain of Octopus vulgaris, cloned a cDNA encoding the precursor protein, and named it oct-GnRH. In the current study, we investigated the expression and distribution of oct-GnRH throughout the central nervous system (CNS) and peripheral organs of Octopus by in situ hybridization on the basis of the cDNA sequence and by immunohistochemistry using a specific antiserum against oct-GnRH. Oct-GnRH mRNA-expressing cell bodies were located in 10 of 19 lobes in the supraesophageal and subesophageal parts of the CNS. Several oct-GnRH-like immunoreactive fibers were seen in all the neuropils of the CNS lobes. The sites of oct-GnRH mRNA expression and the mature peptide distribution were consistent with each other as judged by in situ hybridization and immunohistochemistry. In addition, many immunoreactive fibers were distributed in peripheral organs such as the heart, the oviduct, and the oviducal gland. Modulatory effects of oct-GnRH on the contractions of the heart and the oviduct were demonstrated. The results suggested that, in the context of reproduction, oct-GnRH is a key peptide in the subpedunculate lobe and/or posterior olfactory lobe-optic gland-gonadal axis, an octopus analogue of the hypothalamo-hypophysial-gonadal axis. It may also act as a modulatory factor in controlling higher brain functions such as feeding, memory, movement, maturation, and autonomic functions

Hybrid concepts of the co-operation GETRAG-BOSCH and their realisation in a demonstrator car; Hybridkonzepte der Kooperation GETRAG-BOSCH am Beispiel eines Demonstratorfahrzeugs

Energy Technology Data Exchange (ETDEWEB)

Blankenbach, Bernd; Blessing, Uli Christian [GETRAG KG, Untergruppenbach (Germany); Imaseki, Takashi; Richter, Boyke [Robert Bosch GmbH, Stuttgart (Germany)

2009-07-01

At present, the partners of hybrid co-operation Getrag KG (Untergruppenbach, Federal Republic of Germany) and Robert Bosch GmbH (Stuttgart, Federal Republic of Germany) develop a hybrid demonstrator vehicle. The BMW Mini Clubman with a hybridized double clutch transmission and an electrical rear axle is the fundament. In the case of the six-speed double clutch transmission, the E-machine alternatively can be coupled to one of the two torque split hybrid. Using the demonstrator vehicle, on the one hand the hybrid operating strategy shall be optimized. On the other hand, it is used for presentations whereby the axle-split hybrid variants are presentable. For this vehicle, numerical simulations prove consumption reductions of approximately 6 % (micro hybrid), 18 % (axle split hybrid) and 24 % (torque split hybrid). Aspects of comfort and aspects of driving dynamics already are considered.

Validation of interphase fluorescence in situ hybridization (iFISH for multiple myeloma using CD138 positive cells

Directory of Open Access Journals (Sweden)

Renata Kiyomi Kishimoto

2016-06-01

Full Text Available ABSTRACT BACKGROUND: Multiple myeloma is a plasma cell neoplasm with acquired genetic abnormalities of clinical and prognostic importance. Multiple myeloma differs from other hematologic malignancies due to a high fraction of low proliferating malignant plasma cells and the paucity of plasma cells in bone marrow aspiration samples, making cytogenetic analysis a challenge. An abnormal karyotype is found in only one-third of patients with multiple myeloma and interphase fluorescence in situ hybridization is the most useful test for studying the chromosomal abnormalities present in almost 90% of cases. However, it is necessary to study the genetic abnormalities in plasma cells after their identification or selection by morphology, immunophenotyping or sorting. Other challenges are the selection of the most informative FISH panel and determining cut-off levels for FISH probes. This study reports the validation of interphase fluorescence in situ hybridization using CD138 positive cells, according to proposed guidelines published by the European Myeloma Network (EMN in 2012. METHOD: Bone marrow samples from patients with multiple myeloma were used to standardize a panel of five probes [1q amplification, 13q14 deletion, 17p deletion, t(4;14, and t(14;16] in CD138+ cells purified by magnetic cell sorting. RESULTS: This test was validated with a low turnaround time and good reproducibility. Five of six samples showed genetic abnormalities. Monosomy/deletion 13 plus t(4;14 were found in two cases. CONCLUSION: This technique together with magnetic cell sorting is effective and can be used in the routine laboratory practice. In addition, magnetic cell sorting provides a pure plasma cell population that allows other molecular and genomic studies.

Determining the origin of cells in tissue engineered skin substitutes: a pilot study employing in situ hybridization.

Science.gov (United States)

Weber, Andreas Daniel; Pontiggia, Luca; Biedermann, Thomas; Schiestl, Clemens; Meuli, Martin; Reichmann, Ernst

2011-03-01

Definitive and high-quality coverage of large and, in particular, massive skin defects remains a significant challenge in burn as well as plastic and reconstructive surgery because of donor site shortage. A novel and promising approach to overcome these problems is tissue engineering of skin. Clearly, before eventual clinical application, engineered skin substitutes of human origin must be grafted and then evaluated in animal models. For the various tests to be conducted it is indispensable to be able to identify human cells as such in culture and also to distinguish between graft and recipient tissue after transplantation. Here we describe a tool to identify human cells in vitro and in vivo. In situ hybridization allows for the detection and localization of specific DNA or RNA sequences in morphologically preserved cells in culture or tissue sections, respectively. We used digoxigenin-labeled DNA probes corresponding to human-specific Alu repeats in order to identify human keratinocytes grown in culture together with rat cells, and also to label split and full thickness skin grafts of human origin after transplantation on immuno-incompetent rats. Digoxigenin-labeled DNA probing resulted in an intensive nuclear staining of human cells, both in culture and after transplantation onto recipient animals, while recipient animal cells (rat cells) did not stain. In situ hybridization using primate-specific Alu probes reliably allows distinguishing between cells of human and non-human origin both in culture as well as in histological sections. This method is an essential tool for those preclinical experiments (performed on non-primate animals) that must be conducted before novel tissue engineered skin substitutes might be introduced into clinical practice.

A demonstration of in situ bioremediation of CCL4 at the Hanford Site

International Nuclear Information System (INIS)

Hooker, B.S.; Skeen, R.S.; Truex, M.J.; Peyton, B.M.

1994-11-01

The United States Department of Energy's VOC-Arid Integrated Demonstration Program (VOC/Arid-ID) is developing an in situ bioremediation technology to meet the need for a cost-effective method to clean ground water contaminated with chlorinated solvents, nitrates, or other organic and inorganic contaminants. Currently, a field demonstration of the technology is being conducted at the Hanford site in southeastern Washington state. The goal of this demonstration is to stimulate native denitrifying microorganisms to destroy carbon tetrachloride and nitrate. Contaminants are destroyed by mixing an electron donor (acetate) and an electron acceptor (nitrate) into the aquifer, using a matrix of recirculation wells. This work also evaluates the effectiveness.of applying scale-up techniques developed in the petrochemical industry to bioremediation. The scale-up process is based on combining fluid mixing and transport predictions with numerical descriptions for biological transport and reaction kinetics. This paper focuses on the necessity of this design approach to select nutrient feeding strategies that limit biofouling while actively destroying contaminants

Application of Fluorescence In Situ Hybridization (FISH) Technique for the Detection of Genetic Aberration in Medical Science

Science.gov (United States)

Ratan, Zubair Ahmed; Zaman, Sojib Bin; Haidere, Mohammad Faisal; Runa, Nusrat Jahan; Akter, Nasrin

2017-01-01

Fluorescence in situ hybridization (FISH) is a macromolecule recognition technique, which is considered as a new advent in the field of cytology. Initially, it was developed as a physical mapping tool to delineate genes within chromosomes. The accuracy and versatility of FISH were subsequently capitalized upon in biological and medical research. This visually appealing technique provides an intermediate degree of resolution between DNA analysis and chromosomal investigations. FISH consists of a hybridizing DNA probe, which can be labeled directly or indirectly. In the case of direct labeling, fluorescent nucleotides are used, while indirect labeling is incorporated with reporter molecules that are subsequently detected by fluorescent antibodies or other affinity molecules. FISH is applied to detect genetic abnormalities that include different characteristic gene fusions or the presence of an abnormal number of chromosomes in a cell or loss of a chromosomal region or a whole chromosome. It is also applied in different research applications, such as gene mapping or the identification of novel oncogenes. This article reviews the concept of FISH, its application, and its advantages in medical science.  PMID:28690958

Facile fabrication of ultrathin hybrid membrane for highly flexible supercapacitors via in-situ phase separation of polyethersulfone

Science.gov (United States)

Zhao, Xiaoning; Ran, Fen; Shen, Kuiwen; Yang, Yunlong; Wu, Jiayu; Niu, Xiaoqin; Kong, Lingbin; Kang, Long; Chen, Shaowei

2016-10-01

In this article, a facile method based on in-situ phase-separation was developed for the fabrication of ultrathin hybrid membranes for highly flexible supercapacitors. The structures and morphologies of the prepared electrodes were characterized by scanning electron microscopy (SEM), Fourier-transformed infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA) measurements; and the electrochemical behaviors were examined in 2 M KOH solution. SEM and FTIR characterizations reveal that activated carbon was imbedded into the polymer membrane of polyethersulfone to form a uniform and flexible hybrid membrane. When the thin polymer-carbon membrane (PCM) was used as an electrode material for supercapacitor, a high specific capacitance of 169.4 Fg-1 was obtained at a current density of 0.5 Ag-1 along with good long-term cycle life of 94.6% capacity retention after 2000 charging-discharging cycles. Benefiting from these merits, the as-fabricated PCM//PCM cell shows an excellent electrochemical property. These results suggest a promising route towards the fabrication of highly flexible electrodes for high-performance supercapacitors.

Efficiency of fluorescence in situ hybridization for bacterial cell identification in temporary river sediments with contrasting water content.

Science.gov (United States)

Fazi, Stefano; Amalfitano, Stefano; Pizzetti, Ilaria; Pernthaler, Jakob

2007-09-01

We studied the efficiency of two hybridization techniques for the analysis of benthic bacterial community composition under varying sediment water content. Microcosms were set up with sediments from four European temporary rivers. Wet sediments were dried, and dry sediments were artificially rewetted. The percentage of bacterial cells detected by fluorescence in situ hybridization with fluorescently monolabeled probes (FISH) significantly increased from dry to wet sediments, showing a positive correlation with the community activity measured via incorporation of (3)H leucine. FISH and signal amplification by catalyzed reporter deposition (CARD-FISH) could significantly better detect cells with low activity in dried sediments. Through the application of an optimized cell permeabilization protocol, the percentage of hybridized cells by CARD-FISH showed comparable values in dry and wet conditions. This approach was unrelated to (3)H leucine incorporation rates. Moreover, the optimized protocol allowed a significantly better visualization of Gram-positive Actinobacteria in the studied samples. CARD-FISH is, therefore, proposed as an effective technique to compare bacterial communities residing in sediments with contrasting water content, irrespective of differences in the activity state of target cells. Considering the increasing frequencies of flood and drought cycles in European temporary rivers, our approach may help to better understand the dynamics of microbial communities in such systems.

Intracellular distribution of histone mRNAs in human fibroblasts studied by in situ hybridization

International Nuclear Information System (INIS)

Lawrence, J.B.; Singer, R.H.; Villnave, C.A.; Stein, J.L.; Stein, G.S.

1988-01-01

We have used in situ hybridization to study the intracellular distribution of mRNAs for cell cycle-dependent core and H1 histone proteins in human WI-38 fibroblasts. Because histones are abundant nuclear proteins and histone mRNA expression is tightly coupled to DNA synthesis, it was of interest to determine whether histone mRNAs are localized near the nucleus. Cells were hybridized with tritiated DNA probes specific for either histone H1, histone H4, actin, or poly(A)+ mRNA and were processed for autoradiography. In exponentially growing cultures, the fraction of histone mRNA-positive cells correlated well with the fraction of cells in S phase and was eliminated by hydroxyurea inhibition of DNA synthesis. Within individual cells the label for histone mRNA was widely distributed throughout the cytoplasm and did not appear to be more heavily concentrated near the nucleus. However, histone mRNA appeared to exhibit patchy, nonhomogeneous localization, and a quantitative evaluation confirmed that grain distributions were not as uniform as they were after hybridizations to poly(A)+ mRNA. Actin mRNA in WI-38 cells was also widely distributed throughout the cytoplasm but differed from histone mRNA in that label for actin mRNA was frequently most dense at the outermost region of narrow cell extensions. The localization of actin mRNA was less pronounced but qualitatively very similar to that previously described for chicken embryonic myoblasts and fibroblasts. We conclude that localization of histones in WI-38 cells is not facilitated by localization of histone protein synthesis near the nucleus and that there are subtle but discrete and potentially functional differences in the distributions of histone, actin, and poly(A)+ mRNAs

Correlation between HER2 gene amplification and protein overexpression through fluorescence in situ hybridization and immunohistochemistry in breast carcinoma patients

OpenAIRE

R N Makroo; Mohit Chowdhry; Manoj Kumar; Priyanka Srivastava; Richa Tyagi; Preeti Bhadauria; Sumaid Kaul; Ramesh Sarin; P K Das; Harsh Dua

2012-01-01

Background : In India, the incidence of breast cancer has increased in the urban population, with 1 in every 22 women diagnosed with breast cancer. It is important to know the HER2/neu gene status for a better prognostication of these patients. Aim : The aim of this study was to compare the efficacy of fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) for determining HER2/neu alteration in breast carcinoma. Materials and Methods : A total of 188 histologically proven br...

Super-resolution nanofabrication with metal-ion doped hybrid material through an optical dual-beam approach

International Nuclear Information System (INIS)

Cao, Yaoyu; Li, Xiangping; Gu, Min

2014-01-01

We apply an optical dual-beam approach to a metal-ion doped hybrid material to achieve nanofeatures beyond the optical diffraction limit. By spatially inhibiting the photoreduction and the photopolymerization, we realize a nano-line, consisting of polymer matrix and in-situ generated gold nanoparticles, with a lateral size of sub 100 nm, corresponding to a factor of 7 improvement compared to the diffraction limit. With the existence of gold nanoparticles, a plasmon enhanced super-resolution fabrication mechanism in the hybrid material is observed, which benefits in a further reduction in size of the fabricated feature. The demonstrated nanofeature in hybrid materials paves the way for realizing functional nanostructures

In situ protocol for butterfly pupal wings using riboprobes.

Science.gov (United States)

Ramos, Diane; Monteiro, Antonia

2007-01-01

Here we present, in video format, a protocol for in situ hybridizations in pupal wings of the butterfly Bicyclus anynana using riboprobes. In situ hybridizations, a mainstay of developmental biology, are useful to study the spatial and temporal patterns of gene expression in developing tissues at the level of transcription. If antibodies that target the protein products of gene transcription have not yet been developed, and/or there are multiple gene copies of a particular protein in the genome that cannot be differentiated using available antibodies, in situs can be used instead. While an in situ technique for larval wing discs has been available to the butterfly community for several years, the current protocol has been optimized for the larger and more fragile pupal wings.

Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain

Directory of Open Access Journals (Sweden)

Hauptmann Giselbert

2011-04-01

Full Text Available Abstract Background In recent years, mapping of overlapping and abutting regulatory gene expression domains by chromogenic two-color in situ hybridization has helped define molecular subdivisions of the developing vertebrate brain and shed light on its basic organization. Despite the benefits of this technique, visualization of overlapping transcript distributions by differently colored precipitates remains difficult because of masking of lighter signals by darker color precipitates and lack of three-dimensional visualization properties. Fluorescent detection of transcript distributions may be able to solve these issues. However, despite the use of signal amplification systems for increasing sensitivity, fluorescent detection in whole-mounts suffers from rapid quenching of peroxidase (POD activity compared to alkaline phosphatase chromogenic reactions. Thus, less strongly expressed genes cannot be efficiently detected. Results We developed an optimized procedure for fluorescent detection of transcript distribution in whole-mount zebrafish embryos using tyramide signal amplification (TSA. Conditions for hybridization and POD-TSA reaction were optimized by the application of the viscosity-increasing polymer dextran sulfate and the use of the substituted phenol compounds 4-iodophenol and vanillin as enhancers of POD activity. In combination with highly effective bench-made tyramide substrates, these improvements resulted in dramatically increased signal-to-noise ratios. The strongly enhanced signal intensities permitted fluorescent visualization of less abundant transcripts of tissue-specific regulatory genes. When performing multicolor fluorescent in situ hybridization (FISH experiments, the highly sensitive POD reaction conditions required effective POD inactivation after each detection cycle by glycine-hydrochloric acid treatment. This optimized FISH procedure permitted the simultaneous fluorescent visualization of up to three unique transcripts

Mapping of the human APOB gene to chromosome 2p and demonstration of a two-allele restriction fragment length polymorphism

International Nuclear Information System (INIS)

Huang, L.; Miller, D.A.; Bruns, G.A.P.; Breslow, J.L.

1986-01-01

ApoB is a large glycoprotein with an apparent molecular mass of 550 kDa on NaDodSO 4 /PAGE. Recently, apoB cDNA clones have been isolated from an expression library made with mRNA from a human hepatoma cell line. These clones, which were all 1.5-1.6 kilobases (kb) long and corresponded to the 3' end of apoB mRNA, were used to demonstrate that hepatic apoB mRNA is ≅ 22 kb long. In the current report, a probe derived from one of these cDNA clones, pB8, was used for in situ hybridization experiments to map the human gene for apoB, APOB, to the distal half of the short arm of chromosome 2. This probe was also used to analyze somatic cell hybrids and, in agreement with the in situ hybridization studies, concordancy was demonstrated with chromosome 2. In addition, two hybrids with chromosome 2 translocations that contain only the short arm reacted with the pB8 probe. A third hybrid with a complex rearrangement of chromosome 2, which deleted an interstitial region and the tip of the short arm of chromosome 2, did not react. These data indicate that APOB maps to either 2p21-p23 or 2p24-pter. In further studies, DNA from normal individuals, digested with the restriction endonuclease EcoRI and subjected to Southern blot analysis with the pB8 probe, revealed a two-allele restriction fragment length polymorphism (RFLP). The mapping studies provide the means for understanding the relationship of the APOB locus to others in the human genome, whereas the demonstration of an APOB RFLP increases their ability to assess the role of this locus in determining plasma lipoprotein levels

Pilot-Scale Demonstration of In-Situ Chemical Oxidation ...

Science.gov (United States)

A pilot-scale in situ chemical oxidation (ISCO) demonstration, involving subsurface injections of sodium permanganate (NaMnO4), was performed at the US Marine Corp Recruit Depot (MCRD), site 45 (Parris Island (PI), SC). The ground water was originally contaminated with perchloroethylene (PCE) (also known as tetrachloroethylene), a chlorinated solvent used in dry cleaner operations. High resolution site characterization involved multiple iterations of soil core sampling and analysis. Nested micro-wells and conventional wells were also used to sample and analyze ground water for PCE and decomposition products (i.e., trichloroethyelene (TCE), dichloroethylene (c-DCE, t-DCE), and vinyl chloride (VC)), collectively referred to as chlorinated volatile organic compounds (CVOC). This characterization methodology was used to develop and refine the conceptual site model and the ISCO design, not only by identifying CVOC contamination but also by eliminating uncontaminated portions of the aquifer from further ISCO consideration. Direct-push injection was selected as the main method of NaMnO4 delivery due to its flexibility and low initial capital cost. Site impediments to ISCO activities in the source area involved subsurface utilities, including a high pressure water main, a high voltage power line, a communication line, and sanitary and stormwater sewer lines. Utility markings were used in conjunction with careful planning and judicious selection of injection locations. A

Standardization and optimization of fluorescence in situ hybridization (FISH for HER-2 assessment in breast cancer: A single center experience

Directory of Open Access Journals (Sweden)

Magdalena Bogdanovska-Todorovska

2018-05-01

Full Text Available Accurate assessment of human epidermal growth factor receptor 2 (HER-2 is crucial in selecting patients for targeted therapy. Commonly used methods for HER-2 testing are immunohistochemistry (IHC and fluorescence in situ hybridization (FISH. Here we presented the implementation, optimization and standardization of two FISH protocols using breast cancer samples and assessed the impact of pre-analytical and analytical factors on HER-2 testing. Formalin fixed paraffin embedded (FFPE tissue samples from 70 breast cancer patients were tested for HER-2 using PathVysion™ HER-2 DNA Probe Kit and two different paraffin pretreatment kits, Vysis/Abbott Paraffin Pretreatment Reagent Kit (40 samples and DAKO Histology FISH Accessory Kit (30 samples. The concordance between FISH and IHC results was determined. Pre-analytical and analytical factors (i.e., fixation, baking, digestion, and post-hybridization washing affected the efficiency and quality of hybridization. The overall hybridization success in our study was 98.6% (69/70; the failure rate was 1.4%. The DAKO pretreatment kit was more time-efficient and resulted in more uniform signals that were easier to interpret, compared to the Vysis/Abbott kit. The overall concordance between IHC and FISH was 84.06%, kappa coefficient 0.5976 (p < 0.0001. The greatest discordance (82% between IHC and FISH was observed in IHC 2+ group. A standardized FISH protocol for HER-2 assessment, with high hybridization efficiency, is necessary due to variability in tissue processing and individual tissue characteristics. Differences in the pre-analytical and analytical steps can affect the hybridization quality and efficiency. The use of DAKO pretreatment kit is time-saving and cost-effective.

Synthesis of ZnO nanorods-Au nanoparticles hybrids via in-situ plasma sputtering-assisted method for simultaneous electrochemical sensing of ascorbic acid and uric acid

Energy Technology Data Exchange (ETDEWEB)

Hou, Chao [College of Life Information Science & Instrument Engineering, Hangzhou Dianzi University, Hangzhou 310018 (China); Liu, Hongying, E-mail: liuhongying@hdu.edu.cn [College of Life Information Science & Instrument Engineering, Hangzhou Dianzi University, Hangzhou 310018 (China); State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry & Chemical Engineering, Nanjing University, Nanjing 210093 (China); Zhang, Dan; Yang, Chi [Department of Pharmacy, Nantong University, Nantong 226001 (China); Zhang, Mingzhen [College of Life Information Science & Instrument Engineering, Hangzhou Dianzi University, Hangzhou 310018 (China)

2016-05-05

In this study, ZnO nanorods-Au nanoparticles (ZnO NRs-Au NPs) hybrids were prepared using an in-situ plasma sputtering-assisted method without any template. Characterization results from scanning electron microscopy, high-resolution transmission electron microscopy, and energy dispersive X-ray spectroscopy showed that Au NPs are highly dispersed and tightly anchored on the surface of ZnO NRs. The size and surface coverage of Au NPs were well controlled by plasma sputtering time. Moreover, the hybrids exhibited excellent electrocatalytic properties towards oxidation of ascorbic acid (AA) and uric acid (UA) due to large surface area of Au NPs and ZnO NRs, and thus can be used as electrochemical sensors. Differential pulse voltammetry results showed that AA and UA could be detected simultaneously by ZnO NRs-Au NPs hybrids modified glassy carbon electrode. The linear ranges for AA and UA are 0.1 to 4 mM and 0.01 to 0.4 mM, respectively. The results suggest promising future applications in clinical diagnosis. - Highlights: • ZnO nanorods-Au nanoparticles were synthesized by in-situ plasma sputtering method. • Influence of sputtering time on the formation of Au nanoparticles was studied. • It exhibited a strong electrocatalytic activity toward the oxidation of ascorbic acid and uric acid. • A portable and cheap approach for simultaneous detection of ascorbic acid and uric acid was developed.

Synthesis of ZnO nanorods-Au nanoparticles hybrids via in-situ plasma sputtering-assisted method for simultaneous electrochemical sensing of ascorbic acid and uric acid

International Nuclear Information System (INIS)

Hou, Chao; Liu, Hongying; Zhang, Dan; Yang, Chi; Zhang, Mingzhen

2016-01-01

In this study, ZnO nanorods-Au nanoparticles (ZnO NRs-Au NPs) hybrids were prepared using an in-situ plasma sputtering-assisted method without any template. Characterization results from scanning electron microscopy, high-resolution transmission electron microscopy, and energy dispersive X-ray spectroscopy showed that Au NPs are highly dispersed and tightly anchored on the surface of ZnO NRs. The size and surface coverage of Au NPs were well controlled by plasma sputtering time. Moreover, the hybrids exhibited excellent electrocatalytic properties towards oxidation of ascorbic acid (AA) and uric acid (UA) due to large surface area of Au NPs and ZnO NRs, and thus can be used as electrochemical sensors. Differential pulse voltammetry results showed that AA and UA could be detected simultaneously by ZnO NRs-Au NPs hybrids modified glassy carbon electrode. The linear ranges for AA and UA are 0.1 to 4 mM and 0.01 to 0.4 mM, respectively. The results suggest promising future applications in clinical diagnosis. - Highlights: • ZnO nanorods-Au nanoparticles were synthesized by in-situ plasma sputtering method. • Influence of sputtering time on the formation of Au nanoparticles was studied. • It exhibited a strong electrocatalytic activity toward the oxidation of ascorbic acid and uric acid. • A portable and cheap approach for simultaneous detection of ascorbic acid and uric acid was developed.

Hybrid silica luminescent materials based on lanthanide-containing lyotropic liquid crystal with polarized emission

Energy Technology Data Exchange (ETDEWEB)

Selivanova, N.M., E-mail: natsel@mail.ru [Kazan National Research Technological University, 68 Karl Marx Str., Kazan 420015 (Russian Federation); Vandyukov, A.E.; Gubaidullin, A.T. [A.E. Arbuzov Institute of Organic and Physical Chemistry of the Kazan Scientific Center of the Russian Academy of Sciences, 8 Acad. Arbuzov Str., Kazan 420088 (Russian Federation); Galyametdinov, Y.G. [Kazan National Research Technological University, 68 Karl Marx Str., Kazan 420015 (Russian Federation)

2014-11-14

This paper represents the template method for synthesis of hybrid silica films based on Ln-containing lyotropic liquid crystal and characterized by efficient luminescence. Luminescence films were prepared in situ by the sol–gel processes. Lyotropic liquid crystal (LLC) mesophases C{sub 12}H{sub 25}O(CH{sub 2}CH{sub 2}O){sub 10}H/Ln(NO{sub 3}){sub 3}·6H{sub 2}O/H{sub 2}O containing Ln (III) ions (Dy, Tb, Eu) were used as template. Polarized optical microscopy, X-ray powder diffraction, and FT-IR-spectroscopy were used for characterization of liquid crystal mesophases and hybrid films. The morphology of composite films was studied by the atomic force microscopy method (AFM). The optical properties of the resulting materials were evaluated. It was found that hybrid silica films demonstrate significant increase of their lifetime in comparison with an LLC system. New effects of linearly polarized emission revealed for Ln-containing hybrid silica films. Polarization in lanthanide-containing hybrid composites indicates that silica precursor causes orientation of emitting ions. - Highlights: • We suggest a new simple approach for creating luminescence hybrid silica films. • Ln-containing hybrid silica films demonstrate yellow, green and red emissions. • Tb(III)-containing hybrid film have a high lifetime. • We report effects of linearly polarized emission in hybrid film.

CuO reduction induced formation of CuO/Cu2O hybrid oxides

Science.gov (United States)

Yuan, Lu; Yin, Qiyue; Wang, Yiqian; Zhou, Guangwen

2013-12-01

Reduction of CuO nanowires results in the formation of a unique hierarchical hybrid nanostructure, in which the parent oxide phase (CuO) works as the skeleton while the lower oxide (Cu2O) resulting from the reduction reaction forms as partially embedded nanoparticles that decorate the skeleton of the parent oxide. Using in situ transmission electron microscopy observations of the reduction process of CuO nanowires, we demonstrate that the formation of such a hierarchical hybrid oxide structure is induced by topotactic nucleation and growth of Cu2O islands on the parent CuO nanowires.

Direct fluorescence in situ hybridization on human metaphase chromosomes using quantum dot-platinum labeled DNA probes

Energy Technology Data Exchange (ETDEWEB)

Hwang, Gyoyeon [Chemical Kinomics Research Center, Future Convergence Research Division, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791 (Korea, Republic of); Biological Chemistry, Korea University of Science and Technology, 217, Gajeong-ro, Yuseong-gu, Deajeon (Korea, Republic of); Lee, Hansol [Chemical Kinomics Research Center, Future Convergence Research Division, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791 (Korea, Republic of); Lee, Jiyeon, E-mail: jylee@kist.re.kr [Chemical Kinomics Research Center, Future Convergence Research Division, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791 (Korea, Republic of); Biological Chemistry, Korea University of Science and Technology, 217, Gajeong-ro, Yuseong-gu, Deajeon (Korea, Republic of)

2015-11-13

The telomere shortening in chromosomes implies the senescence, apoptosis, or oncogenic transformation of cells. Since detecting telomeres in aging and diseases like cancer, is important, the direct detection of telomeres has been a very useful biomarker. We propose a telomere detection method using a newly synthesized quantum dot (QD) based probe with oligonucleotide conjugation and direct fluorescence in situ hybridization (FISH). QD-oligonucleotides were prepared with metal coordination bonding based on platinum-guanine binding reported in our previous work. The QD-oligonucleotide conjugation method has an advantage where any sequence containing guanine at the end can be easily bound to the starting QD-Pt conjugate. A synthesized telomeric oligonucleotide was bound to the QD-Pt conjugate successfully and this probe hybridized specifically on the telomere of fabricated MV-4-11 and MOLT-4 chromosomes. Additionally, the QD-telomeric oligonucleotide probe successfully detected the telomeres on the CGH metaphase slide. Due to the excellent photostability and high quantum yield of QDs, the QD-oligonucleotide probe has high fluorescence intensity when compared to the organic dye-oligonucleotide probe. Our QD-oligonucleotide probe, conjugation method of this QD probe, and hybridization protocol with the chromosomes can be a useful tool for chromosome painting and FISH. - Highlights: • We prepared a probe linked between QD and telomeric oligonucleotide with platinum-guanine bonding. • Telomeres were detected by our new telomere probes successfully in three different human metaphase chromosomes. • QDPt-DNA probe has high fluorescence intensity in comparison with organic dye-DNA probe.

Comprehensive Analysis of ETS Family Members in Melanoma by Fluorescence In Situ Hybridization Reveals Recurrent ETV1 Amplification

Science.gov (United States)

Mehra, Rohit; Dhanasekaran, Saravana M; Palanisamy, Nallasivam; Vats, Pankaj; Cao, Xuhong; Kim, Jung H; Kim, David SL; Johnson, Timothy; Fullen, Douglas R; Chinnaiyan, Arul M

2013-01-01

E26 transformation-specific (ETS) transcription factors are known to be involved in gene aberrations in various malignancies including prostate cancer; however, their role in melanoma oncogenesis has yet to be fully explored. We have completed a comprehensive fluorescence in situ hybridization (FISH)-based screen for all 27 members of the ETS transcription factor family on two melanoma tissue microarrays, representing 223 melanomas, 10 nevi, and 5 normal skin tissues. None of the melanoma cases demonstrated ETS fusions; however, 6 of 114 (5.3%) melanomas were amplified for ETV1 using a break-apart FISH probe. For the six positive cases, locus-controlled FISH probes revealed that two of six cases were amplified for the ETV1 region, whereas four cases showed copy gains of the entire chromosome 7. The remaining 26 ETS family members showed no chromosomal aberrations by FISH. Quantitative polymerase chain reaction showed an average 3.4-fold (P value = .00218) increased expression of ETV1 in melanomas, including the FISH ETV1-amplified cases, when compared to other malignancies (prostate, breast, and bladder carcinomas). These data suggest that a subset of melanomas overexpresses ETV1 and amplification of ETV1 may be one mechanism for achieving high gene expression. PMID:23908683

Exploring polycythaemia vera with fluorescence in situ hybridization: additional cryptic 9p is the most frequent abnormality detected.

Science.gov (United States)

Najfeld, Vesna; Montella, Lya; Scalise, Angela; Fruchtman, Steven

2002-11-01

Between 1986 and 2001, 220 patients with polycythaemia vera (PV) were studied using conventional cytogenetics. Of 204 evaluable patients, 52 (25.4%) had clonal abnormalities. The recurrent chromosomal rearrangements were those of chromosome 9 (21.1%), del(20q) (19.2%), trisomy 8 (19.2%), rearrangements of 13q (13.4%), abnormalities of 1q (11.5%), and of chromosomes 5 and 7 (9.6%). Subsequent analysis of 32 patients, performed at follow-up of up to 14.8 years, revealed new clonal abnormalities in five patients and the disappearance of an abnormal clone in four. Eleven patients remained normal up to 11.5 years and seven patients maintained an abnormality for over 10 years. Fifty-three patients were studied retrospectively using interphase fluorescence in situ hybridization (I-FISH), utilizing probes for centromere enumeration of chromosomes 8 and 9, and for 13q14 and 20q12 loci. Conventional cytogenetics demonstrated clonal chromosome abnormalities in 23% of these 53 patients. The addition of I-FISH increased the detection of abnormalities to 29% and permitted clarification of chromosome 9 rearrangements in an additional 5.6% of patients. FISH uncovered rearrangements of chromosome 9 in 53% of patients with an abnormal FISH pattern, which represented the most frequent genomic alteration in this series.

HYBRID SELECTIVE NON-CATALYTIC REDUCTION (SNCR)/SELECTIVE CATALYTIC REDUCTION (SCR) DEMONSTRATION FOR THE REMOVAL OF NOx FROM BOILER FLUE GASES

Energy Technology Data Exchange (ETDEWEB)

Jerry B. Urbas

1999-05-01

The U. S. Department of Energy (DOE), Electric Power Research Institute (EPRI), Pennsylvania Electric Energy Research Council, (PEERC), New York State Electric and Gas and GPU Generation, Inc. jointly funded a demonstration to determine the capabilities for Hybrid SNCR/SCR (Selective Non-Catalytic Reduction/Selective Catalytic Reduction) technology. The demonstration site was GPU Generation's Seward Unit No.5 (147MW) located in Seward Pennsylvania. The demonstration began in October of 1997 and ended in December 1998. DOE funding was provided through Grant No. DE-FG22-96PC96256 with T. J. Feeley as the Project Manager. EPRI funding was provided through agreements TC4599-001-26999 and TC4599-002-26999 with E. Hughes as the Project Manager. This project demonstrated the operation of the Hybrid SNCR/SCR NO{sub x} control process on a full-scale coal fired utility boiler. The hybrid technology was expected to provide a cost-effective method of reducing NO{sub x} while balancing capital and operation costs. An existing urea based SNCR system was modified with an expanded-duct catalyst to provide increased NO{sub x} reduction efficiency from the SNCR while producing increased ammonia slip levels to the catalyst. The catalyst was sized to reduce the ammonia slip to the air heaters to less than 2 ppm while providing equivalent NO{sub x} reductions. The project goals were to demonstrate hybrid technology is capable of achieving at least a 55% reduction in NO{sub x} emissions while maintaining less than 2ppm ammonia slip to the air heaters, maintain flyash marketability, verify the cost benefit and applicability of Hybrid post combustion technology, and reduce forced outages due to ammonium bisulfate (ABS) fouling of the air heaters. Early system limitations, due to gas temperature stratification, restricted the Hybrid NO{sub x} reduction capabilities to 48% with an ammonia slip of 6.1 mg/Nm{sup 3} (8 ppm) at the catalyst inlet. After resolving the stratification

Thermoelectric properties of graphene nanosheets-modified polyaniline hybrid nanocomposites by an in situ chemical polymerization

International Nuclear Information System (INIS)

Lu, Yan; Song, Ying; Wang, Fuping

2013-01-01

A hybrid material of polyaniline protonated with hydrochloric acid and conductive graphene nanosheets (PANi/GNs) has been prepared by an in situ chemical polymerization method. The interactions between PANi and GNs in the hybrid composites are investigated by utilizing XRD, FT-IR, UV–vis and Raman. It is found that the PANi are adsorbed on the surface of the GNs, and the morphology of PANi transforms from twist structure to extended structure after the GNs are introduced. The thermoelectric (TE) properties of PANi/GNs composites have been investigated in the range from 323 K to 453 K. The electrical conductivity and the Seebeck coefficient of PANi/GNs composites are obviously higher than those of the PANi, while the thermal conductivity of the composites still keeps relatively low values even with high GNs content, resulting in the increase in dimensionless figure of merit (ZT). A highest ZT value of 1.95 × 10 −3 has been obtained for the composite containing 30 wt % GNs at 453 K, which is about 70 times higher than that obtained from the PANi. - Highlights: ► PANi adsorbed on the surface of the GNs possesses more extended structure. ► Electrical conductivity and Seebeck coefficient of PANi/GNs composites are superior to those of PANi. ► Thermal conductivity of the composites still keeps relatively low values

A ScaI RFLP demonstrated for the GRO gene on chromosome 4

Energy Technology Data Exchange (ETDEWEB)

Beck, J.S.; Murray, J.C. (Univ. of Iowa Hospitals, Iowa City (USA)); Sager, R. (Dana Farber Cancer Institute, Boston, MA (USA))

1989-11-11

TC870 is a 0.85 kb fragment running from the EcoRI site 200 pb from the 5{prime} end of the human cDNA subcloned into the EcoRI site of pGEM3. ScaI detects a polymorphism with two variable bands of 19 kb and 16 kb. One strong constant band (14 kb) and two fainter constant bands (5 kb and 3 kb) are also present. The polymorphism type is unknown. GRO has been localized to 4q13-4q21 by somatic cell hybrid analysis and in situ hybridization. Codominant segregation and Hardy-Weinberg equilibrium demonstrated in 20 CEPH families. The probe also was assigned to chromosome 4 with significant linkage to ALB, GC, INP10, MT2P1. GRO may be the same gene as MGSA.

Homology of polytene elements between Drosophila and Zaprionus determined by in situ hybridization in Zaprionus indianus.

Science.gov (United States)

Campos, S R C; Rieger, T T; Santos, J F

2007-05-09

The drosophilid Zaprionus indianus due to its economical importance as an insect pest in Brazil deserves more investigation into its genetics. Its mitotic karyotype and a line-drawing map of its polytene chromosomes are already available. This paper presents a photomap of Z. indianus polytene chromosomes, which was used as the reference map for identification of sections marked by in situ hybridization with gene probes. Hybridization signals for Hsp70 and Hsr-omega were detected, respectively, in sections 34B and 32C of chromosome V of Z. indianus, which indicates its homology to the chromosomal arm 3R of Drosophila melanogaster and, therefore, to Muller's element E. The main signal for Hsp83 gene probe hybridization was in section 17C of Z. indianus chromosome III, suggesting its homology to arm 3L of D. melanogaster and to element D of Muller. The Ubi probe hybridized in sections 10C of chromosome II and 17A of chromosome III. Probably the 17A is the polyubiquitin locus, with homology to arm 3L of D. melanogaster and to the mullerian D element, as suggested also by Hsp83 gene location. The Br-C gene was mapped in section 1D, near the tip of the X chromosome, indicating its homology to the X chromosome of D. melanogaster and to mullerian element A. The Dpp gene probe hybridized mainly in the section 32A of chromosome V and, at lower frequencies to other sections, although no signal was observed as expected in the correspondent mullerian B element. This result led to the suggestion of a rearrangement including the Dpp locus in Z. indianus, the secondary signals possibly pointing to related genes of the TGF-beta family. In conclusion, the results indicate that chromosomes X, III, V of Z. indianus are respectively correspondents to elements A, D, and E of Muller. At least chromosome V of Z. indianus seems to share synteny with the 3R arm of D. melanogaster, as indicated by the relative positions of Hsp70 and Hsr-omega, although the Dpp gene indicates a disruption of

Chromogenic in situ hybridization to detect EGFR gene copy number in cell blocks from fine-needle aspirates of non small cell lung carcinomas and lung metastases from colo-rectal cancer

Directory of Open Access Journals (Sweden)

Terrenato Irene

2010-09-01

Full Text Available Abstract Background Several studies demonstrated that epidermal growth factor receptor (EGFR gene copy number (GCN correlates to the response to tyrosine kinase inhibitors in non small cell lung cancer (NSCLC and to anti-EGFR monoclonal antibodies (MoAbs in metastatic colorectal cancer (CRC. In the presence of lung nodules, cytology is often the only possible diagnostic approach. Chromogenic in situ hybridization (CISH is an alternative technique to fluorescence in situ hybridization (FISH, but its feasibility in detecting EGFR GCN in cell blocks from fine-needle aspiration cytology (FNAC of lung nodules has not yet been established. Methods We evaluated the feasibility of CISH on 33 FNAC from 20 primary NSCLC (5 squamous carcinomas, 8 large cell carcinomas and 7 adenocarcinomas and 13 lung metastases from CRC. Results Of the 33 FNAC analyzed by CISH, 27 (82% presented a balanced increase in EGFR gene and chromosome 7 number: 10 cases (30% showed a low polysomy, 15 (45% a high polysomy and 2 (6% NSCLC were amplified. No significant differences between NSCLC and CRC lung metastases were found in relation to disomic or polysomic status. In addition, no correlation between EGFR GCN and EGFR immunohistochemical overexpression was found. Furthermore, we compared CISH results with those obtained by FISH on the same samples and we found 97% overall agreement between the two assays (k = 0.78, p Conclusions Our study shows that CISH is a valid method to detect EGFR GCN in cell blocks from FNAC of primary NSCLC or metastatic CRC to the lung.

Fluorescent in situ hybridization (FISH as a diagnostic tool for Williams-Beuren syndrome

Directory of Open Access Journals (Sweden)

Deise Helena de Souza

2007-01-01

Full Text Available Fluorescent in situ hybridization (FISH with commercial probes covering the elastin gene (ELN was used to determine the frequency of the 7q11.23 deletion in 18 children clinically diagnosed with Williams-Beuren syndrome (WBS. A de novo deletion was detected in 15 of the children (83%. Diagnostic investigation for WBS started late in childhood (median = 5.8 years. All the children showed facial features typical of the syndrome, mental retardation and developmental delay. Over-friendliness was observed in the majority of cases. Clinodactyly of the 5th finger (n = 13, cardiovascular disease (n = 9, loquacity (n = 9, low birthweight (n = 8, and failure to thrive (n = 9 were observed only in those children with the deletion. Respiratory problems (n = 9, though not previously reported in the literature, was a common finding in the group studied. Our results confirmed that FISH is useful in identifying 7q11.23 deletions in cases of WBS. Clinical manifestations were more evident in the deletion-positive children.

A Case of Renal Primitive Neuroectodermal Tumor Confirmed by Fluorescence in situ Hybridization

Directory of Open Access Journals (Sweden)

Toshiki Etani

2015-04-01

Full Text Available Primitive neuroectodermal tumor (PNET is a member of the Ewing's sarcoma family of tumors (ESFT. We report a case of PNET in a 66-year-old male who presented with a large solid tumor within the parenchyma of the middle pole of the left kidney with metastases to the left adrenal gland and right ischium. A fine-needle biopsy was performed and showed a small round cell tumor. Results of immunohistochemical staining suggested this tumor belonged to ESFT. Preoperative VDC-IE (combined vincristine, doxorubicin and cyclophosphamide followed by another combination of ifosfamide and etoposide chemotherapy and left radical nephrectomy and adrenalectomy were performed. The histopathological findings of the resected tumor were similar to those in the biopsy specimen, but the results of AE1/AE3 were different. For the diagnosis, fluorescence in situ hybridization was performed. Split signals of the EWSR1 gene were detected, and transmission electron microscopy showed neuroendocrine granules and microtubules. The final diagnosis of this tumor was PNET of the kidney.

Operations Support of Phase 2 Integrated Demonstration In Situ Bioremediation. Volume 1, Final report: Final report text data in tabular form, Disk 1

Energy Technology Data Exchange (ETDEWEB)

Hazen, T.C. [Westinghouse Savannah River Co., Aiken, SC (United States)

1993-09-01

This project was designed to demonstrate in situ bioremediation of ground water and sediment contaminated with chlorinated solvents. Indigenous microorganisms were stimulated to degrade trichlorethylene (TCE), tetrachloroethylene (PCE) and their daughter products in situ by addition of nutrients to the contaminated aquifer and adjacent vadose zone. The principle carbon/energy source nutrient used in this demonstration was methane (natural gas). In situ biodegradation is a highly attractive technology for remediation because contaminants are destroyed, not simply moved to another location or immobilized, thus decreasing costs, risks, and time, while increasing efficiency, safety, and public and regulatory acceptability. This report describes the preliminary results of the demonstration and provides conclusions only for those measures that the Bioremediation Technical Support Group felt were so overwhelmingly convincing that they do not require further analyses. Though this report is necessarily superficial it does intend to provide a basis for further evaluating the technology and for practitioners to immediately apply some parts of the technology.

Design of an advanced bundle divertor for the Demonstration Tokamak Hybrid Reactor

International Nuclear Information System (INIS)

Yang, T.F.; Lee, A.Y.; Ruck, G.W.; Prevenslik, T.V.; Smeltzer, G.

1979-01-01

The conclusion of this work is that a bundle divertor, using an improved method of designing the magnetic field configuration, is feasible for the Demonstration Tokamak Hybrid Reactor (DTHR) investigated by Westinghouse. The most significant achievement of this design is the reduction in current density (1 kA/cm 2 ) in the divertor coils in comparison to the overall averaged current densities per tesla of field to be nulled for DITE (25 kA/cm 2 ) and for ISX-B 2 (11 kA/cm 2 ). Therefore, superconducting magnets can be built into the tight space available with a sound mechanical structure

Demonstration and field trial of a resilient hybrid NG-PON test-bed

Science.gov (United States)

Prat, Josep; Polo, Victor; Schrenk, Bernhard; Lazaro, Jose A.; Bonada, Francesc; Lopez, Eduardo T.; Omella, Mireia; Saliou, Fabienne; Le, Quang T.; Chanclou, Philippe; Leino, Dmitri; Soila, Risto; Spirou, Spiros; Costa, Liliana; Teixeira, Antonio; Tosi-Beleffi, Giorgio M.; Klonidis, Dimitrios; Tomkos, Ioannis

2014-10-01

A multi-layer next generation PON prototype has been built and tested, to show the feasibility of extended hybrid DWDM/TDM-XGPON FTTH networks with resilient optically-integrated ring-trees architecture, supporting broadband multimedia services. It constitutes a transparent common platform for the coexistence of multiple operators sharing the optical infrastructure of the central metro ring, passively combining the access and the metropolitan network sections. It features 32 wavelength connections at 10 Gbps, up to 1000 users distributed in 16 independent resilient sub-PONs over 100 km. This paper summarizes the network operation, demonstration and field trial results.

Chromosomal instability detected by interphase fluorescence in situ hybridization and its relation to p3 alteration in prostate carcinoma in Saudi patients

International Nuclear Information System (INIS)

Al-Maghrabi, Jaudah A.

2005-01-01

Chromosomal instability (CIN) is a feature of human neoplasm. The p53 mutation has been shown to be associated with CIN in many human dysplastic and neoplastic lesions. The objective of this study was to examine CIN and p53 mutations in prostate carcinoma (Pca) resected from Saudi patients. Testing of p53 alterations using immunohistochemistry was performed on 28 archived prostatic carcinoma specimens containing Pca foci from Saudi patients seen at King Abdul-Aziz University Hospital, Jeddah, Kingdom of Saudi Arabia. Chrosomal instability was evaluated in the same tissues by interphase in situ hybridization (IFISH) using centromere probes for chromosomes 7 and 8. Immunochemistry and IFISH were performed at Princess Margaret Hospital, University Health Network, Toronto, Ontario, Canada in 2001. The p53 immunoreactivity was found in 29% in Pca and 0% in benign epithelium. Interphase in situ hybridization revealed numerical chromosomal alterations in keeping with CIN in 63% of p53 positive and 20% p53 negative Pca. No evidence of CIN was seen in non-neoplastic epithelium. We concluded that CIN as determined by IFISH is present in Pca from Saudi patients similarly to those reported in western countries. The p53 mutation occurs relatively infrequently in Pca and associated with the presence of CIN at least in a subset of Pca. (author)

Chromogenic in situ hybridization is a reliable assay for detection of ALK rearrangements in adenocarcinomas of the lung.

Science.gov (United States)

Schildhaus, Hans-Ulrich; Deml, Karl-Friedrich; Schmitz, Katja; Meiboom, Maren; Binot, Elke; Hauke, Sven; Merkelbach-Bruse, Sabine; Büttner, Reinhard

2013-11-01

Reliable detection of anaplastic lymphoma kinase (ALK) rearrangements is a prerequisite for personalized treatment of lung cancer patients, as ALK rearrangements represent a predictive biomarker for the therapy with specific tyrosine kinase inhibitors. Currently, fluorescent in situ hybridization (FISH) is considered to be the standard method for assessing formalin-fixed and paraffin-embedded tissue for ALK inversions and translocations. However, FISH requires a specialized equipment, the signals fade rapidly and it is difficult to detect overall morphology and tumor heterogeneity. Chromogenic in situ hybridization (CISH) has been successfully introduced as an alternative test for the detection of several genetic aberrations. This study validates a newly developed ALK CISH assay by comparing FISH and CISH signal patterns in lung cancer samples with and without ALK rearrangements. One hundred adenocarcinomas of the lung were included in this study, among them 17 with known ALK rearrangement. FISH and CISH were carried out and evaluated according to the manufacturers' recommendations. For both assays, tumors were considered positive if ≥15% of tumor cells showed either isolated 3' signals or break-apart patterns or a combination of both. A subset of tumors was exemplarily examined by using a novel EML4 (echinoderm microtubule-associated protein-like 4) CISH probe. Red, green and fusion CISH signals were clearcut and different signal patterns were easily recognized. The percentage of aberrant tumor cells was statistically highly correlated (PCISH. On the basis of 86 samples that were evaluable by ALK CISH, we found a 100% sensitivity and 100% specificity of this assay. Furthermore, EML4 rearrangements could be recognized by CISH. CISH is a highly reliable, sensitive and specific method for the detection of ALK gene rearrangements in pulmonary adenocarcinomas. Our results suggest that CISH might serve as a suitable alternative to FISH, which is the current gold

Expression of a serine protease (motopsin PRSS12) mRNA in the mouse brain: in situ hybridization histochemical study.

Science.gov (United States)

Iijima, N; Tanaka, M; Mitsui, S; Yamamura, Y; Yamaguchi, N; Ibata, Y

1999-03-20

Serine proteases are considered to play several important roles in the brain. In an attempt to find novel brain-specific serine proteases (BSSPs), motopsin (PRSS-12) was cloned from a mouse brain cDNA library by polymerase chain reaction (PCR). Northern blot analysis demonstrated that the postnatal 10-day mouse brain contained the most amount of motopsin mRNA. At this developmental stage, in situ hybridization histochemistry showed that motopsin mRNA was specifically expressed in the following regions: cerebral cortical layers II/III, V and VIb, endopiriform cortex and the limbic system, particularly in the CA1 region of the hippocampal formation. In addition, in the brainstem, the oculomotor nucleus, trochlear nucleus, mecencephalic and motor nuclei of trigeminal nerve (N), abducens nucleus, facial nucleus, nucleus of the raphe pontis, dorsoral motor nucleus of vagal N, hypoglossal nucleus and ambiguus nucleus showed motopsin mRNA expression. Expression was also found in the anterior horn of the spinal cord. The above findings strongly suggest that neurons in almost all motor nuclei, particularly in the brainstem and spinal cord, express motopsin mRNA, and that motopsin seems to have a close relation to the functional role of efferent neurons. Copyright 1999 Elsevier Science B.V.

Polymer-Based Black Phosphorus (bP) Hybrid Materials by in Situ Radical Polymerization: An Effective Tool To Exfoliate bP and Stabilize bP Nanoflakes

Science.gov (United States)

2018-01-01

Black phosphorus (bP) has been recently investigated for next generation nanoelectronic multifunctional devices. However, the intrinsic instability of exfoliated bP (the bP nanoflakes) toward both moisture and air has so far overshadowed its practical implementation. In order to contribute to fill this gap, we report here the preparation of new hybrid polymer-based materials where bP nanoflakes (bPn) exhibit a significantly improved stability. The new materials have been prepared by different synthetic paths including: (i) the mixing of conventionally liquid-phase exfoliated bP (in dimethyl sulfoxide, DMSO) with poly(methyl methacrylate) (PMMA) solution; (ii) the direct exfoliation of bP in a polymeric solution; (iii) the in situ radical polymerization after exfoliating bP in the liquid monomer (methyl methacrylate, MMA). This last methodology concerns the preparation of stable suspensions of bPn–MMA by sonication-assisted liquid-phase exfoliation (LPE) of bP in the presence of MMA followed by radical polymerization. The hybrids characteristics have been compared in order to evaluate the bP dispersion and the effectiveness of the bPn interfacial interactions with polymer chains aimed at their long-term environmental stabilization. The passivation of the bPn is particularly effective when the hybrid material is prepared by in situ polymerization. By using this synthetic methodology, the nanoflakes, even if with a gradient of dispersion (size of aggregates), preserve their chemical structure from oxidation (as proved by both Raman and 31P-solid state NMR studies) and are particularly stable to air and UV light exposure. The feasibility of this approach, capable of efficiently exfoliating bP while protecting the bPn, has been then verified by using different vinyl monomers (styrene and N-vinylpyrrolidone), thus obtaining hybrids where the nanoflakes are embedded in polymer matrices with a variety of intriguing thermal, mechanical, and solubility characteristics.

HYBRID SELECTIVE NON-CATALYTIC REDUCTION (SNCR)/SELECTIVE CATALYTIC REDUCTION (SCR) DEMONSTRATION FOR THE REMOVAL OF NOx FROM BOILER FLUE GASES; FINAL

International Nuclear Information System (INIS)

Jerry B. Urbas

1999-01-01

The U. S. Department of Energy (DOE), Electric Power Research Institute (EPRI), Pennsylvania Electric Energy Research Council, (PEERC), New York State Electric and Gas and GPU Generation, Inc. jointly funded a demonstration to determine the capabilities for Hybrid SNCR/SCR (Selective Non-Catalytic Reduction/Selective Catalytic Reduction) technology. The demonstration site was GPU Generation's Seward Unit No.5 (147MW) located in Seward Pennsylvania. The demonstration began in October of 1997 and ended in December 1998. DOE funding was provided through Grant No. DE-FG22-96PC96256 with T. J. Feeley as the Project Manager. EPRI funding was provided through agreements TC4599-001-26999 and TC4599-002-26999 with E. Hughes as the Project Manager. This project demonstrated the operation of the Hybrid SNCR/SCR NO(sub x) control process on a full-scale coal fired utility boiler. The hybrid technology was expected to provide a cost-effective method of reducing NO(sub x) while balancing capital and operation costs. An existing urea based SNCR system was modified with an expanded-duct catalyst to provide increased NO(sub x) reduction efficiency from the SNCR while producing increased ammonia slip levels to the catalyst. The catalyst was sized to reduce the ammonia slip to the air heaters to less than 2 ppm while providing equivalent NO(sub x) reductions. The project goals were to demonstrate hybrid technology is capable of achieving at least a 55% reduction in NO(sub x) emissions while maintaining less than 2ppm ammonia slip to the air heaters, maintain flyash marketability, verify the cost benefit and applicability of Hybrid post combustion technology, and reduce forced outages due to ammonium bisulfate (ABS) fouling of the air heaters. Early system limitations, due to gas temperature stratification, restricted the Hybrid NO(sub x) reduction capabilities to 48% with an ammonia slip of 6.1 mg/Nm(sup 3) (8 ppm) at the catalyst inlet. After resolving the stratification problem

Genetic analysis of tumorigenesis: XXXII. Localization of constitutionally amplified KRAS sequences to Chinese hamster chromosomes X and Y by in situ hybridization.

Science.gov (United States)

Stenman, G; Anisowicz, A; Sager, R

1988-11-01

The KRAS gene is constitutionally amplified in the Chinese hamster. We have mapped the amplified sequences by in situ hybridization to two major sites on the X and Y chromosomes, Xq4 and Yp2. No autosomal site was detected despite a search under relaxed hybridization conditions. KRAS DNA is amplified about 50-fold compared to a human cell line known to have a diploid number of KRAS sequences, whereas mRNA expression is 5- to 10-fold lower than in normal human cells. While mRNA expression levels do not necessarily parallel gene copy number, the low expression level strongly suggests that the amplified sequences are transcriptionally silent. It is suggested that the amplified sequences arose from the original KRAS gene on chromosome 8 and that the KRAS sequences on the Y chromosome arose by X-Y recombination.

Linkage of cDNA expression profiles of mesencephalic dopaminergic neurons to a genome-wide in situ hybridization database

Directory of Open Access Journals (Sweden)

Simon Horst H

2009-01-01

Full Text Available Abstract Midbrain dopaminergic neurons are involved in control of emotion, motivation and motor behavior. The loss of one of the subpopulations, substantia nigra pars compacta, is the pathological hallmark of one of the most prominent neurological disorders, Parkinson's disease. Several groups have looked at the molecular identity of midbrain dopaminergic neurons and have suggested the gene expression profile of these neurons. Here, after determining the efficiency of each screen, we provide a linked database of the genes, expressed in this neuronal population, by combining and comparing the results of six previous studies and verification of expression of each gene in dopaminergic neurons, using the collection of in situ hybridization in the Allen Brain Atlas.

Demonstration, testing, & evaluation of in situ heating of soil. Draft final report, Volume II: Appendices A to E

Energy Technology Data Exchange (ETDEWEB)

Dev, H.; Enk, J.; Jones, D.; Saboto, W.

1996-02-12

This document is a draft final report for US DOE contract entitled, {open_quotes}Demonstration Testing and Evaluation of In Situ Soil Heating,{close_quotes} Contract No. DE-AC05-93OR22160, IITRI Project No. C06787. This report is presented in two volumes. Volume I contains the technical report This document is Volume II, containing appendices with background information and data. In this project approximately 300 cu. yd. of clayey soil containing a low concentration plume of volatile organic chemicals was heated in situ by the application of electrical energy. It was shown that as a result of heating the effective permeability of soil to air flow was increased such that in situ soil vapor extraction could be performed. The initial permeability of soil was so low that the soil gas flow rate was immeasurably small even at high vacuum levels. When scaled up, this process can be used for the environmental clean up and restoration of DOE sites contaminated with VOCs and other organic chemicals boiling up to 120{degrees}to 130{degrees}C in the vadose zone. Although it may applied to many types of soil formations, it is particularly attractive for low permeability clayey soil where conventional in situ venting techniques are limited by low air flow.

Fluorescent in situ hybridization (FISH) on corneal impression cytology specimens (CICS): study of epithelial cell survival after keratoplasty.

Science.gov (United States)

Catanese, Muriel; Popovici, Cornel; Proust, Hélène; Hoffart, Louis; Matonti, Frédéric; Cochereau, Isabelle; Conrath, John; Gabison, Eric E

2011-02-22

To assess corneal epithelial cell survival after keratoplasty. Corneal impression cytology (CIC) was performed on sex-mismatched corneal transplants. Fluorescent in situ hybridization (FISH) with sex chromosome-specific probes was performed to identify epithelial cell mosaicism and therefore allocate the donor or recipient origin of the cells. Twenty-four samples of corneal epithelial cells derived from 21 transplanted patients were analyzed. All patients received post-operative treatment using dexamethasone eye drops, with progressive tapering over 18 months, and nine patients also received 2% cyclosporine eye drops. Out of the 24 samples reaching quality criteria, sex mosaicism was found in 13, demonstrating the presence of donor-derived cells at the center of the graft for at least 211 days post keratoplasty. Kaplan-Meier analysis established a median survival of donor corneal epithelial cells of 385 days. Although not statistically significant, the disappearance of donor cells seemed to be delayed and the average number of persistent cells appeared to be greater when 2% cyclosporine was used topically as an additional immunosuppressive therapy. The combination of corneal impressions and FISH analysis is a valuable tool with negligible side effects to investigate the presence of epithelial cell mosaicism in sex-mismatched donor transplants. Epithelial cells survived at the center of the graft with a median survival of more than one year, suggesting slower epithelial turnover than previously described.

Whole-mount in situ hybridization in the rotifer Brachionus plicatilis representing a basal branch of lophotrochozoans.

Science.gov (United States)

Boell, Louis A; Bucher, Gregor

2008-08-01

In order to broaden the comparative scope of evolutionary developmental biology and to refine our picture of animal macroevolution, it is necessary to establish new model organisms, especially from previously underrepresented groups, like the Lophotrochozoa. We have established the culture and protocols for molecular developmental biology in the rotifer species Brachionus plicatilis Müller (Rotifera, Monogononta). Rotifers are nonsegmented animals with enigmatic basal position within the lophotrochozoans and marked by several evolutionary novelties like the wheel organ (corona), the median eye, and the nonpaired posterior foot. The expression of Bp-Pax-6 is shown using whole-mount in situ hybridization. The inexpensive easy culture and experimental tractability of Brachionus as well as the range of interesting questions to which it holds the key make it a promising addition to the "zoo" of evo-devo model organisms.

Detection of dengue group viruses by fluorescence in situ hybridization

Directory of Open Access Journals (Sweden)

Raquin Vincent

2012-10-01

Full Text Available Abstract Background Dengue fever (DF and dengue hemorrhagic fever (DHF represent a global challenge in public health. It is estimated that 50 to 100 million infections occur each year causing approximately 20,000 deaths that are usually linked to severe cases like DHF and dengue shock syndrome. The causative agent of DF is dengue virus (genus Flavivirus that comprises four distinct serotypes (DENV-1 to DENV-4. Fluorescence in situ hybridization (FISH has been used successfully to detect pathogenic agents, but has not been implemented in detecting DENV. To improve our understanding of DENV infection and dissemination in host tissues, we designed specific probes to detect DENV in FISH assays. Methods Oligonucleotide probes were designed to hybridize with RNA from the broadest range of DENV isolates belonging to the four serotypes, but not to the closest Flavivirus genomes. Three probes that fit the criteria defined for FISH experiments were selected, targeting both coding and non-coding regions of the DENV genome. These probes were tested in FISH assays against the dengue vector Aedes albopictus (Diptera: Culicidae. The FISH experiments were led in vitro using the C6/36 cell line, and in vivo against dissected salivary glands, with epifluorescence and confocal microscopy. Results The three 60-nt oligonucleotides probes DENV-Probe A, B and C cover a broad range of DENV isolates from the four serotypes. When the three probes were used together, specific fluorescent signals were observed in C6/36 infected with each DENV serotypes. No signal was detected in either cells infected with close Flavivirus members West Nile virus or yellow fever virus. The same protocol was used on salivary glands of Ae. albopictus fed with a DENV-2 infectious blood-meal which showed positive signals in the lateral lobes of infected samples, with no significant signal in uninfected mosquitoes. Conclusion Based on the FISH technique, we propose a way to design and use

FISHIS: fluorescence in situ hybridization in suspension and chromosome flow sorting made easy.

Directory of Open Access Journals (Sweden)

Debora Giorgi

Full Text Available The large size and complex polyploid nature of many genomes has often hampered genomics development, as is the case for several plants of high agronomic value. Isolating single chromosomes or chromosome arms via flow sorting offers a clue to resolve such complexity by focusing sequencing to a discrete and self-consistent part of the whole genome. The occurrence of sufficient differences in the size and or base-pair composition of the individual chromosomes, which is uncommon in plants, is critical for the success of flow sorting. We overcome this limitation by developing a robust method for labeling isolated chromosomes, named Fluorescent In situ Hybridization In suspension (FISHIS. FISHIS employs fluorescently labeled synthetic repetitive DNA probes, which are hybridized, in a wash-less procedure, to chromosomes in suspension following DNA alkaline denaturation. All typical A, B and D genomes of wheat, as well as individual chromosomes from pasta (T. durum L. and bread (T. aestivum L. wheat, were flow-sorted, after FISHIS, at high purity. For the first time in eukaryotes, each individual chromosome of a diploid organism, Dasypyrum villosum (L. Candargy, was flow-sorted regardless of its size or base-pair related content. FISHIS-based chromosome sorting is a powerful and innovative flow cytogenetic tool which can develop new genomic resources from each plant species, where microsatellite DNA probes are available and high quality chromosome suspensions could be produced. The joining of FISHIS labeling and flow sorting with the Next Generation Sequencing methodology will enforce genomics for more species, and by this mightier chromosome approach it will be possible to increase our knowledge about structure, evolution and function of plant genome to be used for crop improvement. It is also anticipated that this technique could contribute to analyze and sort animal chromosomes with peculiar cytogenetic abnormalities, such as copy number variations

FISHIS: fluorescence in situ hybridization in suspension and chromosome flow sorting made easy.

Science.gov (United States)

Giorgi, Debora; Farina, Anna; Grosso, Valentina; Gennaro, Andrea; Ceoloni, Carla; Lucretti, Sergio

2013-01-01

The large size and complex polyploid nature of many genomes has often hampered genomics development, as is the case for several plants of high agronomic value. Isolating single chromosomes or chromosome arms via flow sorting offers a clue to resolve such complexity by focusing sequencing to a discrete and self-consistent part of the whole genome. The occurrence of sufficient differences in the size and or base-pair composition of the individual chromosomes, which is uncommon in plants, is critical for the success of flow sorting. We overcome this limitation by developing a robust method for labeling isolated chromosomes, named Fluorescent In situ Hybridization In suspension (FISHIS). FISHIS employs fluorescently labeled synthetic repetitive DNA probes, which are hybridized, in a wash-less procedure, to chromosomes in suspension following DNA alkaline denaturation. All typical A, B and D genomes of wheat, as well as individual chromosomes from pasta (T. durum L.) and bread (T. aestivum L.) wheat, were flow-sorted, after FISHIS, at high purity. For the first time in eukaryotes, each individual chromosome of a diploid organism, Dasypyrum villosum (L.) Candargy, was flow-sorted regardless of its size or base-pair related content. FISHIS-based chromosome sorting is a powerful and innovative flow cytogenetic tool which can develop new genomic resources from each plant species, where microsatellite DNA probes are available and high quality chromosome suspensions could be produced. The joining of FISHIS labeling and flow sorting with the Next Generation Sequencing methodology will enforce genomics for more species, and by this mightier chromosome approach it will be possible to increase our knowledge about structure, evolution and function of plant genome to be used for crop improvement. It is also anticipated that this technique could contribute to analyze and sort animal chromosomes with peculiar cytogenetic abnormalities, such as copy number variations or cytogenetic

Demonstration of hybrid orbital angular momentum multiplexing and time-division multiplexing passive optical network.

Science.gov (United States)

Wang, Andong; Zhu, Long; Liu, Jun; Du, Cheng; Mo, Qi; Wang, Jian

2015-11-16

Mode-division multiplexing passive optical network (MDM-PON) is a promising scheme for next-generation access networks to further increase fiber transmission capacity. In this paper, we demonstrate the proof-of-concept experiment of hybrid mode-division multiplexing (MDM) and time-division multiplexing (TDM) PON architecture by exploiting orbital angular momentum (OAM) modes. Bidirectional transmissions with 2.5-Gbaud 4-level pulse amplitude modulation (PAM-4) downstream and 2-Gbaud on-off keying (OOK) upstream are demonstrated in the experiment. The observed optical signal-to-noise ratio (OSNR) penalties for downstream and upstream transmissions at a bit-error rate (BER) of 2 × 10(-3) are less than 2.0 dB and 3.0 dB, respectively.

Longitudinal fluorescence in situ hybridization reveals cytogenetic evolution in myeloma relapsing after autologous transplantation

Science.gov (United States)

Merz, Maximilian; Jauch, Anna; Hielscher, Thomas; Mai, Elias K.; Seckinger, Anja; Hose, Dirk; Bertsch, Uta; Neben, Kai; Raab, Marc S.; Salwender, Hans; Blau, Igor W.; Lindemann, Hans-Walter; Schmidt-Wolf, Ingo; Scheid, Christof; Haenel, Mathias; Weisel, Katja; Goldschmidt, Hartmut; Hillengass, Jens

2017-01-01

To investigate cytogenetic evolution after upfront autologous stem cell transplantation for newly diagnosed myeloma we retrospectively analyzed fluorescence in situ hybridization results of 128 patients with paired bone marrow samples from the time of primary diagnosis and at relapse. High-risk cytogenetic abnormalities (deletion 17p and/or gain 1q21) occurred more frequently after relapse (odds ratio: 6.33; 95% confidence interval: 1.86–33.42; P<0.001). No significant changes were observed for defined IGH translocations [t(4;14); t(11;14); t(14;16)] or hyperdiploid karyotypes between primary diagnosis and relapse. IGH translocations with unknown partners occurred more frequently at relapse. New deletion 17p and/or gain 1q21 were associated with cytogenetic heterogeneity, since some de novo lesions with different copy numbers were present only in subclones. No distinct baseline characteristics were associated with the occurrence of new high-risk cytogenetic abnormalities after progression. Patients who relapsed after novel agent-based induction therapy had an increased risk of developing high-risk aberrations (odds ratio 10.82; 95% confidence interval: 1.65–127.66; P=0.03) compared to those who were treated with conventional chemotherapy. Survival analysis revealed dismal outcomes regardless of whether high-risk aberrations were present at baseline (hazard ratio, 3.53; 95% confidence interval: 1.53–8.14; P=0.003) or developed at relapse only (hazard ratio, 3.06; 95% confidence interval: 1.09–8.59; P=0.03). Our results demonstrate cytogenetic evolution towards high-risk disease after autologous transplantation and underline the importance of repeated genetic testing in relapsed myeloma (EudraCT number of the HD4 trial: 2004-000944-26). PMID:28495913

Flat mount preparation for observation and analysis of zebrafish embryo specimens stained by whole mount in situ hybridization.

Science.gov (United States)

Cheng, Christina N; Li, Yue; Marra, Amanda N; Verdun, Valerie; Wingert, Rebecca A

2014-07-17

The zebrafish embryo is now commonly used for basic and biomedical research to investigate the genetic control of developmental processes and to model congenital abnormalities. During the first day of life, the zebrafish embryo progresses through many developmental stages including fertilization, cleavage, gastrulation, segmentation, and the organogenesis of structures such as the kidney, heart, and central nervous system. The anatomy of a young zebrafish embryo presents several challenges for the visualization and analysis of the tissues involved in many of these events because the embryo develops in association with a round yolk mass. Thus, for accurate analysis and imaging of experimental phenotypes in fixed embryonic specimens between the tailbud and 20 somite stage (10 and 19 hours post fertilization (hpf), respectively), such as those stained using whole mount in situ hybridization (WISH), it is often desirable to remove the embryo from the yolk ball and to position it flat on a glass slide. However, performing a flat mount procedure can be tedious. Therefore, successful and efficient flat mount preparation is greatly facilitated through the visual demonstration of the dissection technique, and also helped by using reagents that assist in optimal tissue handling. Here, we provide our WISH protocol for one or two-color detection of gene expression in the zebrafish embryo, and demonstrate how the flat mounting procedure can be performed on this example of a stained fixed specimen. This flat mounting protocol is broadly applicable to the study of many embryonic structures that emerge during early zebrafish development, and can be implemented in conjunction with other staining methods performed on fixed embryo samples.

Innovative technology demonstrations

International Nuclear Information System (INIS)

Anderson, D.B.; Hartley, J.N.; Luttrell, S.P.

1992-04-01

Currently, several innovative technologies are being demonstrated at Tinker Air Force Base (TAFB) to address specific problems associated with remediating two contaminated test sites at the base. Cone penetrometer testing (CPT) is a form of testing that can rapidly characterize a site. This technology was selected to evaluate its applicability in the tight clay soils and consolidated sandstone sediments found at TAFB. Directionally drilled horizontal wells have been successfully installed at the US Department of Energy's (DOE) Savannah River Site to test new methods of in situ remediation of soils and ground water. This emerging technology was selected as a method that may be effective in accessing contamination beneath Building 3001 without disrupting the mission of the building, and in enhancing the extraction of contamination both in ground water and in soil. A soil gas extraction (SGE) demonstration, also known as soil vapor extraction, will evaluate the effectiveness of SGE in remediating fuels and TCE contamination contained in the tight clay soil formations surrounding the abandoned underground fuel storage vault located at the SW Tanks Site. In situ sensors have recently received much acclaim as a technology that can be effective in remediating hazardous waste sites. Sensors can be useful for determining real-time, in situ contaminant concentrations during the remediation process for performance monitoring and in providing feedback for controlling the remediation process. A demonstration of two in situ sensor systems capable of providing real-time data on contamination levels will be conducted and evaluated concurrently with the SGE demonstration activities. Following the SGE demonstration, the SGE system and SW Tanks test site will be modified to demonstrate bioremediation as an effective means of degrading the remaining contaminants in situ

Single molecule localization imaging of telomeres and centromeres using fluorescence in situ hybridization and semiconductor quantum dots.

Science.gov (United States)

Wang, Le; Zong, Shenfei; Wang, Zhuyuan; Lu, Ju; Chen, Chen; Zhang, Ruohu; Cui, Yiping

2018-07-13

Single molecule localization microscopy (SMLM) is a powerful tool for imaging biological targets at the nanoscale. In this report, we present SMLM imaging of telomeres and centromeres using fluorescence in situ hybridization (FISH). The FISH probes were fabricated by decorating CdSSe/ZnS quantum dots (QDs) with telomere or centromere complementary DNA strands. SMLM imaging experiments using commercially available peptide nucleic acid (PNA) probes labeled with organic fluorophores were also conducted to demonstrate the advantages of using QDs FISH probes. Compared with the PNA probes, the QDs probes have the following merits. First, the fluorescence blinking of QDs can be realized in aqueous solution or PBS buffer without thiol, which is a key buffer component for organic fluorophores' blinking. Second, fluorescence blinking of the QDs probe needs only one excitation light (i.e. 405 nm). While fluorescence blinking of the organic fluorophores usually requires two illumination lights, that is, the activation light (i.e. 405 nm) and the imaging light. Third, the high quantum yield, multiple switching times and a good optical stability make the QDs more suitable for long-term imaging. The localization precision achieved in telomeres and centromeres imaging experiments is about 30 nm, which is far beyond the diffraction limit. SMLM has enabled new insights into telomeres or centromeres on the molecular level, and it is even possible to determine the length of telomere and become a potential technique for telomere-related investigation.

Facile in situ synthesis of hierarchical porous Ni/Ni(OH)₂ hybrid sponges with excellent electrochemical energy-storage performances for supercapacitors.

Science.gov (United States)

Wang, Wanren; Wang, Wenhua; Wang, Mengjiao; Guo, Xiaohui

2014-09-01

Herein, we report the in situ growth of single-crystalline Ni(OH)2 nanoflakes on a Ni support by using facile hydrothermal processes. The as-prepared Ni/Ni(OH)2 sponges were well-characterized by using X-ray diffraction (XRD), SEM, TEM, and X-ray photoelectron spectroscopy (XPS) techniques. The results revealed that the nickel-skeleton-supported Ni(OH)2 rope-like aggregates were composed of numerous intercrossed single-crystal Ni(OH)2 flake-like units. The Ni/Ni(OH)2 hybrid sponges served as electrodes and displayed ultrahigh specific capacitance (SC=3247 F g(-1)) and excellent rate-capability performance, likely owing to fast electron and ion transport, sufficient Faradic redox reaction, and robust structural integrity of the Ni/Ni(OH)2 hybrid electrode. These results support the promising application of Ni(OH)2 nanoflakes as advanced pseudocapacitor materials. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Preparation of Transparent Bulk TiO2/PMMA Hybrids with Improved Refractive Indices via an in Situ Polymerization Process Using TiO2 Nanoparticles Bearing PMMA Chains Grown by Surface-Initiated Atom Transfer Radical Polymerization.

Science.gov (United States)

Maeda, Satoshi; Fujita, Masato; Idota, Naokazu; Matsukawa, Kimihiro; Sugahara, Yoshiyuki

2016-12-21

Transparent TiO 2 /PMMA hybrids with a thickness of 5 mm and improved refractive indices were prepared by in situ polymerization of methyl methacrylate (MMA) in the presence of TiO 2 nanoparticles bearing poly(methyl methacrylate) (PMMA) chains grown using surface-initiated atom transfer radical polymerization (SI-ATRP), and the effect of the chain length of modified PMMA on the dispersibility of modified TiO 2 nanoparticles in the bulk hybrids was investigated. The surfaces of TiO 2 nanoparticles were modified with both m-(chloromethyl)phenylmethanoyloxymethylphosphonic acid bearing a terminal ATRP initiator and isodecyl phosphate with a high affinity for common organic solvents, leading to sufficient dispersibility of the surface-modified particles in toluene. Subsequently, SI-ATRP of MMA was achieved from the modified surfaces of the TiO 2 nanoparticles without aggregation of the nanoparticles in toluene. The molecular weights of the PMMA chains cleaved from the modified TiO 2 nanoparticles increased with increases in the prolonging of the polymerization period, and these exhibited a narrow distribution, indicating chain growth controlled by SI-ATRP. The nanoparticles bearing PMMA chains were well-dispersed in MMA regardless of the polymerization period. Bulk PMMA hybrids containing modified TiO 2 nanoparticles with a thickness of 5 mm were prepared by in situ polymerization of the MMA dispersion. The transparency of the hybrids depended significantly on the chain length of the modified PMMA on the nanoparticles, because the modified PMMA of low molecular weight induced aggregation of the TiO 2 nanoparticles during the in situ polymerization process. The refractive indices of the bulk hybrids could be controlled by adjusting the TiO 2 content and could be increased up to 1.566 for 6.3 vol % TiO 2 content (1.492 for pristine PMMA).

Diagnostic and Prognostic Utility of Fluorescence In situ Hybridization (FISH) Analysis in Acute Myeloid Leukemia.

Science.gov (United States)

Gonzales, Patrick R; Mikhail, Fady M

2017-12-01

Acute myeloid leukemia (AML) is a hematologic neoplasia consisting of incompletely differentiated hematopoietic cells of the myeloid lineage that proliferate in the bone marrow, blood, and/or other tissues. Clinical implementation of fluorescence in situ hybridization (FISH) in cytogenetic laboratories allows for high-resolution analysis of recurrent structural chromosomal rearrangements specific to AML, especially in AML with normal karyotypes, which comprises approximately 33-50% of AML-positive specimens. Here, we review the use of several FISH probe strategies in the diagnosis of AML. We also review the standards and guidelines currently in place for use by clinical cytogenetic laboratories in the evaluation of AML. Updated standards and guidelines from the WHO, ACMG, and NCCN have further defined clinically significant, recurring cytogenetic anomalies in AML that are detectable by FISH. FISH continues to be a powerful technique in the diagnosis of AML, with higher resolution than conventional cytogenetic analysis, rapid turnaround time, and a considerable diagnostic and prognostic utility.

JC virus chromogenic in situ hybridization in brain biopsies from patients with and without PML.

Science.gov (United States)

Procop, Gary W; Beck, Rose C; Pettay, James D; Kohn, Debra J; Tuohy, Marion J; Yen-Lieberman, Belinda; Prayson, Richard A; Tubbs, Raymond R

2006-06-01

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system caused by the JC polyoma virus. Electron microscopy and immunohistochemistry are the traditional methods of confirming the presence of the virus in brain biopsies from these patients. We studied the brain biopsies from 7 patients with PML and 6 patients without PML with chromogenic in situ hybridization (CISH) for the JC polyoma virus using a commercially available probe. The biopsies from the patients with the PML cases were proven to contain the JC polyoma virus by traditional and molecular methods. The CISH findings were compared with the known state of infection. All (7/7) of the biopsies from patients with PML were positive for the presence of polyoma virus by CISH, whereas the biopsies from patients without PML were uniformly negative. CISH seems to be a useful tool for the detection of the JC virus in brain biopsies from patients with PML, and is more accessible because a commercial probe is available.

Scanning electron microscopy and fluorescent in situ hybridization of experimental Brachyspira (Serpulina) pilosicoli infection in growing pigs

DEFF Research Database (Denmark)

Jensen, Tim Kåre; Møller, Kristian; Boye, Mette

2000-01-01

Two groups of six 8-week-old pigs were challenged with 1X10(9) cfu Brachyspira (Serpulina) pilosicoli or Serpulina intermedia daily for 3 consecutive days to study the pathology of porcine colonic spirochetosis by scanning electron microscopy (SEM) and fluorescent in situ hybridization (FISH......; however, only two pigs developed transient watery diarrhea. S. intermedia was reisolated from four of the inoculated pigs, but clinical signs were not observed. Gross examination of the B. pilosicoli-infected pigs revealed dilated large intestines with a hyperemic mucosa, whereas the large intestines...... of the S. intermedia-inoculated pigs and the control pigs appeared normal. SEM examination of B. pilosicoli-infected pigs revealed degenerated epithelial cells and spirochetal colonization of the colonic mucosa in four pigs. By FISH, B. pilosicoli cells were found colonizing and invading the surface...

Dual color chromogenic in situ hybridization for determination of HER2 status in breast cancer: a large comparative study to current state of the art fluorescence in situ hybridization

Directory of Open Access Journals (Sweden)

Mollerup Jens

2012-02-01

Full Text Available Abstract Background Chromogenic in situ hybridization (CISH is fast becoming a well established technique for easy and sensitive determination of HER2 gene status in breast cancer. However, for the chromogenic method to achieve status as a safe and reliable technique, the method needs to be validated against already known and validated FISH techniques. Methods Here it is reported from a comparative study where HER2 gene status obtained by HER2 CISH pharmDx™ Kit was compared to HER2 gene status obtained by the FDA approved HER2 FISH pharmDx™ Kit and the PathVysion HER-2 DNA probe Kit. The study included 365 formalin fixed and paraffin-embedded invasive breast cancer tissue specimens collected consecutively at a US reference laboratory. Results The data obtained revealed an overall HER2 status concordance of approximately 98% for comparisons of HER2 CISH pharmDx™ Kit to both HER2 FISH pharmDx™ Kit and PathVysion HER-2 DNA Probe Kit. Conclusions The concordance between results obtained using the recently FDA approved HER2 CISH pharmDx™ Kit with previously FDA approved FISH techniques for HER2 gene status determination indicate that the HER2 CISH pharmDx™ Kit is a reliable chromogenic alternative to fluorescence-based methods.

Dual color chromogenic in situ hybridization for determination of HER2 status in breast cancer: a large comparative study to current state of the art fluorescence in situ hybridization

Science.gov (United States)

2012-01-01

Background Chromogenic in situ hybridization (CISH) is fast becoming a well established technique for easy and sensitive determination of HER2 gene status in breast cancer. However, for the chromogenic method to achieve status as a safe and reliable technique, the method needs to be validated against already known and validated FISH techniques. Methods Here it is reported from a comparative study where HER2 gene status obtained by HER2 CISH pharmDx™ Kit was compared to HER2 gene status obtained by the FDA approved HER2 FISH pharmDx™ Kit and the PathVysion HER-2 DNA probe Kit. The study included 365 formalin fixed and paraffin-embedded invasive breast cancer tissue specimens collected consecutively at a US reference laboratory. Results The data obtained revealed an overall HER2 status concordance of approximately 98% for comparisons of HER2 CISH pharmDx™ Kit to both HER2 FISH pharmDx™ Kit and PathVysion HER-2 DNA Probe Kit. Conclusions The concordance between results obtained using the recently FDA approved HER2 CISH pharmDx™ Kit with previously FDA approved FISH techniques for HER2 gene status determination indicate that the HER2 CISH pharmDx™ Kit is a reliable chromogenic alternative to fluorescence-based methods. PMID:22333181

Implementation and importance of fluorescence in situ hybridization (fish) in paraffin tissues for categorization of B-cell lymphoma unclassifiable, with features intermediate between Burkitt lymphoma and diffuse large B-cell lymphoma

International Nuclear Information System (INIS)

Carvajal Cuenca, Alejandra

2011-01-01

The diagnostic criteria have been defined based on the tools that the country has acquired and international guidelines for pure entities: the LB, LDCGB, and the new entity of B lymphoma unclassifiable with features intermediate LDCGB and LB. The fluorescence in situ hybridization for the translocation (8;14) has been implemented in paraffin tissues for proper categorization. A total of 21 cases have been studied: the characteristics of patients, morphology, immunohistochemistry and the presence or absence of the translocation (8;14). Twelve of the cases have been classified as B-cell lymphoma unclassifiable with features intermediate between LDCGB and LB. Furthermore, nine of the cases were classified in LB. Fluorescence in situ hybridization (FISH) has been negative in 5 of the 21 cases. The diagnosis of lymphoma with features bordering between the LB and the LDCGB has been imperative for the survival of the patient and the corresponding treatment [es

Multicenter Evaluation of a New Shortened Peptide Nucleic Acid Fluorescence In Situ Hybridization Procedure for Species Identification of Select Gram-Negative Bacilli from Blood Cultures▿

Science.gov (United States)

Morgan, Margie; Marlowe, Elizabeth; Della-Latta, Phyllis; Salimnia, Hossein; Novak-Weekley, Susan; Wu, Fann; Crystal, Benjamin S.

2010-01-01

A shortened protocol for two peptide nucleic acid fluorescence in situ hybridization (PNA FISH) assays for the detection of Gram-negative bacilli from positive blood cultures was evaluated in a multicenter trial. There was 100% concordance between the two protocols for each assay (368 of 368 and 370 of 370 results) and 99.7% (367 of 368 and 369 of 370 results) agreement with routine laboratory techniques. PMID:20357212

In Silico and Fluorescence In Situ Hybridization Mapping Reveals Collinearity between the Pennisetum squamulatum Apomixis Carrier-Chromosome and Chromosome 2 of Sorghum and Foxtail Millet

OpenAIRE

Sapkota, Sirjan; Conner, Joann A.; Hanna, Wayne W.; Simon, Bindu; Fengler, Kevin; Deschamps, St?phane; Cigan, Mark; Ozias-Akins, Peggy

2016-01-01

Apomixis, or clonal propagation through seed, is a trait identified within multiple species of the grass family (Poaceae). The genetic locus controlling apomixis in Pennisetum squamulatum (syn Cenchrus squamulatus) and Cenchrus ciliaris (syn Pennisetum ciliare, buffelgrass) is the apospory-specific genomic region (ASGR). Previously, the ASGR was shown to be highly conserved but inverted in marker order between P. squamulatum and C. ciliaris based on fluorescence in situ hybridization (FISH) a...

Accuracy Assessment of Interphase Fluorescence In-Situ Hybridization on Uncultured Amniotic Fluid Cells

Directory of Open Access Journals (Sweden)

Hamideh Karimi

2007-01-01

Full Text Available Background: Parental anxiety while waiting for the results of amniocentesis has been investigatedby many authors. It seems that the implementation of faster techniques such as fluorescence in-situhybridization (FISH will have some benefits in reducing this anxiety. Besides the patients' attitudesto choosing this method, gynecologists who are the persons responsible for treatment, must feelcomfortable about prescribing FISH techniques.Materials and Methods: This study, using a simple methodology, was undertaken to evaluate theresults of FISH tests on the amniotic fluid from 40 pregnant women undergoing cesarean surgery.Two sets of probes including X/Y cocktail and 13, 21 and 18 were applied on different slides.Results: The results of FISH tests were compared with the reports of the pediatrician about thehealth condition of the newborn. Complete conformity between the two sets of findings, haveconvinced our gynecologists of the benefit of prescribing this method to reduce the anxiety ofpatients at risk of having abnormal offspring due to chromosomal anuploidies.Conclusion: As has been documented by many authors, conventional chromosome analysis hasgreat advantages over fluorescence in situ hybridization of interphase amniocytes, but reducing theanxiety of parents is a good reason for employing the FISH technique.

Fast ion transport at solid-solid interfaces in hybrid battery anodes

Science.gov (United States)

Tu, Zhengyuan; Choudhury, Snehashis; Zachman, Michael J.; Wei, Shuya; Zhang, Kaihang; Kourkoutis, Lena F.; Archer, Lynden A.

2018-04-01

Carefully designed solid-electrolyte interphases are required for stable, reversible and efficient electrochemical energy storage in batteries. We report that hybrid battery anodes created by depositing an electrochemically active metal (for example, Sn, In or Si) on a reactive alkali metal electrode by a facile ion-exchange chemistry lead to very high exchange currents and stable long-term performance of electrochemical cells based on Li and Na electrodes. By means of direct visualization and ex situ electrodeposition studies, Sn-Li anodes are shown to be stable at 3 mA cm-2 and 3 mAh cm-2. Prototype full cells in which the hybrid anodes are paired with high-loading LiNi0.8Co0.15Al0.05O2(NCA) cathodes are also reported. As a second demonstration, we create and study Sn-Na hybrid anodes and show that they can be cycled stably for more than 1,700 hours with minimal voltage divergence. Charge storage at the hybrid anodes is reported to involve a combination of alloying and electrodeposition reactions.

Cytogenetic, genomic in situ hybridization (GISH) and agronomic ...

African Journals Online (AJOL)

F3 generations of a wheat-Psathyrostachys huashanica intergeneric cross. Their agronomic traits were evaluated in the field and their meiotic behaviors and chromosome composition were analyzed by cytogenetic and GISH (genomic in situ ...

Chromosome translocations in chinese medical X-ray workers analyzed by fluorescence in situ hybridization

International Nuclear Information System (INIS)

Sun Yuanming; Li Jin; Wang Qin; Tang Weisheng; Wang Zhiquan

2002-01-01

Objective: To study long-term radiation effect in occupational workers exposed to low dose X-rays using the method of fluorescence in situ hybridization (FISH). Method: Chromosome translocations of 25 medical X-ray workers were analyzed by FISH with chromosome No. 4 and No. 7 probes according to PAINT (The Protocol for Aberration Identification and Nomenclature Terminology) system. Results: The frequency of genome translocation in X-ray workers was (13.14 ± 1.23)/1000 cells. The rate of complete and incomplete translocation was 1:1.7. According to the calendar year of entry before/after the year of 1965 as the border, the data showed that the incomplete translocation of the after 1965 group was obviously higher than those of the controls (P < 0.01 and P < 0.05, respectively). Conclusion: The chromosome translocation in early Chinese medical X-ray workers is mainly the incomplete one, the frequency of translocation does not dependent on chromosomal DNA content, and incomplete and complete ones increase along with prolongation of working years in their position

Demonstration of Improved Charge Transfer in Graphene/Au Nanorods Plasmonic Hybrids Stabilized by Benzyl Thiol Linkers

Directory of Open Access Journals (Sweden)

Giuseppe Valerio Bianco

2016-01-01

Full Text Available Hybrids based on graphene decorated with plasmonic gold (Au nanostructures are being investigated as possible materials combination to add to graphene functionalities of tunable plasmon resonance and enhanced absorption at selected wavelength in the visible-near-infrared region of the spectrum. Here, we report a solution drop-casting approach for fabricating stable hybrids based on chemical vapor deposition (CVD graphene and Au nanorods, which are able to activate effective charge transfer from graphene. We demonstrate that CVD graphene functionalization by benzyl thiol (BZT provides the linker to strong anchoring, via S-Au bonds, Au nanorods to graphene. Optical measurements by spectroscopic ellipsometry give evidence of the introduction of plasmon resonances at 1.85 and 2.25 eV in the Au nanorods/BZT/graphene hybrids, which enable surface enhanced Raman scattering (SERS detection. Furthermore, an effective electron transfer from graphene to Au nanorods, resulting in an enhancement of p-type doping of graphene with a consequent decrease of its sheet resistance, is probed by Raman spectroscopy and corroborated by electrical measurements.

Automated brightfield dual-color in situ hybridization for detection of mouse double minute 2 gene amplification in sarcomas.

Science.gov (United States)

Zhang, Wenjun; McElhinny, Abigail; Nielsen, Alma; Wang, Maria; Miller, Melanie; Singh, Shalini; Rueger, Ruediger; Rubin, Brian P; Wang, Zhen; Tubbs, Raymond R; Nagle, Raymond B; Roche, Pat; Wu, Ping; Pestic-Dragovich, Lidija

2011-01-01

The human homolog of the mouse double minute 2 (MDM2) oncogene is amplified in about 20% of sarcomas. The measurement of the MDM2 amplification can aid in classification and may provide a predictive value for recently formulated therapies targeting MDM2. We have developed and validated an automated bright field dual-color in situ hybridization application to detect MDM2 gene amplification. A repeat-depleted MDM2 probe was constructed to target the MDM2 gene region at 12q15. A chromosome 12-specific probe (CHR12) was generated from a pα12H8 plasmid. The in situ hybridization assay was developed by using a dinitrophenyl-labeled MDM2 probe and a digoxigenin-labeled CHR12 probe on the Ventana Medical Systems' automated slide-staining platforms. The specificity of the MDM2 and CHR12 probes was shown on metaphase spreads and further validated against controls, including normal human tonsil and known MDM2-amplified samples. The assay performance was evaluated on a cohort of 100 formalin-fixed, paraffin-embedded specimens by using a conventional bright field microscope. Simultaneous hybridization and signal detection for MDM2 and CHR12 showed that both DNA targets were present in the same cells. One hundred soft tissue specimens were stained for MDM2 and CHR12. Although 26 of 29 lipomas were nonamplified and eusomic, MDM2 amplification was noted in 78% of atypical lipomatous tumors or well-differentiated liposarcomas. Five of 6 dedifferentiated liposarcoma cases were amplified for MDM2. MDM2 amplification was observed in 1 of 8 osteosarcomas; 3 showed CHR12 aneusomy. MDM2 amplification was present in 1 of 4 chondrosarcomas. Nine of 10 synovial sarcomas displayed no evidence of MDM2 amplification in most tumor cells. In pleomorphic sarcoma, not otherwise specified (pleomorphic malignant fibrous histiocytoma), MDM2 was amplified in 38% of cases, whereas 92% were aneusomic for CHR12. One alveolar rhabdomyosarcoma and 2 embryonal rhabdomyosarcomas displayed low-level aneusomy

Facile approach to prepare Pt decorated SWNT/graphene hybrid catalytic ink

Energy Technology Data Exchange (ETDEWEB)

Mayavan, Sundar, E-mail: sundarmayavan@cecri.res.in [Centre for Innovation in Energy Research, CSIR–Central Electrochemical Research Institute, Karaikudi 630006, Tamil Nadu (India); Department of Nuclear and Quantum Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701 (Korea, Republic of); Mandalam, Aditya; Balasubramanian, M. [Centre for Innovation in Energy Research, CSIR–Central Electrochemical Research Institute, Karaikudi 630006, Tamil Nadu (India); Sim, Jun-Bo [Department of Nuclear and Quantum Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701 (Korea, Republic of); Choi, Sung-Min, E-mail: sungmin@kaist.ac.kr [Department of Nuclear and Quantum Engineering, Korea Advanced Institute of Science and Technology, Daejeon, 305-701 (Korea, Republic of)

2015-07-15

Highlights: • Pt NPs were in situ synthesized onto CNT–graphene support in aqueous solution. • The as-prepared material was used directly as a catalyst ink without further treatment. • Catalyst ink is active toward methanol oxidation. • This approach realizes both scalable and greener production of hybrid catalysts. - Abstract: Platinum nanoparticles were in situ synthesized onto hybrid support involving graphene and single walled carbon nanotube in aqueous solution. We investigate the reduction of graphene oxide, and platinum nanoparticle functionalization on hybrid support by X-ray photoelectron spectroscopy, Raman spectroscopy, X-ray diffraction, scanning electron microscopy and transmission electron microscopy. The as-prepared platinum on hybrid support was used directly as a catalyst ink without further treatment and is active toward methanol oxidation. This work realizes both scalable and greener production of highly efficient hybrid catalysts, and would be valuable for practical applications of graphene based fuel cell catalysts.

Localization of introduced genes on the chromosomes of transgenic barley, wheat and triticale by fluorescence in situ hybridization

DEFF Research Database (Denmark)

Pedersen, C.; Zimny, J.; Becker, D.

1997-01-01

Using fluorescence in situ hybridization (FISH) we localized introduced genes on metaphase chromosomes of barley, wheat, and triticale transformed by microprojectile bombardment of microspores and scutellar tissue with the pDB1 plasmid containing the uidA and bar genes. Thirteen integration sites...... of single-copy integrations. There was a slight tendency towards the localization of transgenes in distal chromosome regions. Using the GAA-satellite sequence for chromosome banding, the chromosomes containing the inserted genes were identified in most cases. Two barley lines derived from the same...... transformant showed a totally different integration pattern. Southern analysis confirmed that the inserted genes were segregating independently, resulting in different integration patterns among the progeny lines. The application of the FISH technique for the analysis of transgenic plants is discussed....

Y-chromosome status identification suggests a recipient origin of posttransplant non-small cell lung carcinomas: chromogenic in situ hybridization analysis.

Science.gov (United States)

Chen, Wei; Brodsky, Sergey V; Zhao, Weiqiang; Otterson, Gregory A; Villalona-Calero, Miguel; Satoskar, Anjali A; Hasan, Ayesha; Pelletier, Ronald; Ivanov, Iouri; Ross, Patrick; Nadasdy, Tibor; Shilo, Konstantin

2014-05-01

Owing to the need of lifelong immunosuppression, solid-organ transplant recipients are known to have an increased risk of posttransplant malignancies including lung cancer. Posttransplant neoplastic transformation of donor-derived cells giving rise to hematopoietic malignancies, Kaposi sarcoma, and basal cell carcinoma in nongraft tissues has been reported. The goal of this study was to assess the cell origin (donor versus recipient derived) of posttransplant non-small cell lung carcinomas (NSCLCs) in kidney and heart transplant recipients. An institutional database search identified 2557 kidney and heart transplant recipients in 8 consecutive years. Among this cohort, 20 (0.8%) renal and 18 (0.7%) heart transplant recipients developed NSCLC. The study cohort comprised 6 of 38 NSCLCs arising in donor-recipient sex-mismatched transplant patients. The tumor cell origin was evaluated by chromogenic in situ hybridization with Y-chromosome probe on formalin-fixed, paraffin-embedded tissues. Y-chromosome was identified in 97% ± 1% (range from 92% to 99%) of all types of nucleated cells in male control tissues. In all 5 NSCLCs from male recipients of female donor organ, Y-chromosome was identified in 97% ± 2% (range from 92% to 100%) of tumor cells, statistically equivalent to normal control (P recipient of male kidney. These findings suggest a recipient derivation of NSCLC arising in kidney and heart transplant recipients. A combination of histologic evaluation and chromogenic in situ hybridization with Y-chromosome analysis allows reliable determination of tissue origin in sex-mismatched solid-organ transplant recipients and may aid in management of posttransplant malignancy in such cases. Copyright © 2014 Elsevier Inc. All rights reserved.

Frequency of reciprocal translocations and dicentrics induced in human blood lymphocytes by X-irradiation as determined by fluorescence in situ hybridization

International Nuclear Information System (INIS)

Nakano, M.; Nakashima, E.; Pawel, D.; Kodama, Y.; Awa, A.

1993-01-01

This study was designed to test the scoring efficiency of reciprocal translocations and dicentrics induced by X-irradiation in vitro using the fluorescence in situ hybridization (FISH) technique. An excess was found in the frequencies of reciprocal translocations relative to those of dicentrics by measurement with FISH at the first cell division after irradiation (translocation:dicentric ≅ 60:40). However, when the same metaphases were also evaluated sequentially by a conventional staining method, the ratio of about 50:50 was restored. This was due in part to misclassification of certain dicentrics as reciprocal translocations by the FISH technique. (author)

In Situ Production of Graphene-Fiber Hybrid Structures

DEFF Research Database (Denmark)

Akia, Mandana; Cremar, Lee; Chipara, Mircea

2017-01-01

We report a scalable method to obtain a new material where large graphene sheets form webs linking carbon fibers. Film-fiber hybrid nonwoven mats are formed during fiber processing and converted to carbon structures after a simple thermal treatment. This contrasts with multistep methods...... that attempt to mix previously prepared graphene and fibers, or require complicated and costly processes for deposition of graphene over carbon fibers. The developed graphene-fiber hybrid structures have seamless connections between graphene and fibers, and in fact the graphene "veils" extend directly from one...... a capillarity effect that promoted the formation of thin veils, which become graphene sheets upon dehydration by sulfuric acid vapor followed by carbonization (at relatively low temperatures, below 800 °C). These veils extend over several micrometers within the pores of the fiber network, and consist...

Mapping 3D genome architecture through in situ DNase Hi-C.

Science.gov (United States)

Ramani, Vijay; Cusanovich, Darren A; Hause, Ronald J; Ma, Wenxiu; Qiu, Ruolan; Deng, Xinxian; Blau, C Anthony; Disteche, Christine M; Noble, William S; Shendure, Jay; Duan, Zhijun

2016-11-01

With the advent of massively parallel sequencing, considerable work has gone into adapting chromosome conformation capture (3C) techniques to study chromosomal architecture at a genome-wide scale. We recently demonstrated that the inactive murine X chromosome adopts a bipartite structure using a novel 3C protocol, termed in situ DNase Hi-C. Like traditional Hi-C protocols, in situ DNase Hi-C requires that chromatin be chemically cross-linked, digested, end-repaired, and proximity-ligated with a biotinylated bridge adaptor. The resulting ligation products are optionally sheared, affinity-purified via streptavidin bead immobilization, and subjected to traditional next-generation library preparation for Illumina paired-end sequencing. Importantly, in situ DNase Hi-C obviates the dependence on a restriction enzyme to digest chromatin, instead relying on the endonuclease DNase I. Libraries generated by in situ DNase Hi-C have a higher effective resolution than traditional Hi-C libraries, which makes them valuable in cases in which high sequencing depth is allowed for, or when hybrid capture technologies are expected to be used. The protocol described here, which involves ∼4 d of bench work, is optimized for the study of mammalian cells, but it can be broadly applicable to any cell or tissue of interest, given experimental parameter optimization.

[Chromosomal localization of foreign genes in transgenic mice using dual-color fluorescence in situ hybridization].

Science.gov (United States)

Lin, Dan; Gong, Xiu-li; Li, Wei; Guo, Xin-bing; Zhu, Yi-wen; Huang, Ying

2008-02-01

To establish a highly sensitive and specific dual-color fluorescence in situ hybridization (D-FISH) method used for chromosomal localization of foreign genes in double transgenic mice. Two strains of double transgenic mice were used in this experiment, one was integrated with the herpes simplex virus thymidine kinase (HSV-tk) and the enhanced green fluorescence protein (eGFP), the other was with the short hairpin RNA interference(RNAi) and beta(654). Splenic cells cultured in vitro were arrested in metaphase by colchicine and hybridized with digoxigenin-labeled and biotinylated DNA probes, then detected by rhodamine-conjugated avidin and FITC-conjugated anti-digoxigenin. Dual-color fluorescence signals were detected on the same metaphase in both transgenic mice strains. In HSV-tk/eGFP double transgenic mice, strong green fluorescence for HSV-tk and red for eGFP were observed and localized at 2E5-G3 and 8A2-A4 respectively. In beta(654)/RNAi mice, beta(654) was detected as red fluorescence on chromosome 7D3-E2, and RNAi showed random integration on chromosomes. It was detected as green fluorescence on chromosome 12B1 in one mouse, while on 1E2.3-1F and 3A3 in the other. Highly sensitive and specific D-FISH method was established using the self-prepared DNA probes, and chromosomal localization of the foreign genes was also performed in combination with G-banding in double transgenic mice. This technology will facilitate the researches in transgenic animals and gene therapy models.

In Situ Integration of Anisotropic SnO₂ Heterostructures inside Three-Dimensional Graphene Aerogel for Enhanced Lithium Storage.

Science.gov (United States)

Yao, Xin; Guo, Guilue; Ma, Xing; Zhao, Yang; Ang, Chung Yen; Luo, Zhong; Nguyen, Kim Truc; Li, Pei-Zhou; Yan, Qingyu; Zhao, Yanli

2015-12-02

Three-dimensional (3D) graphene aerogel (GA) has emerged as an outstanding support for metal oxides to enhance the overall energy-storage performance of the resulting hybrid materials. In the current stage of the studies, metals/metal oxides inside GA are in uncrafted geometries. Introducing structure-controlled metal oxides into GA may further push electrochemical properties of metal oxide-GA hybrids. Using rutile SnO2 as an example, we demonstrated here a facile hydrothermal strategy combined with a preconditioning technique named vacuum-assisted impregnation for in situ construction of controlled anisotropic SnO2 heterostructures inside GA. The obtained hybrid material was fully characterized in detail, and its formation mechanism was investigated by monitoring the phase-transformation process. Rational integration of the two advanced structures, anisotropic SnO2 and 3D GA, synergistically led to enhanced lithium-storage properties (1176 mAh/g for the first cycle and 872 mAh/g for the 50th cycle at 100 mA/g) as compared with its two counterparts, namely, rough nanoparticles@3D GA and anisotropic SnO2@2D graphene sheets (618 and 751 mAh/g for the 50th cycle at 100 mA/g, respectively). It was also well-demonstrated that this hybrid material was capable of delivering high specific capacity at rapid charge/discharge cycles (1044 mAh/g at 100 mA/g, 847 mAh/g at 200 mA/g, 698 mAh/g at 500 mA/g, and 584 mAh/g at 1000 mA/g). The in situ integration strategy along with vacuum-assisted impregnation technique presented here shows great potential as a versatile tool for accessing a variety of sophisticated smart structures in the form of anisotropic metals/metal oxides within 3D GA toward useful applications.

Intrinsic bacterial biodegradation of petroleum contamination demonstrated in situ using natural abundance, molecular-level 14C analysis

International Nuclear Information System (INIS)

Slater, G.F.; Nelson, R.K.; Kile, B.M.; Reddy, C.M.

2006-01-01

Natural abundance, molecular-level C 14 analysis was combined with comprehensive gas chromatography (GC x GC) to investigate, in situ, the role of intrinsic biodegradation in the loss of petroleum hydrocarbons from the rocky, inter-tidal zone impacted by the Bouchard 120 oil spill. GC x GC analysis indicated accelerated losses of n-alkane components of the residual petroleum hydrocarbons between day 40 and day 50 after the spill. 14 C analysis of bacterial phospholipid fatty acids (PLFA) from the impacted zone on day 44 showed that the polyunsaturated fatty acids attributed to the photoautotrophic component of the microbial community had the same ( 14 C as the local dissolved inorganic carbon (DIG), indicating that this DIG was their carbon source. In contrast there was significant (C depletion in the saturated and mono-unsaturated PLFA indicating incorporation of petroleum carbon. This correlation between the observed accelerated n-alkane losses and microbial incorporation of (C-depleted carbon directly demonstrated, in situ, that intrinsic biodegradation was affecting the petroleum. Since the majority of organic contaminants originate from petroleum feed-stocks, in situ molecular-level 14 C analysis of microbial PLFA can provide insights into the occurrence and pathways of biodegradation of a wide range of organic contaminants. (Author)

Demonstration, testing, ampersand evaluation of in situ heating of soil. Draft final report, Volume II: Appendices A to E

International Nuclear Information System (INIS)

Dev, H.; Enk, J.; Jones, D.; Saboto, W.

1996-01-01

This document is a draft final report for US DOE contract entitled, open-quotes Demonstration Testing and Evaluation of In Situ Soil Heating,close quotes Contract No. DE-AC05-93OR22160, IITRI Project No. C06787. This report is presented in two volumes. Volume I contains the technical report This document is Volume II, containing appendices with background information and data. In this project approximately 300 cu. yd. of clayey soil containing a low concentration plume of volatile organic chemicals was heated in situ by the application of electrical energy. It was shown that as a result of heating the effective permeability of soil to air flow was increased such that in situ soil vapor extraction could be performed. The initial permeability of soil was so low that the soil gas flow rate was immeasurably small even at high vacuum levels. When scaled up, this process can be used for the environmental clean up and restoration of DOE sites contaminated with VOCs and other organic chemicals boiling up to 120 degrees to 130 degrees C in the vadose zone. Although it may applied to many types of soil formations, it is particularly attractive for low permeability clayey soil where conventional in situ venting techniques are limited by low air flow

Multi-objective optimization of in-situ bioremediation of groundwater using a hybrid metaheuristic technique based on differential evolution, genetic algorithms and simulated annealing

Directory of Open Access Journals (Sweden)

Kumar Deepak

2015-12-01

Full Text Available Groundwater contamination due to leakage of gasoline is one of the several causes which affect the groundwater environment by polluting it. In the past few years, In-situ bioremediation has attracted researchers because of its ability to remediate the contaminant at its site with low cost of remediation. This paper proposed the use of a new hybrid algorithm to optimize a multi-objective function which includes the cost of remediation as the first objective and residual contaminant at the end of the remediation period as the second objective. The hybrid algorithm was formed by combining the methods of Differential Evolution, Genetic Algorithms and Simulated Annealing. Support Vector Machines (SVM was used as a virtual simulator for biodegradation of contaminants in the groundwater flow. The results obtained from the hybrid algorithm were compared with Differential Evolution (DE, Non Dominated Sorting Genetic Algorithm (NSGA II and Simulated Annealing (SA. It was found that the proposed hybrid algorithm was capable of providing the best solution. Fuzzy logic was used to find the best compromising solution and finally a pumping rate strategy for groundwater remediation was presented for the best compromising solution. The results show that the cost incurred for the best compromising solution is intermediate between the highest and lowest cost incurred for other non-dominated solutions.

Expanding probe repertoire and improving reproducibility in human genomic hybridization

Science.gov (United States)

Dorman, Stephanie N.; Shirley, Ben C.; Knoll, Joan H. M.; Rogan, Peter K.

2013-01-01

Diagnostic DNA hybridization relies on probes composed of single copy (sc) genomic sequences. Sc sequences in probe design ensure high specificity and avoid cross-hybridization to other regions of the genome, which could lead to ambiguous results that are difficult to interpret. We examine how the distribution and composition of repetitive sequences in the genome affects sc probe performance. A divide and conquer algorithm was implemented to design sc probes. With this approach, sc probes can include divergent repetitive elements, which hybridize to unique genomic targets under higher stringency experimental conditions. Genome-wide custom probe sets were created for fluorescent in situ hybridization (FISH) and microarray genomic hybridization. The scFISH probes were developed for detection of copy number changes within small tumour suppressor genes and oncogenes. The microarrays demonstrated increased reproducibility by eliminating cross-hybridization to repetitive sequences adjacent to probe targets. The genome-wide microarrays exhibited lower median coefficients of variation (17.8%) for two HapMap family trios. The coefficients of variations of commercial probes within 300 nt of a repetitive element were 48.3% higher than the nearest custom probe. Furthermore, the custom microarray called a chromosome 15q11.2q13 deletion more consistently. This method for sc probe design increases probe coverage for FISH and lowers variability in genomic microarrays. PMID:23376933

Morphological spot counting from stacked images for automated analysis of gene copy numbers by fluorescence in situ hybridization.

Science.gov (United States)

Grigoryan, Artyom M; Dougherty, Edward R; Kononen, Juha; Bubendorf, Lukas; Hostetter, Galen; Kallioniemi, Olli

2002-01-01

Fluorescence in situ hybridization (FISH) is a molecular diagnostic technique in which a fluorescent labeled probe hybridizes to a target nucleotide sequence of deoxyribose nucleic acid. Upon excitation, each chromosome containing the target sequence produces a fluorescent signal (spot). Because fluorescent spot counting is tedious and often subjective, automated digital algorithms to count spots are desirable. New technology provides a stack of images on multiple focal planes throughout a tissue sample. Multiple-focal-plane imaging helps overcome the biases and imprecision inherent in single-focal-plane methods. This paper proposes an algorithm for global spot counting in stacked three-dimensional slice FISH images without the necessity of nuclei segmentation. It is designed to work in complex backgrounds, when there are agglomerated nuclei, and in the presence of illumination gradients. It is based on the morphological top-hat transform, which locates intensity spikes on irregular backgrounds. After finding signals in the slice images, the algorithm groups these together to form three-dimensional spots. Filters are employed to separate legitimate spots from fluorescent noise. The algorithm is set in a comprehensive toolbox that provides visualization and analytic facilities. It includes simulation software that allows examination of algorithm performance for various image and algorithm parameter settings, including signal size, signal density, and the number of slices.

The application of fluorescence in situ hybridization (FISH technique for studying the microbial communities in intestinal tissues of white shrimp (Penaeus vannamei

Directory of Open Access Journals (Sweden)

Supamattaya, K.

2005-02-01

Full Text Available Fluorescence in situ hybridization technique is very useful for the evaluation of microbial communities in various environments. It is possible to apply this technique to study the intestinal microflora in white shrimp (Penaeus vannamei. Different fixatives and storage temperature were tested in this technique. It was found that fixation with 10% buffered formalin for 12 hours and changed to 70% ethanol shown positive results when compared to the fixation with Davidson's fixative or RF fixative. The best signaling was obtainedfrom the samples which were stored in -20ºC. By using the DNA probe targeted to the Eubacteria domain (EUB338 probe, 5′-GCT GCC TCC CGT AGG AGT-3′ labeled with fluorescein as a hybridizing probe, it was found that most intestinal microflora were aggregated with the intestinal contents, or dispersed in the lumen. There was not evidence of the attachment of the microflora with the intestinal epithelium in this study.

Characterisation of the NA62 GigaTracker End of Column Demonstrator Hybrid Pixel Detector

Science.gov (United States)

Noy, M.; Aglieri Rinella, G.; Cotta Ramusino, A.; Fiorini, M.; Jarron, P.; Kaplon, J.; Kluge, A.; Martin, E.; Morel, M.; Perktold, L.; Poltorak, K.; Riedler, P.

2011-11-01

The architecture and characterisation of the NA62 GigaTracker End of Column Demonstrator Hybrid Pixel Detector (HPD) are presented. This detector must perform time stamping to 200 ps (RMS) or better, provide 300 μm pitch position information and operate with a dead time of 1% or less for 800 MHz-1 GHz beam rate. The demonstrator HPD Assembly comprises a readout chip with a test column of 45 pixels, alongside other test structures, bump bonded to a p-in-n detector 200 μm in thickness. Validation of the performance of the HPD and the time-over-threshold timewalk compensation mechanism with both beam particles and a high precision laser system was performed and is presented. Confirmation of better than the required time stamping precision has been demonstrated and subsequent work on the design of the full-scale ASIC, dubbed TDCPix, is underway. An overview of the TDCPix architecure is given.

Demonstration, testing, and evaluation of in situ heating of soil. Final report, Volume 2, Appendices A to E

International Nuclear Information System (INIS)

Dev, H.; Enk, J.; Jones, D.; Sabato, W.

1996-01-01

This is a final report presented in two volumes. Volume I contains the technical report and Volume II contains appendices with background information and data. In this project approximately 300 cubic yards of clayey soil containing a low concentration plume of volatile organic chemicals was heated in situ by the application of electrical energy. It was shown that as a result of heating the effective permeability of soil to air flow was increased such that in situ soil vapor extraction could be performed. The initial permeability of soil was so low that the soil gas flow rate was immeasurably small even at high vacuum levels. It was demonstrated that the mass flow rate of the volatile organic chemicals was enhanced in the recovered soil gas as a result of heating. When scaled up, this process can be used for the environmental clean up and restoration of DOE sites contaminated with VOC's and other organic chemicals. Although it may be applied to many types of soil formations, it is particularly attractive for low permeability clayey soil where conventional in situ venting techniques are limited by air flow

Demonstration, testing, and evaluation of in situ heating of soil. Final report, Volume 2, Appendices A to E

Energy Technology Data Exchange (ETDEWEB)

Dev, H.; Enk, J.; Jones, D.; Sabato, W.

1996-04-05

This is a final report presented in two volumes. Volume I contains the technical report and Volume II contains appendices with background information and data. In this project approximately 300 cubic yards of clayey soil containing a low concentration plume of volatile organic chemicals was heated in situ by the application of electrical energy. It was shown that as a result of heating the effective permeability of soil to air flow was increased such that in situ soil vapor extraction could be performed. The initial permeability of soil was so low that the soil gas flow rate was immeasurably small even at high vacuum levels. It was demonstrated that the mass flow rate of the volatile organic chemicals was enhanced in the recovered soil gas as a result of heating. When scaled up, this process can be used for the environmental clean up and restoration of DOE sites contaminated with VOC`s and other organic chemicals. Although it may be applied to many types of soil formations, it is particularly attractive for low permeability clayey soil where conventional in situ venting techniques are limited by air flow.

Innovative technology demonstrations

International Nuclear Information System (INIS)

Anderson, D.B.; Luttrell, S.P.; Hartley, J.N.

1992-08-01

Environmental Management Operations (EMO) is conducting an Innovative Technology Demonstration Program for Tinker Air Force Base (TAFB). Several innovative technologies are being demonstrated to address specific problems associated with remediating two contaminated test sites at the base. Cone penetrometer testing (CPT) is a form of testing that can rapidly characterize a site. This technology was selected to evaluate its applicability in the tight clay soils and consolidated sandstone sediments found at TAFB. Directionally drilled horizontal wells was selected as a method that may be effective in accessing contamination beneath Building 3001 without disrupting the mission of the building, and in enhancing the extraction of contamination both in ground water and in soil. A soil gas extraction (SGE) demonstration, also known as soil vapor extraction, will evaluate the effectiveness of SGE in remediating fuels and TCE contamination contained in the tight clay soil formations surrounding the abandoned underground fuel storage vault located at the SW Tanks Site. In situ sensors have recently received much acclaim as a technology that can be effective in remediating hazardous waste sites. Sensors can be useful for determining real-time, in situ contaminant concentrations during the remediation process for performance monitoring and in providing feedback for controlling the remediation process. Following the SGE demonstration, the SGE system and SW Tanks test site will be modified to demonstrate bioremediation as an effective means of degrading the remaining contaminants in situ. The bioremediation demonstration will evaluate a bioventing process in which the naturally occurring consortium of soil bacteria will be stimulated to aerobically degrade soil contaminants, including fuel and TCE, in situ

Superfund Innovative Technology Evaluation - Demonstration Bulletin: In-Situ Soil Stabilization

Science.gov (United States)

In-situ stabilization technology immobilizes organics and inorganic compounds in wet or dry soils by using reagents (additives) to polymerize with the soils and sludges producing a cement-like mass. Two basic components of this technology are the Geo-Con/DSM Deep Soil Mixing Sy...

Examination of the early infection stages of koi herpesvirus (KHV) in experimentally infected carp, Cyprinus carpio L. using in situ hybridization.

Science.gov (United States)

Monaghan, S J; Thompson, K D; Adams, A; Kempter, J; Bergmann, S M

2015-05-01

Koi herpesvirus (KHV) causes a highly infectious disease afflicting common carp and koi, Cyprinus carpio L. Various molecular and antibody-based detection methods have been used to elucidate the rapid attachment and dissemination of the virus throughout carp tissues, facilitating ongoing development of effective diagnostic approaches. In situ hybridization (ISH) was used here to determine the target tissues of KHV during very early infection, after infecting carp with a highly virulent KHV isolate. Analysis of paraffin-embedded tissues (i.e. gills, skin, spleen, kidney, gut, liver and brain) during the first 8 h and following 10 days post-infection (hpi; dpi) revealed positive signals in skin mucus, gills and gut sections after only 1 hpi. Respiratory epithelial cells were positive as early as 2 hpi. Viral DNA was also detected within blood vessels of various tissues early in the infection. Notable increases in signal abundance were observed in the gills and kidney between 5 and 10 dpi, and viral DNA was detected in all tissues except brain. This study suggests that the gills and gut play an important role in the early pathogenesis of this Alloherpesvirus, in addition to skin, and demonstrates ISH as a useful diagnostic tool for confirmation of acutely infected carp. © 2014 John Wiley & Sons Ltd.

Evaluation of fluorescent in-situ hybridization technique for diagnosis of malaria in Ahero Sub-County hospital, Kenya.

Science.gov (United States)

Kandie, Regina; Ochola, Rachel; Njaanake, Kariuki

2018-01-08

Malaria is a major cause of morbidity and mortality. Treatment of malaria in a timely manner could avert deaths. Treatment ultimately relies on the rapid and accurate diagnosis. Fluorescence in situ hybridization (FISH), a cytogenetic technique based on detection of specific nucleic acid, has the potential to address the limitations of the current diagnostic approaches. This study investigates further the performance of FISH for the diagnosis of malaria in a rural setting in Western Kenya. Blood samples from 302 patients presenting with fever (temperature ≥ 37.5 °C) were examined for malaria using the Giemsa microscopy (GM), rapid diagnostic test (RDT), polymerase chain reaction (PCR) and FISH. The sensitivity and specificity of FISH was 85.6% and 96.2% respectively, while the corresponding values for GM were 82.2% and 100% respectively. RDT and PCR had sensitivities of 91.1% and 98.9%, respectively with their specificities being 89.6 and 100%, respectively. The positive predictive values for RDT, GM, FISH and PCR were 78.8%, 100%, 90.6% and 100%, respectively. The negative predictive values for RDT, GM, FISH and PCR were 96.0%, 93.0%, 94.0% and 99.5%, respectively. Their respective diagnostic accuracies were 90.1%, 94.7% 93.0% and 99.7%. The present study demonstrates that the specificity and reproducibility of FISH assays are high, thus adding to the growing evidence on the potential of the technique as an effective tool for the detection of malaria parasites in remote settings.

Radiation induced wheat-rye chromosomal translocations in triticale. Optimizing the dose using fluorescence in situ hybridization

International Nuclear Information System (INIS)

Ahmad, F.; Comeau, A.; Chen, Q.; Collin, J.; St-Pierre, C.A.

2000-01-01

Fluorescent in situ hybridization (FISH) was utilized to monitor the level of ionizing radiation ( 60 Co source) in their ability to cause intra- and intergeneric chromosomal aberrations in triticale seeds. Seeds were irradiated with 0, 20, 50, 100, 200, 300, 400, 500 and 1000 Gy doses. The root growth of irradiated seeds was greatly inhibited at 200 Gy and above. Various types of aberrations including wheat-rye, wheat-wheat, rye-rye, wheat-rye-wheat, rye-wheat-rye translocations and acentric fragments with or without translocations were observed. There was a consistent increase in proportion of aberrations per cell with an increase in radiation dose. It was concluded that for an optimal level of chromosomal translocation and least number of acentric fragments, a 20 Gy dose was quite sufficient for inducing a desirable level of wheat-rye chromosomal translocations. The excellent efficiency and importance of utilizing FISH in such studies of alien-introgression via chromosomal translocations are discussed. (author)

Radiation induced wheat-rye chromosomal translocations in triticale. Optimizing the dose using fluorescence in situ hybridization

Energy Technology Data Exchange (ETDEWEB)

Ahmad, F. [Brandon Univ., Manitoba (Canada); Comeau, A.; Chen, Q.; Collin, J.; St-Pierre, C.A.

2000-03-01

Fluorescent in situ hybridization (FISH) was utilized to monitor the level of ionizing radiation ({sup 60}Co source) in their ability to cause intra- and intergeneric chromosomal aberrations in triticale seeds. Seeds were irradiated with 0, 20, 50, 100, 200, 300, 400, 500 and 1000 Gy doses. The root growth of irradiated seeds was greatly inhibited at 200 Gy and above. Various types of aberrations including wheat-rye, wheat-wheat, rye-rye, wheat-rye-wheat, rye-wheat-rye translocations and acentric fragments with or without translocations were observed. There was a consistent increase in proportion of aberrations per cell with an increase in radiation dose. It was concluded that for an optimal level of chromosomal translocation and least number of acentric fragments, a 20 Gy dose was quite sufficient for inducing a desirable level of wheat-rye chromosomal translocations. The excellent efficiency and importance of utilizing FISH in such studies of alien-introgression via chromosomal translocations are discussed. (author)

Demonstration of Brachyspira aalborgi lineages 2 and 3 in human colonic biopsies with intestinal spirochaetosis by specific fluorescent in situ hybridization

DEFF Research Database (Denmark)

Jensen, Tim Kåre; Teglbjærg, Peter S.; Lindboe, Christian F.

2004-01-01

of these organisms in human intestinal spirochaetosis. Seventeen human colonic biopsies from Norway and Denmark with intestinal spirochaetosis caused by Brachyspira-like organisms different from the type strain of B. aalborgi (lineage 1) were examined. Application of the probe gave a positive signal in two Norwegian...... biopsies, whereas the 15 other biopsies were hybridization-negative. The positive reaction visualized the spirochaetes as a fluorescent, 3-5 mum-high fringe on the surface epithelium, extending into the crypts. The study verified the presence of B. aalborgi lineages 2 and 3 and identified the bacteria...

Chromogenic in situ hybridization (CISH): a novel alternative in screening archival breast cancer tissue samples for HER-2/neu status.

Science.gov (United States)

Madrid, Manuelito A; Lo, Raymundo W

2004-01-01

Chromogenic in situ hybridization (CISH) is emerging as a practical, cost-effective, and valid alternative to fluorescent in situ hybridization in testing for gene alteration, especially in centers primarily working with immunohistochemistry (IHC). We assessed Her-2/neu alteration using CISH on formalin-fixed paraffin-embedded primary invasive ductal carcinoma tumors in which IHC (CB11 antibody) had previously been performed, and we compared the results with IHC. The 160 selected cases were equally stratified randomly into the four IHC categories (scores of 0, 1+, 2+, and 3+). We also compared age at diagnosis and tumor histologic grade with IHC and CISH Her-2/neu. We were able to perform and evaluate CISH successfully on all cases. The agreement between 3+ IHC and CISH-amplified cases as well as between all IHC and CISH Her-2/neu negative cases was 100%, and the concordance on all positive cases was 72.50%, with an overall agreement of 86.25%. All the discordant cases had 2+ IHC scores. Although we noted Her-2/neu positivity more in premenopausal women, the age at diagnosis was not significantly associated with IHC or CISH results. Similarly, although the small group of well-differentiated tumors was apparently Her-2/neu negative in both tests, no significant association was noted between any tumor histologic grade and either IHC or CISH results. CISH is easily integrated into routine testing in our laboratory. It is a necessary adjunct in determining the subset of non-amplified IHC-positive invasive tumors that will not benefit from trastuzumab therapy. Those cases with 2+ IHC results will be triaged and subjected to CISH. Her-2/neu testing should be done on all breast cancer cases regardless of age at presentation and tumor histologic grade.

Chromogenic in situ hybridization (CISH): a novel alternative in screening archival breast cancer tissue samples for HER-2/neu status

International Nuclear Information System (INIS)

Madrid, Manuelito A; Lo, Raymundo W

2004-01-01

Chromogenic in situ hybridization (CISH) is emerging as a practical, cost-effective, and valid alternative to fluorescent in situ hybridization in testing for gene alteration, especially in centers primarily working with immunohistochemistry (IHC). We assessed Her-2/neu alteration using CISH on formalin-fixed paraffin-embedded primary invasive ductal carcinoma tumors in which IHC (CB11 antibody) had previously been performed, and we compared the results with IHC. The 160 selected cases were equally stratified randomly into the four IHC categories (scores of 0, 1+, 2+, and 3+). We also compared age at diagnosis and tumor histologic grade with IHC and CISH Her-2/neu. We were able to perform and evaluate CISH successfully on all cases. The agreement between 3+ IHC and CISH-amplified cases as well as between all IHC and CISH Her-2/neu negative cases was 100%, and the concordance on all positive cases was 72.50%, with an overall agreement of 86.25%. All the discordant cases had 2+ IHC scores. Although we noted Her-2/neu positivity more in premenopausal women, the age at diagnosis was not significantly associated with IHC or CISH results. Similarly, although the small group of well-differentiated tumors was apparently Her-2/neu negative in both tests, no significant association was noted between any tumor histologic grade and either IHC or CISH results. CISH is easily integrated into routine testing in our laboratory. It is a necessary adjunct in determining the subset of non-amplified IHC-positive invasive tumors that will not benefit from trastuzumab therapy. Those cases with 2+ IHC results will be triaged and subjected to CISH. Her-2/neu testing should be done on all breast cancer cases regardless of age at presentation and tumor histologic grade

Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

International Nuclear Information System (INIS)

Rosa, Fabíola E; Silveira, Sara M; Silveira, Cássia GT; Bérgamo, Nádia A; Neto, Francisco A Moraes; Domingues, Maria AC; Soares, Fernando A; Caldeira, José RF; Rogatto, Silvia R

2009-01-01

HER-2 gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). These procedures permit correlation between HER-2 expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic in situ hybridization) is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas. To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH. The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+ immunostaining cases, which showed high-level amplification (15.4%) and HER-2 transcript overexpression (20%). Moreover, 2+ immunostaining cases presented nonamplified status (50%) by CISH and HER-2 downexpression (38.5%) by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between HER-2 downexpression and the involvement of less than four lymph nodes (P = 0.0350). Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the HER-2 gene

Comparative mapping of DNA probes derived from the V{sub k} immunoglobulin gene regions on human and great ape chromosomes by fluorescence in situ hybridization

Energy Technology Data Exchange (ETDEWEB)

Arnold, N.; Wienberg, J.; Ermert, K. [Universitaet Muenchen (Germany)] [and others

1995-03-01

Fluorescence in situ hybridization (FISH) of cosmid clones of human V{sub K} gene regions to human and primate chromosomes contributed to the dating of chromosome reorganizations in evolution. A clone from the K locus at 2p11-p12 (cos 106) hybridized to the assumed homologous chromosome bands in the chimpanzees Pan troglodytes (PTR) and P. paniscus (PPA), the Gorilla gorilla (GGO), and the orangutan Pongo Pygmaeus (PPY). Human and both chimpanzees differed from gorilla and orangutan by the mapping of cos 170, a clone derived from chromosome 2cen-q11.2; the transposition of this orphon to the other side of the centromere can, therefore, be dated after the human/chimpanzee and gorilla divergence. Hybridization to homologous bands was also found with a cosmid clone containing a V{sub K}I orphon located on chromosome 1 (cos 115, main signal at 1q31-q32), although the probe is not fully unique. Also, a clone derived from the orphon V{sub K} region on chromosome 22q11 (cos 121) hybridized to the homologous bands in the great apes. This indicates that the orphons on human chromosomes 1 and 22 had been translocated early in primate evolution. 18 refs., 2 figs.

Scoping and sensitivity analyses for the Demonstration Tokamak Hybrid Reactor (DTHR)

International Nuclear Information System (INIS)

Sink, D.A.; Gibson, G.

1979-03-01

The results of an extensive set of parametric studies are presented which provide analytical data of the effects of various tokamak parameters on the performance and cost of the DTHR (Demonstration Tokamak Hybrid Reactor). The studies were centered on a point design which is described in detail. Variations in the device size, neutron wall loading, and plasma aspect ratio are presented, and the effects on direct hardware costs, fissile fuel production (breeding), fusion power production, electrical power consumption, and thermal power production are shown graphically. The studies considered both ignition and beam-driven operations of DTHR and yielded results based on two empirical scaling laws presently used in reactor studies. Sensitivity studies were also made for variations in the following key parameters: the plasma elongation, the minor radius, the TF coil peak field, the neutral beam injection power, and the Z/sub eff/ of the plasma

Interphase fluorescent in situ hybridization deletion analysis of the 9p21 region and prognosis in childhood acute lymphoblastic leukaemia (ALL)

DEFF Research Database (Denmark)

Kuchinskaya, Ekaterina; Heyman, Mats; Nordgren, Ann

2011-01-01

Interphase fluorescent in situ hybridization (FISH) was applied on diagnostic BM smears from 519 children with acute lymphoblastic leukaemia (ALL) in order to establish the frequency and prognostic importance of 9p21 deletion in children enrolled in the Nordic Society of Paediatric Haematology...... and Oncology (NOPHO) - 2000 treatment protocol. Among the patients, 452 were diagnosed with B-cell precursor (BCP)-ALL and 66 with T-ALL. A higher incidence of 9p21 deletions was found in T-ALL (38%) compared to BCP-ALL (15·7%). Homozygous deletions were found in 19·7% of T-ALL and 4·0% of BCP-ALL; hemizygous...

Monosomy 3 by chromogenic in situ hybridization (CISH) in Iranian patients with uveal melanoma.

Science.gov (United States)

Naseripour, Masood; Mehrazma, Mitra; Pourmatin, Rama; Kashkouli, Mohsen Bahmani; Sedaghat, Ahad; Gheytanchi, Elmira

2015-01-01

The aim of this study was to investigate the rate of monosomy 3 by CISH technique in Iranian patients with uveal melanoma (UM) and its correlation with clinical and histopathological features. Archival formalin fixed, paraffin-embedded material from 50 patients who had undergone enucleation for large uveal melanomas was obtained. Monosomy of chromosome 3 alteration by chromogenic in situ hybridization (CISH) was investigated. Clinical and histopathological features of tumors were collected. The patients had a mean age of 56.6±7.6 years. Mean basal diameter and thickness of tumors were 14.1 mm and 10.2 mm, respectively. Four patients (8%) were identified to harbor monosomy of chromosome 3. In the mean follow-up of 5.3 years (range, 3.2-9.5 y), only one case with monosomy 3 died of UM metastasis. The most common type of cellularity was mixed cell (86%). There was not any statistically significant correlation between monosomy of chromosome 3 and type of cellularity, ciliary body involvement, and largest basal diameter. The low rate of monosomy chromosome 3 and the consequent low rate of mortality may be indicative of good prognosis in Iranian patients with uveal melanoma.

Genetic Relationships among Hylocereus and Selenicereus Vine Cacti (Cactaceae): Evidence from Hybridization and Cytological Studies

Science.gov (United States)

TEL-ZUR, NOEMI; ABBO, SHAHAL; BAR-ZVI, DUDY; MIZRAHI, YOSEF

2004-01-01

• Background and Aims Hylocereus and Selenicereus are native to tropical and sub-tropical America. Based on its taxonomic status and crossability relations it was postulated that H. megalanthus (syn. S. megalanthus) is an allotetraploid (2n = 4x = 44) derived from natural hybridization between two closely related diploid taxa. The present work aimed at elucidating the genetic relationships between species of the two genera. • Methods Crosses were performed and the putative hybrids were analysed by chromosome counts and morphological traits. The ploidy level of hybrids was confirmed by fluorescent in situ hybridization (FISH) of rDNA sites. Genomic in situ hybridization (GISH) was used in an attempt to identify the putative diploid genome donors of H. megalanthus and an artificial interploid hybrid. • Key Results Reciprocal crosses among four diploid Hylocereus species (H. costaricensis, H. monacanthus (syn. H. polyrhizus), H. undatus and Hylocereus sp.) yielded viable diploid hybrids, with regular chromosome pairing. Reciprocal crosses between these Hylocereus spp. and H. megalanthus yielded viable triploid, pentaploid, hexaploid and aneuploid hybrids. Morphological and phenological traits confirm the hybrid origin. In situ detection of rDNA sites was in accord with the ploidy status of the species and hybrid studied. GISH results indicated that overall sequence composition of H. megalanthus is similar to that of H. ocamponis and S. grandiflorus. High sequence similarity was also found between the parental genomes of H. monacanthus and H. megalanthus in one triploid hybrid. • Conclusions The ease of obtaining partially fertile F1 hybrids and the relative sequence similarity (in GISH study) suggest close genetic relationships among the taxa analysed. PMID:15329334

In Situ Formation of Carbon Nanomaterials on Bulk Metallic Materials

Directory of Open Access Journals (Sweden)

J. Y. Xu

2014-01-01

Full Text Available Carbon nanomaterials were synthesized in situ on bulk 316L stainless steel, pure cobalt, and pure nickel by hybrid surface mechanical attrition treatment (SMAT. The microstructures of the treated samples and the resulted carbon nanomaterials were investigated by SEM and TEM characterizations. Different substrates resulted in different morphologies of products. The diameter of carbon nanomaterials is related to the size of the nanograins on the surface layer of substrates. The possible growth mechanism was discussed. Effects of the main parameters of the synthesis, including the carbon source and gas reactant composition, hydrogen, and the reaction temperature, were studied. Using hybrid SMAT is proved to be an effective way to synthesize carbon nanomaterials in situ on surfaces of metallic materials.

Application of fluorescence in situ hybridization technique in the diagnosis of acute promyelocytic leukemia with abnormal immunophenotype

International Nuclear Information System (INIS)

Chen Leilei; Sun Xuemei; Chen Junhao; Zhang Le

2005-01-01

To evaluate the utilization of fluorescence in situ hybridization (FISH) technique in the diagnosis of acute promyelocytic leukemia(APL) with abnormal immunophenotype, flow cytometry was used to detect the immunophenotype of mononuclear cells in APL patients and PML/RARα fusion gene was detected by FISH. The mononuclear cells of several APL patients showed abnormal immunophenotype: CD13 + , CD33 + , CD34 - , HLA-DR + and PML/RARα fusion gene was also detected, which was different from the regular result of APL: HLA- DR - , PML/RARα + . Therefore, the detection of immunophenotype in APL patients should not be regarded as the sole accurate target for diagnosing leukemia. FISH ,associated with traditional FAB classification, is a simple, rapid, accurate and direct method. It can be used to help confirm the diagnosis, to guide the formulation of a reasonable chemotherapy scheme and to supervise the efficacy of the treatment in patients with leukemia. (authors)

SUPERSENSITIVE IN SITU HYBRIDIZATION BY TYRAMIDE SIGNAL AMPLIFICATION AND NANOGOLD SILVER STAINING: THE CONTRIBUTION OF AUTOMETALLOGRAPHY AND CATALYZED REPORTER DEPOSITION TO THE REJUVENATION OF IN SITU HYBRIDIZATION.

Energy Technology Data Exchange (ETDEWEB)

TUBBS,R.R.PETTAY,J.GROGAN,T.CHEUNG,A.L.M.POWELL,R.D.HAINFELD,J.HAUSER-KRONBERGER,C.HACKER,G.W.

2002-04-17

It is peculiar that in situ hybridization (ISH), a technique with many similarities to immunohistochemistry (IHC), has not enjoyed the phenomenal growth in both basic research and clinical applications as has its sister technique IHC. Since the late 1970s, when immunoperoxidase techniques began to be applied to routine diagnostic material and to numerous research applications, there has been a natural evolution of the IHC procedure. Namely, only a few primary antibodies were available commercially at the onset, and only one indirect and the peroxidase-antiperoxidase (PAP) technique detection systems were in place. With the advent of avidin-biotin detection systems and monoclonal antibodies, and a viable commercial market, extraordinary growth of the procedure's applications in clinical research and diagnostic pathology occurred during the subsequent two decades. Today, IHC is automated and widely used for research purposes and, to a large extent, has become a routine diagnostic ''special stain'' in most clinical laboratories. During the same period, ISH enjoyed very little growth in both research and diagnostic applications. What has accounted for this lack of maturation of the technique? The success of IHC is part of the reason measuring a gene's encoded protein routinely and inexpensively, particularly as automation evolved, rendered IHC a more viable choice in many instances. Inherent comparative sensitivity of the procedures has also clearly been a factor. Unfortunately, the chromogenic procedures in place are often insufficiently sensitive to detect the relatively low amounts of DNA and RNA levels at which the clinical utility is to be found.

In Situ Synthesis of Reduced Graphene Oxide and Gold Nanocomposites for Nanoelectronics and Biosensing

Directory of Open Access Journals (Sweden)

Chen Peng

2011-01-01

Full Text Available Abstract In this study, an in situ chemical synthesis approach has been developed to prepare graphene–Au nanocomposites from chemically reduced graphene oxide (rGO in aqueous media. UV–Vis absorption, atomic force microscopy, scanning electron microscopy, transmission electron microscopy, and Raman spectroscopy were used to demonstrate the successful attachment of Au nanoparticles to graphene sheets. Configured as field-effect transistors (FETs, the as-synthesized single-layered rGO-Au nanocomposites exhibit higher hole mobility and conductance when compared to the rGO sheets, promising its applications in nanoelectronics. Furthermore, we demonstrate that the rGO-Au FETs are able to label-freely detect DNA hybridization with high sensitivity, indicating its potentials in nanoelectronic biosensing.

Comparative cytogenetic characterization of primary canine melanocytic lesions using array CGH and fluorescence in situ hybridization.

Science.gov (United States)

Poorman, Kelsey; Borst, Luke; Moroff, Scott; Roy, Siddharth; Labelle, Philippe; Motsinger-Reif, Alison; Breen, Matthew

2015-06-01

Melanocytic lesions originating from the oral mucosa or cutaneous epithelium are common in the general dog population, with up to 100,000 diagnoses each year in the USA. Oral melanoma is the most frequent canine neoplasm of the oral cavity, exhibiting a highly aggressive course. Cutaneous melanocytomas occur frequently, but rarely develop into a malignant form. Despite the differential prognosis, it has been assumed that subtypes of melanocytic lesions represent the same disease. To address the relative paucity of information about their genomic status, molecular cytogenetic analysis was performed on the three recognized subtypes of canine melanocytic lesions. Using array comparative genomic hybridization (aCGH) analysis, highly aberrant distinct copy number status across the tumor genome for both of the malignant melanoma subtypes was revealed. The most frequent aberrations included gain of dog chromosome (CFA) 13 and 17 and loss of CFA 22. Melanocytomas possessed fewer genome wide aberrations, yet showed a recurrent gain of CFA 20q15.3-17. A distinctive copy number profile, evident only in oral melanomas, displayed a sigmoidal pattern of copy number loss followed immediately by a gain, around CFA 30q14. Moreover, when assessed by fluorescence in situ hybridization (FISH), copy number aberrations of targeted genes, such as gain of c-MYC (80 % of cases) and loss of CDKN2A (68 % of cases), were observed. This study suggests that in concordance with what is known for human melanomas, canine melanomas of the oral mucosa and cutaneous epithelium are discrete and initiated by different molecular pathways.

Three-dimensional graphene-polypyrrole hybrid electrochemical actuator

Science.gov (United States)

Liu, Jia; Wang, Zhi; Zhao, Yang; Cheng, Huhu; Hu, Chuangang; Jiang, Lan; Qu, Liangti

2012-11-01

The advancement of mechanical actuators benefits from the development of new structural materials with prominent properties. A novel three-dimensional (3D) hydrothermally converted graphene and polypyrrole (G-PPy) hybrid electrochemical actuator is presented, which is prepared via a convenient hydrothermal process, followed by in situ electropolymerization of pyrrole. The 3D pore-interconnected G-PPy pillar exhibits strong actuation responses superior to pure graphene and PPy film. In response to the low potentials of +/-0.8 V, the saturated strain of 3D G-PPy pillar can reach a record of 2.5%, which is more than 10 times higher than that of carbon nanotube film and about 3 times that of unitary graphene film under an applied potential of +/-1.2 V. Also, the 3D G-PPy actuator exhibits high actuation durability with high operating load as demonstrated by an 11 day continuous measurement. Finally, a proof-of-concept application of 3D G-PPy as smart filler for on/off switch is also demonstrated, which indicates the great potential of the 3D G-PPy structure developed in this study for advanced actuator systems.The advancement of mechanical actuators benefits from the development of new structural materials with prominent properties. A novel three-dimensional (3D) hydrothermally converted graphene and polypyrrole (G-PPy) hybrid electrochemical actuator is presented, which is prepared via a convenient hydrothermal process, followed by in situ electropolymerization of pyrrole. The 3D pore-interconnected G-PPy pillar exhibits strong actuation responses superior to pure graphene and PPy film. In response to the low potentials of +/-0.8 V, the saturated strain of 3D G-PPy pillar can reach a record of 2.5%, which is more than 10 times higher than that of carbon nanotube film and about 3 times that of unitary graphene film under an applied potential of +/-1.2 V. Also, the 3D G-PPy actuator exhibits high actuation durability with high operating load as demonstrated by an 11 day

Seven-hour fluorescence in situ hybridization technique for enumeration of Enterobacteriaceae in food and environmental water sample.

Science.gov (United States)

Ootsubo, M; Shimizu, T; Tanaka, R; Sawabe, T; Tajima, K; Ezura, Y

2003-01-01

A fluorescent in situ hybridization (FISH) technique using an Enterobacteriaceae-specific probe (probe D) to target 16S rRNA was improved in order to enumerate, within a single working day, Enterobacteriaceae present in food and environmental water samples. In order to minimize the time required for the FISH procedure, each step of FISH with probe D was re-evaluated using cultured Escherichia coli. Five minutes of ethanol treatment for cell fixation and hybridization were sufficient to visualize cultured E. coli, and FISH could be performed within 1 h. Because of the difficulties in detecting low levels of bacterial cells by FISH without cultivation, a FISH technique for detecting microcolonies on membrane filters was investigated to improve the bacterial detection limit. FISH with probe D following 6 h of cultivation to grow microcolonies on a 13 mm diameter membrane filter was performed, and whole Enterobacteriaceae microcolonies on the filter were then detected and enumerated by manual epifluorescence microscopic scanning at magnification of x100 in ca 5 min. The total time for FISH with probe D following cultivation (FISHFC) was reduced to within 7 h. FISHFC can be applied to enumerate cultivable Enterobacteriaceae in food (above 100 cells g-1) and environmental water samples (above 1 cell ml-1). Cultivable Enterobacteriaceae in food and water samples were enumerated accurately within 7 h using the FISHFC method. A FISHFC method capable of evaluating Enterobacteriaceae contamination in food and environmental water within a single working day was developed.

Synergistic Enhancement of Microwave Absorption Using Hybridized Polyaniline@helical CNTs with Dual Chirality.

Science.gov (United States)

Tian, Xin; Meng, Fanbin; Meng, Fanchen; Chen, Xiangnan; Guo, Yifan; Wang, Ying; Zhu, Wenjun; Zhou, Zuowan

2017-05-10

In this study, we designed a dual-chirality hierarchical structure to achieve a synergistically enhanced effect in microwave absorption via the hybridization of nanomaterials. Herein, polyaniline (PANi) nanorods with tunable chirality are grown on helical carbon nanotubes (HCNTs), a typical nanoscale chiral structure, through in situ polymerization. The experimental results show that the hierarchical hybrids (PANi@HCNTs) exhibit distinctly dual chirality and a significant enhancement in electromagnetic (EM) losses compared to those of either pure PANi or HCNTs. The maximum reflection loss of the as-prepared hybrids can reach -32.5 dB at 8.9 GHz. Further analysis demonstrates that combinations of chiral acid-doped PANi and coiled HCNTs with molecular and nanoscale chirality lead to synergistic effects resulting from the dual chirality. The so-called cross-polarization may result in additional interactions with induced EM waves in addition to multiscaled relaxations from functional groups and interfacial polarizations, which can benefit EM absorption.

High-Temperature Tensile and Tribological Behavior of Hybrid (ZrB2+Al3Zr)/AA5052 In Situ Composite

Science.gov (United States)

Gautam, G.; Kumar, N.; Mohan, A.; Gautam, R. K.; Mohan, S.

2016-09-01

During service life, components such as piston, cylinder blocks, brakes, and discs/drums, have to work under high-temperature conditions. In order to have appropriate material for such applications high-temperature studies are important. Hybrid (ZrB2+Al3Zr)/AA5052 in situ composite has been investigated from ambient to 523 K (250 °C) at an interval of 50 deg. (ZrB2+Al3Zr)/AA5052 in situ composite has been fabricated by the direct melt reaction of AA5052 alloy with zirconium and boron salts. Microstructure studies show refinement in the grain size of base alloy on in situ formation of reinforcement particles. Al3Zr particles are observed in rectangular and polyhedron shapes. It is observed from the tensile studies that ultimate tensile strength, yield strength, and percentage elongation decrease with increase in test temperature. Similar kind of behavior is also observed for flow curve properties. The tensile results have also been correlated with fractography. Wear and friction results indicate that the wear rate increases with increase in normal load, whereas coefficient of friction shows decreasing trend. With increasing test temperature, wear rate exhibits a typical phenomenon. After an initial increase, wear rate follows a decreasing trend up to 423 K (150 °C), and finally a rapid increase is observed, whereas coefficient of friction increases continuously with increase in test temperature. The mechanisms responsible for the variation of wear and friction with different temperatures have been discussed in detail with the help of worn surfaces studies under scanning electron microscope (SEM) & 3D-profilometer and debris analysis by XRD.

Polyaniline-coated halloysite nanotubes via in-situ chemical polymerization

Energy Technology Data Exchange (ETDEWEB)

Zhang Long [State Key Laboratory of Applied Organic Chemistry and Institute of Polymer Science and Engineering, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000 (China); Wang Tingmei [State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000 (China); Liu Peng [State Key Laboratory of Applied Organic Chemistry and Institute of Polymer Science and Engineering, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou 730000 (China)], E-mail: pliu@lzu.edu.cn

2008-12-30

Polyaniline coated halloysite nanotubes (PANI/HNTs) were prepared by the in-situ soapless emulsion polymerization of the anilinium chloride adsorbed halloysite nanotubes (HNTs), obtained by the dispersion of HNTs in acidic aqueous solution of aniline with magnetic stirring and ultrasonic irradiation, by using ammonium persulfate (APS) as oxidant. The effect of the acidities of the polymerizing media on the crystal structure of the nanotubes was investigated with X-ray diffraction (XRD) technique. The surface conducting coatings of the hybrids were characterized with X-ray photoelectron spectroscopy (XPS). The morphological analyses showed that the polyaniline coated halloysite nanotubes via the in-situ chemical oxidation polymerization with ultrasonic irradiation had the better well-defined structures, by the transmission electron microscopy (TEM). The conductivities of the PANI/HNTs hybrids increased with the increasing of the amounts of HCl dopant added in the emulsion polymerization.

Polyaniline-coated halloysite nanotubes via in-situ chemical polymerization

International Nuclear Information System (INIS)

Zhang Long; Wang Tingmei; Liu Peng

2008-01-01

Polyaniline coated halloysite nanotubes (PANI/HNTs) were prepared by the in-situ soapless emulsion polymerization of the anilinium chloride adsorbed halloysite nanotubes (HNTs), obtained by the dispersion of HNTs in acidic aqueous solution of aniline with magnetic stirring and ultrasonic irradiation, by using ammonium persulfate (APS) as oxidant. The effect of the acidities of the polymerizing media on the crystal structure of the nanotubes was investigated with X-ray diffraction (XRD) technique. The surface conducting coatings of the hybrids were characterized with X-ray photoelectron spectroscopy (XPS). The morphological analyses showed that the polyaniline coated halloysite nanotubes via the in-situ chemical oxidation polymerization with ultrasonic irradiation had the better well-defined structures, by the transmission electron microscopy (TEM). The conductivities of the PANI/HNTs hybrids increased with the increasing of the amounts of HCl dopant added in the emulsion polymerization.

Locked Nucleic Acid-Based In Situ Hybridization Reveals miR-7a as a Hypothalamus-Enriched MicroRNA with a Distinct Expression Pattern

DEFF Research Database (Denmark)

Herzer, S; Silahtaroglu, A; Meister, B

2012-01-01

, a part of the brain that controls vital bodily functions, we employed locked nucleic acid (LNA) - fluorescent in situ hybridization (FISH). The expression pattern of the mature miRNAs miR-7a, miR-7b, miR-137 and miR-153 in mouse brain tissue sections was investigated. Whereas all studied miRNAs were......R-7a expression was particularly prominent in the subfornical organ, suprachiasmatic, paraventricular, periventricular, supraoptic, dorsomedial and arcuate nuclei. Identical expression patterns for miR-7a was seen in mouse and rat hypothalamus. By combining LNA-FISH with immunohistochemistry...

Experimental demonstration of synergy between electron cyclotron and lower hybrid current drive on Tore Supra

International Nuclear Information System (INIS)

Artaud, J.F.; Giruzzi, G.; Dumont, R.J.; Imbeaux, F.; Bibet, P.; Bouquey, F.; Clary, J.; Ekedahl, A.; Hoang, G.T.; Lennholm, M.; Magne, R.; Segui, J.L.

2004-01-01

Non-inductive current drive (CD) has two main applications in tokamaks: sustainment of a substantial fraction of the toroidal plasma current necessary for the plasma confinement and control of the plasma stability and transport properties by appropriate shaping of the current density profile. For the first kind of applications, lower hybrid (LH) waves are known to provide the highest efficiency (defined as the ratio of the driven current to the injected wave power), although with limited control capability. Conversely, electron cyclotron (EC) waves drive highly localized currents, and are therefore particularly suited for control purposes, but their CD efficiency is much lower than that of LH waves (typically, an order of magnitude in present day experiments). Various calculations have demonstrated an interesting property: the current driven by the simultaneous use of the two waves, I(LH+EC), can be significantly larger than the sum I(LH)+I(EC) of the currents separately driven by the two waves in the same plasma conditions. This property, called synergy effect. The peculiar experimental conditions attainable on the Tore Supra tokamak have allowed the first experimental demonstration of the synergy between EC and LH current drive. The significant improvement of the electron cyclotron current drive (ECCD) efficiency in the presence of low hybrid current drive (LHCD), predicted by kinetic theory and confirmed by stationary experiments on Tore Supra, opens up the possibility of using ECCD as an efficient current profile control tool in LHCD plasmas

In situ permeable flow sensors at the Savannah River Integrated Demonstration: Phase 2 results

International Nuclear Information System (INIS)

Ballard, S.

1994-08-01

A suite of In Situ Permeable Flow Sensors was deployed at the site of the Savannah River Integrated Demonstration to monitor the interaction between the groundwater flow regime and air injected into the saturated subsurface through a horizontal well. One of the goals of the experiment was to determine if a groundwater circulation system was induced by the air injection process. The data suggest that no such circulation system was established, perhaps due to the heterogeneous nature of the sediments through which the injected gas has to travel. The steady state and transient groundwater flow patterns observed suggest that the injected air followed high permeability pathways from the injection well to the water table. The preferential pathways through the essentially horizontal impermeable layers appear to have been created by drilling activities at the site

Augmented In Situ Subsurface Bioremediation Process™BIO-REM, Inc. - Demonstration Bulletin

Science.gov (United States)

The Augmented In Situ Subsurface Bioremediation Process™ developed by BIO-REM, Inc., uses microaerophilic bacteria and micronutrients (H-10) and surface tension depressants/penetrants for the treatment of hydrocarbon contaminated soils and groundwater. The bacteria utilize hydroc...

Rapid detection of chromosome rearrangement in medical diagnostic X-ray workers by using fluorescence in situ hybridization and study on dose estimation

International Nuclear Information System (INIS)

Wang Zhiquan; Sun Yuanming; Li Jin

1998-01-01

Objective: Biological doses were estimated for medical diagnostic X-ray workers. Methods: Chromosome rearrangements in X-ray workers were analysed by fluorescence in situ hybridization (FISH) with composite whole chromosome paintings number 4 and number 7. Results: The frequency of translocation in medical diagnostic X-ray workers was much higher than that in control group (P<0.01). The biological doses to individual X-ray workers were calculated by their translocation frequency. The translocation frequencies of both FISH and G-banding were in good agreement. Conclusion: The biological doses to X-ray workers are estimated by FISH first when their dosimetry records are not documented

Demonstration of a collimated in situ method for determining depth distributions using gamma-ray spectrometry

CERN Document Server

Benke, R R

2002-01-01

In situ gamma-ray spectrometry uses a portable detector to quantify radionuclides in materials. The main shortcoming of in situ gamma-ray spectrometry has been its inability to determine radionuclide depth distributions. Novel collimator designs were paired with a commercial in situ gamma-ray spectrometry system to overcome this limitation for large area sources. Positioned with their axes normal to the material surface, the cylindrically symmetric collimators limited the detection of un attenuated gamma-rays from a selected range of polar angles (measured off the detector axis). Although this approach does not alleviate the need for some knowledge of the gamma-ray attenuation characteristics of the materials being measured, the collimation method presented in this paper represents an absolute method that determines the depth distribution as a histogram, while other in situ methods require a priori knowledge of the depth distribution shape. Other advantages over previous in situ methods are that this method d...

Human cDNA mapping using fluorescence in situ hybridization. Progress report, April 1, 1992--December 31, 1992

Energy Technology Data Exchange (ETDEWEB)

Korenberg, J.R.

1993-03-04

Genetic mapping is approached using the techniques of high resolution fluorescence in situ hybridization (FISH). This technology and the results of its application are designed to rapidly generate whole genome as tool box of expressed sequence to speed the identification of human disease genes. The results of this study are intended to dovetail with and to link the results of existing technologies for creating backbone YAC and genetic maps. In the first eight months, this approach generated 60--80% of the expressed sequence map, the remainder expected to be derived through more long-term, labor-intensive, regional chromosomal gene searches or sequencing. The laboratory has made significant progress in the set-up phase, in mapping fetal and adult brain and other cDNAs, in testing a model system for directly linking genetic and physical maps using FISH with small fragments, in setting up a database, and in establishing the validity and throughput of the system.

Investigating the Inverse Square Law with the Timepix Hybrid Silicon Pixel Detector: A CERN [at] School Demonstration Experiment

Science.gov (United States)

Whyntie, T.; Parker, B.

2013-01-01

The Timepix hybrid silicon pixel detector has been used to investigate the inverse square law of radiation from a point source as a demonstration of the CERN [at] school detector kit capabilities. The experiment described uses a Timepix detector to detect the gamma rays emitted by an [superscript 241]Am radioactive source at a number of different…

Oriented Arrays of Graphene in a Polymer Matrix by in situ Reduction of Graphite Oxide Nanosheets

KAUST Repository

Ansari, Seema; Kelarakis, Antonios; Estevez, Luis; Giannelis, Emmanuel P.

2010-01-01

Graphite oxide-Nafion hybrids with a high degree of alignment are cast from aqueous solution in the absence of any external field and reduced in situ by exposure to hydrazine to produce graphene-Nafion hybrids. Dramatic enhancement of electrical

Distribution of herpes simplex virus types 1 and 2 genomes in human spinal ganglia studied by PCR and in situ hybridization.

Science.gov (United States)

Obara, Y; Furuta, Y; Takasu, T; Suzuki, S; Suzuki, H; Matsukawa, S; Fujioka, Y; Takahashi, H; Kurata, T; Nagashima, K

1997-06-01

Clinical data indicate that the recurring herpes simplex virus (HSV) from oro-labial lesions is HSV subtype 1 and that the virus from genital lesions is HSV-2. This suggests that HSV-1 and HSV-2 reside in latent forms in the trigeminal ganglia and sacral ganglia, respectively. However, the distribution of latent HSV-1 and HSV-2 infections in human spinal ganglia has not been fully examined. This report concerns the application of polymerase chain reaction (PCR) and in situ hybridization (ISH) to such a study. By using PCR and employing the respective primers, HSV-1 and HSV-2 DNAs were detected in 207 of 524 samples from 262 spinal ganglia (from the cervical to the sacral ganglia) examined on both sides. The percentages of HSV-1 and HSV-2 detected in a given set of ganglia were similar, indicating an absence of site preference. By ISH, few but positive hybridization signals were detected evenly in sacral ganglia sections. The data suggest that regional specificity of recurrent HSV infections is not due to regional distribution of latent virus, but that local host factors may be important for recurrences.

Automated processing of fluorescence in-situ hybridization slides for HER2 testing in breast and gastro-esophageal carcinomas.

Science.gov (United States)

Tafe, Laura J; Allen, Samantha F; Steinmetz, Heather B; Dokus, Betty A; Cook, Leanne J; Marotti, Jonathan D; Tsongalis, Gregory J

2014-08-01

HER2 fluorescence in-situ hybridization (FISH) is used in breast and gastro-esophageal carcinoma for determining HER2 gene amplification and patients' eligibility for HER2 targeted therapeutics. Traditional manual processing of the FISH slides is labor intensive because of multiple steps that require hands on manipulation of the slides and specifically timed intervals between steps. This highly manual processing also introduces inter-run and inter-operator variability that may affect the quality of the FISH result. Therefore, we sought to incorporate an automated processing instrument into our FISH workflow. Twenty-six cases including breast (20) and gastro-esophageal (6) cancer comprising 23 biopsies and three excision specimens were tested for HER2 FISH (Pathvysion, Abbott) using the Thermobrite Elite (TBE) system (Leica). Up to 12 slides can be run simultaneously. All cases were previously tested by the Pathvysion HER2 FISH assay with manual preparation. Twenty cells were counted by two observers for each case; five cases were tested on three separate runs by different operators to evaluate the precision and inter-operator variability. There was 100% concordance in the scoring between the manual and TBE methods as well as among the five cases that were tested on three runs. Only one case failed due to poor probe hybridization. In total, seven cases were positive for HER2 amplification (HER2:CEP17 ratio >2.2) and the remaining 19 were negative (HER2:CEP17 ratio <1.8) utilizing the 2007 ASCO/CAP scoring criteria. Due to the automated denaturation and hybridization, for each run, there was a reduction in labor of 3.5h which could then be dedicated to other lab functions. The TBE is a walk away pre- and post-hybridization system that automates FISH slide processing, improves work flow and consistency and saves approximately 3.5h of technologist time. The instrument has a small footprint thus occupying minimal counter space. TBE processed slides performed

Human cDNA mapping using fluorescence in situ hybridization. Progress report, April 1--December 31, 1992

Energy Technology Data Exchange (ETDEWEB)

Korenberg, J.R.

1993-12-31

The ultimate goal of this proposal is to create a cDNA map of the human genome. Mapping is approached using the techniques of high resolution fluorescence in situ hybridization (FISH). This technology and the results of its application are designed to rapidly generate whole genome as tool box of expressed sequence to speed the identification of human disease genes. The results of this study are intended to dovetail with and to link the results of existing technologies for creating backbone YAC and genetic maps. In the first eight months, this approach will generate 60--80% of the expressed sequence map, the remainder expected to be derived through more long-term, labor-intensive, regional chromosomal gene searches or sequencing. The laboratory has made significant progress in the set-up phase, in mapping fetal and adult brain and other cDNAs, in testing a model system for directly linking genetic and physical maps using FISH with small fragments, in setting up a database, and in establishing the validity and throughput of the system.

Analysis of methanogenic activity in a thermophilic-dry anaerobic reactor: Use of fluorescent in situ hybridization

International Nuclear Information System (INIS)

Montero, B.; Garcia-Morales, J.L.; Sales, D.; Solera, R.

2009-01-01

Methanogenic activity in a thermophilic-dry anaerobic reactor was determined by comparing the amount of methane generated for each of the organic loading rates with the size of the total and specific methanogenic population, as determined by fluorescent in situ hybridization. A high correlation was evident between the total methanogenic activity and retention time [-0.6988Ln(x) + 2.667] (R 2 0.8866). The total methanogenic activity increased from 0.04 x 10 -8 mLCH 4 cell -1 day -1 to 0.38 x 10 -8 mLCH 4 cell -1 day -1 while the retention time decreased, augmenting the organic loading rates. The specific methanogenic activities of H 2 -utilizing methanogens and acetate-utilizing methanogens increased until they stabilised at 0.64 x 10 -8 mLCH 4 cell -1 day -1 and 0.33 x 10 -8 mLCH 4 cell -1 day -1 , respectively. The methanogenic activity of H 2 -utilizing methanogens was higher than acetate-utilizing methanogens, indicating that maintaining a low partial pressure of hydrogen does not inhibit the acetoclastic methanogenesis or the anaerobic process

Fluorescence in situ hybridization in combination with the comet assay and micronucleus test in genetic toxicology

Directory of Open Access Journals (Sweden)

Hovhannisyan Galina G

2010-09-01

Full Text Available Abstract Comet assay and micronucleus (MN test are widely applied in genotoxicity testing and biomonitoring. While comet assay permits to measure direct DNA-strand breaking capacity of a tested agent MN test allows estimating the induced amount of chromosome and/or genome mutations. The potential of these two methods can be enhanced by the combination with fluorescence in situ hybridization (FISH techniques. FISH plus comet assay allows the recognition of targets of DNA damage and repairing directly. FISH combined with MN test is able to characterize the occurrence of different chromosomes in MN and to identify potential chromosomal targets of mutagenic substances. Thus, combination of FISH with the comet assay or MN test proved to be promising techniques for evaluation of the distribution of DNA and chromosome damage in the entire genome of individual cells. FISH technique also permits to study comet and MN formation, necessary for correct application of these methods. This paper reviews the relevant literature on advantages and limitations of Comet-FISH and MN-FISH assays application in genetic toxicology.

Modeling the effects of forest management on in situ and ex situ longleaf pine forest carbon stocks

Science.gov (United States)

C.A. Gonzalez-Benecke; L.J. Samuelson; T.A. Martin; W.P. Cropper Jr; Kurt Johnsen; T.A. Stokes; John Butnor; P.H. Anderson

2015-01-01

Assessment of forest carbon storage dynamics requires a variety of techniques including simulation models. We developed a hybrid model to assess the effects of silvicultural management systems on carbon (C) budgets in longleaf pine (Pinus palustris Mill.) plantations in the southeastern U.S. To simulate in situ C pools, the model integrates a growth and yield model...

Final Report for the Demonstration of Plasma In-situ Vitrification at the 904-65G K-Reactor Seepage Basin

Energy Technology Data Exchange (ETDEWEB)

Blundy, R.F. [Westinghouse Savannah River Company, AIKEN, SC (United States); Zionkowki, P.G.

1997-12-22

The In-situ Vitrification (ISV) process potentially offers the most stable waste-form for containment of radiologically contaminated soils while minimizing personnel contamination. This is a problem that is extensive, and at the same time unique, to the US Department of Energy`s (DOE) Weapons Complex. An earlier ISV process utilized joule heating of the soil to generate the subsurface molten glass product. However previous test work has indicated that the Savannah river Site soils (SRS) may not be entirely suitable for vitrification by joule heating due to their highly refractory nature. The concept of utilizing a plasma torch for soil remediation by in-situ vitrification has recently been developed, and laboratory test work on a 100 kW unit has indicated a potentially successful application with SRS soils. The Environmental Restoration Division (ERD) of Westinghouse Savannah River Company (WSRC) conducted the first field scale demonstration of this process at the (904-65G) K-Reactor Seepage Basin in October 1996 with the intention of determining the applicability and economics of the process for remediation of a SRS radioactive seepage basin. The demonstration was successful in completing three vitrification runs, including two consecutive runs that fused together adjacent columns of glass to form a continuous monolith. This report describes the demonstration, documents the engineering data that was obtained, summarizes the process economics and makes recommendations for future development of the process and equipment.

Final Report for the Demonstration of Plasma In-situ Vitrification at the 904-65G K-Reactor Seepage Basin

International Nuclear Information System (INIS)

Blundy, R.F.; Zionkowki, P.G.

1997-01-01

The In-situ Vitrification (ISV) process potentially offers the most stable waste-form for containment of radiologically contaminated soils while minimizing personnel contamination. This is a problem that is extensive, and at the same time unique, to the US Department of Energy's (DOE) Weapons Complex. An earlier ISV process utilized joule heating of the soil to generate the subsurface molten glass product. However previous test work has indicated that the Savannah river Site soils (SRS) may not be entirely suitable for vitrification by joule heating due to their highly refractory nature. The concept of utilizing a plasma torch for soil remediation by in-situ vitrification has recently been developed, and laboratory test work on a 100 kW unit has indicated a potentially successful application with SRS soils. The Environmental Restoration Division (ERD) of Westinghouse Savannah River Company (WSRC) conducted the first field scale demonstration of this process at the (904-65G) K-Reactor Seepage Basin in October 1996 with the intention of determining the applicability and economics of the process for remediation of a SRS radioactive seepage basin. The demonstration was successful in completing three vitrification runs, including two consecutive runs that fused together adjacent columns of glass to form a continuous monolith. This report describes the demonstration, documents the engineering data that was obtained, summarizes the process economics and makes recommendations for future development of the process and equipment

Mechanisms of induction of chromosomal aberrations and their detection by fluorescence in situ hybridization

International Nuclear Information System (INIS)

Natarajan, A.T.

2002-01-01

Recently introduced fluorescence in situ hybridization (FISH) technique employing chromosome specific DNA libraries as well as region specific DNA probes (e.g., centromere, telomere) have helped to analyse chromosomal aberrations in great detail and thus have given some new insights into the mechanisms of induction of chromosomal aberrations. The relative proportion of induction of translocations and dicentrics by ionising radiation was studied in human, mice and Chinese hamster cells. Many of the studies point to the differences between the mechanisms of induction of dicentrics and translocations. Preliminary results obtained in our laboratory using arm specific probes for human chromosomes 1 and 3 indicate that the aberrations between the arms appear to be more than expected on a random basis. By employing telomeric probes the frequencies of interstitial deletions were found to be high and similar to the frequencies of dicentrics both in human and mouse lymphocytes. A recent study with human chromosome specific probes clearly shows variation of sensitivity of chromosomes for the induction of exchange aberrations. Radiation response studies with Chinese hamster cells using telomeric probes, suggest that telomeric sequences, especially interstitial ones appear to be an important factor in the origin of both spontaneous and induced chromosomal aberrations

Considerations in the use of fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy to characterize rumen methanogens and define their spatial distributions.

Science.gov (United States)

Valle, Edith R; Henderson, Gemma; Janssen, Peter H; Cox, Faith; Alexander, Trevor W; McAllister, Tim A

2015-06-01

In this study, methanogen-specific coenzyme F420 autofluorescence and confocal laser scanning microscopy were used to identify rumen methanogens and define their spatial distribution in free-living, biofilm-, and protozoa-associated microenvironments. Fluorescence in situ hybridization (FISH) with temperature-controlled hybridization was used in an attempt to describe methanogen diversity. A heat pretreatment (65 °C, 1 h) was found to be a noninvasive method to increase probe access to methanogen RNA targets. Despite efforts to optimize FISH, 16S rRNA methanogen-specific probes, including Arch915, bound to some cells that lacked F420, possibly identifying uncharacterized Methanomassiliicoccales or reflecting nonspecific binding to other members of the rumen bacterial community. A probe targeting RNA from the methanogenesis-specific methyl coenzyme M reductase (mcr) gene was shown to detect cultured Methanosarcina cells with signal intensities comparable to those of 16S rRNA probes. However, the probe failed to hybridize with the majority of F420-emitting rumen methanogens, possibly because of differences in cell wall permeability among methanogen species. Methanogens were shown to integrate into microbial biofilms and to exist as ecto- and endosymbionts with rumen protozoa. Characterizing rumen methanogens and defining their spatial distribution may provide insight into mitigation strategies for ruminal methanogenesis.

Field demonstration of in situ grouting of radioactive solid waste burial trenches with polyacrylamide

International Nuclear Information System (INIS)

Spalding, B.P.; Fontaine, T.A.

1990-01-01

Demonstrations of in situ grouting with polyacrylamide were carried out on two undisturbed burial trenches and one dynamically compacted burial trench in Solid Waste Storage Area (SWSA) 6 at Oak Ridge National Laboratory (ORNL). The injection of polyacrylamide was achieved quite facilely for the two undisturbed burial trenches which were filled with grout, at typical pumping rates of 95 L/min, in several batches injected over several days. The compacted burial trench, however, failed to accept grout at more than 1.9 L/min even when pressure was applied. Thus, it appears that burial trenches, stabilized by dynamic compaction, have a permeability too low to be considered groutable. The water table beneath the burial trenches did not respond to grout injections indicating a lack of hydrologic connection between fluid grout and the water table which would have been observed if the grout failed to set. Because grout set times were adjusted to less than 60 min, the lack of hydrologic connection was not surprising. Postgrouting penetration testing revealed that the stability of the burial trenches was increased from 26% to 79% that measured in the undisturbed soil surrounding the trenches. In situ permeation tests on the grouted trenches indicated a significant reduction in hydraulic conductivity of the trench contents from a mean of 2.1 x 10 -3 to 1.85 x 10 -5 cm/s. Preliminary observations indicated that grouting with polyacrylamide is an excellent method for both improved stability and hydrologic isolation of radioactive waste and its incidental hazardous constituents

Parental exposure to environmental concentrations of diuron leads to aneuploidy in embryos of the Pacific oyster, as evidenced by fluorescent in situ hybridization.

Science.gov (United States)

Barranger, Audrey; Benabdelmouna, Abdellah; Dégremont, Lionel; Burgeot, Thierry; Akcha, Farida

2015-02-01

Changes in normal chromosome numbers (i.e. aneuploidy) due to abnormal chromosome segregation may arise either spontaneously or as a result of chemical/radiation exposure, particularly during cell division. Coastal ecosystems are continuously subjected to various contaminants originating from urban, industrial and agricultural activities. Genotoxicity is common to several families of major environmental pollutants, including pesticides, which therefore represent a potential important environmental hazard for marine organisms. A previous study demonstrated the vertical transmission of DNA damage by subjecting oyster genitors to short-term exposure to the herbicide diuron at environmental concentrations during gametogenesis. In this paper, Fluorescent in situ hybridization (FISH) was used to further characterize diuron-induced DNA damage at the chromosomal level. rDNA genes (5S and 18-5.8-28S), previously mapped onto Crassostrea gigas chromosomes 4, 5 and 10, were used as probes on the interphase nuclei of embryo preparations. Our results conclusively show higher aneuploidy (hypo- or hyperdiploidy) level in embryos from diuron-exposed genitors, with damage to the three studied chromosomal regions. This study suggests that sexually developing oysters are vulnerable to diuron exposure, incurring a negative impact on reproductive success and oyster recruitment. Copyright © 2014 Elsevier B.V. All rights reserved.

Persistence of translocations detected by means of fluorescence in situ hybridization in peripheral lymphocytes of accidentally exposed radiation workers

International Nuclear Information System (INIS)

Pressl, S.; Stephan, G.

1997-01-01

The translocation frequency in lymphocytes of radiation workers accidentally exposed a number of years earlier was determined by means of fluorescence in situ hybridization. Chromosomes 2, 4 and 8 were painted, and simultaneously the centromeres. The genomic frequency of translocations is between 1.7 and 17.9 per 1000 cells. This variation is not significantly different from the level in healthy control subjects. Therefore, no radiation exposure could be detected retrospectively. On the other hand, the frequency of dicentrics in these radiation workers measured by means of fluorescence plus Giemsa staining shortly after the exposure was significantly increased, and whole body doses between 0.2 and 0.3 Gy could be calculated. Consequently, it would seem that dicentrics measured shortly after an exposure are a more sensitive indicator than translocations which are determined years later. (author)

Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

Directory of Open Access Journals (Sweden)

Soares Fernando A

2009-03-01

Full Text Available Abstract Background HER-2 gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC and fluorescence in situ hybridization (FISH. These procedures permit correlation between HER-2 expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic in situ hybridization is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas. Methods To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH. Results The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+ immunostaining cases, which showed high-level amplification (15.4% and HER-2 transcript overexpression (20%. Moreover, 2+ immunostaining cases presented nonamplified status (50% by CISH and HER-2 downexpression (38.5% by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between HER-2 downexpression and the involvement of less than four lymph nodes (P = 0.0350. Conclusion Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the HER-2 gene.

Fluorescence in situ hybridization as adjunct to cytology improves the diagnosis and directs estimation of prognosis of malignant pleural effusions

Directory of Open Access Journals (Sweden)

Han Jingquan

2012-11-01

Full Text Available Abstract Background The identification of malignant cells in effusions by conventional cytology is hampered by its limited sensitivity and specificity. The aim of this study was to investigate the value of fluorescence in situ hybridization (FISH as adjuncts to conventional cytologic examination in patients with malignant pleural effusions. Methods We conducted a retrospective cohort study of 93 inpatients with pleural effusions (72 malignant pleural effusions metastatic from 11 different organs and 21 benign over 23 months. All the patients came from Chinese northeast areas. Aspirated pleural fluid underwent cytologic examination and fluorescence in situ hybridization (FISH for aneuploidy. We used FISH in single-colour or if appropriate in dual-colour evaluation to detect chromosomal aberrations (chromosomes 7, 11, and 17 in effusion cells as markers of malignancy, to raise the diagnostic yield and identified the efficiency by diagnostic biopsy. Predominant cytogenetic anomalies and patterns of intratumor cytogenetic heterogeneity were brought in relation to overall survival rate. Results Cytology alone confirmed malignant pleural effusions in 45 of 72 patients (sensitivity 63%, whereas FISH alone positively identified 48 of 72 patients (sensitivity 67%. Both tests had high specificity in predicting benign effusions. If cytology and FISH were considered together, they exhibited 88% sensitivity and 94.5% specificity in discriminating benign and malignant effusions. Combined, the two assays were more sensitive than either test alone. Although the positive predictive value of each test was 94.5%, the negative predictive value of cytology and FISH combined was 78%, better than 47% and 44% for FISH and cytology alone, respectively. There was a significantly prolonged survival rate for patients with aneuploidy for chromosome 17. Conclusions FISH in combination with conventional cytology is a highly sensitive and specific diagnostic tool for detecting

High photoresponse of individual WS2 nanowire-nanoflake hybrid materials

Science.gov (United States)

Asres, Georgies Alene; Järvinen, Topias; Lorite, Gabriela S.; Mohl, Melinda; Pitkänen, Olli; Dombovari, Aron; Tóth, Geza; Spetz, Anita Lloyd; Vajtai, Robert; Ajayan, Pulickel M.; Lei, Sidong; Talapatra, Saikat; Kordas, Krisztian

2018-06-01

van der Waals solids have been recognized as highly photosensitive materials that compete conventional Si and compound semiconductor based devices. While 2-dimensional nanosheets of single and multiple layers and 1-dimensional nanowires of molybdenum and tungsten chalcogenides have been studied, their nanostructured derivatives with complex morphologies are not explored yet. Here, we report on the electrical and photosensitive properties of WS2 nanowire-nanoflake hybrid materials we developed lately. We probe individual hybrid nanostructured particles along the structure using focused ion beam deposited Pt contacts. Further, we use conductive atomic force microscopy to analyze electrical behavior across the nanostructure in the transverse direction. The electrical measurements are complemented by in situ laser beam illumination to explore the photoresponse of the nanohybrids in the visible optical spectrum. Photodetectors with responsivity up to ˜0.4 AW-1 are demonstrated outperforming graphene as well as most of the other transition metal dichalcogenide based devices.

Construction of g-C_3N_4/Al_2O_3 hybrids via in-situ acidification and exfoliation with enhanced photocatalytic activity

International Nuclear Information System (INIS)

Wang, Xiao-jing; Liu, Chao; Li, Xu-li; Li, Fa-tang; Li, Yu-pei; Zhao, Jun; Liu, Rui-hong

2017-01-01

Highlights: • Ultrathin g-C_3N_4/Al_2O_3 hybrids are prepared via in-situ reaction. • The structure modification role of in-situ formed HNO_3 for g-C_3N_4 is found. • The ultrathin g-C_3N_4 nanosheets are formed by the acidified melamine and Al(OH)_3. • In-situ calcination of melamine and Al(OH)_3 benefits the contact of C_3N_4 and Al_2O_3. • The activity of g-C_3N_4/Al_2O_3 is 16.6 times that of pristine g-C_3N_4 in degrading RhB. - Abstract: Homogeneous ultrathin g-C_3N_4 nanosheets/Al_2O_3 heterojunctions are synthesized using melamine and Al(NO_3)_3 via in-situ reaction and the following thermal polymerization approach. The in-situ reaction between melamine and Al(NO_3)_3 results in the existence of HNO_3-acidified melamine and Al(OH)_3 aggregates via the hydrolysis of Al(NO_3)_3. After thermal polymerization, the aggregates are converted to g-C_3N_4/Al_2O_3 composites. The thermal polymerization of acidified melamine and the support effect of aluminum hydroxide for g-C_3N_4 during the calcination process lead to highly dispersed amrophous Al_2O_3 on ultrathin g-C_3N_4 nanosheets, which is beneficial for the separation of photogenerated electron-hole pairs in the heterojunction. The degradation rate for Rhodamine B (RhB) over the most activie sample is 16.6 times than that of pristine g-C_3N_4 under visible light irradiation, which can be attributed to the high specific surface area, highly dispersion of amorphous Al_2O_3 on ultrathin g-C_3N_4 nanosheet, and the effective electrons transfer from g-C_3N_4 to the amorphous Al_2O_3.

Detection of group B streptococci in Lim broth by use of group B streptococcus peptide nucleic acid fluorescent in situ hybridization and selective and nonselective agars.

Science.gov (United States)

Montague, Naomi S; Cleary, Timothy J; Martinez, Octavio V; Procop, Gary W

2008-10-01

The sensitivity, specificity, and positive and negative predictive values for the detection of group B streptococci from Lim enrichment broth with sheep blood agar (SBA), with selective Streptococcus agar (SSA), and by a peptide nucleic acid fluorescent in situ hybridization (PNA FISH) assay were as follows: for culture on SBA, 68.4%, 100%, 100%, and 87.9%, respectively; for culture on SSA, 85.5%, 100%, 100%, and 94.1%, respectively; and for the PNA FISH assay, 97.4%, 98.3%, 96.1%, and 98.9%, respectively.

Haematoxylin and eosin staining identifies medium to large bacterial aggregates with a reliable specificity: A comparative analysis of follicular bacterial aggregates in axillary biopsies using peptide nucleic acid-fluorescence in situ hybridization and haematoxylin and eosin staining.

Science.gov (United States)

Ring, Hans Christian; Theut Riis, Peter; Bay, Lene; Kallenbach, Klaus; Bjarnsholt, Thomas; Jemec, Gregor B E

2017-10-01

Although peptide nucleic acid (PNA), fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM) are the reference tools in the study of bacterial aggregates/biofilms, it may also be rather time-consuming. This study aimed to investigate the sensitivity and specificity between bacterial aggregates identified by haematoxylin and eosin (HE) staining vs bacterial aggregates in corresponding PNA-FISH samples. Axillary biopsies were obtained in 24 healthy controls. HE-stained and PNA-FISH samples were investigated using traditional light microscopy and CLSM, respectively. The data demonstrate that HE staining identifies large bacterial aggregates (>10 μm) with a sensitivity of 0.43 and specificity of 1. The methods, however, are not equivalent as demonstrated by a McNemar's test (P=.04). Where bacterial aggregates >10 μm in diameter, HE staining may offer a rapid and practical low-cost tool to evaluate bacterial aggregates. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Localization of pig Na[sup +], K[sup +]-ATPase [alpha] and [beta] subunit genes to chromosome 4 by radioactive in situ hybridization

Energy Technology Data Exchange (ETDEWEB)

Lahbib-Mansais, Y.; Yerle, M.; Dalens, M.; Chevalet, C.; Gellin, J. (Centre de Recherches de Toulouse (France))

1993-01-01

Two genes coding for Na[sup +],K[sup +] -ATPase [alpha] and [beta] subunits are localized on pig chromosome 4, to the q1.6[yields]q2.3 and 1.3[yields]q2.1 regions, respectively, by radioactive in situ hybridization. According to nucleotide and amino acid sequence comparisons with different human isoforms of Na[sup +] ,K[sup +]-ATPase, these pig [alpha] and [beta] ATPase genes show strong homologies with human [alpha]1 and [beta] subunit ATPase genes, respectively. These results are discussed with respect to comparative mapping data of conserved genes in mammalian species. We showed that the pig cDNA probes encoding ATPase [alpha] and, [beta] genes reveal DNA polymorphism in Meishan an Large White pigs. 35 refs., 4 figs., 2 tabs.

Fluorescence in situ hybridization and qPCR to detect Merkel cell polyomavirus physical status and load in Merkel cell carcinomas.

Science.gov (United States)

Haugg, Anke M; Rennspiess, Dorit; zur Hausen, Axel; Speel, Ernst-Jan M; Cathomas, Gieri; Becker, Jürgen C; Schrama, David

2014-12-15

The Merkel cell polyomavirus (MCPyV) is detected in 80% of Merkel cell carcinomas (MCC). Clonal integration and tumor-specific mutations in the large T antigen are strong arguments that MCPyV is a human tumor virus. However, the relationship between viral presence and cancer induction remains discussed controversially. Since almost all studies on virus prevalence are based on PCR techniques, we performed MCPyV fluorescence in situ hybridization (FISH) on MCC to gain information about the quality of the viral presence on the single cell level. MCPyV-FISH was performed on tissue microarrays containing 62 formalin-fixed and paraffin-embedded tissue samples including all tumor grades of 42 patients. The hybridization patterns were correlated to the qPCR data determined on corresponding whole tissue sections. Indeed, MCPyV-FISH and qPCR data were highly correlated, i.e. 83% for FISH-positive and 93% for FISH-negative cores. Accordingly, the mean of the qPCR values of all MCPyV-positive cores differed significantly from the mean of the negative cores (p = 0.0076). Importantly, two hybridization patterns were definable in the MCPyV-FISH: a punctate pattern (85%) indicating viral integration, which correlated with a moderate viral abundance and a combination of the punctate with a diffuse pattern (15%), suggesting a possible coexistence of integrated and episomal virus which was associated with very high viral load and VP1 expression. Thus, MCPyV-FISH adds important information on the single cell level within the histomorphological context and could therefore be an important tool to further elucidate MCPyV related carcinogenesis. © 2014 UICC.

One-step route to a hybrid TiO2/TixW1−xN nanocomposite by in situ selective carbothermal nitridation

Directory of Open Access Journals (Sweden)

Zoë Schnepp, Martin J Hollamby, Masahiko Tanaka, Yoshitaka Matsushita, Yoshio Katsuya and Yoshio Sakka

2012-01-01

Full Text Available Metal oxide/nitride nanocomposites have many existing and potential applications, e.g. in energy conversion or ammonia synthesis. Here, a hybrid oxide/nitride nanocomposite (anatase/TixW1−xN was synthesized by an ammonia-free sol–gel route. Synchrotron x-ray diffraction, complemented with electron microscopy and thermogravimetric analysis, was used to study the structure, composition and mechanism of formation of the nanocomposite. The nanocomposite contained nanoparticles (<5 nm diameter of two highly intermixed phases. This was found to arise from controlled nucleation and growth of a single oxide intermediate from the gel precursor, followed by phase separation and in situ selective carbothermal nitridation. Depending on the preparation conditions, the composition varied from anatase/TixW1−xN at low W content to an isostructural mixture of Ti-rich and W-rich TixW1−xN at high W content. In situ selective carbothermal nitridation offers a facile route to the synthesis of nitride-oxide nanocomposites. This conceptually new approach is a significant advance from previous methods, which generally require ammonolysis of a pre-synthesized oxide.

Biofilm formed from organic-inorganic hybrid tri-ureasil PPO for transdermal drug delivery system

Energy Technology Data Exchange (ETDEWEB)

Molina, Eduardo F.; Jesus, Natana Aparecida; Oliveira, Pollyana Francielli; Furtado, Ricardo A.; Tavares, Denise Crispim, E-mail: eduardo.molina@unifran.edu.br [Universidade de Franca (UNIFRAN), SP (Brazil)

2016-07-01

Full text: In this work we evaluated the viability of the tri-ureasil PPO hybrid as biofilm forming for release of active substances such as lignans. The samples were characterized by X-ray diffraction (XRD) and infrared (FTIR). The swelling degree and the influence of the catalyst on time of formation of a hybrid biofilm were evaluated. The cytotoxicity of the materials were evaluated using the XTT colorimetric assay where GM07492A strain was treated with different concentrations of the hybrid. The time of film formation depends on the quantity of the catalyst used in the synthesis. By varying the catalyst quantity during the synthesis, a good flexible film can be obtained, which is easy to be coated on the skin surface and in situ formed a very thin and comfortable film with an aesthetical appearance. Moreover, the hybrid films were colorless and transparent. The toxicity/viability of all samples has also been studied using normal human cells for future applications. The hybrid matrices did not significantly reduce cell viability, demonstrating that siloxane-polyether materials were biocompatible. All the materials presenting a amorphous structure (XRD) and the characteristic bands of vibrations (FTIR) of the polymer chain do not change after incorporation of lignans. (author)

Biofilm formed from organic-inorganic hybrid tri-ureasil PPO for transdermal drug delivery system

International Nuclear Information System (INIS)

Molina, Eduardo F.; Jesus, Natana Aparecida; Oliveira, Pollyana Francielli; Furtado, Ricardo A.; Tavares, Denise Crispim

2016-01-01

Full text: In this work we evaluated the viability of the tri-ureasil PPO hybrid as biofilm forming for release of active substances such as lignans. The samples were characterized by X-ray diffraction (XRD) and infrared (FTIR). The swelling degree and the influence of the catalyst on time of formation of a hybrid biofilm were evaluated. The cytotoxicity of the materials were evaluated using the XTT colorimetric assay where GM07492A strain was treated with different concentrations of the hybrid. The time of film formation depends on the quantity of the catalyst used in the synthesis. By varying the catalyst quantity during the synthesis, a good flexible film can be obtained, which is easy to be coated on the skin surface and in situ formed a very thin and comfortable film with an aesthetical appearance. Moreover, the hybrid films were colorless and transparent. The toxicity/viability of all samples has also been studied using normal human cells for future applications. The hybrid matrices did not significantly reduce cell viability, demonstrating that siloxane-polyether materials were biocompatible. All the materials presenting a amorphous structure (XRD) and the characteristic bands of vibrations (FTIR) of the polymer chain do not change after incorporation of lignans. (author)

Detection of Group B Streptococci in Lim Broth by Use of Group B Streptococcus Peptide Nucleic Acid Fluorescent In Situ Hybridization and Selective and Nonselective Agars▿

Science.gov (United States)

Montague, Naomi S.; Cleary, Timothy J.; Martinez, Octavio V.; Procop, Gary W.

2008-01-01

The sensitivity, specificity, and positive and negative predictive values for the detection of group B streptococci from Lim enrichment broth with sheep blood agar (SBA), with selective Streptococcus agar (SSA), and by a peptide nucleic acid fluorescent in situ hybridization (PNA FISH) assay were as follows: for culture on SBA, 68.4%, 100%, 100%, and 87.9%, respectively; for culture on SSA, 85.5%, 100%, 100%, and 94.1%, respectively; and for the PNA FISH assay, 97.4%, 98.3%, 96.1%, and 98.9%, respectively. PMID:18667597

Fluorescent nanodiamond and lanthanide labelled in situ hybridization for the identification of RNA transcripts in fixed and CLARITY-cleared central nervous system tissues (Conference Presentation)

Science.gov (United States)

Parker, Lindsay M.; Staikopoulos, Vicky; Cordina, Nicole M.; Sayyadi, Nima; Hutchinson, Mark R.; Packer, Nicolle H.

2016-03-01

Despite significant advancement in the methodology used to conjugate, incorporate and visualize fluorescent molecules at the cellular and tissue levels, biomedical imaging predominantly relies on the limitations of established fluorescent molecules such as fluorescein, cyanine and AlexaFluor dyes or genetic incorporation of fluorescent proteins by viral or other means. These fluorescent dyes and conjugates are highly susceptible to photobleaching and compete with cellular autofluorescence, making biomedical imaging unreliable, difficult and time consuming in many cases. In addition, some proteins have low copy numbers and/or poor antibody recognition, further making detection and imaging difficult. We are developing better methods for imaging central nervous system neuroinflammatory markers using targeted mRNA transcripts labelled with fluorescent nanodiamonds or lanthanide chelates. These tags have increased signal and photostability and can also discriminate against tissue/cell autofluorescence. Brains and spinal cords from BALB/c mice with a chronic constriction model of neuropathic pain (neuroinflammation group) or that have undergone sham surgeries (control group) were collected. A subset of brains and spinal cords were perfused and fixed with paraformaldehyde (n=3 sham and n=3 pain groups) prior to sectioning and in situ hybridization using nanodiamond or lanthanide chelate conjugated complementary RNA probes. Another subset of brains and spinal cords from the same cohort of animals were perfused and processed for CLARITY hydrogel based clearing prior to in situ hybridization with the same probes. We will present our findings on the photostability, sensitivity and discrimination from background tissue autofluorescence of our novel RNA probes, compared to traditional fluorophore tags.

Highly photoluminescent and photostable CdSe quantum dot-nylon hybrid composites for efficient light conversion applications

Energy Technology Data Exchange (ETDEWEB)

Yuan Ying; Riehle, Frank-Stefan [Freiburg Materials Research Centre (FMF), University of Freiburg, Stefan-Meier-Str. 21, D-79104 Freiburg (Germany); Department of Microsystems Engineering (IMTEK), Georg Koehler Allee 103, University of Freiburg, D-79110 Freiburg (Germany); Nitschke, Roland [Life Imaging Center, Centre of Systems Biology, University of Freiburg Habsburgerstr. 49, D-79104 Freiburg (Germany); Centre for Biological Signalling Studies (BIOSS), University of Freiburg (Germany); Krueger, Michael, E-mail: michael.krueger@fmf.uni-freiburg.de [Freiburg Materials Research Centre (FMF), University of Freiburg, Stefan-Meier-Str. 21, D-79104 Freiburg (Germany); Department of Microsystems Engineering (IMTEK), Georg Koehler Allee 103, University of Freiburg, D-79110 Freiburg (Germany)

2012-02-15

Highlights: Black-Right-Pointing-Pointer A novel in situ synthesis approach for highly luminescent CdSe core QDs-nylon hybrid materials. Black-Right-Pointing-Pointer Potential applications for light and energy conversion are demonstrated. Black-Right-Pointing-Pointer Three dimensional structures out of this hybrid material are available. - Abstract: Highly photoluminescent hexadecylamine (HDA) capped core CdSe quantum dots (QDs) with fluorescent quantum yields (QYs) up to 60% were synthesized using a hot injection method and directly incorporated into nylon polymer. For the incorporation of crude CdSe QDs into nylon a simple reproducible and upscalable one pot approach was developed without the need of further purification steps. The photoluminescence (PL) properties of the core QDs and the resulting QD-polymer hybrid composites were investigated and compared. Red emitting hybrid materials exhibit a QY of 60% with a high potential for applications in direct light and energy conversion. The hybrid materials could be successfully utilized as LED conversion layers. By avoiding exposure to oxygen the hybrid films can be kept for a month without detecting a significant decrease in luminescence. Various three dimensional structures are easily available opening doors for further applications such as novel materials for fluorescence standard development in laser scanning microscopy (LSM).

Genetic basis to hybrid inviability is more complex than hybrid male sterility in Caenorhabditis nematodes.

Science.gov (United States)

Bundus, Joanna D; Wang, Donglin; Cutter, Asher D

2018-04-07

Hybrid male sterility often evolves before female sterility or inviability of hybrids, implying that the accumulation of divergence between separated lineages should lead hybrid male sterility to have a more polygenic basis. However, experimental evidence is mixed. Here, we use the nematodes Caenorhabditis remanei and C. latens to characterize the underlying genetic basis of asymmetric hybrid male sterility and hybrid inviability. We demonstrate that hybrid male sterility is consistent with a simple genetic basis, involving a single X-autosome incompatibility. We also show that hybrid inviability involves more genomic compartments, involving diverse nuclear-nuclear incompatibilities, a mito-nuclear incompatibility, and maternal effects. These findings demonstrate that male sensitivity to genetic perturbation may be genetically simple compared to hybrid inviability in Caenorhabditis and motivates tests of generality for the genetic architecture of hybrid incompatibility across the breadth of phylogeny.

Investigations of inorganic and hybrid inorganic-organic nanostructures

Science.gov (United States)

Kam, Kinson Chihang

This thesis focuses on the exploratory synthesis and characterization of inorganic and hybrid inorganic-organic nanomaterials. In particular, nanostructures of semiconducting nitrides and oxides, and hybrid systems of nanowire-polymer composites and framework materials, are investigated. These materials are characterized by a variety of techniques for structure, composition, morphology, surface area, optical properties, and electrical properties. In the study of inorganic nanomaterials, gallium nitride (GaN), indium oxide (In2O3), and vanadium dioxide (VO2) nanostructures were synthesized using different strategies and their physical properties were examined. GaN nanostructures were obtained from various synthetic routes. Solid-state ammonolysis of metastable gamma-Ga2O 3 nanoparticles was found to be particularly successful; they achieved high surface areas and photoluminescent study showed a blue shift in emission as a result of surface and size defects. Similarly, In2O3 nanostructures were obtained by carbon-assisted solid-state syntheses. The sub-oxidic species, which are generated via a self-catalyzed vapor-liquid-solid mechanism, resulted in 1D nanostructures including nanowires, nanotrees, and nanobouquets upon oxidation. On the other hand, hydrothermal methods were used to obtain VO2 nanorods. After post-thermal treatment, infrared spectroscopy demonstrated that these nanorods exhibit a thermochromic transition with temperature that is higher by ˜10°C compared to the parent material. The thermochromic behavior indicated a semiconductor-to-metal transition associated with a structural transformation from monoclinic to rutile. The hybrid systems, on the other hand, enabled their properties to be tunable. In nanowire-polymer composites, zinc oxide (ZnO) and silver (Ag) nanowires were synthesized and incorporated into polyaniline (PANI) and polypyrrole (PPy) via in-situ and ex-situ polymerization method. The electrical properties of these composites are

Preparation of zeolite-A/chitosan hybrid composites and their bioactivities and antimicrobial activities

International Nuclear Information System (INIS)

Yu, Liang; Gong, Jie; Zeng, Changfeng; Zhang, Lixiong

2013-01-01

Zeolite-A/chitosan hybrid composites with zeolite contents of 20–55 wt.% were prepared by in situ transformation of silica/chitosan mixtures in a sodium aluminate alkaline solution through impregnation–gelation–hydrothermal synthesis. The products were characterized by X-ray diffraction, diffuse reflectance infrared Fourier transform spectroscopy, scanning electron microscopy, thermogravimetric analysis, and mercury penetration porosimetry. Their in vitro bioactivities were examined using as-synthesized and Ca 2+ -exchanged hybrid composites in simulated body fluid (SBF) for hydroxyapatite (HAP) growth. Their antimicrobial activities for Escherichia coli (E. coli) in trypticase soy broth (TSB) were evaluated using Ag + -exchanged hybrid composites. The zeolite-A/chitosan hybrid composites could be prepared as various shapes, including cylinders, plates and thin films. They possessed macropores with pore sizes ranging from 100 to 300 μm and showed compressive mechanical strength as high as 3.2 MPa when the zeolite content was 35 wt.%. Fast growth on the Ca 2+ -exchanged hybrid composites was observed with the highest weight gain of 51.4% in 30 days. The 35 wt.% Ag + -exchanged hybrid composite showed the highest antimicrobial activity, which could reduce the 9 × 10 6 CFU mL −1 E. coli concentration to zero within 4 h of incubation time with the Ag + -exchanged hybrid composite amount of 0.4 g L −1 . The bioactivity and antimicrobial activity could be combined by ion-exchanging the composites first with Ca 2+ and then with Ag + . These zeolite-A/chitosan hybrid composites have potential applications on tissue engineering and antimicrobial food packaging. - Graphical abstract: Zeolite A/chitosan hybrid composites were prepared by in situ transformation of precursors in the chitosan matrix, which possess macroporous structures and exhibit superior bioactivity and antimicrobial activity and potential biomedical application. Highlights: • Zeolite A

Preparation of zeolite-A/chitosan hybrid composites and their bioactivities and antimicrobial activities

Energy Technology Data Exchange (ETDEWEB)

Yu, Liang; Gong, Jie [State Key Laboratory of Materials-Oriented Chemical Engineering, College of Chemistry and Chemical Engineering, Nanjing University of Technology, Nanjing 210009 (China); Zeng, Changfeng [College of Mechanic and Power Engineering, Nanjing University of Technology, Nanjing 210009 (China); Zhang, Lixiong, E-mail: lixiongzhang@yahoo.com [State Key Laboratory of Materials-Oriented Chemical Engineering, College of Chemistry and Chemical Engineering, Nanjing University of Technology, Nanjing 210009 (China)

2013-10-15

Zeolite-A/chitosan hybrid composites with zeolite contents of 20–55 wt.% were prepared by in situ transformation of silica/chitosan mixtures in a sodium aluminate alkaline solution through impregnation–gelation–hydrothermal synthesis. The products were characterized by X-ray diffraction, diffuse reflectance infrared Fourier transform spectroscopy, scanning electron microscopy, thermogravimetric analysis, and mercury penetration porosimetry. Their in vitro bioactivities were examined using as-synthesized and Ca{sup 2+}-exchanged hybrid composites in simulated body fluid (SBF) for hydroxyapatite (HAP) growth. Their antimicrobial activities for Escherichia coli (E. coli) in trypticase soy broth (TSB) were evaluated using Ag{sup +}-exchanged hybrid composites. The zeolite-A/chitosan hybrid composites could be prepared as various shapes, including cylinders, plates and thin films. They possessed macropores with pore sizes ranging from 100 to 300 μm and showed compressive mechanical strength as high as 3.2 MPa when the zeolite content was 35 wt.%. Fast growth on the Ca{sup 2+}-exchanged hybrid composites was observed with the highest weight gain of 51.4% in 30 days. The 35 wt.% Ag{sup +}-exchanged hybrid composite showed the highest antimicrobial activity, which could reduce the 9 × 10{sup 6} CFU mL{sup −1}E. coli concentration to zero within 4 h of incubation time with the Ag{sup +}-exchanged hybrid composite amount of 0.4 g L{sup −1}. The bioactivity and antimicrobial activity could be combined by ion-exchanging the composites first with Ca{sup 2+} and then with Ag{sup +}. These zeolite-A/chitosan hybrid composites have potential applications on tissue engineering and antimicrobial food packaging. - Graphical abstract: Zeolite A/chitosan hybrid composites were prepared by in situ transformation of precursors in the chitosan matrix, which possess macroporous structures and exhibit superior bioactivity and antimicrobial activity and potential biomedical

Assignment of the human gene for pregnancy-associated plasma protein A (PAPPA) to 9q33.1 by fluorescence in situ hybridization to mitotic and meiotic chromosomes

DEFF Research Database (Denmark)

Silahtaroglu, A N; Tümer, Z; Kristensen, Torsten

1993-01-01

Low levels of pregnancy-associated plasma protein A (PAPPA) during the first trimester has been suggested as a biochemical indicator of pregnancies with aneuploid fetuses. Furthermore, the complete absence of PAPPA in pregnancies associated with Cornelia de Lange syndrome (CL) has suggested...... a causal connection between PAPPA and the development of CL. We have assigned the locus for PAPPA to chromosome region 9q33.1 on mitotic and meiotic chromosomes by fluorescence in situ hybridization, using a 3.7-kb partial PAPPA cDNA probe...

Fluorescence in situ hybridization of TP53 for the detection of chromosome 17 abnormalities in myelodysplastic syndromes.

Science.gov (United States)

Sánchez-Castro, Judit; Marco-Betés, Víctor; Gómez-Arbonés, Xavier; García-Cerecedo, Tomás; López, Ricard; Talavera, Elisabeth; Fernández-Ruiz, Sara; Ademà, Vera; Marugan, Isabel; Luño, Elisa; Sanzo, Carmen; Vallespí, Teresa; Arenillas, Leonor; Marco Buades, Josefa; Batlle, Ana; Buño, Ismael; Martín Ramos, María Luisa; Blázquez Rios, Beatriz; Collado Nieto, Rosa; Vargas, Ma Teresa; González Martínez, Teresa; Sanz, Guillermo; Solé, Francesc

2015-01-01

Conventional G-banding cytogenetics (CC) detects chromosome 17 (chr17) abnormalities in 2% of patients with de novo myelodysplastic syndromes (MDS). We used CC and fluorescence in situ hybridization (FISH) (LSI p53/17p13.1) to assess deletion of 17p in 531 patients with de novo MDS from the Spanish Group of Hematological Cytogenetics. FISH detected - 17 or 17p abnormalities in 13 cases (2.6%) in whom no 17p abnormalities were revealed by CC: 0.9% of patients with a normal karyotype, 0% in non-informative cytogenetics, 50% of patients with a chr17 abnormality without loss of 17p and 4.7% of cases with an abnormal karyotype not involving chr17. Our results suggest that applying FISH of 17p13 to identify the number of copies of the TP53 gene could be beneficial in patients with a complex karyotype. We recommend using FISH of 17p13 in young patients with a normal karyotype or non-informative cytogenetics, and always in isolated del(17p).

In-situ preparation of poly(ethylene oxide)/Li3PS4 hybrid polymer electrolyte with good nanofiller distribution for rechargeable solid-state lithium batteries

Science.gov (United States)

Chen, Shaojie; Wang, Junye; Zhang, Zhihua; Wu, Linbin; Yao, Lili; Wei, Zhenyao; Deng, Yonghong; Xie, Dongjiu; Yao, Xiayin; Xu, Xiaoxiong

2018-05-01

Nano-sized fillers in a polymer matrix with good distribution can play a positive role in improving polymer electrolytes in the aspects of ionic conductivity, mechanical property and electrochemical performance of Li-ion cells. Herein, polyethylene oxide (PEO)/Li3PS4 hybrid polymer electrolyte is prepared via a new in-situ approach. The ionic conductivities of the novel hybrid electrolytes with variable proportions are measured, and the optimal electrolyte of PEO-2%vol Li3PS4 presents a considerable ionic conductivity of 8.01 × 10-4 S cm-1 at 60 °C and an electrochemical window up to 5.1 V. The tests of DSC and EDXS reveal that the Li3PS4 nanoparticles with better distribution, as active fillers scattering in the PEO, exhibit a positive effect on the transference of lithium ion and electrochemical interfacial stabilities. Finally, the assembled solid-state LiFePO4/Li battery presents a decent cycling performance (80.9% retention rate after 325 cycles at 60 °C) and excellent rate capacities with 153, 143, 139 and 127 mAh g-1 at the discharging rate of 0.1 C, 0.2 C, 0.5 C and 1 C at 60 °C. It is fully proved that it is an advanced strategy to preparing the new organic/inorganic hybrid electrolytes for lithium-ion batteries applications.

In situ hybridization for the detection of rust fungi in paraffin embedded plant tissue sections.

Science.gov (United States)

Ellison, Mitchell A; McMahon, Michael B; Bonde, Morris R; Palmer, Cristi L; Luster, Douglas G

2016-01-01

Rust fungi are obligate pathogens with multiple life stages often including different spore types and multiple plant hosts. While individual rust pathogens are often associated with specific plants, a wide range of plant species are infected with rust fungi. To study the interactions between these important pathogenic fungi and their host plants, one must be able to differentiate fungal tissue from plant tissue. This can be accomplished using the In situ hybridization (ISH) protocol described here. To validate reproducibility using the ISH protocol, samples of Chrysanthemum × morifolium infected with Puccinia horiana, Gladiolus × hortulanus infected with Uromyces transversalis and Glycine max infected with Phakopsora pachyrhizi were tested alongside uninfected leaf tissue samples. The results of these tests show that this technique clearly distinguishes between rust pathogens and their respective host plant tissues. This ISH protocol is applicable to rust fungi and potentially other plant pathogenic fungi as well. It has been shown here that this protocol can be applied to pathogens from different genera of rust fungi with no background staining of plant tissue. We encourage the use of this protocol for the study of plant pathogenic fungi in paraffin embedded sections of host plant tissue.

Elucidating the Burden of HIV in Tissues Using Multiplexed Immunofluorescence and In Situ Hybridization: Methods for the Single-Cell Phenotypic Characterization of Cells Harboring HIV In Situ.

Science.gov (United States)

Vasquez, Joshua J; Hussien, Rajaa; Aguilar-Rodriguez, Brandon; Junger, Henrik; Dobi, Dejan; Henrich, Timothy J; Thanh, Cassandra; Gibson, Erica; Hogan, Louise E; McCune, Joseph; Hunt, Peter W; Stoddart, Cheryl A; Laszik, Zoltan G

2018-02-01

Persistent tissue reservoirs of HIV present a major barrier to cure. Defining subsets of infected cells in tissues is a major focus of HIV cure research. Herein, we describe a novel multiplexed in situ hybridization (ISH) (RNAscope) protocol to detect HIV-DNA (vDNA) and HIV-RNA (vRNA) in formalin-fixed paraffin-embedded (FFPE) human tissues in combination with immunofluorescence (IF) phenotyping of the infected cells. We show that multiplexed IF and ISH (mIFISH) is suitable for quantitative assessment of HIV vRNA and vDNA and that multiparameter IF phenotyping allows precise identification of the cellular source of the ISH signal. We also provide semi-quantitative data on the impact of various tissue fixatives on the detectability of vDNA and vRNA with RNAscope technology. Finally, we describe methods to quantitate the ISH signal on whole-slide digital images and validation of the quantitative ISH data with quantitative real-time PCR for vRNA. It is our hope that this approach will provide insight into the biology of HIV tissue reservoirs and to inform strategies aimed at curing HIV.

Conventional and fluorescence in situ hybridization analysis of three-way complex BCR-ABL rearrangement in a chronic myeloid leukemia patient

Directory of Open Access Journals (Sweden)

Ganguly Bani

2007-01-01

Full Text Available Chromosomal analysis was carried out in bone marrow sample of an 11-year-old girl suspected of myeloproliferative disorder. Conventional G-banding study detected a complex three-way translocation involving 7, 9 and 22, which has resulted in the formation of a variant Philadelphia chromosome causing rearrangement of abl and bcr genes in 87% cells. Fluorescence in situ hybridization (FISH confirmed the fusion of bcr-abl oncogene. Thus the bone marrow karyotype was observed as 46,XX (13% / 46,XX,t(7;9;22(q11;q34;q11 (87%. Hyperdiploidy was present in two cells. In this study, both conventional cytogenetic and FISH diagnosis proved to be significant to identify the variant nature of the Philadelphia chromosome and hyperdiploid condition for introduction of a suitable treatment regimen and estimation of life expectancy of the young girl.

Graphene oxide chemically decorated with hybrid Ag-Ru/chitosan nanoparticles: fabrication and properties

OpenAIRE

Veerapandian, Murugan; Neethirajan, Suresh

2015-01-01

Hybridization of distinct materials into a single nanoplatform is relevant to advance material’s properties for functional application such as biosensor platform. We report the synthesis and characterization of nanosheets of graphene oxide decorated with hybrid nanoparticles of silver-ruthenium bipyridine complex (Ag@[Ru(bpy)3]2+) core and chitosan shell. Hybrid nanoparticles were first obtained through a sequential wet-chemical approach using in situ reduction, electrostatic and coordination...

Pre-implantation genetic screening using fluorescence in situ hybridization in couples of Indian ethnicity: Is there a scope?

Directory of Open Access Journals (Sweden)

Shailaja Gada Saxena

2014-01-01

Full Text Available Context: There is a high incidence of numerical chromosomal aberration in couples with repeated in vitro fertilization (IVF failure, advanced maternal age, repeated unexplained abortions, severe male factor infertility and unexplained infertility. Pre-implantation genetic screening (PGS, a variant of pre-implantation genetic diagnosis, screens numerical chromosomal aberrations in couples with normal karyotype, experiencing poor reproductive outcome. The present study includes the results of the initial pilot study on 9 couples who underwent 10 PGS cycles. Aim: The aim of the present study was to evaluate the beneficial effects of PGS in couples with poor reproductive outcome. Settings and Design: Data of initial 9 couples who underwent 10 PGS for various indications was evaluated. Subjects and Methods: Blastomere biopsy was performed on cleavage stage embryos and subjected to two round fluorescence in situ hybridization (FISH testing for chromosomes 13, 18, 21, X and Y as a two-step procedure. Results: Six of the 9 couples (10 PGS cycles conceived, including a twin pregnancy in a couple with male factor infertility, singleton pregnancies in a couple with secondary infertility, in three couples with adverse obstetric outcome in earlier pregnancies and in one couple with repeated IVF failure. Conclusion: In the absence of availability of array-comparative genomic hybridization in diagnostic clinical scenario for PGS and promising results with FISH based PGS as evident from the current pilot study, it is imperative to offer the best available services in the present scenario for better pregnancy outcome for patients.

In situ gold nanoparticles formation: contrast agent for dental optical coherence tomography

Science.gov (United States)

Braz, Ana K. S.; Araujo, Renato E. de; Ohulchanskyy, Tymish Y.; Shukla, Shoba; Bergey, Earl J.; Gomes, Anderson S. L.; Prasad, Paras N.

2012-06-01

In this work we demonstrate the potential use of gold nanoparticles as contrast agents for the optical coherence tomography (OCT) imaging technique in dentistry. Here, a new in situ photothermal reduction procedure was developed, producing spherical gold nanoparticles inside dentinal layers and tubules. Gold ions were dispersed in the primer of commercially available dental bonding systems. After the application and permeation in dentin by the modified adhesive systems, the dental bonding materials were photopolymerized concurrently with the formation of gold nanoparticles. The gold nanoparticles were visualized by scanning electron microscopy (SEM). The SEM images show the presence of gold nanospheres in the hybrid layer and dentinal tubules. The diameter of the gold nanoparticles was determined to be in the range of 40 to 120 nm. Optical coherence tomography images were obtained in two- and three-dimensions. The distribution of nanoparticles was analyzed and the extended depth of nanosphere production was determined. The results show that the OCT technique, using in situ formed gold nanoparticles as contrast enhancers, can be used to visualize dentin structures in a non-invasive and non-destructive way.

Hydraulic Hybrid Vehicles

Science.gov (United States)

EPA and the United Parcel Service (UPS) have developed a hydraulic hybrid delivery vehicle to explore and demonstrate the environmental benefits of the hydraulic hybrid for urban pick-up and delivery fleets.

Core-shell LiFePO4 /carbon-coated reduced graphene oxide hybrids for high-power lithium-ion battery cathodes.

Science.gov (United States)

Ha, Sung Hoon; Lee, Yun Jung

2015-01-26

Core-shell carbon-coated LiFePO4 nanoparticles were hybridized with reduced graphene (rGO) for high-power lithium-ion battery cathodes. Spontaneous aggregation of hydrophobic graphene in aqueous solutions during the formation of composite materials was precluded by employing hydrophilic graphene oxide (GO) as starting templates. The fabrication of true nanoscale carbon-coated LiFePO4 -rGO (LFP/C-rGO) hybrids were ascribed to three factors: 1) In-situ polymerization of polypyrrole for constrained nanoparticle synthesis of LiFePO4 , 2) enhanced dispersion of conducting 2D networks endowed by colloidal stability of GO, and 3) intimate contact between active materials and rGO. The importance of conducting template dispersion was demonstrated by contrasting LFP/C-rGO hybrids with LFP/C-rGO composites in which agglomerated rGO solution was used as the starting templates. The fabricated hybrid cathodes showed superior rate capability and cyclability with rates from 0.1 to 60 C. This study demonstrated the synergistic combination of nanosizing with efficient conducting templates to afford facile Li(+) ion and electron transport for high power applications. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Identification of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa in Blood Cultures: a Multicenter Performance Evaluation of a Three-Color Peptide Nucleic Acid Fluorescence In Situ Hybridization Assay▿

Science.gov (United States)

Della-Latta, Phyllis; Salimnia, Hossein; Painter, Theresa; Wu, Fann; Procop, Gary W.; Wilson, Deborah A.; Gillespie, Wendy; Mender, Alayna; Crystal, Benjamin S.

2011-01-01

A multicenter evaluation was undertaken to evaluate the performance of a new three-color peptide nucleic acid fluorescence in situ hybridization assay that identifies isolates directly from blood cultures positive for Gram-negative bacilli (GNB). The assay correctly identified 100% (186/186) of the Escherichia coli isolates, 99.1% (109/110) of the Klebsiella pneumoniae isolates, and 95.8% (46/48) of the Pseudomonas aeruginosa isolates in this study. Negative assay results were correctly obtained for 162 of 165 other GNB (specificity, 98.2%). PMID:21490185

In situ growth of lamellar ZnTiO3 nanosheets on TiO2 tubular array with enhanced photocatalytic activity.

Science.gov (United States)

Cai, Yunyu; Ye, Yixing; Tian, Zhenfei; Liu, Jun; Liu, Yishu; Liang, Changhao

2013-12-14

We report a self-sacrificed in situ growth design toward preparation of ZnTiO3-TiO2 heterojunction structure. Highly reactive zinc oxide colloidal particles derived by laser ablation in liquids can react with TiO2 nanotubes to form a lamellar ZnTiO3 nanosheet structure in a hydrothermal-treatment process. Such hybrid structural product was characterized by X-ray diffraction, scanning and transmission electron microscopy, UV-vis diffuse reflection spectroscopy and X-ray photoelectron spectroscopy. The enhanced photocatalytic activity of the hybrid structure toward degradation of methyl orange (MO) and pentachlorophenol (PCP) molecules was demonstrated and compared with single phase TiO2, as a result of the efficient separation of light excited electrons and holes at the hetero-interfaces in the two semiconductors.

In situ vitrification demonstration for the stabilization of buried wastes at the Oak Ridge National Laboratory

International Nuclear Information System (INIS)

Jacobs, G.K.; Spalding, B.P.; Carter, J.G.; Koegler, S.S.

1987-01-01

A demonstration of In Situ Vitrification (ISV) technology for the stabilization of radioactively contaminated soil sites at the Oak Ridge National Laboratory (ORNL) was successfully completed during July 1987. This demonstration is the first application of the ISV process not performed at the Hanford Site, where the technology was developed and patented by Pacific Northwest Laboratory (PNL). The joint ORNL-PNL pilot-scale demonstration was performed on a 3/8-scale trench (2 m deep x 1 m wide x 10 m long) that was constructed to simulate a typical seepage trench used for liquid low-level radioactive waste disposal at ORNL from 1951 to 1966. In the ISV process, electrodes are inserted around a volume of contaminated soil, power is applied to the electrodes, and the entire mass is melted from the surface of the soil down through the contaminated zone, thus making a glassy-to-microcrystalline waste form that incorporates the contaminants. Gases produced during the melting are collected, treated, monitored, and released through an off-gas process trailer. In the ORNL demonstration, a 25-t mass of melted rock approximately 1.2 m thick x 2.1 m wide x 4.9 m long was formed during 110 h of operation that consumed approximately 29 MWh of power. Data obtained on the operational performance of the test and waste-form durability will be used to assess the feasibility of applying the ISV technology to an actual waste trench

Hybrid carbon nanomaterials for electrochemical detection of biomolecules

International Nuclear Information System (INIS)

Laurila, Tomi

2015-01-01

Electrochemical detection of different biomolecules in vivo is a promising path towards in situ monitoring of human body and its functions. However, there are several major obstacles, such as sensitivity, selectivity and biocompatiblity, which must be tackled in order to achieve reliably and safely operating sensor devices. Here we show that by utilizing hybrid carbon materials as electrodes to detect two types of neurotransmitters, dopamine and glutamate, several advantages over commonly used electrode materials can be achieved. In particular, we will demonstrate here that it is possible to combine the properties of different carbon allotropes to obtain hybrid materials with greatly improved electrochemical performance. Three following examples of the approach are given: (i) diamond-like carbon (DLC) thin film electrodes with different layer thicknesses, (ii) multi-walled carbon nanotubes grown directly on top of DLC and (iii) carbon nanofibres synthesized on top of DLC thin films. Detailed structural and electrochemical characterization is carried out to rationalize the reasons behind the observed behvior. In addition, results from the atomistic simulations are utilized to obtain more information about the properties of the amorphous carbon thin films. (paper)

SEAMIST trademark in-situ instrumentation and vapor sampling system applications in the Sandia Mixed Waste Landfill Integrated Demonstration Program

International Nuclear Information System (INIS)

Lowry, W.E.; Dunn, S.D.; Cremer, S.C.; Williams, C.

1994-01-01

The SEAMIST trademark inverting membrane deployment system has been used successfully at the Mixed Waste Landfill Integrated Demonstration (MWLID) for multipoint vapor sampling/pressure measurement/permeability measurement/sensor integration demonstrations and borehole lining. Several instruments were deployed inside the SEAMIST trademark lined boreholes to detect metals, radionuclides, moisture, and geologic variations. The liner protected the instruments from contamination, maintained support of the uncased borehole wall, and sealed the total borehole from air circulation. The current activities have included the installation of three multipoint vapor sampling systems and sensor integration systems in 100-foot-deep vertical boreholes. A long term pressure monitoring program has recorded barometric pressure effects at depth with relatively high spatial resolution. The SEAMIST trademark system has been integrated with a variety of hydrologic and chemical sensors for in-situ measurements, demonstrating its versatility as an instrument deployment system which allows easy emplacement and removal. Standard SEAMIST trademark vapor sampling systems were also integrated with state-of-the-art VOC analysis technologies (automated GC, UV laser fluorometer). The results and status of these demonstration tests are presented

Fluorescence photooxidation with eosin: a method for high resolution immunolocalization and in situ hybridization detection for light and electron microscopy

Science.gov (United States)

1994-01-01

A simple method is described for high-resolution light and electron microscopic immunolocalization of proteins in cells and tissues by immunofluorescence and subsequent photooxidation of diaminobenzidine tetrahydrochloride into an insoluble osmiophilic polymer. By using eosin as the fluorescent marker, a substantial improvement in sensitivity is achieved in the photooxidation process over other conventional fluorescent compounds. The technique allows for precise correlative immunolocalization studies on the same sample using fluorescence, transmitted light and electron microscopy. Furthermore, because eosin is smaller in size than other conventional markers, this method results in improved penetration of labeling reagents compared to gold or enzyme based procedures. The improved penetration allows for three-dimensional immunolocalization using high voltage electron microscopy. Fluorescence photooxidation can also be used for high resolution light and electron microscopic localization of specific nucleic acid sequences by in situ hybridization utilizing biotinylated probes followed by an eosin-streptavidin conjugate. PMID:7519623

Fluorescence in situ hybridization (FISH screening for the 22q11.2 deletion in patients with clinical features of velocardiofacial syndrome but without cardiac anomalies

Directory of Open Access Journals (Sweden)

Paula Sandrin-Garcia

2007-01-01

Full Text Available The velocardiofacial syndrome (VCFS, a condition associated with 22q11.2 deletions, is characterized by a typical facies, palatal anomalies, learning disabilities, behavioral disturbances and cardiac defects. We investigated the frequency of these chromosomal deletions in 16 individuals with VCFS features who presented no cardiac anomalies, one of the main characteristics of VCFS. Fluorescent in situ hybridization (FISH with the N25 (D22S75; 22q11.2 probe revealed deletions in ten individuals (62%. Therefore, even in the absence of cardiac anomalies testing for the 22q11.2 microdeletions in individuals showing other clinical features of this syndrome is recommended.

Overview of ERA Integrated Technology Demonstration (ITD) 51A Ultra-High Bypass (UHB) Integration for Hybrid Wing Body (HWB)

Science.gov (United States)

Flamm, Jeffrey D.; James, Kevin D.; Bonet, John T.

2016-01-01

The NASA Environmentally Responsible Aircraft Project (ERA) was a ve year project broken into two phases. In phase II, high N+2 Technical Readiness Level demonstrations were grouped into Integrated Technology Demonstrations (ITD). This paper describes the work done on ITD-51A: the Vehicle Systems Integration, Engine Airframe Integration Demonstration. Refinement of a Hybrid Wing Body (HWB) aircraft from the possible candidates developed in ERA Phase I was continued. Scaled powered, and unpowered wind- tunnel testing, with and without acoustics, in the NASA LARC 14- by 22-foot Subsonic Tunnel, the NASA ARC Unitary Plan Wind Tunnel, and the 40- by 80-foot test section of the National Full-Scale Aerodynamics Complex (NFAC) in conjunction with very closely coupled Computational Fluid Dynamics was used to demonstrate the fuel burn and acoustic milestone targets of the ERA Project.

Automated image analysis for quantitative fluorescence in situ hybridization with environmental samples.

Science.gov (United States)

Zhou, Zhi; Pons, Marie Noëlle; Raskin, Lutgarde; Zilles, Julie L

2007-05-01

When fluorescence in situ hybridization (FISH) analyses are performed with complex environmental samples, difficulties related to the presence of microbial cell aggregates and nonuniform background fluorescence are often encountered. The objective of this study was to develop a robust and automated quantitative FISH method for complex environmental samples, such as manure and soil. The method and duration of sample dispersion were optimized to reduce the interference of cell aggregates. An automated image analysis program that detects cells from 4',6'-diamidino-2-phenylindole (DAPI) micrographs and extracts the maximum and mean fluorescence intensities for each cell from corresponding FISH images was developed with the software Visilog. Intensity thresholds were not consistent even for duplicate analyses, so alternative ways of classifying signals were investigated. In the resulting method, the intensity data were divided into clusters using fuzzy c-means clustering, and the resulting clusters were classified as target (positive) or nontarget (negative). A manual quality control confirmed this classification. With this method, 50.4, 72.1, and 64.9% of the cells in two swine manure samples and one soil sample, respectively, were positive as determined with a 16S rRNA-targeted bacterial probe (S-D-Bact-0338-a-A-18). Manual counting resulted in corresponding values of 52.3, 70.6, and 61.5%, respectively. In two swine manure samples and one soil sample 21.6, 12.3, and 2.5% of the cells were positive with an archaeal probe (S-D-Arch-0915-a-A-20), respectively. Manual counting resulted in corresponding values of 22.4, 14.0, and 2.9%, respectively. This automated method should facilitate quantitative analysis of FISH images for a variety of complex environmental samples.

Correlation between Photoluminescence and Carrier Transport and a Simple In Situ Passivation Method for High-Bandgap Hybrid Perovskites.

Science.gov (United States)

Stoddard, Ryan J; Eickemeyer, Felix T; Katahara, John K; Hillhouse, Hugh W

2017-07-20

High-bandgap mixed-halide hybrid perovskites have higher open-circuit voltage deficits and lower carrier diffusion lengths than their lower-bandgap counterparts. We have developed a ligand-assisted crystallization (LAC) technique that introduces additives in situ during the solvent wash and developed a new method to dynamically measure the absolute intensity steady-state photoluminescence and the mean carrier diffusion length simultaneously. The measurements reveal four distinct regimes of material changes and show that photoluminescence brightening often coincides with losses in carrier transport, such as in degradation or phase segregation. Further, the measurements enabled optimization of LAC on the 1.75 eV bandgap FA 0.83 Cs 0.17 Pb(I 0.66 Br 0.34 ) 3 , resulting in an enhancement of the photoluminescence quantum yield (PLQY) of over an order of magnitude, an increase of 80 meV in the quasi-Fermi level splitting (to 1.29 eV), an increase in diffusion length by a factor of 3.5 (to over 1 μm), and enhanced open-circuit voltage and short-circuit current from photovoltaics fabricated from the LAC-treated films.

DNA hybridization sensing for cytogenetic analysis

DEFF Research Database (Denmark)

Kwasny, Dorota; Dapra, Johannes; Brøgger, Anna Line

2013-01-01

are rearrangements between two chromosome arms that results in two derivative chromosomes having a mixed DNA sequence. The current detection method is a Fluorescent In situ Hybridization, which requires a use of expensive, fluorescently labeled probes that target the DNA sequences of two chromosomes involved...... in the translocation (Kwasny et al., 2012). We have developed a new double hybridization assay that allows for sorting of the DNA chromosomal fragments into separate compartment, moreover allowing for detection of the translocation. To detect the translocation it is necessary to determine that the two DNA sequences...... forming a derivative chromosome are connected, which is achieved by two subsequent hybridization steps. The first example of the translocation detection was presented on lab-on-a-disc using fluorescently labeled DNA fragments, representing the derivative chromosome (Brøgger et al., 2012). To allow...

Fluctuation localization imaging-based fluorescence in situ hybridization (fliFISH) for accurate detection and counting of RNA copies in single cells

Energy Technology Data Exchange (ETDEWEB)

Cui, Yi; Hu, Dehong; Markillie, Lye Meng; Chrisler, William B.; Gaffrey, Matthew J.; Ansong, Charles; Sussel, Lori; Orr, Galya

2017-10-04

Quantitative gene expression analysis in intact single cells can be achieved using single molecule- based fluorescence in situ hybridization (smFISH). This approach relies on fluorescence intensity to distinguish between true signals, emitted from an RNA copy hybridized with multiple FISH sub-probes, and background noise. Thus, the precision in smFISH is often compromised by partial or nonspecific binding of sub-probes and tissue autofluorescence, limiting its accuracy. Here we provide an accurate approach for setting quantitative thresholds between true and false signals, which relies on blinking frequencies of photoswitchable dyes. This fluctuation localization imaging-based FISH (fliFISH) uses blinking frequency patterns, emitted from a transcript bound to multiple sub-probes, which are distinct from blinking patterns emitted from partial or nonspecifically bound sub-probes and autofluorescence. Using multicolor fliFISH, we identified radial gene expression patterns in mouse pancreatic islets for insulin, the transcription factor, NKX2-2, and their ratio (Nkx2-2/Ins2). These radial patterns, showing higher values in β cells at the islet core and lower values in peripheral cells, were lost in diabetic mouse islets. In summary, fliFISH provides an accurate, quantitative approach for detecting and counting true RNA copies and rejecting false signals by their distinct blinking frequency patterns, laying the foundation for reliable single-cell transcriptomics.

A conceptual demonstration of freeze desalination-membrane distillation (FD-MD) hybrid desalination process utilizing liquefied natural gas (LNG) cold energy.

Science.gov (United States)

Wang, Peng; Chung, Tai-Shung

2012-09-01

The severe global water scarcity and record-high fossil oil price have greatly stimulated the research interests on new desalination technologies which can be driven by renewable energy or waste energy. In this study, a hybrid desalination process comprising freeze desalination and membrane distillation (FD-MD) processes was developed and explored in an attempt to utilize the waste cold energy released from re-gasification of liquefied natural gas (LNG). The concept of this technology was demonstrated using indirect-contact freeze desalination (ICFD) and direct-contact membrane distillation (DCMD) configurations. By optimizing the ICFD operation parameters, namely, the usage of nucleate seeds, operation duration and feed concentration, high quality drinkable water with a low salinity ∼0.144 g/L was produced in the ICFD process. At the same time, using the optimized hollow fiber module length and packing density in the DCMD process, ultra pure water with a low salinity of 0.062 g/L was attained at a condition of high energy efficiency (EE). Overall, by combining FD and MD processes and adopting the optimized operation parameters, the hybrid FD-MD system has been successfully demonstrated. A high total water recovery of 71.5% was achieved, and the water quality obtained met the standard for drinkable water. In addition, with results from specific energy calculation, it was proven that the hybrid process is an energy-saving process and utilization of LNG cold energy could greatly reduce the total energy consumption. Copyright © 2012 Elsevier Ltd. All rights reserved.

Role of chromogenic in situ hybridization (CISH) in the evaluation of HER2 status in breast carcinoma: comparison with immunohistochemistry and FISH.

Science.gov (United States)

Li-Ning-T, Elsa; Ronchetti, Ruben; Torres-Cabala, Carlos; Merino, Maria J

2005-10-01

We report our experience with Chromogenic in Situ Hybridization (CISH) for the evaluation of HER2 amplification on 55 cases of formalin-fixed, paraffin-embedded invasive breast carcinomas of different histology. All the results were corrected for chromosome 17 aneusomy and compared with immunohistochemistry (IHC); a subset of cases was compared to FISH. Thirty-one of 32 cases in which FISH and CISH were performed yielded the same results. CISH and IHC showed a good concordance in the 0/1+ and 3+ category, while a poor agreement with weakly protein overexpression was confirmed. Chromosome 17 analysis was necessary in cases with a low number of HER2 gene copies. CISH is a useful tool to evaluate breast cancer HER2 status that can be easily implemented in a laboratory of surgical pathology.

Fluorescence In Situ Hybridization with Peptide Nucleic Acid Probes for Rapid Identification of Candida albicans Directly from Blood Culture Bottles

Science.gov (United States)

Rigby, Susan; Procop, Gary W.; Haase, Gerhard; Wilson, Deborah; Hall, Geraldine; Kurtzman, Cletus; Oliveira, Kenneth; Von Oy, Sabina; Hyldig-Nielsen, Jens J.; Coull, James; Stender, Henrik

2002-01-01

A new fluorescence in situ hybridization (FISH) method that uses peptide nucleic acid (PNA) probes for identification of Candida albicans directly from positive-blood-culture bottles in which yeast was observed by Gram staining (herein referred to as yeast-positive blood culture bottles) is described. The test (the C. albicans PNA FISH method) is based on a fluorescein-labeled PNA probe that targets C. albicans 26S rRNA. The PNA probe is added to smears made directly from the contents of the blood culture bottle and hybridized for 90 min at 55°C. Unhybridized PNA probe is removed by washing of the mixture (30 min), and the smears are examined by fluorescence microscopy. The specificity of the method was confirmed with 23 reference strains representing phylogenetically related yeast species and 148 clinical isolates covering the clinically most significant yeast species, including C. albicans (n = 72), C. dubliniensis (n = 58), C. glabrata (n = 5), C. krusei (n = 2), C. parapsilosis (n = 4), and C. tropicalis (n = 3). The performance of the C. albicans PNA FISH method as a diagnostic test was evaluated with 33 routine and 25 simulated yeast-positive blood culture bottles and showed 100% sensitivity and 100% specificity. It is concluded that this 2.5-h method for the definitive identification of C. albicans directly from yeast-positive blood culture bottles provides important information for optimal antifungal therapy and patient management. PMID:12037084

Expression of μ, κ, and δ opioid receptor messenger RNA in the human CNS: a 33P in situ hybridization study

International Nuclear Information System (INIS)