Search results for: fluorescent pseudomonad

Authors: Martin De Biasio, Raimund Leitner, Franz G. Wuertz, Sergey Verzakov, Pierre J. Elbischger

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Breast carcinoma is the most common form of cancer in women. Multicolour fluorescent in-situ hybridisation (m-FISH) is a common method for staging breast carcinoma. The interpretation of m-FISH images is complicated due to two effects: (i) Spectral overlap in the emission spectra of fluorochrome marked DNA probes and (ii) tissue autofluorescence. In this paper hyper-spectral images of m-FISH samples are used and spectral unmixing is applied to produce false colour images with higher contrast and better information content than standard RGB images. The spectral unmixing is realised by combinations of: Orthogonal Projection Analysis (OPA), Alterating Least Squares (ALS), Simple-to-use Interactive Self-Modeling Mixture Analysis (SIMPLISMA) and VARIMAX. These are applied on the data to reduce tissue autofluorescence and resolve the spectral overlap in the emission spectra. The results show that spectral unmixing methods reduce the intensity caused by tissue autofluorescence by up to 78% and enhance image contrast by algorithmically reducing the overlap of the emission spectra.

Keywords: breast carcinoma, hyperspectral imaging, m-FISH, spectral unmixing

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Authors: Giwrgos S. Gkarmpounis, Aikaterini G. Rokkou, Marios N. Moschakis

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Increased energy consumption in the academic buildings, creates the need to implement energy saving measures and to take advantage of the renewable energy sources to cover the electrical needs of those buildings. An Academic Library will be used as a case study. With the aid of RETScreen software that takes into account the energy consumptions and characteristics of the Library Building, it is proved that measures such as the replacement of fluorescent lights with led lights, the installation of outdoor shading, the replacement of the openings and Building Management System installation, provide a high level of energy savings. Moreover, given the available space of the building and the climatic data, the installation of a photovoltaic system of 100 kW can also cover a serious amount of the building energy consumption, unlike a wind system that seems uncompromising. Lastly, HOMER software is used to compare the use of a photovoltaic system against a wind system in order to verify the results that came up from the RETScreen software concerning the renewable energy sources.

Keywords: Energy saving measures, homer software, renewable energy sources, RETScreen software, energy efficiency and quality.

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Authors: Andreas Heindl, Alexander K. Seewald, Martin Schepelmann, Radu Rogojanu, Giovanna Bises, Theresia Thalhammer, Isabella Ellinger

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Bone remodeling occurs by the balanced action of bone resorbing osteoclasts (OC) and bone-building osteoblasts. Increased bone resorption by excessive OC activity contributes to malignant and non-malignant diseases including osteoporosis. To study OC differentiation and function, OC formed in in vitro cultures are currently counted manually, a tedious procedure which is prone to inter-observer differences. Aiming for an automated OC-quantification system, classification of OC and precursor cells was done on fluorescence microscope images based on the distinct appearance of fluorescent nuclei. Following ellipse fitting to nuclei, a combination of eight features enabled clustering of OC and precursor cell nuclei. After evaluating different machine-learning techniques, LOGREG achieved 74% correctly classified OC and precursor cell nuclei, outperforming human experts (best expert: 55%). In combination with the automated detection of total cell areas, this system allows to measure various cell parameters and most importantly to quantify proteins involved in osteoclastogenesis.

Keywords: osteoclasts, machine learning, ellipse fitting.

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Authors: C. Sinescu, M. Negrutiu, M. Romînu, C. Haiduc, E. Petrescu, M. Leretter, A.G. Podoleanu

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Optical Coherence Tomography (OCT) combined with the Confocal Microscopy, as a noninvasive method, permits the determinations of materials defects in the ceramic layers depth. For this study 256 anterior and posterior metal and integral ceramic fixed partial dentures were used, made with Empress (Ivoclar), Wollceram and CAD/CAM (Wieland) technology. For each investigate area 350 slices were obtain and a 3D reconstruction was perform from each stuck. The Optical Coherent Tomography, as a noninvasive method, can be used as a control technique in integral ceramic technology, before placing those fixed partial dentures in the oral cavity. The purpose of this study is to evaluate the capability of En face Optical Coherence Tomography (OCT) combined with a fluorescent method in detection and analysis of possible material defects in metalceramic and integral ceramic fixed partial dentures. As a conclusion, it is important to have a non invasive method to investigate fixed partial prostheses before their insertion in the oral cavity in order to satisfy the high stress requirements and the esthetic function.

Keywords: Ceramic Fixed Partial Dentures, Material Defects, En face Optical Coherence Tomography, Fluorescence.

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Authors: Tzyy Rong Jinn, Nguyen Tiep Khac, Tzong Yuan Wu

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Highly pathogenic avian influenza (HPAI) H5N1 viruses have created demand for a cost-effective vaccine to prevent a pandemic of the disease. Here, we report that Trichoplusia ni (T. ni) larvae can act as a cost-effective bioreactor to produce recombinant HA5 (rH5HA) proteins as an potential effective vaccine for chickens. To facilitate the recombinant virus identification, virus titer determination and access the infected larvae, we employed the internal ribosome entry site (IRES) derived from Perina nuda virus (PnV, belongs to insect picorna like Iflavirus genus) to construct a bi-cistronic baculovirus expression vector that can express the rH5HA protein and enhanced green fluorescent protein (EGFP) simultaneously. Western blot analysis revealed that the 70 kDa rH5HA protein and partially cleaved products (40 kDa H5HA1) were generated in T. ni larvae infected with recombinant baculovirus carrying the H5HA gene. These data suggest that the baculovirus-larvae recombinant protein expression system could be a cost-effective platform for H5N1 vaccine production.

Keywords: Avian Influenza, baculovirus, hemagglutinin, Trichoplusia ni larvae

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Authors: Stela Papa, Klementina Puto, Migena Pllaha

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HCV is a hepatotropic RNA virus, transmitted primarily via the blood route, which causes progressive disease such as chronic hepatitis, liver cirrhosis, or hepatocellular carcinoma. HCV nowadays is a global healthcare problem. A variety of immunoassays including old and new technologies are being applied to detect HCV in our country. These methods include Immunochromatography assays (ICA), Fluorescence immunoassay (FIA), Enzyme linked fluorescent assay (ELFA), and Enzyme linked immunosorbent assay (ELISA) to detect HCV antibodies in blood serum, which lately is being slowly replaced by more sensitive methods such as rapid automated analyzer chemiluminescence immunoassay (CLIA). The aim of this study is to estimate HCV infection in carriers and chronic acute patients and to evaluate the use of new diagnostic methods. This study was realized from September 2016 to May 2018. During this study period, 2913 patients were analyzed for the presence of HCV by taking samples from their blood serum. The immunoassays performed were ICA, FIA, ELFA, ELISA, and CLIA assays. Concluding, 82% of patients taken in this study, resulted infected with HCV. Diagnostic methods in clinical laboratories are crucial in the early stages of infection, in the management of chronic hepatitis and in the treatment of patients during their disease.

Keywords: CLIA, ELISA, hepatitis C virus, immunoassay.

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Authors: Amanda M. Tadini, Houssam Hajjoul, Gustavo Nicolodelli, Stéphane Mounier, Célia R. Montes, Débora M. B. P. Milori

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Soil organic matter (SOM) plays an important role in maintaining soil productivity and accounting for the promotion of biological diversity. The main components of the SOM are the humic substances which can be fractionated according to its solubility in humic acid (HA), fulvic acids (FA) and humin (HU). The determination of the chemical properties of organic matter as well as its interaction with metallic species is an important tool for understanding the structure of the humic fractions. Fluorescence spectroscopy has been studied as a source of information about what is happening at the molecular level in these compounds. Specially, soils of Amazon region are an important ecosystem of the planet. The aim of this study is to understand the molecular and structural composition of HA samples from Spodosol of Amazonia using the fluorescence Emission-Excitation Matrix (EEM) and Time Resolved Fluorescence Spectroscopy (TRFS). The results showed that the samples of HA showed two fluorescent components; one has a more complex structure and the other one has a simpler structure, which was also seen in TRFS through the evaluation of each sample lifetime. Thus, studies of this nature become important because it aims to evaluate the molecular and structural characteristics of the humic fractions in the region that is considered as one of the most important regions in the world, the Amazon.

Keywords: Amazonian soil, characterization, fluorescence, humic acid, lifetime.

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Authors: Makoto Hasegawa, Seika Tokumitsu

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Spectra of light beams emitted from white-color LED torches are different from those of conventional electric torches. In order to confirm if white-color LED torches can be used as light sources for popular sunset color demonstrations in spite of such differences, spectra of travelled light beams and scattered light beams with each of a white-color LED torch (composed of a blue LED and yellow-color fluorescent material) and a conventional electric torch as a light source were measured and compared with each other in a 50 cm-long water tank for sunset color demonstration experiments. Suspension liquid was prepared from acryl-emulsion and tap-water in the water tank, and light beams from the white-color LED torch or the conventional electric torch were allowed to travel in this suspension liquid. Sunset-like color was actually observed when the white-color LED torch was used as the light source in sunset color demonstrations. However, the observed colors when viewed with naked eye look slightly different from those obtainable with the conventional electric torch. At the same time, with the white-color LED, changes in colors in short to middle wavelength regions were recognized with careful observations. From those results, white-color LED torches are confirmed to be applicable as light sources in sunset color demonstrations, although certain attentions have to be paid. Further advanced classes will be successfully performed with white-color LED torches as light sources.

Keywords: Blue sky demonstration, sunset color demonstration, white LED torch.

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Authors: Srinivasan Aruchamy, Partha Bhattacharjee, Goutam Sanyal

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Optic disk segmentation plays a key role in the mass screening of individuals with diabetic retinopathy and glaucoma ailments. An efficient hardware-based algorithm for optic disk localization and segmentation would aid for developing an automated retinal image analysis system for real time applications. Herein, TMS320C6416DSK DSP board pixel intensity based fractal analysis algorithm for an automatic localization and segmentation of the optic disk is reported. The experiment has been performed on color and fluorescent angiography retinal fundus images. Initially, the images were pre-processed to reduce the noise and enhance the quality. The retinal vascular tree of the image was then extracted using canny edge detection technique. Finally, a pixel intensity based fractal analysis is performed to segment the optic disk by tracing the origin of the vascular tree. The proposed method is examined on three publicly available data sets of the retinal image and also with the data set obtained from an eye clinic. The average accuracy achieved is 96.2%. To the best of the knowledge, this is the first work reporting the use of TMS320C6416DSK DSP board and pixel intensity based fractal analysis algorithm for an automatic localization and segmentation of the optic disk. This will pave the way for developing devices for detection of retinal diseases in the future.

Keywords: Color retinal fundus images, Diabetic retinopathy, Fluorescein angiography retinal fundus images, Fractal analysis.

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Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast.

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